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  • General Chemistry  (3,401)
  • Biochemistry and Biotechnology  (743)
  • SPACE VEHICLES  (620)
  • 1995-1999  (3,206)
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  • 1996  (2,456)
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  • 1
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 49 (1996), S. 204-216 
    ISSN: 0006-3592
    Keywords: expanded bed adsorption ; bakers' yeast ; G6PDH ; STREAMLINE ion exchange adsorbents ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The use of expanded beds of STREAMLINE ion exchange adsorbents for the direct extraction of an intracellular enzyme glucose-6-phosphate dehydrogenase (G6PDH) from unclarified yeast cell homogenates has been investigated. It has been demonstrated that such crude feedstocks can be applied to the bed without prior clarification steps. The purification of G6PDH from an unclarified yeast homogenate was chosen as a model system containing the typical features of a direct extraction technique. Optimal conditions for the purification were determined in small scale, packed bed experiments conducted with clarified homogenates. Results from these experiments were used to develop a preparative scale separation of G6PDH in a STREAMLINE 50 EBA apparatus. The use of an on-line rotameter for measuring and controlling the height of the expanded bed when operated in highly turbid feedstocks was demonstrated. STREAMLINE DEAE has been shown to be successful in achieving isolation of G6PDH from an unclarified homogenate with a purification factor of 12 and yield of 98% in a single step process. This ion exchange adsorbent is readily cleaned using simple cleaning-in-place procedures without affecting either adsorption or the bed expansion properties of the adsorbent after many cycles of operation. The ability of combining clarification, capture, and purification in a single step will greatly simplify downstream processing flowsheets and reduce the costs of protein purification. © 1996 John Wiley & Sons, Inc.
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 49 (1996), S. 259-265 
    ISSN: 0006-3592
    Keywords: hepatocytes ; lactose-derivatized polystyrene ; polystyrene ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Hepatocytes isolated from male Fisher 344VF rats were cultured on two substrates, collagen I and a lactose-derivatized polystyrene (PS-lactose), to compare morphological and functional differences. Hepatocyte morphology changed dramatically depending upon the substrate, shown through actin cytoskeletal staining and scanning electron microscopy. Functional assays performed included albumin secretion, reduced glutathione content, UDP-glucuronosyl transferase, and cytochrome P4501A1 activity. The presence of dexamethasone and dimethylsulfoxide (DMSO) in the media was required for the maintenance of several differentiated functions for cells cultured on collagen. In general, cells cultured on the PS-lactose substrate showed a much slower loss of function over the same period of time. The maintenance of differentiated function of cells on PS-lactose was enhanced with the addition of dexamethasone and DMSO. This is the first report of a culture system in which hepatocytes, cultured on a polymer substrate without additional protein coatings or media additives, have been able to maintain differentiated functions for up to 1 week. © 1996 John Wiley & Sons, Inc.
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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 49 (1996), S. 290-299 
    ISSN: 0006-3592
    Keywords: proteins, modified ; partitioning in aqueous system ; thaumatin ; β-lactoglobulin ; BSA ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Relatively conservative modifications of three proteins were carried out to alter their surface properties. The protein properties modified were hydrophobicity and charge. This was done by acylation of amino groups with anhydrides. For the hydrophobic modification experiments, two proteins (β-lactoglobulin and bovine serum albumin [BSA]) and four anhydrides (hexanoic, butyric, succinic, acetic) were used. For the modification of surface charge the protein thaumatin was selected and various proportions of the free amino groups were blocked with acetic anhydride to give a series of proteins with differing isoelectric points. Detailed characterization and purification of selected modified proteins was carried out including molecular weight measurements and conformational analysis. The criteria used for selecting the modified proteins for subsequent investigation of their partitioning in aqueous two-phase systems (ATPS) is described. With a judicious choice of starting material it was found that limited chemical modifications to proteins could effectively alter surface hydrophobicity or charge almost independently, with little effect on other molecular properties. It appears, however, that the method for chemical modification and the reaction conditions must also be carefully controlled. © 1996 John Wiley & Sons, Inc.
