ALBERT

Description: Quercetin shows interesting pharmacological effects, but its use in topical applications is limited by its low skin permeability and solubility. In this work, the synthesis of highly lipophilic quercetin esters with oleic, linoleic and linolenic acid useful as topical quercetin prodrugs is reported. Partial OH esterification is advisable to maintain the antioxidant activity of these compounds; tetraesters and triesters can be achieved by modulating the reaction conditions utilized for the total esterification of quercetin. The chemical structures of the esters were proven by spectroscopic techniques; quantum chemical NMR calculation were mandatory to unequivocally assign the free position in triesters. Finally, the antioxidant activity of all the synthesized compounds was determined by the 2,2-diphenyl-1-picryl-hydrazyl method and by 2,2-azinobis(3-ethyl-benzothiazoline-6-sulfonic acid) assay.

Description: Clostridium acetobutylicum has been considered as an attractive platform host for biorefinery due to its metabolic diversity. Considering its capability to overproduce butanol through butyrate, it was thought that butyric acid can also be efficiently produced by this bacterium through metabolic engineering. The pta-ctfB -deficient C. acetobutylicum CEKW, in which genes encoding phosphotransacetylase and CoA-transferase were knocked out, was assessed for its potential as a butyric acid producer in fermentations with four controlled pH values at 5.0, 5.5, 6.0, and 6.4. Butyric acid could be best produced by fermentation of the CEKW at pH 6.0, resulting in the highest titer of 26.6 g/l, which is 6.4 times higher than that obtained with the wild type. However, due to the remaining solventogenic ability of the CEKW, 3.6 g/l solvents were also produced. Thus, the CEKW was further engineered by knocking out the adhE1 -encoding aldehyde/alcohol dehydrogenase to prevent solvent production. Batch fermentation of the resulting C. acetobutylicum HCEKW at pH 6.0 showed increased butyric acid production to 30.8 g/l with a ratio of butyric-to-acetic acid (BA/AA) of 6.6 g/g and a productivity of 0.72 g/l/h from 86.9 g/l glucose, while negligible solvent (0.8 g/l ethanol only) was produced. The butyric acid titer, BA/AA ratio, and productivity obtained in this study were the highest values reported for C. acetobutylicum , and the BA/AA ratio and productivity were also comparable to those of native butyric acid producer Clostridium tyrobutyricum . These results suggested that the simultaneous deletion of the pta-ctfB-adhE1 in C. acetobutylicum resulted in metabolic switch from biphasic to acidogenic fermentation, which enhanced butyric acid production.

Description: Acetic- and butyric-capped oleic estolide 2-ethylhexyl (2-EH) esters were synthesized in a perchloric acid catalyzed (0.05 equiv) one-pot process from industrial 90 % oleic acid and either acetic or butyric fatty acids at two different ratios. This was directly followed by the esterification process incorporated into an in-situ second step to provide a low viscosity estolide ester functional fluid. The monoestolide and polyestolides were separated via vacuum distillation (6–13 Pa) at 240–250 °C. The physical properties of these materials were followed throughout the synthetic process and are reported. The final low viscosity acetic- and butyric-capped monoestolide 2-EH esters had viscosities of 19.9 and 24.2 cSt at 40 °C and 4.8 and 5.5 cSt at 100 °C with viscosity indexes (VI) of 161 and 163, respectively. Both monoestolide esters displayed excellent pour points (PPs). The PPs of the two were as follows: acetic-capped estolide 2-EH ester PP = −45 °C and butyric-capped estolide 2-EH ester PP = −27 °C. The biodegradable short-capped oleic estolide 2-EH esters had excellent low temperature properties and should perform well in low viscosity applications.

Description: Disruption of spatiotemporal behavior of intracellular signaling cascades including tumor necrosis factor alpha-related apoptosis-inducing ligand (TRAIL)-mediated signaling in prostate cancer has gained tremendous attention in the past few years. There is an increasing effort in translating the emerging information about TRAIL-mediated signaling obtained through experimental and preclinical data to clinic. Fascinatingly, novel targeting approaches are being developed to enhance the tissue- or subcellular-specific delivery of drugs with considerable focus on prostate cancer. These applications have the potential to revolutionize prostate cancer therapeutic strategies and include the accumulation of drugs in target tissue as well as the selection of internalizing ligands for enhanced receptor-mediated uptake of drugs. In this mini-review, we outline outstanding developments in therapeutic strategies based on the regulation and/or targeting of TRAIL pathway for the treatment of prostate cancer. Moreover, microRNAs (miRNAs), with potential transcriptional and posttranscriptional regulation of gene expression, will be presented for their potential in prostate cancer treatment. Emphasis has been given to the use of delivery approaches, especially based on nanotechnology. Considerably, enhanced information regarding miRNA regulation of TRAIL-mediated signaling in prostate cancer cells may provide potential biomarkers for the characterization of patients as responders and nonresponders of TRAIL-based therapy and could provide rationalized basis for combination therapies with TRAIL death receptor-targeting drugs.

Description: The deterging effect and wetting capacity of some widely used commercial surfactants and demulsifiers are examined and studied from the standpoint of the oilfield treatment of crude oil. The deterging effect of surfactants of various structures is shown to be selective in washing off disperse particles of mechanical paraffin, resin, and asphaltene impurities from the surface. New methods are proposed for determining the deterging effect of surfactacts in oil-bearing disperse systems. The mechanism of breakdown of the structure of disperse particles surrounded by a hydrocarbon adsorption layer under the action of detergents is described. The studies identified a number of surfactants that have an intrinsically pronounced deterging effect.

Description: A method has been developed for determination of the thermooxidative stability of lubricating oils by substitution of single-stage extraction of mechanical impurities from oxidized oil in a Soxhlet apparatus for stages wherein the oxidized oil is held in a solvent, filtered, and additionally eluted of residue with solvent. The precision indexes of the proposed method meet specifications GOST 11063–77, GOST 23652–79, and GOST 6370–83.

Description: The development of cellulase-based bioprocess is afflicted by the processing efficiency of enzymes. To address this issue, a method based on artificial oil bodies (AOBs) was proposed to integrate production and immobilization of recombinant cellulase. First, the heterologous endoglucanase ( celA ), cellobiohydrolase ( celK ), and β-glucosidase ( gls ) genes were individually fused with oleosin, a structural protein of plant seed oils. After expression in Escherichia coli , each fusion protein of insolubility was mixed together with plant oils. AOBs were assembled by subjecting the mixture to sonication. Consequently, active CelA, CelK, and Gls were resumed and co-immobilized on AOBs surface. Finally, the assembly condition (including the protein ratio) and the reaction condition were further optimized by response surface methodology. The resulting AOBs-bound cellulase remained stable for 4 cycles of cellulose–hydrolyzed reactions. Overall, the result shows a promise of this proposed approach for processing recombinant cellulase, which may provide a facile method to investigate optimum combination of cellulase components towards various cellulosic materials.

Description: The membrane and solute diffusion properties of Cladophora cellulose and polypyrrole (PPy) functionalized Cladophora cellulose were analyzed to investigate the feasibility of using electroactive membranes in hemodialysis. The membranes were characterized with scanning electron microscopy, ζ-potentiometry, He-pycnometry, N 2 gas adsorption, and Hg porosimetry. The diffusion properties across the studied membranes for three model uremic toxins, i.e. creatinine, vitamin B12 and bovine serum albumin, were also analyzed. The characterization work revealed that the studied membranes present an open structure of weakly negatively charged nanofibers with an average pore size of 21 and 53 nm for pristine cellulose and PPy-Cladophora cellulose, respectively. The results showed that the diffusion of uremic toxins across the PPy-Cladophora cellulose membrane was faster than through pure cellulose membrane, which was related to the higher porosity and larger average pore size of the former. Since it was found that the average pore size of the membranes was larger than the hydrodynamic radius of the studied model solutes, it was concluded that these types of membranes are favorable to expand the Mw spectrum of uremic toxins to also include conditions associated with accumulation of large pathologic proteins during hemodialysis. The large average pore size of the composite membrane could also be exploited to ensure high-fluxes of solutes through the membrane while simultaneously extracting ions by an externally applied electric current.

Description: Thirty-four strains of docosahexaenoic acid (DHA)-producing microorganisms were newly isolated from brackish areas in Japan. These strains showing various compositions of fatty acids. Especially, the fatty acids produced by one of the strains, named D31, had a high DHA content (over 60 % of the total fatty acids) and the simple fatty acid composition (16:0, 18:0, 18:1 and DHA without any other polyunsaturated acids). Although most oleaginous microorganisms accumulate DHA as triacylglycerol, the strain D31 accumulated DHA mainly as a polar lipid (79.4 % of total DHA), especially as phosphatidylcholine (71.4 % of polar DHA). This strain D31 was identified as a related species of Crypthecodinium cohnii on the basis of phylogenetic analysis. Crypthecodinium sp. D31 showed high DHA productivity when cultivated in a medium containing glycerol as the carbon source and a mixture of yeast extract and polypeptone as the nitrogen sources, with a salinity that was equivalent to 50 % of that of seawater and a pH in the acidic range (〈pH 6.0). Crypthecodinium sp. D31 is considered as a promising producer of high-purity DHA-containing phospholipids.

Description: CLA-rich soy oil (CLARSO) has been produced by linoleic acid isomerization in soy oil TAG. The objective was to determine the physicochemical properties of the novel CLARSO relative to conventional refined bleached deodorized soy oil (RBDSO). Iodine value decreased in CLARSO samples despite unsaturation being unchanged, probably because conjugated double bonds in triacylglycerol (TAG) of CLARSO impede the complete addition of iodine. Thermal analysis by differential scanning calorimetry showed the melting point temperature increased with the increase in the CLARSO CLA concentration, and melting point broadening occurred in CLA contained samples. Dynamic viscosity of CLARSO conducted from 4 to 44 °C increased with the increase in CLA concentration, relative to RBDSO. Greatest viscosity differences occurred at the lower temperatures. Refractive indices and density were not greatly affected. The change in physical properties was attributed to the increased intermolecular hydrophobic interaction force due to conjugated double bonds and trans , trans isomers, relative to RBDSO. This may provide hard fat characteristics that may be useful for use in producing margarine or shortenings. Furthermore, the higher viscosity at refrigerator temperatures may be useful in salad oil and refrigerated sauces to enable suspension of particles and maintain a rich, thicker texture.

Description: Radiofrequency (RF) ablation (RFA) is a minimally invasive treatment for colorectal-cancer liver metastases (CLM) in selected nonsurgical patients. Unlike surgical resection, RFA is not followed by routine pathological examination of the target tumor and the surrounding liver tissue. The aim of this study was the evaluation of apoptotic events after RFA. Specifically, we evaluated YO-PRO-1 (YP1), a green fluorescent DNA marker for cells with compromised plasma membrane, as a potential, early marker of cell death. YP1 was applied on liver tissue adherent on the RF electrode used for CLM ablation, as well as on biopsy samples from the center and the margin of the ablation zone as depicted by dynamic CT immediately after RFA. Normal pig and mouse liver tissues were used for comparison. The same samples were also immunostained for fragmented DNA (TUNEL assay) and for active mitochondria (anti-OxPhos antibody). YP1 was also used simultaneously with propidium iodine (PI) to stain mouse liver and samples from ablated CLM. Following RFA of human CLM, more than 90 % of cells were positive for YP1. In nonablated, dissected pig and mouse liver however, we found similar YP1 signals (93.1 % and 65 %, respectively). In samples of intact mouse liver parenchyma, there was a significantly smaller proportion of YP1 positive cells (22.7 %). YP1 and PI staining was similar for ablated CLM. However in dissected normal mouse liver there was initial YP1 positivity and complete absence of the PI signal and only later there was PI signal. Conclusion: This is the first time that YP1 was applied in liver parenchymal tissue (rather than cell culture). The results suggest that YP1 is a very sensitive marker of early cellular events reflecting an early and widespread plasma membrane injury that allows YP1 penetration into the cells.

Description: Vitamin D receptor polymorphisms may predispose that not all individuals could have benefits from the nutritional supplementation of 25-hydroxyvitamin D. Furthermore, vitamin D-related cardiovascular effects may also be influenced by soy isoflavones considered endocrine regulators of cardiovascular homeostasis. To find possible gene–diet interactions by evaluating individualized lipid metabolism benefits from an increase in soy and 25-hydroxyvitamin D intake, 106 healthy individuals, genotyped for vitamin D receptor (VDR) gene polymorphism rs1544410 (BsmI) were randomly assigned to either no intake, to daily 250 mL or 500 mL of a 25-hydroxyvitamin D supplemented SB for 2 months. The soybean beverage induced differences in cardiovascular risk factors (lipid profile, blood pressure, TNFα and MCP-1), as well as vitamin D metabolites in a dose-gene-dependent relation. Thus, VDR BsmI polymorphism affected individual response being the GG genotype the ones that showed dose-dependent manner responsiveness in the reduction in total cholesterol, LDL and triglycerides in comparison with the AA/AG genotype. These differences were associated with increased plasma levels of 1α,25-dyhydroxyvitamin D3 in the carriers of the GG genotype. It was concluded that metabolic response to 25-hydroxyvitamin D and soybean supplementation is dependent on VDR BsmI GG genotype due to a higher conversion rate from vitamin D precursors.

Description: Separation strategies based on size-selective precipitation of DNA fragments with polyethylene glycol (PEG) have been used for achieving desired DNA interval in automated sample preparation for next-generation sequencing. By varying PEG concentration, DNA fragments of different sizes can be precipitated onto surfaces of carboxyl-coated paramagnetic particles selectively, and therefore, the desired DNA interval can be obtained. However, one of the crucial points in this approach is to determine the critical PEG concentration for DNA fragment of a certain size. The aim of this work was to develop a convenient and reliable method for accurately determining the critical PEG concentration. In our method, at a fixed concentration of sodium chloride (NaCl), recovered DNA samples obtained with different PEG concentrations were directly quantified, and their concentrations as a function of the PEG concentration were fitted by the logistic function. The critical PEG value was easily and accurately determined from the fitted logistic function. The repeatability and stability of the critical PEG value were assessed, showing an excellent reliability of the method. Based on this method, critical PEG values of different-size DNA fragments were determined at different NaCl concentrations. The effectiveness of the method was also demonstrated by selective precipitation of DNA fragments.

