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  • Articles  (5,186)
  • Springer  (5,186)
  • 2010-2014  (5,186)
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  • Articles  (5,186)
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  • Springer  (5,186)
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  • 1
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    Simultaneous oral immunization of mice with live attenuated Escherichia coli expressing LT 192 -STa 13 and LT 192 -STb fusion immunogen, respectively, for polyvalent vaccine candidate (2014)
    Springer
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    Publication Date: 2014-12-31
    Description: Previous epidemiological study showed that most of the porcine enterotoxin Escherichia coli (ETEC) strains harbor multiple enterotoxins but lack any of the fimbriae or non-fimbrial adhesion genes. Therefore, effective ETEC vaccines need to aim directly at all the enterotoxin antigens. The objective of this study was to evaluate the simultaneous immune effect of two live attenuated E. coli strains expressing LT R192G -STa A13Q and LT R192G -STb fusion immunogen, respectively. The results showed that both local mucosal and systemic immune responses against LT, STa, STb, and F41 were induced in BALB/c mice immunized orally with the recombinant E. coli strains ER - A and ER - B simultaneously. In addition, results of cellular immune responses showed that stimulation index (SI) values of immunized mice were significantly higher than control mice ( P  〈 0.05) and a marked shift toward type-2 helper T lymphocyte (Th 2) immunity. Moreover, the induced antibodies demonstrated neutralizing effects on LT, STa, and STb producing E. coli infection. These data indicated that the use of recombinant E. coli ER - A and ER - B could be a valuable strategy for future polyvalent vaccine development of ETEC.
    Print ISSN: 0175-7598
    Electronic ISSN: 1432-0614
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 2
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    One-pot production of fructooligosaccharides by a Saccharomyces cerevisiae strain expressing an engineered invertase (2014)
    Springer
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    Publication Date: 2014-12-31
    Description: We describe a simple, efficient process for the production of 6-kestose, a trisaccharide with well-documented prebiotic properties. A key factor is the use of a yeast transformant expressing an engineered version of Saccharomyces invertase with enhanced transfructosylating activity. When the yeast transformant was grown with 30 % sucrose as the carbon source, 6-kestose accumulated up to ca. 100 g/L in the culture medium. The 6-kestose yield was significantly enhanced (up to 200 g/L) using a two-stage process carried out in the same flask. In the first stage, the culture was grown in 30 % sucrose at physiological temperature (30 °C) to allow overexpression of the invertase. In the second stage, sucrose was added to the culture at high concentration (60 %) and the temperature shifted to 50 °C. In both cases, 6-kestose was synthesized with high specificity, representing more than 95 % of total FOS.
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    Electronic ISSN: 1432-0614
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 3
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    Continuous nitrous oxide abatement in a novel denitrifying off-gas bioscrubber (2014)
    Springer
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    Publication Date: 2014-12-31
    Description: The potential of a bioscrubber composed of a packed bed absorption column coupled to a stirred tank denitrification bioreactor (STR) was assessed for 95 days for the continuous abatement of a diluted air emission of N 2 O at different liquid recycling velocities. N 2 O removal efficiencies of up to 40 ± 1 % were achieved at the highest recirculation velocity (8 m h −1 ) at an empty bed residence time of 3 min using a synthetic air emission containing N 2 O at 104 ± 12 ppm v . N 2 O was absorbed in the packed bed column and further reduced in the STR at efficiencies 〉80 % using methanol as electron donor. The long-term operation of the bioscrubber suggested that the specialized N 2 O degrading community established was not able to use N 2 O as nitrogen source. Additional nitrification assays showed that the activated sludge used as inoculum was not capable of aerobically oxidizing N 2 O to nitrate or nitrite, regardless of the inorganic carbon concentration tested. Denitrification assays confirmed the ability of non-acclimated activated sludge to readily denitrify N 2 O at a specific rate of 3.9 mg N 2 O g VSS h -1 using methanol as electron donor. This study constitutes, to the best of our knowledge, the first systematic assessment of the continuous abatement of N 2 O in air emission. A characterization of the structure of the microbial population in the absorption column by DGGE-sequencing revealed a high microbial diversity and the presence of heterotrophic denitrifying methylotrophs.