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 49 (1996), S. 309-315 
    ISSN: 0006-3592
    Keywords: surface charge ; proteins, modified ; partitioning in aqueous system ; thaumatin ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A series of charge-modified thaumatins with different values of surface charge were partitioned in aqueous two-phase systems (ATPS) to study the effect of surface charge as a single property on partitioning. Electrophoretic mobility of the proteins in titration curves was used as a measure of surface charge. Four modified proteins derived from thaumatin with the following values of isoelectric point: 8.70, 8.15, 5.60, and 4.50 were used for partitioning. The resolution of the systems in terms of protein surface charge was calculated. Partitioning of modified thaumatins in PEG 4000/dextran systems with phosphate buffer, Tris buffer, NaCl, KCl, and sulfate salts was carried out. Among the sulfate salts tested, the addition of 50 mM Li2SO4 to the system buffered with phosphate gave the highest value of resolution for differences in surface protein charge (RSPC). It shows a decrease in the value of K (partition coefficient) with an increase in the protein's charge. The addition of 100 mM KCl to the system promoted the opposite effect on the RSPC value. Charge-modified proteins were partitioned in PEG/salt systems to investigate the ability of these systems for resolving differences in surface charge. The PEG/citrate system seemed to have almost no ability for resolving proteins on the basis of surface charge differences; PEG/phosphate systems had some capability for resolving differently charged proteins. The more negative proteins tended to have higher values of K than the more positively charged fractions. The use of charge-modified proteins allowed the investigation of the effect of protein surface charge on partitioning in aqueous two-phase systems independently from other protein parameters as they were prepared from a common parent protein thaumatin. This technique provides an interesting novel tool to investigate the effect of protein surface charge on partitioning in ATPS taking protein charge as an independent parameter. © 1996 John Wiley & Sons, Inc.
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  • 5
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 49 (1996), S. 348-354 
    ISSN: 0006-3592
    Keywords: oxygenator ; NMR spectroscopy ; organ perfusion ; mammalian cell culture ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A compact, reusable membrane oxygenator has been constructed for the perfusion of cultured cells and isolated organs. While the oxygenator was designed to be compatible with nuclear magnetic resonance (NMR) spectroscopy studies, it can also be used for any experiment which requires warming and oxygenation of perfusates. For the NMR studies, the oxygenator can be positioned at the opening of the magnet bore which allows oxygenation and warming of the perfusate immediately prior to delivery to the tissue, therefore eliminating problems with heat or oxygen loss which may occur with the long perfusion lines. © 1996 John Wiley & Sons, Inc.
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  • 6
    ISSN: 0006-3592
    Keywords: c-fos protein ; endothelium ; hemodynamics ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The c-fos protein belongs to a family of transcriptional cofactors that can complex with proteins of the Jun family and activate mRNA transcription from gene promoters containing an activator protein 1 (AP-1) binding element. The shear stress inducibility of the c-fos protein was studied in human and animal cell lines of vastly different origins. Primary human umbilical vein endothelial cells (HUVEC), bovine aortic endothelial cells (BAEC, passage 2-14), HeLa cells, and Chinese hamster ovary (CHO) cells were subjected to steady laminar shear stress using a parallel plate flow apparatus. After 1 h of flow exposure at 25 dyn/cm2, the c-fos levels in nuclei of shear stress HUVEC, BAEC, HeLa, and CHO were 5.4 ± 2.0 (n = 3), 2.25 ± 1.38 (n = 6), 2.14 ± 0.07 (n = 8), 1.92 ± 0.58 (n = 2) times higher, respectively, than in matched stationary controls. Flow exposure at 4 dyn/cm2 caused no enhancement of c-fos levels in any of the cell lines tested, but caused significant reduction in c-fos expression in the HeLa cells. The c-fos induction by shear stress could be blocked by pharmacological agents. For example, the flow induction of the c-fos protein levels was blocked by 50% with the preincubation of HUVEC with a protein kinase C inhibitor, H7 (10 μM) and blocked completely in HeLa cells preincubated with the phospholipase C inhibitor, neomycin (5 mM). The minimum time of shear stress exposure required to induce the c-fos protein expression in HeLa cells was found to be as low as 1 min. By Northern analysis, the c-fos mRNA levels were found to be elevated in BAEC, CHO, and HeLa cells exposed to 25 dyn/cm2 for 30 min. These studies indicate that c-fos induction is a consistent genetic response in a variety of mammalian cells that may alter cellular phenotype in mechanical environments. © 1996 John Wiley & Sons, Inc.