Description: A recombinant oleate hydratase from Lysinibacillus fusiformis converted ricinoleic acid to a product, whose chemical structure was identified as the novel compound 10,12-dihydroxystearic acid by gas chromatograph/mass spectrometry, Fourier transform infrared, and nuclear magnetic resonance analysis. The reaction conditions for the production of 10,12-dihydroxystearic acid were optimized as follows: pH 6.5, 30 °C, 15 g l −1 ricinoleic acid, 9 mg ml −1 of enzyme, and 4 % ( v / v ) methanol. Under the optimized conditions, the enzyme produced 13.5 g l −1 10,12-dihydroxystearic acid without detectable byproducts in 3 h, with a conversion of substrate to product of 90 % ( w / w ) and a productivity of 4.5 g l −1  h −1 . The emulsifying activity of 10,12-dihydroxystearic acid was higher than that of oleic acid, ricinoleic acid, stearic acid, and 10-hydroxystearic acid, indicating that 10,12-dihydroxystearic acid can be used as a biosurfactant.

Description: For better understanding of the adhesive properties of different fractions of cottonseed protein, cottonseed meals from both glanded and glandless cotton varieties were separated into several fractions. Each meal was sequentially extracted with water and 1 M NaCl solution, or with phosphate buffer and NaCl solution. Adhesives were prepared from the recovered fractions and hot-pressed onto maple veneer strips and tested for their properties. The adhesive strength of the water- and buffer-washed solid fractions (i.e., the un-extractable residues of the meals) from the glanded seed ranged from 1.32 to 1.62 MPa and were unchanged or increased compared with the adhesive strength of the original meal that varied from 0.98 and 1.49 MPa. Soaking the wood specimens bonded at 80 °C revealed that the water resistance of these water- and buffer-washed adhesives was significantly improved in that they exhibited no delamination during soaking compared with the meal adhesive that showed some delamination (20–30 % of the samples). Furthermore, the water resistance of these fractions with wet shear strength around 1.5 MPa was comparable to that of cottonseed protein isolate (〉90 % protein) when the joints were bonded at 100 °C. The preparations from glandless cottonseed meals showed similar adhesive performances. Additional extraction of the meals with NaCl solution reduced adhesive performance. The results suggest that water- or buffer-washed cottonseed meal fractions can be used as wood adhesives and would be less costly to prepare than cottonseed protein isolates.

Description: In this study, seeds from the safflower variety called “Dinçer” were roasted and microwaved before oil extraction by cold pressing. Some physico-chemical analyses (moisture, ash, oil content and color) were performed in safflower meals. Physico-chemical properties (refractive index, viscosity, turbidity, specific gravity, color, free acidity, peroxide value, iodine number), nutritional components (total phenolics, antioxidant capacity, tocopherol content), sterol composition and fatty acid composition of produced oils were also determined. Volatile components of the oils were detected by solid-phase microextraction/gas chromatography–mass spectrometry technique. Quantitative descriptive analysis was accomplished with trained panelists by 11 definition terms. Cold pressing yielded less oil than solvent extraction, but oil quality was superior and a refining process was not required. There was no significant difference between samples for fatty acid composition and some physico-chemical parameters. Whereas, microwave treatment caused a decrease in oil turbidity, free acidity, α-tocopherol and some sterol contents and an enhancement in total phenolic content, antioxidant capacity and peroxide value. Moreover, microwave treatment led to an increased nutty aroma in the oil. In contrast, isot pepper aroma was decreased by microwave treatment. This study provides very important information about the safflower oils for the first time in the literature.

Description: Gills cells of the freshwater mussel Lasmigona costata and the seawater clam Mesodesma mactroides were isolated (mussel: chemical dissociation; clam: mechanical dissociation) and fractionated (Percoll gradient) into Fractions I and II. Mitochondrial dyes (DASPEI: mussel; MitoTracker ® : clam) and Na + , K + -ATPase activity measurement were used to distinguish between cells of Fractions I and II. For mussel and clam, 80.5 ± 1.5 and 48.3 ± 3.2 % of cells were in Fraction II, respectively. For both species, cells of Fraction II had higher fluorescence emission and higher enzyme activity than those of Fraction I, being characterized as ‘cells rich in mitochondria’. Cells of Fraction II were kept in saline solutions approximating the ionic composition of hemolymph either under control conditions (no Cu addition) or exposed (3 h) to copper (Cu: 5, 9 and 20 μg Cu/L). Cell viability and Cu and Na + content were measured. For both species, Cu content was higher and Na + content was lower in cells exposed to 20 μg Cu/L. Furthermore, a strong negative correlation was observed between cell Na + and Cu content in the two bivalve species, indicating a possible competition between Cu and Na + for ion-transporting mechanisms or binding sites at gill cells of Fraction II. Considering that Cu is an ionoregulatory toxicant in aquatic invertebrates, these preliminary toxicological data support the idea of using isolated gill cells rich in mitochondria to study the mechanisms underlying the acute toxicity of waterborne Cu in freshwater and marine bivalves.

Description: Bacillus amyloliquefaciens FZB42 has been shown to stimulate plant growth and to suppress the growth of plant pathogenic organisms including nematodes. However, the mechanism underlying its effect against nematodes remains unknown. In this study, we screened a random mutant library of B. amyloliquefaciens FZB42 generated by the mariner transposon Tn YLB-1 and identified a mutant strain F5 with attenuated nematicidal activity. Reversible polymerase chain reaction revealed that three candidate genes RAMB_007470 , yhdY , and prkA that were disrupted by the transposon in strain F5 potentially contributed to its decreased nematicidal activity. Bioassay of mutants impaired in the three candidate genes demonstrated that directed deletion of gene RBAM_007470 resulted in loss of nematicidal activity comparable with that of the F5 triple mutant. RBAM_007470 has been reported as being involved in biosynthesis of plantazolicin, a thiazole/oxazole-modified microcin with hitherto unknown function. Electrospray ionization time-of-flight mass spectrometry (ESI-TOF-MS) analyses of surface extracts revealed that plantazolicin bearing a molecular weight of 1,354 Da was present in wild-type B. amyloliquefaciens FZB42, but absent in the Δ RABM_007470 mutant. Furthermore, bioassay of the organic extract containing plantazolicin also showed a moderate nematicidal activity. We conclude that a novel gene RBAM_007470 and its related metabolite are involved in the antagonistic effect exerted by B. amyloliquefaciens FZB42 against nematodes.

Description: Syngas fermentation is a promising route for resource recovery. Acetate is an important industrial chemical product and also an attractive precursor for liquid biofuels production. This study demonstrated high fraction acetate production from syngas (H 2 and CO 2 ) in a hollow-fiber membrane biofilm reactor, in which the hydrogen utilizing efficiency reached 100 % during the operational period. The maximum concentration of acetate in batch mode was 12.5 g/L, while the acetate concentration in continuous mode with a hydraulic retention time of 9 days was 3.6 ± 0.1 g/L. Since butyrate concentration was rather low and below 0.1 g/L, the acetate fraction was higher than 99 % in both batch and continuous modes. Microbial community analysis showed that the biofilm was dominated by Clostridium spp., such as Clostridium ljungdahlii and Clostridium drakei , the percentage of which was 70.5 %. This study demonstrates a potential technology for the in situ utilization of syngas and valuable chemical production.

Description: Lignosulfonates(LSs), by-products from chemical pulping processes, are low-value products with limited dispersion properties. The ability of commercially available horseradish peroxidase (HRP) to polymerize LS macromolecules and improve the dispersion properties of LSs was investigated. The polymerization of LSs proceeded efficiently under mild reaction conditions in an aqueous solution with HRP/H 2 O 2 . Gel permeation chromatography showed a significant increase in weight-average molecular weight ( M w ) of sulfonated kraft lignin and sodium lignosulfonate (NaLS) by 8.5-fold and 4.7-fold, respectively. The mechanism of polymerization was investigated by elemental analysis, surface charge measurement, headspace gas chromatography, infrared spectroscopy (IR), and hydrogen nuclear magnetic resonance spectrometry ( 1 H-NMR). The functional group measurements indicated that HRP incubation did not reduce the sulfonic group content. However, it decreased the phenolic and methoxyl group contents. As the phenolic group content decreased, M w increased as a power function. The polymerization was proposed to involve the random coupling of phenoxy radical intermediates. The radicals coupled with each other to form different inter-unit linkages, most of which were the β-O-4’ type, as the 1 H-NMR spectra indicated. Moreover, the HRP/H 2 O 2 incubation induced a significant improvement in the adsorption and dispersion properties of LSs. Therefore, the HRP/H 2 O 2 incubation is a promising approach for industrial applications of LSs.

Description: Inspired by the surface structure of lotus leaves, different types of superhydrophobic cellulosic materials with contact angle (CA) of higher than 150° are currently provided. However, fabrication of these surfaces in a facile one-step coating process is one of the challenging issues. This paper describes a facile method to sonochemically synthesize superhydrophobic organic–inorganic hybrid coatings on cotton fabric by an alkaline-catalyzed co-hydrolysis and co-condensation of tetraethylorthosilicate and alkyltrialkoxysilanes. The influence of alkyl chain length (methyl, octyl, hexadecyl) of silane and reaction time was investigated. Surface structure of the fabrics was investigated by SEM, EDS, FTIR spectroscopies, and reflectance spectrophotometry. Wettability properties were studied by measuring water CA, shedding angle (SHA) and resistance to wetting by a series of ethanol–water mixtures of different surface tensions. The results showed that the treated fabrics were coated with a homogeneous thin nano-scaled coating of hybrid silica nano-particles. The fabrics demonstrated CA of higher than 150°, SHA in the range of 6–24° and different stickiness to water droplets. The fabrics treated by silanes with longer alkyl chain length and at higher reaction time revealed better water repellency. The coatings were nearly transparent, could not affect the color of the fabrics and had high stability against repeated washing. In addition, mechanical properties of the fabrics were not substantially affected.

Description: Composites of cellulose acetate and polysiloxane were prepared using 3-isocyanatepropyltriethoxysilane, as a coupling agent. The structure, the thermal and dynamic-mechanical behaviors, and the morphology of the obtained composites were investigated. The composites showed phase separation which was confirmed by the presence of siloxane micro- and nano-domains dispersed in the cellulose acetate matrix, with good interfacial adhesion between the phases. The results demonstrated that the incorporation of a polysiloxane phase on a cellulose acetate matrix caused a decrease in the glass transition temperature, storage modulus and hardness. The proposed methodology was seen to be convenient for the preparation of cellulose acetate/polysiloxane composites with useful properties.

Description: Marine microalga Nannochloropsis oculata possesses nutrients valuable for human health. In this study, we added freeze-dried N. oculata powder to soybean oil and observed a remarkable inhibition in oil oxidation. The amount of microalgae powder added was positively correlated to the increase in oil stability. The addition of 5.0 % ( w / w ) microalgae powder increased the oil stability index (OSI) values of soybean oil more than twofold at the tested temperatures 120 and 130 °C. N . oculata contains high levels of both phenolic compounds and α-tocopherols that could be the contributors to such an increase of the OSI. Two methods were conducted to assay the active ingredients released from microalgae: one employed three solvent systems to extract the microalgae and the other was the soybean oil added with microalgae. Analyses of free radical scavenging and reducing power suggested that the phenolic compounds dominated the antioxidation activities in soybean oil when it was infused with the microalgae powder. Our results suggest that N. oculata could potentially be used as an additive in cooking oil to increase the shelf life and nutritional value of the oil and to reduce the production of free radicals from lipid oxidation when the oil is used at high-temperature cooking processes.

Description: The actinomycete Streptomyces platensis produces two compounds that display antibacterial activity: platensimycin and platencin. These compounds were discovered by the Merck Research Laboratories, and a complex insoluble production medium was reported. We have used this medium as our starting point in our studies. In a previous study, we developed a semi-defined production medium, i.e., PM5. In the present studies, by varying the concentration of the components of PM5, we were able to develop a superior semi-defined medium, i.e., PM6, which contains a higher concentration of lactose. Versions of PM6, containing lower concentrations of all components, were also found to be superior to PM5. The new semi-defined production media contain dextrin, lactose, MOPS buffer, and ammonium sulfate in different concentrations. We determined antibiotic production capabilities using agar diffusion assays and chemical assays via thin-layer silica chromatography and high-performance liquid chromatography. We reduced crude nutrient carryover from the seed medium by washing the cells with distilled water. Using these semi-defined media, we determined that addition of the semi-defined component soluble starch stimulated antibiotic production and that it and dextrin could both be replaced with glucose, resulting in the chemically defined medium, PM7.

Description: Pichia pastoris is widely used as a host system for heterologous protein expression in both academia and industry. Production is typically accomplished by a fed-batch induction process that is known to have negative impacts on cell physiology that impose limits on both protein yields and quality. We have analysed recombinant protein production in chemostat cultures to understand the physiological responses associated with methanol-induced production of two human lysozyme variants with different degrees of misfolding by P . pastoris . Confounding variables associated with nutrient stress or growth-rate are minimised during steady-state growth in chemostats. Comparison of transcriptome-level data obtained during the non-inducing and inducing steady states identified changes in expression of only about 1 % of the genome during production of either an amyloidogenic human lysozyme variant prone to intracellular aggregation (I56T) or a misfolded but secretable variant (T70N), indicating near-complete acclimation to their production. A marked, but temporary, stress response involving both the unfolded protein response (UPR) and ER-associated degradation pathway was observed during the transient between steady states, particularly following induction of the T70N variant synthesis, and was accompanied by changes in expression of around 50 antisense transcripts. The results suggest that optimal heterologous protein production could best be achieved by a continuous process that minimises the number of methanol-induced transients experienced by the cultures. The processing of HAC1 mRNA required for the UPR was found to be constitutive in the culture conditions used, even in the absence of recombinant protein induction.