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    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 4
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    Fundamental role of cobalamin biosynthesis in the developmental growth of Streptomyces coelicolor A3 (2) (2014)
    Springer
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    Publication Date: 2014-12-31
    Description: Cobalamin (Cbl) (synonym, vitamin B12) is the cobalt-containing cofactor produced only by some prokaryotes. Streptomyces is an effective Cbl producer. To study the role of Cbl production in Streptomyces , a knockout mutant for Cbl biosynthesis ( cob ) was generated in Streptomyces coelicolor A3 (2). The growth of the mutant was similar to that of the wild type in a rich medium, but inhibited in minimal medium, suggesting the involvement of Cbl in some step of primary metabolism. Methionine synthesis catalyzed by MetH, the Cbl-dependent methionine synthase, is a candidate. However, supplementing the minimal medium with methionine did not rescue the growth of the cob mutant, indicating that the availability of Cbl affects another primary function. Transcriptional analysis confirmed that the mutant induced metE encoding an alternative Cbl-independent methionine synthase, probably due to the Cbl-dependent riboswitch mechanism. The cob mutant produced low levels of pigment antibiotics and formed fewer aerial mycelium and spores in a rich medium, suggesting that a Cbl-dependent mechanism controls development. A similar developmental defect was observed for a knockout mutant for SCO4800, encoding the putative Cbl-dependent isobutyryl-CoA mutase (Icm) small subunit. Since the knockout of the Icm large subunit (SCO5415) did not affect the developmental phenotype, SCO4800 likely regulates development independently from SCO5415. Effective Cbl production is fundamental to the diverse functions underlying the complex developmental life cycle of S. coelicolor A3 (2).
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    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 5
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    Fermentative production of 1-propanol from sugars using wild-type and recombinant Shimwellia blattae (2014)
    Springer
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    Publication Date: 2014-12-31
    Description: Shimwellia blattae is an enteric bacterium and produces endogenous enzymes that convert 1,2-propanediol (1,2-PD) to 1-propanol, which is expected to be used as a fuel substitute and a precursor of polypropylene. Therefore, if S. blattae could be induced to generate its own 1,2-PD from sugars, it might be possible to produce 1-propanol from sugars with this microorganism. Here, two 1,2-PD production pathways were constructed in S. blattae , resulting in two methods for 1-propanol production with the bacterium. One method employed the L-rhamnose utilization pathway, in which L-rhamnose is split into dihydroxyacetone phosphate and 1,2-PD. When wild-type S. blattae was cultured with L-rhamnose, an accumulation of 1,2-PD was observed. The other method for producing 1,2-PD was to introduce an engineered 1,2-PD production pathway from glucose into S. blattae . In both cases, the produced 1,2-PD was then converted to 1-propanol by 1,2-PD converting enzymes, whose production was induced by the addition of glycerol.
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    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 6
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    Cloning, expression, and purification of insect ( Sitophilus oryzae ) alpha-amylase, able to digest granular starch, in Yarrowia lipolytica host (2014)
    Springer
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    Publication Date: 2014-12-31
    Description: Raw-starch-digesting enzymes (RSDE) are of major importance for industrial applications, as their usage greatly simplifies the starch processing pipeline. To date, only microbial RSDE have gained considerable attention, since only microbial production of enzymes meets industrial demands. In this study, α-amylase from rice weevil ( Sitophilus oryzae ), the major rice pest, was cloned and expressed in Yarrowia lipolytica Po1g strain. The enzyme was secreted into the culture medium, and the peak activity (81 AU/L) was reached after only 29 h of culturing in 5-L bioreactors. Through simple purification procedure of ammonium sulfate precipitation and affinity chromatography, it was possible to purify the enzyme to apparent homogeneity (25-fold purification factor, at 5 % yield). The optimal conditions for the α-amylase activity were pH 5.0 and a temperature of 40 °C. The α-amylase studied here did not show any obligate requirement for Ca 2+ ions. The recombinant α-amylase appeared to efficiently digest granular starch from pea, amaranth, waxy corn, and waxy rice.
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    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 7
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    Involvement of the NADH oxidase-encoding noxA gene in oxidative stress responses in Corynebacterium glutamicum (2014)
    Springer
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    Publication Date: 2014-12-31
    Description: Corynebacterium glutamicum ORF NCgl0328, designated noxA , encodes an NADH oxidase enzyme. The noxA gene, which was preferentially expressed in the log growth phase, was found to be under the control of the whcA , whcB , and whcE genes, which play regulatory roles in cells under oxidative stress. While noxA transcription was minimal in whcE- deleted mutant cells (Δ whcE ) during growth, its transcription was maximal even in the stationary phase in Δ whcA cells. The transcription levels of noxA in Δ whcB and whcB -overexpressing cells were comparable to the levels only in the log growth phase in Δ whcA and whcA -overexpressing cells, respectively. Direct binding of purified WhcA to the promoter region of noxA was observed in vitro. The DNA-protein interaction was only possible in the presence of the reducing agent dithiothreitol. A noxA -deleted mutant strain and a strain overexpressing the noxA gene (P 180 - noxA ) were established, and these strains were found to exhibit defective cell growth. The Δ noxA and P 180 - noxA strains were sensitive to the redox-cycling oxidant menadione, suggesting a role of noxA in redox balancing. Accordingly, the purified NoxA enzyme exhibited NADH-oxidizing activity. Taken together, these data show that noxA plays a role in oxidative stress responses and also that the gene is under direct control of the WhcA protein, which was shown to be a regulatory DNA-binding protein. Furthermore, the involvement and roles of the whcA , whcB , and whcE genes in regulating the expression of noxA were demonstrated.