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 49 (1996), S. 412-420 
    ISSN: 0006-3592
    Keywords: Amycolatopsis orientalis ; vancomycin production ; chemostat culture ; phosphate inhibition ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Production of the glycopeptide antibiotic vancomycin by two Amycolatopsis orientalis strains was examined in batch shake flask culture in a semidefined medium with peptone as the nitrogen source. Different growth and production profiles were observed with the two strains; specific production (Yp/x) was threefold higher with strain ATCC 19795 than with strain NCIMB 12945. A defined medium with amino acids as the nitrogen source was developed by use of the Plackett-Burman statistical screening method. This technique identified certain amino acids (glycine, phenylalanine, tyrosine, and arginine) that gave significant increased specific production, whereas phosphate was identified as inhibitory for high specific vancomycin production. Experiments made with the improved medium and strain ATCC 19795 showed that vancomycin production kinetics were either growth dissociated or growth associated, depending on the amino acid concentration. In chemostat culture at a constant dilution rate (0.087 h-1), specific vancomycin production rate (qvancomycin) decreased linearly as the medium phosphate concentration was increased from 2 to 8 mM. In both phosphate and glucose limited chemostats, qvancomycin was a function of specific growth rate; the maximum value was observed at D = 0.087 h-1 (52% of the maximum specific growth rate). Under phosphate limited growth conditions, qvancomycin was threefold higher (0.37 mg/g dry weight/h) than under glucose limitation (0.12 mg/g dry weight/h). © 1996 John Wiley & Sons, Inc.
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  • 8
    Electronic Resource
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 50 (1996), S. 36-48 
    ISSN: 0006-3592
    Keywords: insect cell culture ; Sf-9 cells ; respiration ; bioreactor ; on-line monitoring ; baculovirus expression vector system ; recombinant proteins ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Respiration rates in Spodoptera frugiperda (Sf-9) cell bioreactor cultures were successfully measured on-line using two methods: The O2 uptake rate (OUR) was determined using gas phase pO2 values imposed by a dissolved oxygen controller and the CO2 evolution rate (CER) was measured using an infrared detector. The measurement methods were accurate, reliable, and relatively inexpensive. The CER was routinely determined in bioreactor cultures used for the production of several recombinant proteins. Simple linear relationships between viable cell densities and both OUR and CER in exponentially growing cultures were used to predict viable cell density. Respiration measurements were also used to follow the progress of baculoviral infections in Sf-9 cultures. Infection led to increases in volumetric and per-cell respiration rates. The relationships between respiration and several other culture parameters, including viable cell density, cell protein, cell volume, glucose consumption, lactate production, viral titer, and recombinant β-galactosidase accumulation, were examined. The extent of the increase in CER following infection and the time postinfection at which maximum CER was attained were negatively correlated with the multiplicity of infection (MOI) at multiplicities below the level required to infect all the cells in a culture. Delays in the respiration peak related to the MOI employed were correlated with delays in the peak in recombinant protein accumulation. DO levels in the range 5-100% did not exert any major effects on viable cell densities, CER, or product titer in cultures infected with a baculovirus expressing recombinant β-galactosidase. © 1996 John Wiley & Sons, Inc.
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  • 9
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 50 (1996) 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 10
    Electronic Resource
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 50 (1996), S. 169-183 
    ISSN: 0006-3592
    Keywords: liposomes ; biotin ; aggregation kinetics ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The aggregation of biotinylated phospholipid vesicles (liposomes) cross-linked by antibiotin IgG was studied experimentally and theoretically. The liposomes were either low density liposomes that contained 0.4 mol% biotinylated phospholipid (≈100 exposed biotin molecules per liposome), or high density liposomes that contained 2.7 mol% biotinylated phospholipid (≈1000 exposed biotin molecules per liposome). The solution turbidity and mean particle size measured by quasi-elastic light scattering (QLS) were monitored throughout the aggregation. Three different lots of antibiotin antibodies, each with different association constants and binding heterogeneities, were used. The antibody binding characteristics affected the aggregation rates. The aggregation kinetics were analyzed using a model based on the Smoluchowski theory of aggregation, fractal concepts of aggregate microstructure, and Rayleigh and Mie light scattering theory. The experimental conditions of liposome concentration, protein concentration, and ligand density under which aggregation occurred correlated well with calculated sticking probabilities based on isotherms describing the adsorption of antibiotin antibody to the liposomes. These results are compared with prior observations made when avidin was used as the cross-linking protein. © 1996 John Wiley & Sons, Inc.
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  • 11
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 50 (1996) 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 12
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 50 (1996), S. 211-216 
    ISSN: 0006-3592
    Keywords: microgravity ; bioprocessing ; sedimentation ; turbulence ; collagenase ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effect of a quiescent microgravity fluid environment on the activity of collagenase directed at demineralized bone fragments was investigated over a period of 10 days. Enzyme treatment resulted in greater mass loss in microgravity, with nearly three times the loss of mass during Space Shuttle mission STS-62 compared to the stationary ground control. Clinorotation enhanced the loss of mass relative to a stationary control, but this increase was still significantly less than the increase with exposure to microgravity. This suggests the detrimental influence of turbulence on the enzyme function and the benefit of using microgravity to provide both low turbulence and uniformity of unequally dense materials within the reaction chamber. The results are considered for their general applicability to a variety of bioprocessing applications that may be enhanced in microgravity. © 1996 John Wiley & Sons, Inc.