Description: The Aminobacter sp. strain MSH1 has potential for pesticide bioremediation because it degrades the herbicide metabolite 2,6-dichlorobenzamide (BAM). Production of the BAM-degrading bacterium using aerobic bioreactor fermentation was investigated. A mineral salt medium limited for carbon and with an element composition similar to the strain was generated. The optimal pH and temperature for strain growth were determined using shaker flasks and verified in bioreactors. Glucose, fructose, and glycerol were suitable carbon sources for MSH1 ( μ  = 0.1 h −1 ); slower growth was observed on succinate and acetic acid ( μ  = 0.01 h −1 ). Standard conditions for growth of the MSH1 strain were defined at pH 7 and 25 °C, with glucose as the carbon source. In bioreactors (1 and 5 L), the specific growth rate of MSH1 increased from μ  = 0.1 h −1 on traditional mineral salt medium to μ  = 0.18 h −1 on the optimized mineral salt medium. The biomass yield under standard conditions was 0.47 g dry weight biomass/g glucose consumed. An investigation of the catabolic capacity of MSH1 cells harvested in exponential and stationary growth phases showed a degradation activity per cell of about 3 × 10 −9  μg BAM h −1 . Thus, fast, efficient, large-scale production of herbicide-degrading Aminobacter was possible, bringing the use of this bacterium in bioaugmentation field remediation closer to reality.

Description: We performed differential scanning calorimetry (DSC) and synchrotron radiation X-ray diffraction (SR-XRD) experiments of polymorphic structures and binary mixing characteristics of the enantiomers of 1-oleoyl-2,3-dipalmitoyl- sn -glycerol (S-OPP) and 1,2-dipalmitoyl-3-oleoyl- sn -glycerol (R-PPO). In the two enantiomers, oleic and palmitic acid moieties are asymmetrically connected at the sn -1 and sn -3 positions of the glycerol groups, with palmitic acid moiety at the sn -2 position. Pure enantiomer samples (〉99 %) were synthesized and employed throughout this study. The following results were obtained. (1) A basic feature of the mixture systems of S-OPP and R-PPO is of a eutectic nature due to different polymorphic structures of two enantiomers and the racemic compound of PPO ( rac -PPO). (2) Polymorphic behavior of S-OPP and R-PPO was quite similar, both having α-2 and β′-3, whereas rac -PPO contained α rac -3, β′ rac -2, and β′ rac -3. The DSC measurements showed that the melting points of β′-3 (S-OPP = 35.3 °C and R-PPO = 34.9 °C) were higher than that of β′ rac -3 (31.0 °C). β was not crystallized in the pure enantiomers, and rac -PPO. (3) α rac -3 was crystallized at low cooling rates (~2 °C/min), whereas α-2 of the two enantiomers was crystallized only with very rapid cooling (~10 °C/min). (4) Triple-chain-length structures were formed in α rac -3, β′ S -3 (=β′ R -3), and β′ rac -3; α-2 with a double-chain-length structure was formed in both enantiomers. These results indicate the importance of the relationship between subcell packing and glycerol conformation in the polymorphism and mixing characteristics of asymmetric unsaturated–saturated-saturated mixed-acid triacylglycerols.

Description: Among oil compounds, fatty acids, tocopherols and xanthophylls (lutein and zeaxanthin) are of special interest due to their nutritional properties. The identification and quantification of these compounds in pecan nuts ( Carya illinoinensis ) could therefore be very useful to produce functional foods rich in compounds of this type. This paper reports studies on their accumulation and the effect of ripening on the content of these high value-added compounds. The total lipid content increased during the ripening. Saturated and polyunsaturated fatty acids decreased significantly, whereas, monounsaturated fatty acids increased during the ripening of pecan nut fruit. Maximum levels of total tocopherol (279.53 mg/kg oil) and xanthophyll (6.18 mg/kg oil) were detected at 20th weeks after the flowering date. These amounts decreased gradually as ripening advances. The early stages of pecan ripening seem to have nutritional and pharmaceutical interests. These results may be useful for evaluating the pecan nut quality and determining the optimal period when the pecans accumulated the maximum of these nutritional and healthy compounds.

Description: Soy sauce is a traditional condiment manufactured by natural inoculation and mixed culture fermentation. As is well known, it is the microbial community that plays an important role in the formation of its flavors. However, to date, its dynamic changes during the long period of fermentation process are still unclear, intensively constraining the improvement and control of the soy sauce quality. In this work, we revealed the dynamic changes of the microbial community by combining a cultured dependent method and a cultured independent method of polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis. Results indicated that the two methods verified and complemented each other in profiling microbial community, and that significant dynamics of the microbial community existed during the fermentation process, especially the strong inhibition of the growth of most of the microbes when entering into the mash stage from the koji stage. In the analysis of bacterial community, Staphylococcus and Bacillus were found to be the dominant bacteria and detected in the whole fermentation process. Kurthia and Klebsiella began to appear in the koji stage and then fade away in the early stage of the mash fermentation. In the analysis of fungal community, Aspergillus sojae and Zygosaccharomyces rouxii were found to be the dominant fungi in the koji and mash fermentation, respectively. It was clearly shown that when A. sojae decreased and disappeared in the middle stage of the mash fermentation, Z. rouxii appeared and increased at the meantime. Aspergillus parasiticus , Trichosporon ovoides and Trichosporon asahii also appeared in the koji and the early period of the mash fermentation and disappeared thereafter. Similar to Z. rouxii , Millerozyma farinosa and Peronospora farinosa were also found spontaneously which appeared in the mid-late period of the mash fermentation. The principal component analysis suggested that the microbial community underwent significant changes in the early period of the fermentation and, thereafter, tended to the stabilization in the mid-late periods. This study gave us important clues to understand the fermentation process and can serve as a foundation for improving the quality of soy sauce in the future.

Description: The effect of surface hydrophobicity and side-chain variation on xyloglucan adsorption onto cellulose microfibrils (CMF) is investigated via molecular dynamics simulations. A molecular model of CMF with (100), (010), (1–10), (110) and (200) crystal faces was built. We considered xylogluco-oligosaccharides (XGO) with three repeating units, namely (XXXG) 3, (XXLG) 3 , and (XXFG) 3 (where each (1,4)-β- d -glucosyl residue in the backbone is given a one-letter code according to its substituents: G = β- d -Glc; X = α- d -Xyl-(1,6)-β- d -Glc; L = β- d -Gal-(1,2)-α- d -Xyl-(1,6)-β- d -Glc; F = α- l -Fuc-(1,2)-β- d -Gal-(1,2)-α- d -Xyl-(1,6)-β- d -Glc). Our work shows that (XXXG) 3 binds more favorably to the CMF (100) and (200) hydrophobic surfaces than to the (110), (010) and (1–10) hydrophilic surfaces. The origin of this behavior is attributed to the topography of hydrophobic CMF surface, which stabilizes (XXXG) 3 in flat conformation. In contrast, on the rough hydrophilic CMF surface (XXXG) 3 adopts a less favorable random-coil conformation to facilitate more hydrogen bonds with the surface. Extending the xyloglucan side chains from (XXXG) 3 to (XXLG) 3 hinders their stacking on the CMF hydrophobic surface. For (XXFG) 3 , the interaction with the hydrophobic surface is as strong as (XXXG) 3 . All three XGOs have similar binding to the hydrophilic surface. Steered molecular dynamics simulation was performed on an adhesive model where (XXXG) 3 was sandwiched between two CMF hydrophobic surfaces. Our analysis suggests that this sandwich structure might help provide mechanical strength for plant cell walls. Our study relates to a recently revised model of primary cell walls in which extensibility is largely determined by xyloglucan located in limited regions of tight contact between CMFs.

Description: The aims of the present study were to prepare hydroxypropylmethyl cellulose (HPMC)-based porous matrix tablets for gastroretentive drug delivery and to characterize their physicochemical properties. Gabapentin (GBP) was used as a model drug. Paste containing GBP, HPMC and water was molded and freeze-dried to prepare freeze-dried gastroretentive matrix tablet (FD-GRT). In vitro drug release and erosion studies were also performed. Although FD-GRT exhibited porous structure, they had good tablet strength and friability. Density of FD-GRT ranged from 0.402 to 0.509 g/cm 3 and thus they could float on the medium surface without any lag time. FD-GRT was remained floated until the entire matrix erosion or end of drug release during in vitro release test. Release behavior of GBP could be modulated by the amount and the viscosity grade of HPMC. However, large amount and high viscosity of HPMC caused trouble in molding prior to freeze-drying. Addition of ethylcellulose could retard the release rate of GBP, with relatively low increase in viscosity of paste. Since pores generated by freeze drying imparted buoyancy for gastric retention to FD-GRT, additional materials for buoyancy was not necessary and FD-GRT had no lag time for buoyancy due to low density. Therefore it could be a promising tool for gastroretentive drug delivery.

Description: Introductory material first describes electron density approaches and demonstrates visualization of electron lone pairs and bonding as concentrations of electron density. Then it focuses on the application of Bader’s Quantum Theory of Atoms-in-Molecules (AIM) to cellulose models. The purpose of the work is to identify the various interactions that stabilize cellulose structure. AIM analysis aids study of non-covalent interactions, especially those for which geometric criteria are not well established. The models were in the form of pairs of cellotriose molecules, methylated at the O1 and O4 ends. Based on the unit cell of cellulose Iβ, there were corner–corner, and center–center pairs that correspond to (200) sheets, and corner–center pairings that corresponded to (1–10) and (110) stacks. AIM analysis (or charge-density topology analysis) was applied before and after minimization in vacuum and in continuum solvation. Besides the conventional O–H···O hydrogen bonds, all of which were known from geometric criteria, C–H···O hydrogen bonds (some previously reported), and some O···O and H···H interactions were found. Non-covalent bonds in the (200) sheets were maintained in all calculations with the exception of a weak, bifurcated O6–H···O2′′ bond that was not found in the corner–corner pair model and did not survive minimization. Nor did the O6···O4 interactions on the reducing ends of the triosides. Pairs of molecules along the (110) plane had an equal number (12) of non-covalent bonds compared to the pairs along the (1–10) plane, but the AIM parameters indicated the bonds between the pairs in the (110) plane were weaker. Intra-molecular O–H···O hydrogen bonds survived in these minimized pairs, but the relative chain alignments usually did not.

Description: To improve antibody production in Chinese hamster ovary (CHO) cells, the humanized antibody-producing CHO DP-12-SF cell line was transfected with the gene encoding activating transcription factor 4 (ATF4), a central factor in the unfolded protein response. Overexpression of ATF4 significantly enhanced the production of antibody in the CHO DP-12-SF cell line. The specific IgG production rate of in the ATF4 -overexpressing CHO-ATF4-16 cells was approximately 2.4 times that of the parental host cell line. Clone CHO-ATF4-16 did not show any change in growth rate compared with the parental cells or mock-transfected CHO-DP12-SF cells. The expression levels of mRNAs encoding both the antibody heavy and light chains in the CHO-ATF4-16 clone were analyzed. This analysis showed that ATF4 overexpression improved the total production and specific production rate of antibody without affecting the mRNA transcription level. These results indicate that ATF4 overexpression is a promising method for improving recombinant IgG production in CHO cells.

Description: Galactosidases are widespread enzymes that are used for manifold applications, including production of prebiotics, biosynthesis of different transgalactosylated products, improving lactose tolerance and in various analytical approaches. The nature of these applications often require galactosidases to be present in a purified form with clearly defined properties, including precisely determined substrate specificities, low sensitivity to inhibitors, and high efficiency and stability under distinct conditions. In this study, we present the recombinant expression and purification of two previously uncharacterized β-galactosidases from Aspergillus nidulans as well as one β-galactosidase from Aspergillus niger . All enzymes were active toward p -nitrophenyl-β- d -galactopyranoside as substrate and displayed similar temperature and pH optima. The purified recombinant galactosidases digested various complex substrates containing terminal galactose β-1,4 linked to either N -acetylglucosamine or fucose, such as N -glycans derived from bovine fibrin and Caenorhabditis elegans . In our comparative study of the recombinant galactosidases with the commercially available galactosidase from Aspergillus oryzae , all enzymes also displayed various degrees of activity toward complex oligosaccharides containing β-1,3-linked terminal galactose residues. All recombinant enzymes were found to be robust in the presence of various organic solvents, temperature variations, and freeze/thaw cycles and were also tested for their ability to synthesize galactooligosaccharides. Furthermore, the use of fermentors considerably increased the yield of recombinant galactosidases. Taken together, we demonstrate that purified recombinant galactosidases from A. niger and from A. nidulans are suitable for various glycobiological and biotechnological applications.

Description: A Lolium perenne ice-binding protein ( Lp IBP) demonstrates superior ice recrystallization inhibition (IRI) activity and has proposed applications in cryopreservation, food texturing, as well as in being a “green” gas hydrate inhibitor. Recombinant production of Lp IBP has been previously conducted in bacterial and yeast systems for studies of protein characterization, but large-scale applications have been hitherto limited due to high production costs. In this work, a codon-optimized Lp IBP was recombinantly expressed and secreted in a novel one-step vector system from the nuclear genome of the green microalga Chlamydomonas reinhardtii . Both mixotrophic and photoautotrophic growth regimes supported Lp IBP expression, indicating the feasibility of low-cost production using minimal medium, carbon dioxide, and light energy as input. In addition, multiple growth and bioproduct extraction cycles were performed by repetitive batch cultivation trials, demonstrating the potential for semi-continuous production and biomass harvesting. Concentrations of recombinant protein reached in this proof of concept approach were sufficient to demonstrate IRI activity in culture media without additional purification or concentration, with activity further verified by thermal hysteresis and morphology assays. The incorporation of the recombinant Lp IBP into a model gas hydrate offers the promise that algal production may eventually find application as a “green” hydrate inhibitor.