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    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 8
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    Antimicrobial compounds from seaweeds-associated bacteria and fungi (2014)
    Springer
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    Publication Date: 2014-12-31
    Description: In recent decade, seaweeds-associated microbial communities have been significantly evaluated for functional and chemical analyses. Such analyses let to conclude that seaweeds-associated microbial communities are highly diverse and rich sources of bioactive compounds of exceptional molecular structure. Extracting bioactive compounds from seaweed-associated microbial communities have been recently increased due to their broad-spectrum antimicrobial activities including antibacterial, antifungal, antiviral, anti-settlement, antiprotozoan, antiparasitic, and antitumor. These allelochemicals not only provide protection to host from other surrounding pelagic microorganisms, but also ensure their association with the host. Antimicrobial compounds from marine sources are promising and priority targets of biotechnological and pharmaceutical applications. This review describes the bioactive metabolites reported from seaweed-associated bacterial and fungal communities and illustrates their bioactivities. Biotechnological application of metagenomic approach for identifying novel bioactive metabolites is also dealt, in view of their future development as a strong tool to discover novel drug targets from seaweed-associated microbial communities.
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    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 9
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    Cell-specific delivery of messenger RNA and microRNA by recombinant MS2 virus-like particles carrying cell-penetrating peptide (2014)
    Springer
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    Publication Date: 2014-12-31
    Description: Messenger RNA (mRNA) and microRNA have great therapeutic potential in cancer and other diseases. However, their instability and low in vivo delivery efficiency have limited their application. Recombinant MS2 bacteriophage-based virus-like particles (VLPs) can protect mRNA and microRNA from the rapid degradation by exogenous RNase, based on their ability to package specific exogenous RNA with pac site . The delivery of mRNA and microRNA in mammalian cells can be significantly improved by inserting cell-penetrating peptide (CPP) between amino acid residues 15 and 16 of the second subunit of the MS2 coat protein single-chain dimer. Unlike the other delivery approaches for mRNA and microRNA (viral or nonviral vectors), recombinant MS2 VLPs carrying CPP and mRNA or microRNA were expressed from the one-plasmid double-expression system in Saccharomyces cerevisiae with high efficiency and were purified easily. They were also nuclease-resistant, nonreplicative, noninfectious, and nontoxic. MS2 VLPs carrying CPP penetrated cells of the cervical cancer cell line HeLa, lung cancer cell line A549, and acute myeloid leukemia cell line Kasumi-1 and delivered high concentrations of mRNAs/pre-microRNA into the cells, which were translated into mature products within 24 h. Moreover, it was observed that the VLPs carrying microRNA-29b and TAT peptide induced the apoptosis of HeLa cells, A549 cells, and Kasumi-1 cells at a concentration of 3.7 pM. Therefore, recombinant MS2 VLPs can simultaneously be used as a targeted delivery vector for both RNA and peptides, due to their abilities to package specific exogenous RNA with the help of the pac site and display peptides.
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    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 10
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    Toward metabolic engineering in the context of system biology and synthetic biology: advances and prospects (2014)
    Springer
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    Publication Date: 2014-12-31
    Description: Metabolic engineering facilitates the rational development of recombinant bacterial strains for metabolite overproduction. Building on enormous advances in system biology and synthetic biology, novel strategies have been established for multivariate optimization of metabolic networks in ensemble, spatial, and dynamic manners such as modular pathway engineering, compartmentalization metabolic engineering, and metabolic engineering guided by genome-scale metabolic models, in vitro reconstitution, and systems and synthetic biology. Herein, we summarize recent advances in novel metabolic engineering strategies. Combined with advancing kinetic models and synthetic biology tools, more efficient new strategies for improving cellular properties can be established and applied for industrially important biochemical production.
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    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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