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  • 13
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    Biotechnology and Bioengineering 50 (1996), S. 430-437 
    ISSN: 0006-3592
    Keywords: cartilage ; tissue regeneration ; chondrocytes ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In the last 5 to 10 years, tissue engineering has revolutionized the way in which medical researchers and clinicians are thinking of and, in some cases, actually treating diseases involving tissue damage and destruction. One such disease, osteoarthritis, results from progressive degeneration of articular cartilage, which has a limited ability to repair itself. With tissue engineering, scientists are now able to regenerate cartilage in vitro from isolated mature chondrocytes. While the regeneration process is still not fully understood, enough has been learned that physicians are already implanting cultured chondrocytes into humans and other animals in the hopes of effecting joint repair. One aspect which has not been fully explored is the effect of mechanical stress on developing and implanted cartilage, especially over the long term. This article will review in brief what is now known about the mechanical factors affecting cartilage regeneration in vitro and what still remains to be determined for optimum tissue engineering of cartilage constructs. © 1996 John Wiley & Sons, Inc.
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  • 14
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    Biotechnology and Bioengineering 50 (1996), S. 443-451 
    ISSN: 0006-3592
    Keywords: osteoblast ; migration ; poly(αhydroxy esters) ; poly(DL-lactic-co-glycolic acid) ; PLGA ; biodegradable polymers ; tissue engineering ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: We investigated the migration of rat calvaria osteoblast populations on poly(α-hydroxy ester) films for up to 14 days to determine effects of substrate composition and culture conditions on the migratory characteristics of osteoblasts. Initial osteoblast culture conditions included cell colonies formed by seeding a high (84,000 cells/cm2) or low (42,000 cells/cm2) density of isolated osteoblasts on the polymer films, and bone tissue cultures formed by plating bone chips directly on the substrates. High density osteoblast colonies cultured and allowed to migrate and proliferate radially on 85:15 poly(DL-lactic-co-glycolic acid) (PLGA) films, 75:25 PLGA films, and tissue culture polystyrene controls demonstrated that the copolymer ratio in the polymer films did not affect the rate of increase in substrate surface area (or culture area) covered by the growing cell colony. However, the rate of increase in culture area was dependent on the initial osteoblast seeding density. Initial cell colonies formed with a lower osteoblast seeding density on 75:25 PLGA resulted in a lower rate of increase in culture area, specifically 4.9 ± 0.3 mm2/day, versus 14.1 ± 0.7 mm2/day for colonies seeded with a higher density of cells on the same polymer films. The proliferation rate for osteoblasts in the high and low density seeded osteoblast colonies did not differ, whereas the proliferation rate for the osteoblasts arising from the bone chips was lower than either of these isolated cell colonies. Confocal and light microscopy revealed that the osteoblast migration occurred as a monolayer of individual osteoblasts and not a calcified tissue front. These results demonstrated that cell seeding conditions strongly affect the rates of osteoblast migration and proliferation on biodegradable poly(α-hydroxy esters). © 1996 John Wiley & Sons, Inc.
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  • 15
    ISSN: 0006-3592
    Keywords: bone marrow ; hematopoiesis ; perfusion ; culture optimization ; stroma ; stem cells ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Hematopoiesis, the formation of mature blood cells from stem (LTC-IC) and progenitor (CFU-GM) cells in the bone marrow, is a complex tissue-forming process that leads to many important physiological functionalities. Consequently, a functioning ex vivo hematopoietic system has a variety of basic scientific and clinical uses. The design and operation of such a system presents the tissue engineer with challenges and choices. In this study, three culture variables were used to control ex vivo human hematopoiesis. Systematic variation of inoculum density (ID), medium exchange interval (MEI), and the use of preformed stroma (PFS) showed that (1) all three variables significantly influenced culture performance, (2) the three variables interacted strongly, and (3) the variables could be manipulated to achieve the optimization of different performance criteria. Donor-to-donor variability in culture performance was great at low ID but was minimized at higher ID. PFS had a large positive effect on cell and CFU-GM output at low ID, but had minimal effect at higher ID. In fact, PFS caused a decrease in LTC-IC output at high ID. The effects of PFS indicated that stromal cell elements became more limiting than proliferative cell elements as ID was reduced.In cultures without PFS, maximum cell output was obtained with high ID using a short MEI, whereas the greatest cell expansion ratio was obtained at low ID with an intermediate MEI. Maximum CFU-GM output was obtained from cultures with high ID using a short to intermediate MEI, whereas the greatest CFU-GM expansion ratio was obtained at intermediate ID with an intermediate MEI. The addition of PFS altered the locations of these maxima. In general, PFS moved the maxima to lower ID, and culture output became more sensitive to MEI. Therefore, the optimization of one performance criterion always resulted in a decline of the others. This study demonstrates that ex vivo tissue function is sensitive to many culture variables in an interactive fashion and that systematic multivariable studies are required to characterize tissue function. Once the effects of individual variables and their interactions are known, this knowledge can be used to optimize tissue performance with respect to desired criteria. © 1996 John Wiley & Sons, Inc.