Description: The interest in biofertilizers is increasing and so is the potential for their use in sustainable agriculture. However, many of the products that are currently available worldwide are often of very poor quality, resulting in the loss of confidence from farmers. The formulation of an inoculant is a crucial multistep process that should result in one or several strains of microorganisms included in a suitable carrier, providing a safe environment to protect them from the often harsh conditions during storage and ensuring survival and establishment after introduction into soils. One of the key issues in formulation development and production is the quality control of the products, at each stage of the process. This review presents the different components and the major steps involved in the formulation of good quality biofertilizers, including the techniques used to assess the quality of the products following production. The quality of currently available inoculants is also reviewed, emphasizing the need for better quality control systems worldwide.

Description: A correlation is established experimentally between the content of oxidation products determined by IR spectroscopy and the acid number of lubricating oils used in gas-pumping units. A new express method is proposed fro determining acid number.

Description: We have studied the melting points and the hexagonal—rhombic transition temperatures in the solid state of the crude oil paraffin P-1 (mixed Ozek-Suat oil) and its narrow fractions. We have identified for the first time a correlation between the phase transition temperatures for the narrow fractions and the corresponding temperatures for the predominant high-melting and low-melting n-alkanes in these fractions.

Description: Crude oil acids, isolated from high-acid crude oil, were decarboxylated in a continuous flow-through reactor in the presence of nanosized titanium dioxide. We studied the effect of the reaction temperature on the acid number of the product and the acid conversion. The product obtained, with low acidity and high calorific value and representing a mixture of naphthenic hydrocarbons, is proposed as a component of jet fuels.

Description: We use the Aspen HYSYS software to simulate the process of amine sweetening of natural gas with high acid gas content. We analyze the effect of the lean amine (absorbent) temperature on the acid gas content in the sweetened gas and saturated amine solution (the rich amine). We found that the optimal temperature for the regenerated amine solution (lean amine), at which the maximum sweetening of the gas is achieved with minimum amine circulation rate, is within the range 38°C-45°C.

Description: We describe a statistical method for determining the fracture parameters and the vertical and lateral distribution of these parameters. We examine the correlation between the fracture properties and the filtration characteristics: permeability and deliverability. The grid method is an accurate and efficient tool that makes it possible to describe the surface density of fractures in the core. The parameters determined from the core can be used to calibrate the results from interpretation of well logging data, in order to sufficiently accurately determine the nature of the change in the parameters across the entire wellbore. We have established that the permeability and deliverability of the deposits clearly depends on the fracture spacing and the fracture surface density within the Dabei block of the Kuqa depression.

Description: A major by-product of biodiesel production is waste glycerol, which has numerous potential applications. In this study, we isolated a novel bacterium capable of producing cellulose from waste glycerol, and identified it as a novel strain (named NEDO-01) of Gluconacetobacter intermedius . Scanning electron microscopy revealed that the morphology of the pellicle produced by NEDO-01 was similar to that of cellulose produced by Gluconacetobacter hansenii ATCC23769. Furthermore, X-ray diffraction and solid-state nuclear magnetic resonance spectroscopic analyses suggested that cellulose produced by NEDO-01 had molecular and crystalline structures similar to those of cellulose produced by ATCC23769. After the optimization of cultivation conditions, NEDO-01 mediated the one-step production of nanofibrillated bacterial cellulose (NFBC) from waste glycerol in a medium supplemented with carboxymethyl cellulose. Transmission electron microscopic analysis revealed that the NFBC was composed of relatively uniform fibers with diameters of approximately 20 nm. NFBC was produced as uniform water suspensions, the yield of which was 3.4 g/L from cultivation in 7.5 L medium in a 10-L jar fermenter. The bioconversion of waste glycerol to NFBC, which has superior fluidity, moldability, and miscibility, has a wide variety of applications, including potential uses in the medical and materials engineering fields.

Description: The potential content and properties of oil distillate fractions of heavy crude oil from the Ashal’chinskoe field are analyzed. The properties of the lubricants obtained from these fractions are studied. Theoretical possibility of use of this type of hydrocarbon feedstock for production of commercial motor oils and conserving lubricants is shown.

Description: Quaternary ammonium salts and N -halamines are widely used as biocides in antimicrobial coatings, and have been extensively studied over the past two decades. In this work, 5,5-dimethyl-3-(3′-triethoxysilylpropyl)hydantoin (SPH), and 3-(trimethoxysilylpropyl) octadecyl dimethyl ammonium chloride (SPODA) were synthesized and coated onto cotton fibers using a pad-dry process (PD) and the traditional pad-dry-cure process (PDC). The coated cotton swatches were characterized by FT-IR and SEM. The quaternary ammonium salt showed a relatively lower inactivating bacteria efficacy than did the N -halamine compounds. The chlorinated swatches coated with both SPH and SPODA using the PD process could inactivate about 7 logs of the Staphylococci aureus within 5–10 min and 7 logs of Escherichia coli O157:H7 within 10–30 min, respectively. The addition of quats in N -halamine coatings improved antimicrobial activity against Gram-negative bacteria E. coli O157:H7. However, this result was not observed when the PDC process was applied in coatings because of the increasing hydrophobicity of the coated samples under high coating temperature.

Description: Glycerides obtained from the glycerolysis of soybean oil with crude glycerol were acylated with maleic anhydride at 80–100 °C. Both uncatalyzed and catalyzed reactions with 2-methylimidazole (2-MI) were evaluated. Formation of maleated glycerides was confirmed by 1 H-NMR and FTIR analyses. Consumption of maleic anhydride as a function of time was followed by acid value titration. High conversion (ca. 95 %) of hydroxyl groups was obtained at 90 °C in 60 min without catalyst. Under these conditions, a product with 1.9 maleate moieties per glyceride molecule was obtained. Catalyzed reactions afforded lower conversions of hydroxyl groups and lower maleate yields compared to uncatalyzed reactions.

Description: Sunflower oil was used for deep frying of potatoes at 170 ± 5 °C and for 8 h per day for 5 days in a fryer with an automatic oil filtration system. Three different frying operations were performed: operation (OP)-1, OP-2 and OP-3; that correspond to the oil unfiltered at the end of each frying day, the oil filtered through the fryer's own filter (passive filtration) and the oil firstly subjected to passive filtration and then filtered through a polyethersulfone membrane modified with hexamethyldisiloxane via radio frequency plasma (75 W-5 min, discharge power–time), respectively. The performance of each operation was investigated in terms of free fatty acids (FFA), conjugated dienoic acids (CD), TOTOX value, total polar content (TPC), Hunter color, viscosity, fatty acid composition, and tocopherol content. The results showed that OP-3 could decrease FFA, CD, TOTOX, TPC, L*a*b* value, viscosity and linoleic acid (18:2)/palmitic acid (16:0) ratio in 29.6, 11.7, 25, 30.8, 6.1*11.3*20.8*, 7.8, 12.2 %, respectively, compared to the unfiltered oil (OP-1). Regenerated oil from OP-3 had a frying life approximately 17 h more than oils from both OP-1 and OP-2.

Description: A potent antioxidant, anti-inflammatory and anti-mutagenic agent; 4-vinyl-2,6-dimethoxyphenol (canolol) was obtained from canola meal in a significant yield via alkaline (NaOH)/enzymatic (ferulic acid esterase) hydrolysis followed by microwave-assisted decarboxylation. The hydrolysis was carried out either through using canola meal directly as a substrate or by using the 70 % aqueous methanolic extract filtrates. The hydrolyzed extracts underwent RP-HPLC analysis which showed that 81.0 and 94.8 % of the total phenolics were hydrolyzed to sinapic acid after the alkaline hydrolysis of the meal and the methanolic extracts, respectively. The enzymatic hydrolysis showed lower conversion rates (49.5 and 58.3 %). The hydrolyzed extracts were consequently decarboxylated using 8-diazabicyclo[5.4.0]undec-7-ene under microwave irradiation at different conditions. The HPLC profiling of decarboxylated extracts showed that using microwave at 300 W of microwave power for 12 min brought the highest sinapic acid conversion to canolol (58.3 %) yielding 4.2 mg canolol from each gram of canola meal suggesting that the process could be commercially economical.

Description: In the margarine and butter industry, sustainable and more efficient refrigerants, such as CO 2 , are introduced to industrial scraped surface heat exchangers, allowing an increased capacity compared to conventional use of NH 3 . The effect of such changes in capacity and a varied rotational speed was studied in relation to the structural behavior of puff pastry butter during 4 weeks of isothermal storage at 20 °C. The physical properties of the fat crystal network were studied in detail at several length scales by combining X-ray diffraction with differential scanning calorimetry, confocal laser scanning microscopy, LR-NMR and rheology. Our data shows that a high capacity combined with high rotational speed decreases the brittleness of puff pastry butter after 7 days of storage. This effect is, however, diminished after 28 days of storage. Likewise, changes in capacity and rotational speed are shown to induce no microstructural and polymorphic differences after 28 days of storage. However, the degree of work softening is related to the manufacturing conditions: a high capacity and a high rotational speed increase the ability of the puff pastry butter to resist structural breakdown during working. With this being the only observed difference; a wide operational window exists on an industrial level to produce puff pastry butter with similar structural behavior.

Description: Soy protein adhesives with a high solid content (28–39 %) were extracted from soy flour slurry modified with sodium bisulfite (NaHSO 3 ) at different concentrations. 11S-dominated soy protein fractions (SP 5.4) and 7S-dominated soy protein fractions (SP 4.5) were precipitated at pH 5.4 and pH 4.5, respectively. The objective of this work was to study the effects of NaHSO 3 on adhesion and physicochemical properties of soy protein. The adhesion performance of NaHSO 3 -modified SP 4.5 was better than SP 5.4; the wet strength of these two fractions was from 2.5 to 3.2 MPa compared with 1.6 MPa of control soy protein isolate. SDS-PAGE results revealed the reducing effects of NaHSO 3 on soy protein. The isoelectric pH of soy protein decreased as NaHSO 3 increased due to the induced extra negative charges (RS-SO 3 − ) on the protein surface. The rheological properties of soy protein adhesives were improved significantly. Unmodified samples SP 5.4 and SP 4.5 had clay-like properties and extremely high viscosity, respectively; with 2–8 g/L NaHSO 3 modification, both SP 5.4 and SP 4.5 had a viscous cohesive phase with good flowability. Overall, NaHSO 3 -modified soy protein adhesives in our study have many advantages over the traditional soy protein isolate adhesive such as better adhesion performance, higher solid content but with good flowability and longer shelf life.

Description: Fiber properties (fiber swelling ability, crystal structure of cellulose, fiber surface morphology, and etc.) of eucalyptus kraft pulp with different contents of carboxyl group in Na-form were studied. There was a direct proportional relationship between water retention value and carboxyl content of pulp. When the carboxyl content increased from 35.6 to 315.7 mmol/kg, tensile index and burst index increased by 56.1 and 117.8 %, respectively, and crystallinity of cellulose decreased by 11.8 %. Environmental scanning electron microscope showed that more fibrillation was observed on the carboxymethylated fiber surface, compared with the control sample. The results from Fourier transform infrared spectra analysis suggested that the relative intensity of the band at 1,633/cm was increased after carboxymethylation treatment, which showed that the carboxyl content increased. The increase in the carboxyl content not only could increase the fiber strength properties, but also could increase the recycling times of the fiber.

Description: A large number of molds serve as producer strains for the industrial production of pharmaceuticals, foods, or organic chemicals. To optimize strains for production processes, conventional strain development programs use random mutagenesis and, more recently, recombinant technologies to generate microbial strains with novel and advantageous properties. The recent detection of mating type genes in fungal production strains and the discovery of cryptic sexuality in presumably asexual fungi open up novel strategies for generating progeny with new, as yet unobserved properties. Mating type genes, which can be considered as “sex genes,” not only direct sexual development but also regulate a broad range of fungal secondary metabolites. In addition, they control hyphal morphology, which has a direct impact on production processes that are often conducted in huge fermenter tanks. Here, we survey the occurrence and function of mating type genes that have been discovered in a wide range of industrial fungal producer strains. The possibility to obtain progeny from industrial producers by sexual mating provides an exciting alternative to conventional strain improvement programs aiming to generate optimized recombinant production strains.

Description: Endo-1,4-β-xylanases (EC 3.2.1.8) hydrolyze the 1,4-β-D-xylosidic linkages in xylans, the most abundant hemicellulose in plant cell walls. Xylanase enzymes have numerous industrial applications, including the manufacturing of animal feed, bread, juice and wine, pulp and paper, and biofuels. In this study, two glycosyl hydrolase family 10 members designated Gt Xyn10A and Gt Xyn10B and two glycosyl hydrolase family 11 members, Op Xyn11A and Cc Xyn11C, were functionally expressed and subjected to biochemical characterization. The K M , V max , and k cat values of the four xylanases, determined using birchwood xylan, ranged from 0.27 to 1.1 mg/mL, 130 to 980 μmol/min/mg, and 109 to 344 s −1 , respectively, where Op Xyn11A gave the highest and Gt Xyn10B the lowest values for all three parameters. Substrate specificity studies and analysis of the products released during the degradation of xylo-oligosaccharides and three types of xylan revealed significant differences in catalytic properties, particularly between Op Xyn11A and the other xylanases and between the family 10 and the family 11 xylanases. Molecular modeling suggests that the unique substrate specificity of Op Xyn11A can be attributed to the presence of a serine rather that an asparagine or aspartate residue at the +1 substrate binding site. Additionally, all four xylanases exhibited biochemical characteristics of interest for various commercial applications.