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  • 16
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    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 50 (1996) 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 17
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    Biotechnology and Bioengineering 51 (1996), S. 410-421 
    ISSN: 0006-3592
    Keywords: lysozyme ; thermal stability ; 1H NMR ; conformational flexibility ; melting temperature ; PEG ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The reversible folding destabilization of hen lysozyme has been confirmed by a melting temperature (Tm) decrease in aqueous poly(ethylene glycol) (PEG). The percent denatured, extracted from the histidine 15 C2H (H15 C2H) native and denatured peak areas from 500-MHz one-dimensional proton nuclear magnetic resonance (1D 1H NMR) spectra in D2O, was analyzed through denaturation temperatures at 0% and 20% (w/w) PEG 1000. The lysozyme (3.5 mM) Tm decreased by 4.2°C and 7.1°C in 20% (w/w) PEG 1000 at pH 3.8 and 3.0, respectively. The Tm decreased with increasing lysozyme concentration. Additionally, the temperature-induced resonance migrations of 17 protons from 8 residues indicate that the native lysozyme structure undergoes temperature-induced conformational changes. The changes were essentially identical in both 0% and 20% (w/w) PEG 1000 at both pH 3.0 and 3.8. This small, local restructuring of the hydrophobic box region may be a manifestation of temperature-dependent solution hydrophobicity, whereas active-site cleft fluctuations may be due to the inherent active-site flexibility. The lysozyme structure in PEG at 35°C was determined to be essentially native from the 1H nuclear Overhauser effect spectroscopy (NOESY) fingerprint regions. Additionally, lysozyme chemical shifts, from 1D spectra, in PEG 200, 300, and 1000 at 35°C and various concentrations were essentially identical, further confirming that the conformation remains native in various PEG solutions. © 1996 John Wiley & Sons, Inc.
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  • 18
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    Biotechnology and Bioengineering 51 (1996), S. 375-383 
    ISSN: 0006-3592
    Keywords: cellulase ; enzyme recycling ; enzyme adsorption ; lignocellulosic hydrolysis ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Past technoeconomic modeling work has identified the relatively large contribution that enzymatic hydrolysis adds to the total cost of producing ethanol from lignocellulosic substrates. This cost was primarily due to the high concentration of enzyme and long incubation time that was required to obtain complete hydrolysis. Although enzyme and substrate concentration and end-product inhibition influenced the rate of hydrolysis, the effect was less pronounced during the initial stages of hydrolysis. During this time most of the cellulases were adsorbed onto the unhydrolyzed residue. By recycling the cellulases adsorbed to the residual substrate remaining after an initial 24 h, a high rate of hydrolysis, with low overall residence time and minimal cellulase input, could be achieved for several rounds of enzyme recycle. A comparison of the front end (pretreatment, fractionation, and hydrolysis) of a softwood/hardwood to ethanol process indicated that the lignin associated with the softwood-derived cellulose stream limited the number of times the cellulose containing residue could be recycled. © 1996 John Wiley & Sons, Inc.
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  • 19
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    Biotechnology and Bioengineering 51 (1996), S. 399-409 
    ISSN: 0006-3592
    Keywords: cell damage ; cell culture ; bubble aeration ; agitation ; bubble coalescence and breakup ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: It has been established that the forces resulting from bubbles rupturing at the free air (gas)/liquid surface injure animal cells in agitated and/or sparged bioreactors. Although it has been suggested that bubble coalescence and breakup within agitated and sparged bioreactors (i.e., away from the free liquid surface) can be a source of cell injury as well, the evidence has been indirect. We have carried out experiments to examine this issue. The free air/liquid surface in a sparged and agitated bioractor was eliminated by completely filling the 2-L reactor and allowing sparged bubbles to escape through an outlet tube. Two identical bioreactors were run in parallel to make comparisons between cultures that were oxygenated via direct air sparging and the control culture in which silicone tubing was used for bubble-free oxygenation. Thus, cell damage from cell-to-bubble interactions due to processes (bubble coalescence and breakup) occurring in the bulk liquid could be isolated by eliminating damage due to bubbles rupturing at the free air/liquid surface of the bioreactor. We found that Chinese hamster ovary (CHO) cells grown in medium that does not contain shear-protecting additives can be agitated at rates up to 600 rpm without being damaged extensively by cell-to bubble interactions in the bulk of the bioreactor. We verified this using both batch and high-density perfusion cultures. We tested two impeller designs (pitched blade and Rushton) and found them not to affect cell damage under similar operational conditions. Sparger location (above vs. below the impeller) had no effect on cell damage at higher agitation rates but may affect the injury process at lower agitation intensities (here, below 250 rpm). In the absence of a headspace, we found less cell damage at higher agitation intensities (400 and 600 rpm), and we suggest that this nonintuitive finding derives from the important effect of bubble size and foam stability on the cell damage process. © 1996 John Wiley & Sons, Inc.