Description: Hand, foot, and mouth disease (HFMD) has caused significant morbidity and mortality in the Asia-Pacific regions, particularly in infants and young children. Coxsackievirus A16 (CA16) represents one of the major causative agents for HFMD, and the development of a safe and effective vaccine preventing CA16 infections has become a public health priority. In this study, we have developed a yeast system for the production of virus-like particles (VLPs) for CA16 by co-expressing P1 and 3CD of CA16 in Saccharomyces cerevisiae . These VLPs exhibit similarity in both protein composition and morphology as empty particles from CA16-infected cells. Immunization with CA16 VLPs in mice potently induced CA16-specific IgG and neutralization antibodies in a dose-dependent manner. IgG subclass isotyping revealed that IgG1 and lgG2b were dominantly induced by VLPs. Meanwhile, cytokine profiling demonstrated that immunization with VLPs significantly induced the secretion of IFN-γ, indicating potent cellular immune response. Furthermore, in vivo challenge experiments showed that passive immunization with anti-VLPs sera conferred full protection against lethal CA16 challenge in neonate mice. Taken together, our data demonstrated that VLPs produced in yeast might have the potential to be further developed as a vaccine candidate against HFMD.

Description: Studies were carried out to assess the utility of the cellular and extracellular constituents of Bacillus megaterium for the flotation of sphalerite and galena minerals. Based on the flotation results on the individual minerals, it was observed that sphalerite was preferentially floated compared to galena. A maximum selectivity index (SI) value of 11.7 was achieved in the presence of the soluble fraction of the thermolysed cells, which was higher than that obtained with the intact cells (SI of 6.5) and the insoluble fraction of the thermolysed cells (SI of 9.6). The results of the various enzymatic treatment tests revealed that extracellular DNA played a vital role in the selective flotation of sphalerite. A noteworthy finding was that the single-stranded DNA (ssDNA) had a higher biocollector capacity vis-à-vis the double-stranded DNA (dsDNA), leading to better flotation efficiency. About 95 % recovery of sphalerite could be achieved from the mineral mixture by the combined addition of the ssDNA with the non-DNA components of the bacterial cells, resulting in a maximum SI of 19.1. Calcium and phosphate components of the nutrient media were found to be essential for better selectivity of separation of sphalerite. The mechanisms of microbe–mineral interaction are discussed.

Description: While the use of anaerobic digestion to generate methane as a source of bioenergy is increasing worldwide, our knowledge of the microbial communities that perform biomethanation is very limited. Using next-generation sequencing, bacterial population profiles were determined in three full-scale mesophilic anaerobic digesters operated on dairy farms in the state of Vermont (USA). To our knowledge, this is the first report of a metagenomic analysis on the bacterial population of anaerobic digesters using dairy manure as their main substrate. A total of 20,366 non-chimeric sequence reads, covering the V1-V2 hypervariable regions of the bacterial 16S rRNA gene, were assigned to 2,176 operational taxonomic units (OTUs) at a genetic distance cutoff value of 5 %. Based on their limited sequence identity to validly characterized species, the majority of OTUs identified in our study likely represented novel bacterial species. Using a naïve Bayesian classifier, 1,624 anaerobic digester OTUs could be assigned to 16 bacterial phyla, while 552 OTUs could not be classified and may belong to novel bacterial taxonomic groups that have yet to be described. Firmicutes , Bacteroidetes , and Chloroflexi were the most highly represented bacteria overall, with Bacteroidetes and Chloroflexi showing the least and the most variation in abundance between digesters, respectively. All digesters shared 132 OTUs, which as a “core” group represented 65.4 to 70.6 % of sequences in individual digesters. Our results show that bacterial populations from microbial communities of anaerobic manure digesters can display high levels of diversity despite sharing a common core substrate.

Description: The discovery of Dehalococcoides mccartyi reducing perchloroethene and trichloroethene (TCE) to ethene was a key landmark for bioremediation applications at contaminated sites. D. mccartyi -containing cultures are typically grown in batch-fed reactors. On the other hand, continuous cultivation of these microorganisms has been described only at long hydraulic retention times (HRTs). We report the cultivation of a representative D. mccartyi -containing culture in continuous stirred-tank reactors (CSTRs) at a short, 3-d HRT, using TCE as the electron acceptor. We successfully operated 3-d HRT CSTRs for up to 120 days and observed sustained dechlorination of TCE at influent concentrations of 1 and 2 mM TCE to ≥97 % ethene, coupled to the production of 10 12 D. mccartyi cells L culture −1 . These outcomes were possible in part by using a medium with low bicarbonate concentrations (5 mM) to minimize the excessive proliferation of microorganisms that use bicarbonate as an electron acceptor and compete with D. mccartyi for H 2 . The maximum conversion rates for the CSTR-produced culture were 0.13 ± 0.016, 0.06 ± 0.018, and 0.02 ± 0.007 mmol Cl − L culture −1 h −1 , respectively, for TCE, cis -dichloroethene, and vinyl chloride. The CSTR operation described here provides the fastest laboratory cultivation rate of high-cell density Dehalococcoides cultures reported in the literature to date. This cultivation method provides a fundamental scientific platform for potential future operations of such a system at larger scales.

Description: Long-term fructose consumption has been shown to evoke leptin resistance, to elevate triglyceride levels and to induce insulin resistance and hepatic steatosis. Autophagy has been suggested to function in processes such as lipid storage in adipose tissue and inflammation in liver. Autophagy and the leptin system have also been suggested to regulate each other. This study aimed to identify the changes caused by fetal undernourishment and postnatal fructose diet in the gene expression of leptin, its receptors (LEPR-a, LEPR-b, LEPR-c, LEPR-e and LEPR-f) and autophagy genes in the white adipose tissue (WAT) and liver of adult male rats in order to clarify the mechanism behind the metabolic alterations. The data clearly revealed that the long-term postnatal fructose diet decreased leptin levels ( p  〈 0.001), LEPR ( p  〈 0.001), especially LEPR-b ( p  = 0.011) and LEPR-f ( p  = 0.005), as well as SOCS3 ( p  〈 0.001), ACC ( p  = 0.006), ATG7 ( p  〈 0.001), MAP1LC3β ( p  〈 0.001) and LAMP2 ( p  = 0.004) mRNA expression in WAT. Furthermore, LEPR ( p  〈 0.001), especially LEPR-b ( p  = 0.001) and LEPR-f ( p  〈 0.001), ACC ( p  = 0.010), ATG7 ( p  = 0.024), MAP1LC3β ( p  = 0.003) and LAMP2 ( p  〈 0.001) mRNA expression in the liver was increased in fructose-fed rats. In addition, the LEPR expression in liver and MAP1LC3β expression in WAT together explained 55.7 % of the variation in the plasma triglyceride levels of the rats ( R adj. 2  = 0.557, p  〈 0.001). These results, together with increased p62 levels in WAT ( p  〈 0.001), could indicate decreased adipose tissue lipid storing capacity as well as alterations in liver metabolism which may represent a plausible mechanism through which fructose consumption could disturb lipid metabolism and result in elevated triglyceride levels.

Description: κ-Carrageenases exhibit apparent distinctions in gene sequence, molecular weight, enzyme properties, and posttranslational processes. In this study, a new κ-carrageenase gene named cgk Z was cloned from the marine bacterium Zobellia sp. ZM-2. The gene comprised an open reading frame of 1,638 bp and encoded 545 amino acids. The natural signal peptide of κ-carrageenase was used successfully for the secretory production of the recombinant enzyme in Escherichia coli . A posttranslational process that removes an amino acid sequence of about 20 kDa from the C-terminal end of κ-carrageenase was first discovered in E. coli . An increase in enzyme activity by 167.3 % in the presence of 5 mM DTT was discovered, and Na + at a certain concentration range was positively correlated with enzyme activity. The κ-carrageenase production of E. coli was 9.0 times higher than that of ZM-2. These results indicate the potential use of the enzyme in the biotechnological industry.

Description: Fifty-five bacterial isolates were obtained from surface-sterilized nodules of woody and shrub legumes growing in Ethiopia: Crotalaria spp., Indigofera spp., and Erythrina brucei , and the food legumes soybean and common bean. Based on partial 16S rRNA gene sequence analysis, the majority of the isolates were identified as Gram-negative bacteria belonging to the genera Achromobacter , Agrobacterium , Burkholderia , Cronobacter , Enterobacter , Mesorhizobium , Novosphingobium , Pantoea , Pseudomonas , Rahnella , Rhizobium , Serratia , and Variovorax . Seven isolates were Gram-positive bacteria belonging to the genera Bacillus , Paenibacillus , Planomicrobium , and Rhodococcus . Amplified fragment length polymorphism (AFLP) fingerprinting showed that each strain was genetically distinct. According to phylogenetic analysis of recA , glnII , rpoB , and 16S rRNA gene sequences, Rhizobium , Mesorhizobium , and Agrobacterium were further classified into six different genospecies: Agrobacterium spp., Agrobacterium radiobacter , Rhizobium sp., Rhizobium phaseoli , Mesorhizobium sp., and putative new Rhizobium species. The strains from R . phaseoli , Rhizobium sp. IAR30, and Mesorhizobium sp. ERR6 induced nodules on their host plants. The other strains did not form nodules on their original host. Nine endophytic bacterial strains representing seven genera, Agrobacterium , Burkholderia , Paenibacillus , Pantoea , Pseudomonas , Rhizobium , and Serratia , were found to colonize nodules of Crotalaria incana and common bean on co-inoculation with symbiotic rhizobia. Four endophytic Rhizobium and two Agrobacterium strains had identical nifH gene sequences with symbiotic Rhizobium strains, suggesting horizontal gene transfer. Most symbiotic and nonsymbiotic endophytic bacteria showed plant growth-promoting properties in vitro, which indicate their potential role in the promotion of plant growth when colonizing plant roots and the rhizosphere.

Description: Escherichia coli BA002, in which the ldhA and pflB genes are deleted, cannot utilize glucose anaerobically due to the inability to regenerate NAD + . To restore glucose utilization, overexpression of nicotinic acid phosphoribosyltransferase (NAPRTase) encoded by the pncB gene, a rate-limiting enzyme of NAD(H) synthesis pathway, resulted in a significant increase in cell mass and succinate production under anaerobic conditions. However, a high concentration of pyruvate accumulated. Thus, co-expression of NAPRTase and the heterologous pyruvate carboxylase (PYC) of Lactococcus lactis subsp. cremoris NZ9000 in recombinant E . coli BA016 was investigated. The total concentration of NAD(H) was 9.8-fold higher in BA016 than in BA002, and the NADH/NAD + ratio decreased from 0.60 to 0.04. Under anaerobic conditions, BA016 consumed 17.50 g l −1 glucose and produced 14.08 g l −1 succinate with a small quantity of pyruvate. Furthermore, when the reducing agent dithiothreitol or reduced carbon source sorbitol was added, the cell growth and carbon source consumption rate of BA016 was reasonably enhanced and succinate productivity increased.

Description: 1,2-Benzenedicarboxaldehyde-3,4,5-trihydroxy-6-methyl (flavipin) was found to be antagonistic against nematodes and fungi. Here we demonstrated that flavipin is a potent antioxidant in vitro and in vivo, which has great potential in the therapy for free radical-associated diseases. Therefore, flavipin-producing bio-source was screened from 80 endophytes in Ginkgo biloba . Seven endophytic fungi were able to synthesize antioxidant substances and identified by ITS rDNA sequences. Among them, Chaetomium globosum CDW7 was a remarkable producer of flavipin. The fermentation parameters of CDW7 were then optimized for high flavipin production. Cultured under the optimal condition (25 °C, 100/250 mL flask, 12 discs/flask, 150 rpm, pH 6.5) for 14 days, CDW7 was able to synthesize flavipin at a production of 315.5 mg/L. In addition, flavipin output was positively correlated to antioxidant activities of crude extracts with a correlation coefficient of 0.8235, indicating that flavipin was the major antioxidant component of CDW7's metabolites. These data demonstrated that CDW7 was a highly yielded bio-source of antioxidant flavipin.

Description: Two bacterial hosts expressing cloned aromatic oxygenases were used to catalyze the oxidation and polymerization of indole and related substrates, creating mixtures of indigoid compounds comprised of novel dimers and trimers. Crude extracts and purified compounds were tested for their ability to inhibit the growth of Gram-positive organisms, in general, and Mycobacterium tuberculosis (TB), in particular. Of the 74 compounds tested against M. tuberculosis , ~66 % had minimum inhibitory concentrations (MIC) of 5 μg/ml or less. The most effective antibiotic found was designated SAB-P1, a heterodimer of indole and anthranil, which had a MIC of 0.16 μg/ml, and did not inhibit kidney cells (IC 50 ) at concentrations of 〉8 μg/ml. Combinatorial biocatalysis was used to create a series of halogenated derivatives of SAB-P1 with a wider therapeutic window. None of the derivatives had MIC values that were superior to SAB-P1, but some had a wider therapeutic window because of decreased kidney cell toxicity. Generally, the indigoid dimers that were effective against TB appeared to be specific for TB. Some of the trimers generated, however, had a broader spectrum of activity inhibiting not only TB (MIC = 1.1 μg/ml) but also the growth of Mycobacterium smegmatis MC2 155, Bacillus cereus , Enterococcus faecalis , Staphylococcus epidermidis , Bacillus subtilis 168, and Clostridium acetobutylicum . The structure of two of the novel dimers (SAB-C4 and SAB-P1) and a trimer (SAB-R1) were solved using X-ray crystallography.

Description: This paper demonstrates a way to utilize the rheological properties of high consistency microfibrillated and nanofibrillated cellulose (MFC and NFC) based furnishes for improved dewatering. This is relevant to a new manufacturing platform that is being developed to form composite webs from suitable mixtures of MFC or NFC, traditional pulp fibres and pigments. The studied furnishes were evaluated in the consistencies range of 5–15 % with an MCR 300 rheometer and an immobilization cell. This setup enables us to characterize the rheology of the samples before and during the dewatering process. Classical rheological methods are used to characterise MFC and NFC furnishes. Yield stress as an indicator of the flocculated network strength was found to increase with the consistencies, following the increase in elastic moduli, which indicated a gel-like strongly flocculated matrix. The shear thinning properties of furnishes are observed to follow the Oswald’s rheological model on a wide range of shear rates. It was found that when the MFC and NFC furnishes were dewatered under vacuum conditions, the final solids content was increased with application of shear. This behaviour is more pronounced for furnishes which contained the more swollen NFC (higher WRV, i.e. higher zeta potential). This effect is further exemplified by the change of the complex and dynamic viscosities during the dewatering. The shear rate, the fibre content, and the furnish consistencies were also found to influence the dewatering rate.

Description: The compartmentalized anaerobic reactor (CAR) is a patent novel high-rate reactor, which consists of three compartments. The reactor has a great potential for application due to its many advantages. In this work, the microbial consortium, spatial distribution, and their relationship with performance of CAR were investigated by means of polymerase chain reaction, denaturing gradient gel electrophoresis, and fluorescence in situ hybridization. The results showed that the predominant archaea were Methanobacterium , Methanosaeta , and Methanospirillum , and the predominant bacteria were Firmicutes , Deltaproteobacteria , Spirochaetes , Actinobacteria , and Gammaproteobacteria in the microbial consortium. The methanogenic archaea (MA), the hydrogen-producing acetogenic bacteria (HAB), and the hydrolytic fermentative bacteria (HFB) were found to be predominant in the upper, middle, and bottom compartments, respectively. The results revealed that the granular sludge took on a stratified microbial structure. The HFB, HAB, and MA were located in the outer shell, middle layer, and core, respectively. The microbial populations from the bottom compartment were relatively homogeneous in the granular sludge, and from the middle and upper compartments, they were relatively heterogeneous in the granular sludge. The microbial consortia and their spatial distribution were in accordance with the organic loading rate and chemical components in the three compartments.

Description: We constructed beta-glucosidase (BGL)-displaying Corynebacterium glutamicum , and direct l -lysine fermentation from cellobiose was demonstrated. After screening active BGLs, Sde1394, which is a BGL from Saccharophagus degradans , was successfully displayed on the C. glutamicum cell surface using porin as an anchor protein, and cellobiose was directly assimilated as a carbon source. The optical density at 600 nm of BGL-displaying C. glutamicum grown on cellobiose as a carbon source reached 23.5 after 48 h of cultivation, which was almost the same as that of glucose after 24 h of cultivation. Finally, Sde1394-displaying C. glutamicum produced 1.08 g/l of l -lysine from 20 g/l of cellobiose after 4 days of cultivation, which was about threefold higher than the amount of produced l -lysine using BGL-secretory C. glutamicum strains (0.38 g/l after 5 days of cultivation). This is the first report on amino acid production using cellobiose as a carbon source by BGL-expressing C. glutamicum .

Description: The risk of antibody-dependent enhancement (ADE) of dengue virus (DENV) infection is a major obstacle for the development of dengue vaccine candidates. Here, we described a novel approach for assessment of ADE by measuring DENV nonstructural protein 1 (NS1) production in culture supernatants with Fcγ receptor-expressing K562 cells in ELISA format (ELISA-ADE). Enhancing activities quantified by measurement of kinetics of NS1 production were in a good agreement with the results of the virus titration assay. In conjunction with the previously established enzyme-linked immunospot-based micro-neutralization test (ELISPOT-MNT) in 96-well format, the observable dose–response profiles of enhancing and neutralizing activities against all four DENV serotypes were produced with two flaviviral envelope cross-reactive monoclonal antibodies and four primary DENV-1-infected human sera. The simple high-throughput ELISA-ADE assay offers advantages for quantitative measurement of infection enhancement that can potentially be applied to large-scale seroepidemiological studies of DENV infection and vaccination.

Description: Increasing concerns over limited petroleum resources and associated environmental problems are motivating the development of efficient cell factories to produce chemicals, fuels, and materials from renewable resources in an environmentally sustainable economical manner. Bacillus spp., the best characterized Gram-positive bacteria, possesses unique advantages as a host for producing microbial enzymes and industrially important biochemicals. With appropriate modifications to heterologous protein expression and metabolic engineering, Bacillus species are favorable industrial candidates for efficiently converting renewable resources to microbial enzymes, fine chemicals, bulk chemicals, and fuels. Here, we summarize the recent advances in developing Bacillus spp. as a cell factory. We review the available genetic tools, engineering strategies, genome sequence, genome-scale structure models, proteome, and secretion pathways, and we list successful examples of enzymes and industrially important biochemicals produced by Bacillus spp. Furthermore, we highlight the limitations and challenges in developing Bacillus spp. as a robust and efficient production host, and we discuss in the context of systems and synthetic biology the emerging opportunities and future research prospects in developing Bacillus spp. as a microbial cell factory.

Description: Glucosamine (GlcN), an amino sugar, is a compound derived from substitution of a hydroxyl group of a glucose molecule with an amino group. GlcN and its acetylated derivative, N -acetylglucosamine (GlcNAc), have been widely used in food, cosmetics, and pharmaceutical industries and are currently produced by acid hydrolysis of chitin (a linear polymer of GlcNAc) extracted from crab and shrimp shells. Microbial fermentation by filamentous fungi or recombinant Escherichia coli , as an alternative method for the production of GlcN and GlcNAc, is attracting increasing attention because it is an environmentally friendly process. Although the microbial production of GlcN and GlcNAc is hampered by low yield and high production cost, considerable advances have been made in recent years. Here we review the applications, commercial market, and production of GlcN and GlcNAc, with emphasis on the metabolic and process engineering strategies used to improve GlcN and GlcNAc production by recombinant microbes.

Description: Steryl ferulates (SFs) are ferulic acid esters of phytosterols and/or triterpene alcohols which have potential as frying oil antioxidants. The objective of this study was to evaluate the anti-polymerization and antioxidant activity at frying temperatures of corn steryl ferulates (CSFs), rice steryl ferulates (oryzanol), and a mixture of CSFs with oryzanol, alone and with tocopherols. Antioxidant activity was measured by the reduction of polymerized triacylglycerol formation, and loss of olefinic and bisallylic protons from fatty acid double bonds by 1 H NMR. CSFs and oryzanol slowed the oxidation and polymerization of soybean oil triacylglycerols heated to 180 °C more effectively than a mixture of alpha and gamma tocopherols. CSFs were more effective at preventing polymerization than oryzanol, but when oryzanol was combined with tocopherols, they all had similar antioxidant activity. In addition, tocopherols had a protective effect on SFs. Corn SFs were degraded more quickly during heating than oryzanol, however, the phytosterol constituents of corn SFs, sitostanol and campestanol, were actually more resistant to degradation compared to the phytosterol constituents of rice SFs. Results demonstrate that corn and rice SFs may be effective antioxidants for use in frying oils, and that their activity is enhanced in the presence of tocopherols.

Description: Cotton fabrics with antibacterial properties were prepared by the treatment with 3,3′4,4′-benzophenone tetracarboxylic dianhydride (BPTCD) in a combined process of shaking immersion in dyeing machine and pad-dry-cure. Environmentally-benign choline chloride (ChCl)-based deep eutectic solvents (DESs) were mainly examined as treatment media instead of using organic solvent. The results revealed that cotton fabrics treated with BPTCD in urea-ChCl DES showed a strong ester carbonyl peak in fourier transform infrared (FTIR) analysis, indicating fixation of BPTCD on cotton cellulose. Detailed characterizations of the BPTCD-treated cotton were carried out by FTIR, thermogravimetric analysis, scanning electron microscopy, dye staining, and evaluation of hydrophilicity and strength. The treated fabrics demonstrated a high level of antibacterial characteristics before and after UV irradiation. This indicated that addition of ChCl could enhance antibacterial activity of cotton before UV irradiation. Therefore, use of ChCl-based DES along with BPTCD incorporation provided environmentally-acceptable and economically-feasible treatment process for preparation of novel antibacterial cotton.

Description: In this work, the successful coupling of enzymatic oxidation and aldol addition reactions for the synthesis of a Cbz-aminopolyol from a Cbz-amino alcohol was achieved for the first time in a multienzymatic one-pot system. The two-step cascade reaction consisted of the oxidation of Cbz-ethanolamine to Cbz-glycinal catalyzed by chloroperoxidase from the fungus Caldariomyces fumago and aldol addition of dihydroxyacetone phosphate to Cbz-glycinal catalyzed by rhamnulose-1-phosphate aldolase expressed as a recombinant enzyme in Escherichia coli , yielding (3 R ,4 S )-5-{[(benzyloxy)carbonyl]amino}-5-deoxy-1- O -phosphonopent-2-ulose. Tools of enzymatic immobilization, reactor configurations, and modification of the reaction medium were applied to highly increase the production of the target compound. While the use of soluble enzymes yielded only 23.6 % of Cbz-aminopolyol due to rapid enzyme inactivation, the use of immobilized ones permitted an almost complete consumption of Cbz-ethanolamine, reaching Cbz-aminopolyol yields of 69.1 and 71.9 % in the stirred-tank and packed-bed reactor, respectively. Furthermore, the reaction production was 18-fold improved when it was catalyzed by immobilized enzymes in the presence of 5 % ( v / v ) dioxane, reaching a value of 86.6 mM of Cbz-aminopoliol (31 g/L).

Description: Rod-like cellulose nanowhiskers and spherical cellulose nanoparticles were prepared from wood-pulp-derived cellulose powder by mechanical refining processes such as high-pressure homogenization (HPH) and ball-milling (BM). The nanowhiskers obtained by the HPH method were found to be 200–500 nm long and 11–16 nm wide. The diameters of the nanoparticles were in the range 40–200 nm, depending on the BM time, and were reduced to 25–50 nm after extra HPH. By adjusting the BM time, cellulose nanoparticles having different polymorphs with similar morphologies were prepared. The X-ray diffraction patterns revealed the recrystallization of cellulose I (1 h of BM time) or cellulose II (4–8 h of BM time) in ball-milled nanoparticles after water washing and solvent exchange treatments. The nanowhisker widths derived from the specific surface areas (SSA) by adsorption methods such as Congo red dye, nitrogen, and water vapor, sorptions were in agreement with those obtained from transmission electron microscopy and atomic force microscopy images. Similar SSA values were obtained for micro- and nano-scale cellulose materials using water vapor adsorption methods, and the SSAs of nanoparticles obtained by different adsorption methods are also discussed.

Description: The complete genome of Acinetobacter oleivorans DR1 contains AqsR and AqsI genes, which are LuxR and LuxI homolog, respectively. In a previous study, we demonstrated that quorum sensing (QS) signals play an important role in biofilm formation and hexadecane biodegradation. However, the regulation of genes controlled by the QS system in DR1 remains unexplored. We constructed an aqsR mutant and performed RNA sequencing analysis to understand the QS system. A total of 353 genes were differentially expressed during the stationary phase of wild-type cells compared to that of the aqsR mutant. AqsR appears to be an exceptionally important regulator because knockout of aqsR affected global gene expression. Genes involved in posttranslational modification, chaperones, cell wall structure, secondary metabolites biosynthesis, and stress defense were highly upregulated only in the wild type. Among upregulated genes, both the AOLE_03905 (putative surface adhesion protein) and the AOLE_11355 (L-asparaginase) genes have putative LuxR binding sites at their promoter regions. Soluble AqsR proteins were successfully purified in Escherichia coli harboring both aqsR and aqsI . Comparison of QS signals in an AqsI–AqsR co-overexpression strain with N -acyl homoserine lactone standards showed that the cognate N -acyl homoserine lactone binding to AqsR might be 3OH C12HSL. Our electrophoretic mobility shift assays with purified AqsR revealed direct binding of AqsR to those promoter regions. Our data showed that AqsR functions as an important regulator and is associated with several phenotypes, such as hexadecane utilization, biofilm formation, and sensitivity to cumene hydroperoxide.

Description: A membrane-bound, pyrroloquinoline quinone (PQQ)-dependent alcohol dehydrogenase (ADH) was purified from Frateuria aurantia LMG 1558 T . Although F . aurantia belongs to a group of γ-Proteobacteria, the characteristics of its PQQ-ADH were similar to the enzyme characteristics of the typical high-acetic acid-resistant bacterium Gluconacetobacter europaeus from the group of α-Proteobacteria. The PQQ-dependent ADH was solubilized from the membranes and purified after anionic, cationic, and affinity chromatography with specific activity of 117 U/mg. The purified enzyme was estimated to be composed of two subunits of ca. 72 and 45 kDa, as judged by SDS-polyacrylamide gel electrophoresis. The purified enzyme had maximum activity at pH 4.5 and showed the highest substrate specificity to ethanol, isoamyl alcohol, 1-butanol, and 1-propanol. The deduced sequences of cloned genes adhA and adhB encoding subunits I and II of PQQ-ADH showed 80 % amino acid (AA) identity to AdhA and 68 % AA identity to AdhB of Ga . europaeus V3 (LMG 18494). Because of the high similarity between genes encoding subunits I and II of PQQ-ADH and its homologous genes found in a distantly related taxonomic group of acetic acid bacteria, the results suggest the possibility of horizontal gene transfer between these two groups of genera.