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  • 20
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    Biotechnology and Bioengineering 51 (1996), S. 434-438 
    ISSN: 0006-3592
    Keywords: polyphosphate ; Escherichia coli ; phosphate starvation ; gene expression ; heterologous ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effect of intracellular polyphosphate on the phosphate-starvation response in Escherichia coli was studied by genetically manipulating the intracellular polyphosphate levels and by performing phosphate shifts on the genetically engineered strains. Strains that produced large quantities of polyphosphate and were able to degrade it induced the phosphate-starvation response to a lesser extent than wild-type strains, whereas strains that were unable to degrade a large intracellular polyphosphate pool induced the phosphate-starvation response to a greater extent than wild-type strains. These results have important implications for expression of heterologous genes under control of the phoA promoter. © 1996 John Wiley & Sons, Inc.
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  • 21
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    Biotechnology and Bioengineering 51 (1996), S. 458-465 
    ISSN: 0006-3592
    Keywords: concentric-cylinder shear device ; rotor/stator homogenization ; shear ; shear rate ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Shear is present in almost all bioprocesses and high shear is associated with processes involving agitation and emulsification. The purpose of this study is to investigate the effect of high shear and high shear rate on proteins. Two concentric cylinder-based shear systems were used. One was a closed concentric-cylinder shear device (CCSD) and the other was a homogenizer with a rotor/stator assembly. Mathematical modeling of these systems allowed calculation of the shear rate and shear. The CCSD generated low shear rates (a few hundred s-1), whereas the homogenizer could generate very high shear rates (〉 105 s-1). High shear could be achieved in both systems by increasing the processing time. Recombinant human growth hormone (rhGH) and recombinant human deoxyribonuclease (rhDNase) were used as the model proteins in this study. It was found that neither high shear nor high shear rate had a significant effect on protein aggregation. However, a lower melting temperature and enthalpy were detected for highly sheared rhGH by using scanning microcalorimetry, presumably due to some changes in protein's conformation. Also, SDS-PAGE indicated the presence of low molecular-weight fragments, suggesting that peptide bond breakage occurred due to high shear. rhDNase was relatively more stable than rhGH under high shear. No conformational changes and protein fragments were observed. © 1996 John Wiley & Sons, Inc.
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  • 22
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    Biotechnology and Bioengineering 51 (1996), S. 494-499 
    ISSN: 0006-3592
    Keywords: cell metabolism ; baculovirus ; insect cells ; recombinant protein OSF-2 ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The properties of Sf9 and Tn5 insect cells were analyzed comparatively under serum-free culture conditions. Sf9 cells in SF900II medium apparently utilized sucrose as a primary nutrient both before and after virus infection, yielding small amounts of lactate and ammonia. Tn5 cells in Excell 401 medium consumed all the nutrients examined, including sucrose. The productivity of a recombinant glycoprotein, OSF-2, by Tn5 cells, was moderate in both monolayer and spinner cultures, but the ability to secrete it was compromised in the former case. Relative to the Tn5 cultures, Sf9 produced 30-fold more OSF-2 in either culture mode. © 1996 John Wiley & Sons, Inc.
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  • 23
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    Biotechnology and Bioengineering 51 (1996), S. 538-543 
    ISSN: 0006-3592
    Keywords: NMR imaging ; biosorption ; alginate ; shrinking core model ; Laminaria ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In this contribution, an NMR imaging study of heavy metal absorption in alginate, immobilized-cell biosorbents, and kombu (Laminaria japonica) algal biomass is presented. This method provides the good possibility of directly monitoring the time evolution of the spatial distribution of the ions in the materials. From these results, we demonstrate that rare earth ions are absorbed with a steep reaction front that can be described very well with a modified shrinking core model, while copper ions are absorbed with a more diffuse front.