Description: Enantioselective oxidation of racemic phenyl-1,2-ethanediol into ( R )-(−)-mandelic acid by a newly isolated Brevibacterium lutescens CCZU12-1 was demonstrated. It was found that optically active ( R )-(−)-mandelic acid ( e.e.p  〉 99.9 %) is produced leaving the other enantiomer ( S )-(+)-phenyl-1,2-ethanediol intact. Using fed-batch method, a total of 172.9 mM ( R )-(−)-mandelic acid accumulated in the reaction mixture after the seventh feed. Moreover, oxidation of phenyl-1,2-ethanediol using calcium alginate-entrapped resting cells was carried out in the aqueous system, and efficient biocatalyst recycling was achieved as a result of cell immobilization in calcium alginate, with a product-to-biocatalyst ratio of 27.94 g ( R )-(−)-mandelic acid g −1 dry cell weight cell after 16 cycles of repeated use.

Description: It is becoming increasingly apparent that responsiveness to dietary fat composition is heterogeneous and dependent on the genetic make-up of the individual. The aim of this study was to evidence a genotype-related differential effect of n-3 long-chain polyunsaturated fatty acids (n-3 LC-PUFA) on the modulation of hepatic genes involved in cholesterol metabolism. Fourteen spontaneously hypertensive (SH) rats, which present a naturally occurring variation in the gene encoding for sterol responsive element binding protein 1 (SREBP-1), contributing to their inherited variation in lipid metabolism, and 14 Wistar-Kyoto (WK) rats were fed a control diet or an n-3 LC-PUFA enriched diet for 90 days. Plasma lipid profile, total lipid fatty acid composition in plasma and liver, and the expression of SREBP-1 and 2, 3-hydroxy-3-methyl-glutaryl-CoA reductase, low-density lipoprotein receptor, and acyl-CoA:cholesterol acyltransferase 2 encoding genes and proteins were determined. The positive effect of the enriched diet on the serum lipid profile, particularly on total cholesterol and triglyceride level, was clearly evidenced in both WK and SH rats, but n-3 LC-PUFA acted through a different modulation of gene and protein expression that appeared related to the genetic background. Our study evidences a different transcriptional effect of specific nutrients related to genetic variants.

Description: The bacterium Bacillus amyloliquefaciens anti-CA isolated from mangrove system was found to be able to actively kill Candida albicans isolated from clinic. The bacterial strain anti-CA could produce high level of bioactive substance, amylase and protease in the cheap medium containing 2.0 % soybean meal, 2.0 % wheat flour, pH 6.5 within 26 h. After purification, the main bioactive substance was confirmed to be a cyclic lipopeptide containing a heptapeptide, L-Asp→L-Leu→L-Leu→L-Val→L-Val→L-Glu→L-Leu and a 3-OH fatty acid (15 carbons). In addition to C. albicans , the purified lipopeptide can also kill many yeast strains including Metschnikowia bicuspidata , Candida tropicalis , Yarrowia lipolytica and Saccharomyces cerevisiae. After treated by the purified lipopeptide, both the whole cells and protoplasts of C. albicans were destroyed.

Description: The purpose of the present study was to investigate the osteoprotective effects of soybean oil (SbO) and sesame oil (SO) in ovarictomized (OVX) rats. The results indicated that the OVX rats exhibited a significant decrease in Ca and P level in both serum and bone, the activities of the antioxidant enzymes SOD and CAT and the antioxidant biomarker GSH accompanied with a marked increase in the oxidative stress markers MDA and PC, the inflammatory indices (TNF-α, CRP levels, WBCs counts and ACP activity) in, both, bone and serum. Supplementating the diet of the OVX rats with SbO (15 % w/w) or SO (10 % w/w) for 2 months to resulted in modulation of the alterations in all tested parameters and succeeded to restore minerals, antioxidant enzymes, antioxidant biomarkers, oxidative stress markers, inflammatory indices, and WBCs counts. It could be concluded that the consumption of diets supplemented with SbO or SO might be useful for preventing bone loss caused by estrogen deficiency in ovariectomy status.

Description: The epothilones, compounds with anticancer mechanisms similar to that of paclitaxel (Taxol), are produced by strains of the myxobacterium Sorangium cellulosum , and the gene cluster responsible for epothilone biosynthesis is organised as a large operon. In this work, we showed that the 440-bp promoter regions of the operons from eight S. cellulosum strains have 94.27 % DNA sequence identity and 50 % variability in promoter activity in Escherichia coli . A primer extension analysis revealed two transcriptional start sites (TSSs) at 246 (TSS1) and 193 bp (TSS2) upstream of the translation start site (TLS), respectively. Promoter truncation determined that the basal promoter from the So0157-2 strain is located within a 264-bp region containing weak promoter activity; whereas in the 38-bp region upstream, the 264-bp promoter was required for the strong promoter activity, which was dramatically increased by 11-fold in average. There was a conserved stem–loop structure between TSS2 and the TLS, which was identified in E. coli as a negative regulatory element. In addition, the upstream non-conserved 357-bp non-coding region contributes to the promoter activity, increasing it by 1.5-fold. In conclusion, the expression of the epothilone operon non-coding region in E. coli is regulated by a double promoter (with −35 and −10 regions and two distinct TSSs), a stem–loop structure, and a distal non-coding region.

Description: Bacterial community compositions were characterized using denaturing gradient gel electrophoresis analysis of bacterial 16S rRNA gene in the sediments of the Pearl River estuary. Sequencing analyses of the excised bands indicated that Gram-negative bacteria, especially Gammaproteobacteria , were dominant in the Pearl River estuary. The diversity of polycyclic aromatic hydrocarbon ring-hydroxylating dioxygenase (PAH-RHD) gene in this estuary was then assessed by clone library analysis. The phylogenetic analyses showed that all PAH-RHD gene sequences of Gram-negative bacteria (PAH-RHD [GN] ) were closely related to the nagAc gene described for Ralstonia sp. U2 or nahAc gene for Pseudomonas sp. 9816–4, while the PAH-RHD gene sequences of Gram-positive bacteria (PAH-RHD [GP] ) at sampling site A1 showed high sequence similarity to the nidA gene from Mycobacterium species. Meanwhile, molecular diversity of the two functional genes was higher at the upstream of this region, while lower at the downstream. Redundancy analysis indicated that environmental factors, such as NH 4 -N, ∑PAHs, pH, SiO 3 -Si, and water depth, affected the distribution of the PAH-RHD [GN] gene in the Pearl River estuary.

Description: In order to provide a more suitable response to public health concerns, we improved the detection of infectious human adenoviruses in water by optimising the commonly used integrated cell culture–PCR method. Risk evaluation studies seek for rapid detection of infectious adenoviruses, including the enteric types 40 and 41 that are considered as the second most common agents of gastroenteritis in children next to rotaviruses. The here-employed 293A cell line used for infectious status assessment showed its ability to multiply adenoviruses including type 41. Two modifications were moreover applied to the workflow for viral detection. The first occurred at the nucleic acid extraction step performed directly on all infected cells, while the second was the application of real-time quantitative PCR as detection tool. All adaptations led to a 3-day reduction of the response delay and an improved sensitivity especially for the enteric adenoviral types. The infectious status of laboratory strain types 2 and 41 was demonstrated by a more than 2-log 10 increase in genome quantity. These conclusions were confirmed and reinforced by the analysis of water samples applying the improved assay. Naturally occurring infectious adenoviruses were detected in wastewater and river water, within 2 days. Types belonging to the species human adenoviruses C and type 31 were observed, but the most frequently identified type was 41 (71 % of identified sequences, n  = 34). This highlights the usefulness of our method for a wide range of types, and especially for the most prevalent and public health-relevant enteric adenoviruses.

Description: In this paper, nanofibrillated cellulose/carboxymethyl cellulose (CMC) composite films were prepared using tape casting. The obtained transparent films showed shear induced partial alignment of fibrils along the casting direction, resulting in birefringence in cross polarized light. The carboxyl groups of CMC could be further utilized to create ionic crosslinking by treatment with glycidyl trimethyl ammonium chloride (GTMA). The GTMA treated composite films had improved mechanical properties both in wet and dry state. The chemical composition and morphologies of composites were analyzed with X-ray photoelectron spectroscopy, elemental analysis, scanning electron microscopy and wide-angle X-ray scattering.

Description: Dietary phosphatidylinositol (PI) can be synthesized via phospholipase D (PLD)-catalyzed transphosphatidylation of phosphatidylcholine (PC), abundant in soy lecithin, with myo-inositol. However, a generated mixture of phospholipid (PL) classes poses a challenge for analysis. Our current work on Streptomyces PLD engineering requires a robust analytical method for profiling of PI and related PLs derived from the transphosphatidylation reactions. Therefore, we optimized an HPLC-based method with charged aerosol detector (CAD) for PL quantification. PLs were separated on a normal phase silica column by a gradient elution system using two solvents containing chloroform/methanol/1 M formic acid–triethylamine buffer in different ratios. Retention times of the PL standards and LC–MS under identical conditions were used to identity PL classes. PL standards gave linear response in 100- and 10-fold (lyso-PI) concentration range. The method provided a simple, sensitive, repeatable, and precise analysis of PI, PC, phosphatidylethanolamine, phosphatidic acid, and lyso forms of PC and PI. Compared to the similar existing method, introduction of CAD provided a three- to fivefold decrease at the lower end and a two- to fivefold increase at the upper end of the dynamic range. High precision, high sensitivity, and low limits of detection and quantification further underline the benefits of CAD in PL analysis.

Description: Frying is a popular practice because of its unique sensory characteristics and low cost. The high temperature reached with this cooking method alters molecules present in the oil. The deterioration of the oil depends primarily on its chemical composition. The aim of this study was to evaluate the thermal stability of high oleic sunflower oil (HOSO), sunflower oil (SO) and mixed oil (MIX) during deep frying of French fries. Octanoic acid and unsaturated fatty acid (UFA)/saturated fatty acid (SFA) ratio showed a good correlation with total polar compounds (TPC) for all frying samples analyzed. HOSO and MIX were characterized by reduced levels of thermal degradation, while SO resulted in the highest values of oxidation products (highest TPC values). SO was also the oil more retained by the food matrix, whereas MIX was the least absorbed. HOSO and MIX, having a high oleic acid content (77.58 and 59.92 %, respectively) and a low linoleic acid content (13.42 and 25.70 %, respectively), showed the best characteristics for the frying process.

Description: Acrolein, which is an irritating and off-flavor compound formed during heating of vegetable oils, was estimated by the gas–liquid chromatography (GLC). Several vegetable oils such as high-oleic sunflower, perilla, rapeseed, rice bran, and soybean oils were heated at 180 °C for 480 min and then the concentration of acrolein in the head space gas was determined by GLC. The formation of acrolein was greatest in perilla oil among the tested oils, while it was much lower in rice bran oil and high-oleic sunflower oil. There was a good correlation between the level of acrolein and linolenate (18:3n-3) in the vegetable oils. To investigate the formation of acrolein from linolenate, methyl oleate, methyl linoleate, and methyl linolenate were also heated at 180 °C, and the amounts of acrolein formed from them were determined by GLC. The level of acrolein was the greatest in methyl linolenate. Acrolein was also formed from methyl linoleate, but not from methyl oleate. Acrolein in vegetable oils may be formed from polyunsaturated fatty acids, especially linolenic acid but not from glycerol backbone in triacylglycerols.

Description: Proteins with a Zn(II) 2 Cys 6 domain, Cys-X 2 -Cys-X 6 -Cys-X 5-12 -Cys-X 2 -Cys-X 6-9 -Cys (hereafter, referred to as the C6 domain), form a subclass of zinc finger proteins found exclusively in fungi and yeast. Genome sequence databases of Saccharomyces cerevisiae and Candida albicans have provided an overview of this family of genes. Annotation of this gene family in most fungal genomes is still far from perfect and refined bioinformatic algorithms are urgently needed. Aspergillus flavus is a saprophytic soil fungus that can produce the carcinogenic aflatoxin. It is the second leading causative agent of invasive aspergillosis. The 37-Mb genome of A . flavus is predicted to encode 12,000 proteins. Two and a half percent of the total proteins are estimated to contain the C6 domain, more than twofold greater than those estimated for yeast, which is about 1 %. The variability in the spacing between cysteines, C 3 -C 4 and C 5 -C 6 , in the zinc cluster enables classification of the domains into distinct subgroups, which are also well conserved in Aspergillus nidulans . Sixty-six percent (202/306) of the A . flavus C6 proteins contain a specific transcription factor domain, and 7 % contain a domain of unknown function, DUF3468. Two A . nidulans C6 proteins containing the DUF3468 are involved in asexual conidiation and another two in sexual differentiation. In the anamorphic A . flavus , a homolog of the latter lacks the C6 domain. A . flavus being heterothallic and reproducing mainly through conidiation appears to have lost some components involved in homothallic sexual development. Of the 55 predicted gene clusters thought to be involved in production of secondary metabolites, only about half have a C6-encoding gene in or near the gene clusters. The features revealed by the A . flavus C6 proteins likely are common for other ascomycete fungi.

Description: A range of isolates of Pseudomonas aeruginosa from widely different environmental sources were examined for their ability to synthesise rhamnolipid biosurfactants. No significant differences in the quantity or composition of the rhamnolipid congeners could be produced by manipulating the growth conditions. Sequences for the rhamnolipid genes indicated low levels of strain variation, and the majority of polymorphisms did lead to amino acid sequence changes that had no evident phenotypic effect. Expression of the rhlB and rhlC rhamnosyltransferase genes showed a fixed sequential expression pattern during growth, and no significant up-regulation could be induced by varying producer strains or growth media. The results indicated that rhamnolipids are highly conserved molecules and that their gene expression has a rather stringent control. This leaves little opportunity to manipulate and greatly increase the yield of rhamnolipids from strains of P. aeruginosa for biotechnological applications.

Description: Streptococcus alactolyticus strain FGM, isolated from chicken cecum, was used to increase the extract yield of polysaccharides during Astragalus membranaceus fermentation. It was previously demonstrated that polysaccharides from fermented A . membranaceus by S . alactolyticus had some similar properties to those from A . membranaceus in terms of its ability to help heal hepatic fibrosis in rats and modulate immunopotentiation of broiler chicken. However, methods to increase the yield of the polysaccharides during fermentation of A . membranaceus are not well understood. In this paper, we investigated the involvement of uridine diphosphate (UDP)-glucose 4-epimerase ( galE ) and glucan-1,6-α-glucosidase ( dexB ) during A . membranaceus fermentation through real-time reverse transcription quantitative PCR. The galE and dexB genes of S . alactolyticus were cloned by homology-based cloning and the genome walking method for the first time, and the 3D structure of dexB was analyzed by Swiss-PdbViewer 4.0.1 software. The expression of both the galE and dexB genes in A . membranaceus fermentation was studied using the determined ideal reference gene ldh for transcript normalization. The results showed that these two genes were both highly induced and peaked after 12 h of fermentation. The expression level of galE was stepwise increased from 48 to 72 h, while dexB transcripts were markedly increased at 60 h and decreased by 72 h. These data suggested that dexB and galE of S . alactolyticus strain FGM were involved in the regulation of A . membranaceus fermentation and they might play some roles in the increase of polysaccharides.

Description: Rhodococcus erythropolis WZ010 was capable of producing optically pure (2 S ,3 S )-2,3-butanediol in alcoholic fermentation. The gene encoding an acetoin(diacetyl) reductase from R. erythropolis WZ010 (ReADR) was cloned, overexpressed in Escherichia coli , and subsequently purified by Ni-affinity chromatography. ReADR in the native form appeared to be a homodimer with a calculated subunit size of 26,864, belonging to the family of the short-chain dehydrogenase/reductases. The enzyme accepted a broad range of substrates including aliphatic and aryl alcohols, aldehydes, and ketones. It exhibited remarkable tolerance to dimethyl sulfoxide (DMSO) and retained 53.6 % of the initial activity after 4 h incubation with 30 % ( v / v ) DMSO. The enzyme displayed absolute stereospecificity in the reduction of diacetyl to (2 S ,3 S )-2,3-butanediol via ( S )-acetoin. The optimal pH and temperature for diacetyl reduction were pH 7.0 and 30 °C, whereas those for (2 S ,3 S )-2,3-butanediol oxidation were pH 9.5 and 25 °C. Under the optimized conditions, the activity of diacetyl reduction was 11.9-fold higher than that of (2 S ,3 S )-2,3-butanediol oxidation. Kinetic parameters of the enzyme showed lower K m values and higher catalytic efficiency for diacetyl and NADH in comparison to those for (2 S ,3 S )-2,3-butanediol and NAD + , suggesting its physiological role in favor of (2 S ,3 S )-2,3-butanediol formation. Interestingly, the enzyme showed higher catalytic efficiency for ( S )-1-phenylethanol oxidation than that for acetophenone reduction. ReADR-catalyzed asymmetric reduction of diacetyl was coupled with stereoselective oxidation of 1-phenylethanol, which simultaneously formed both (2 S ,3 S )-2,3-butanediol and ( R )-1-phenylethanol in great conversions and enantiomeric excess values.

Description: Regioselective glycosylation of flavonoids cannot be easily achieved due to the presence of several hydroxyl groups in flavonoids. This hurdle could be overcome by employing uridine diphosphate-dependent glycosyltransferases (UGTs), which use nucleotide sugars as sugar donors and diverse compounds including flavonoids as sugar acceptors. Quercetin rhamnosides contain antiviral activity. Two quercetin diglycosides, quercetin 3- O -glucoside-7- O -rhamnoside and quercetin 3,7- O -bisrhamnoside, were synthesized using Escherichia coli expressing two UGTs. For the synthesis of quercetin 3- O -glucoside-7- O -rhamnoside, AtUGT78D2, which transfers glucose from UDP-glucose to the 3-hydroxyl group of quercetin, and AtUGT89C1, which transfers rhamnose from UDP-rhamnose to the 7-hydroxyl group of quercetin 3- O -glucoside, were transformed into E. coli . Using this approach, 67 mg/L of quercetin 3- O -glucoside-7- O -rhamnoside was synthesized. For the synthesis of quercetin 3,7- O -bisrhamnoside, AtUGT78D1, which transfers rhamnose to the 3-hydroxy group of quercetin, and AtUGT89C1 were used. The RHM2 gene from Arabidopsis thaliana was coexpressed to supply the sugar donor, UDP-rhamnose. E. coli expressing AtUGT78D1 , AtUGT89C1 , and RHM2 was used to obtain 67.4 mg/L of quercetin 3,7- O -bisrhamnoside.

Description: Biological soil disinfestations (BSDs) were developed separately in Japan and in The Netherlands as an alternative to chemical fumigations. In Japan, it was developed based on the knowledge of irrigated paddy rice and upland crop rotation system that was rather tolerant of soil-borne disease development. The methods consist of application of easily decomposable organic matter, irrigation, and covering the soil surface with plastic film, thereby inducing anaerobic (reductive) soil conditions and suppressing many soil-borne pests including fungi, bacteria, nematodes, and weeds. The methods are widely used by organic farmers in the area where residences and agricultural fields are intermingled. To note one advantage of these methods, maintenance of soil suppressiveness to Fusarium wilt of tomato was suggested, while soil treated with chloropicrin became conducive to the disease. Suppression of soil-borne fungal pathogens by BSDs might be attributed to anaerobicity and high temperature, organic acids generated, and metal ions released into soil water. Contributions of respective factors to suppression of respective pathogens might be diverse. Presumably, these factors might vary on the fungal community structure in BSD-treated soil. These factors also work in paddy fields. Therefore, the BSDs developed in Japan are probably a method to raise the efficacy of paddy–upland rotation through intensive organic matter application and through maintenance of a strongly anaerobic (reductive) soil condition.

Description: The human host cell line, F2N78, is a new somatic hybrid cell line designed for therapeutic antibody production. To verify its potential as a human host cell line, recombinant F2N78 cells that produce antibody against rabies virus (rF2N78) were cultivated at different culture pH (6.8, 7.0, 7.2, 7.4, and 7.6) and temperatures (33.0 °C and 37.0 °C). Regardless of the culture temperature, the highest specific growth rate was obtained at a pH of 7.0–7.4. Lowering the culture temperature from 37.0 °C to 33.0 °C suppressed cell growth while allowing maintenance of high cell viability for a longer period. However, it did not enhance antibody production because specific antibody productivity did not increase at 33.0 °C. The highest maximum antibody concentration was obtained at 37.0 °C and pH 6.8. The N-linked glycosylation of the antibody was affected by the culture pH rather than the temperature. Nevertheless, G1F was dominant and G2F occupied a larger portion than G0F in all culture conditions. Compared to the same antibody produced from recombinant CHO cells, the antibody produced from rF2N78 cells has more galactose capping and was more similar to human plasma IgG. Taken together, the results obtained here demonstrate the potential of F2N78 as an alternative human host cell line for therapeutic antibody production.

Description: Microbial enhanced oil recovery (MEOR) refers to the process of using bacterial activities for more oil recovery from oil reservoirs mainly by interfacial tension reduction and wettability alteration mechanisms. Investigating the impact of these two mechanisms on enhanced oil recovery during MEOR process is the main objective of this work. Different analytical methods such as oil spreading and surface activity measurements were utilized to screen the biosurfactant-producing bacteria isolated from the brine of a specific oil reservoir located in the southwest of Iran. The isolates identified by 16S rDNA and biochemical analysis as Enterobacter cloacae (Persian Type Culture Collection (PTCC) 1798) and Enterobacter hormaechei (PTCC 1799) produce 1.53 g/l of biosurfactant. The produced biosurfactant caused substantial surface tension reduction of the growth medium and interfacial tension reduction between oil and brine to 31 and 3.2 mN/m from the original value of 72 and 29 mN/m, respectively. A novel set of core flooding tests, including in situ and ex situ scenarios, was designed to explore the potential of the isolated consortium as an agent for MEOR process. Besides, the individual effects of wettability alteration and IFT reduction on oil recovery efficiency by this process were investigated. The results show that the wettability alteration of the reservoir rock toward neutrally wet condition in the course of the adsorption of bacteria cells and biofilm formation are the dominant mechanisms on the improvement of oil recovery efficiency.

Description: Bioaugmentation with degrading bacteria is an effective method to improve the treatment of refractory industrial wastewater; nevertheless there were controversial opinions about the fate of inoculated bacteria and microbial community dynamics. In this study, two lab-scale sequencing batch reactors filled with modified zeolite were used to treat a coking wastewater with pyridine and quinoline shock load, and a bacterial consortium containing three degrading strains was added in one reactor for bioaugmentation. During 120-day operation, the bioaugmented reactor removed over 99 % pyridine, 99 % quinoline, 85 % TOC, 65 % COD, and 95 % NO 3 − -N with higher resistance to the shock load than the non-bioaugmented reactor. Based on the terminal restriction fragment length polymorphism of 16S rDNA, bacterial community diversity increased in the bioaugmented reactor. Principal component analysis revealed that, to cope with the shock load, the indigenous bacterial community recovered to the initial structure by acclimatizing itself constantly to the inhospitable environment; but bioaugmentation accelerated the shift of whole bacterial community, resulting in a far different structure from the initial one. Canonical correspondence analysis indicated that the environmental parameters of pyridine, quinoline, TOC, and NO 3 − -N had close negative correlations with bioaugmentation; and NH 3 -N and COD were the main parameters to impact on the bacterial community changes and treatment efficiency.

Description: Pseudomonads are metabolically versatile microbes that employ complex regulatory networks to control gene expression, particularly with respect to carbon and nitrogen metabolism. The aim of this study was to characterise the regulatory networks that control pyrimidine metabolism (hydantoin-hydrolysing activity) in Pseudomonas putida strain RU-KM3 S , focussing on transcriptional activation of dihydropyrimidinase (Dhp) and β-ureidopropionase (Bup), encoding dhp and bup , respectively. The two genes are arranged divergently on the chromosome and are separated by ORF1, encoding a putative transporter, which lies upstream of and in the same orientation as bup . The results from this study reveal that pyrimidine metabolism, as a function of Bup and Dhp activity in P . putida RU-KM3 S , is controlled by a complex regulatory network including several global pathways in addition to induction by the substrate. Three major control pathways act at the level of transcriptional and include: (1) induction of transcriptional activation in the presence of hydantoin, (2) carbon catabolite repression mediated via a pathway independent of Crc and (3) quorum sensing that does not require a putative lux box located upstream of the dhp transcriptional start. Finally, the data suggest a minor role for the global regulators Anr, Vfr and Crc, likely through regulation of the activity of transcription factors interacting directly with the bup /ORF1– dhp promoter.

Description: γ-Decalactone is an industrially interesting peach-like aroma compound that can be produced biotechnologically through the biotransformation of ricinoleic acid. Castor oil (CO) is the raw material most used as the ricinoleic acid source. The effect of different CO concentrations on the γ-decalactone production by Yarrowia lipolytica was investigated in batch processing, and 30 g L −1 was found to be the optimal oil concentration. Under these conditions, cells were able to produce lipase but at low activity levels, which might limit ricinoleic acid release and consequently, the γ-decalactone production rate. Thus, the enzymatic hydrolysis of CO by commercial lipases was studied under different operating conditions. Lipozyme TL IM was found to be the most efficient and the optimal hydrolysis conditions were pH 8 and 27 °C. The use of hydrolyzed CO in the aroma production allowed a decrease in the lag phase for γ-decalactone secretion.

Description: Three phosphonate derivatives of methyl oleate (MeO) were chemically synthesized in a radical chain reaction and their physical and tribological properties investigated. The phosphonates differed from each other in the structure of the alkoxy groups attached to the phosphorous, which were as follows: methoxy, ethoxy, and n -butoxy. Phosphonylation eliminated the unsaturation in MeO and also introduced branching. The phosphonate oils had higher density and viscosity than MeO, which was attributed to the contribution of the heavier phosphorus atom in their structures, and to their higher molecular weights, respectively. The phosphonates also displayed improved oxidation stability and cold flow properties, which were attributed to the elimination of the double bond and the introduction of branching in their molecular structures, respectively. Tribological investigations were conducted using 4-Ball anti-wear (AW) and extreme pressure (EP) methods. Neat phosphonates displayed lower AW coefficients of friction and wear scar diameters than MeO. The improved AW results were attributed to the higher viscosity of the phosphonates, since the AW test is conducted in the mixed film regime. The phosphonates had no effect on EP weld point (WP) as neat oils or as 5 % additives in petroleum-based oils. In soy base oil, 5 % phosphonate additives displayed smaller improvement in WP than zinc dialkyl dithiophosphate (ZDDP). It is proposed that this lack of EP characteristics could be due to the high dissociation energy of C–P bonds in the phosphonates, compared to, for example, S–P bonds in ZDDP.

Description: The present study presents the antioxidant activity of sesamin in canola oil compared with that of butylated hydroxytoluene (BHT) by monitoring the oxygen consumption and the decrease in linoleic acid and α-linolenic acid. The oxidation of canola oil was conducted at 35, 60, 90, 120 and 180 °C with addition of 50–400 ppm sesamin. Results from the oxygen consumption test showed that sesamin dose-dependently inhibited the oxidation of canola oil at concentrations of 50–200 ppm at temperatures of 60–180 °C, however, sesamin lost its antioxidant activity at a low temperature of 35 °C. The fatty acid analysis also demonstrated that sesamin at 50, 100 and 200 ppm dose-dependently prevented the oxidation of linoleic acid and α-linolenic acid in canola oil. Both the oxygen consumption and the fatty acid analysis demonstrated sesamin was less effective than BHT as an antioxidant at temperatures of 60–180 °C. It was therefore concluded that sesamin could prevent the lipid oxidation of frying fats and oil, however, its antioxidant activity was not as potent as that of BHT.