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  • 24
    ISSN: 0006-3592
    Keywords: oxidoreductase ; chiral alcohol ; racemic resolution ; membrane reactor ; continuous extraction ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Oxidations of alcohols by alcohol dehydrogenases often suffer from low conversions and slow reaction rates due to severe product inhibition. This can be overcome by continuous product extraction, because only the concentrations, but not the kinetic parameters, can be changed. As a consequence, it is favorable to apply a differential circulation reactor with continuous product extraction, where only a small amount of product is formed per cycle. The product is then directly extracted using a microporous hydrophobic hollow fiber membrane. This results in an increase of the relative activity of the dehydrogenase at a given conversion. The reaction investigated is the kinetic resolution of racemic 1-phenyl-1,2-ethanediol by glycerol dehydrogenase (GDH). The resulting oxidation product, 2-hydroxyacetophenone, causes a strong product inhibition. Additionally, it reacts in a chemical reaction with the cofactor lowering its active concentration. Because the GDH needs β-nicotinamide adenine dinucleotide (NAD+) as a cofactor, lactate dehydrogenase is used to regenerate NAD+ from NADH by reducing pyruvate to (L)-lactate. A conversion of 50% with respect to the racemate and an enantiomeric excess 〉99% of the (S)-enantiomer was reached.
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  • 25
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    Biotechnology and Bioengineering 51 (1996), S. 581-590 
    ISSN: 0006-3592
    Keywords: microfiber ; graft polymerization ; DNA immobilization ; immunoadsorbent ; DNA ; anti-DNA antibody ; systemic lupus erythematosus ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Immobilization of DNA to the surface of poly(ethylene terephthalate) (PET) microfibers with a high specific surface area of 0.83 m2/g was carried out to give the fiber surface an affinity for anti-DNA antibody. Following ozone oxidation, the microfibers were subjected to graft polymerization of monomers including acrylic acid, methacryloyloxyethyl phosphate, N,N-dimethylaminoethyl methacrylate, N-vinylformamide, and glycidyl methacrylate. Calf thymus DNA was immobilized to the grafted fiber surface through either covalent binding or polyion complexation with the grafted polymer chains. The highest surface density of DNA immobilized (0.6 μg/cm2) was obtained when DNA was immobilized through formation of phosphodiester linkage between the hydroxyl group of DNA and the phosphate group in grafted poly(methacryloyloxyethyl phosphate) using 1,1-carbonyldiimidazole, or through polyion complexation between the anionic DNA and the cationic grafted poly(N,N-dimethylaminoethyl methacrylate) chains. Batch adsorption of anti-DNA antibody to the grafted PET fibers with and without DNA immobilized on their surface was conducted with serum obtained from systemic lupus erythematosus model mice. The DNA-immobilized PET fibers exhibited a higher adsorption capacity and specificity than the others. In addition, the DNA-immobilized fibers effectively adsorbed human anti-DNA antibody.
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    Biotechnology and Bioengineering 12 (1970) 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 27
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    Biotechnology and Bioengineering 12 (1970), S. 51-61 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A model of oxygen absorption in microbiological systems of zero order reaction rate is proposed. The partial differential equation was solved to predict the profile of the oxygen concentration boundary layer next to a gas-liquid interface. Generally speaking, the presence of microbial cells always helps to increase the oxygen absorption rate over that of physical absorption. Only when the microbiological reaction is slow as judged by the fact that the reaction time, tr, is much larger than the diffusion time, tD, can one rightfully approximate the oxygen absorption in microbiological suspensions by physical absorption.
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    Biotechnology and Bioengineering 12 (1970), S. 85-92 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Glucoamylase bound to DEAE-cellulose in 0.05 M sodium acetate, pH 4.0, is active in the conversion of starch to glucose. The activity of the DEAE-cellulose-bound enzyme ranges from 16 to 55% of the activity of the free enzyme. Binding of the enzyme narrows the pH optimum to approximately 4.0 and lowers the temperature optimum to 40-50°C as compared to a 60°C temperature optimum for the free enzyme. Concentrations of acetate buffer above 0.1 M disrupt the DEAE-cellulose-enzyme complex. Columns were used with some success for the continuous conversion of starch. Pretreatment of the starch with α-amylase and clarification were necessary to prevent blocking of the column. Columns maintained activity for more than 3 weeks of continuous operation.
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    Biotechnology and Bioengineering 12 (1970), S. 541-560 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A continuous symbiotic algal-bacterial system was developed consisting essentially of a mixed Chlorella-activated sludge culture which would efficiently remove nutrients from wastewater under aerobic conditions without supplementary aeration. Oxygen decline data were fitted to a mathematical model used to predict respiratory rates, photosynthetic oxygenation, and steady-state oxygen concentrations. Stable relative biological populations and a dissolved oxygen concentration of about 2 mg/1 were maintained during steady-state operation with daily harvesting of excess biomass. Respiratory and physiological relationships indicated that the carbon dioxide-oxygen balance is a primary control that governs the steady-state operation of a symbiotic algal-bacterial culture. The close association of the algae and bacteria resulted in an algal-bacterial floc with settled rapidly yielding a clear supernatant.
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    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
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    Notes: Dialysis was attempted as a means to alleviate the product-activated controls presumed to limit the formation of threonine by an auxotrophic mutant of Escherichia coli strain W. The occurrence of inhibition rather than enhancement of yields by dialysis was traced to the fact that threonine actually was not inhibiting its own synthesis. Instead, α,ε-diaminopimelic acid became depleted, but the imbalance could not be corrected by exogenous replacement.
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    Biotechnology and Bioengineering 12 (1970), S. 641-644 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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    Biotechnology and Bioengineering 12 (1970), S. 771-801 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
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    Biotechnology and Bioengineering 12 (1970) 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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    Biotechnology and Bioengineering 12 (1970), S. 873-887 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The removal of cellular material from fermentation broths is of importance in many fermentation processes. The present work compares the performance of recently developed polyelectrolyte flocculating additives with traditionally available additives. Objectives are to establish criteria for the choice of a flocculating additive and establish optimum conditions for the formation of stable, fast settling floe, and for quantitative separation of cellular material from the medium. Fermentation broths of actively growing Candida intermedia were used to evaluate the effectiveness of fifty commercial flocculating additives at different dosages and pH values. Certain strong anionic and strong cationic polyelectrolytes and mineral hydrocolloids were found to be most effective in their enhancement of settling rates. Some differences in behavior exist between glucose grown cells, hydrocarbon grown cells, and washed cells in buffer suspension. Flocculation of cells from fermentation broths is concluded to be highly dependent upon adsorbed material. A high charge density to interact or compete with adsorbed material and a solubility in the adsorbed material are important factors in choosing an additive for a given application. The fluid mechanics of a flocculating suspension is an important variable since low shear does not provide adequate contacting between cells for floe formation and high shear leads to floe breakup. An apparatus was constructed to grow floe under constant fluid mechanical conditions both in laminar and turbulent flow regimes. Turbulent shear was found to be very important in forming large, compact floe in cases where irreversible ionic bridging is the mechanism as for the strong anionic polyelectrolyte, polystyrene sulfonate. Adequate mixing is required to disperse the flocculating additive, but the level of turbulence is relatively unimportant in cases where reversible colloidal bridging is the mechanism as for the mineral hydrocolloid, bentonite.
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    Biotechnology and Bioengineering 12 (1970), S. 947-959 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In order to isolate proteins from microalgae, yeasts and bacteria, cell disintegration in a special ball-mill was performed. The degree of disintegration of the different microorganisms was compared. The dependence of disintegration on bead size and on the ratio between the volume of suspension and the volume of glass beads was also investigated. Nondisintegrated and disintegrated cells were extracted with sodium hydroxide and the amount of extractable nitrogen and the amount of nitrogen precipitable at pH 4.0 were determined. The dependence of yield on the sodium hydroxide concentration, extraction time, and temperature was studied. When extracting undisintegrated cells, very low yields were obtained and the nitrogen extracted was mostly nonproteinous. For disintegrated cells high yields were obtained. An optimum was found after extraction with 0.3-0.5% sodium hydroxide; at pH 11.0-11.5. The precipitate obtained represented 60-70% of the cell nitrogen. The nitrogen content of the precipitate was 12-14% of the dry weight.
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    Biotechnology and Bioengineering 12 (1970), S. 1081-1098 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Optimization methods based on the continuous maximum principle and the calculus of variations were used to calculate optimum temperature profiles for batch penicillin fermentations. These methods were first applied to several general models to develop effective techniques for the numerical solution of the equations. Subsequently, these methods were applied to two particular models, derived from experimental data, and the optimum temperature profiles were determined. The results indicated that an improvement, in penicillin yield of about 15% was possible if the optimum temperature profiles were followed.
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    Biotechnology and Bioengineering 12 (1970), S. 1-17 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The precipitation of proteins with heteropolyacids has been studied for the purpose of large scale primary purification. A precipitate will form if the pH of the reaction between purified ovalbumin, hemoglobin, trypsin, pepsin, bovine serum albumin, ovomucoid, gelatin or ribonuclease and tungstrophosphoric, tungstosilicic or molybdosilicic acid is close to the isoelectric point of the protein and does not cause the dissociation of the heteropolyacid. Below the isoelctric point, the percent precipitation depends on the conformational changes of the protein. The precipitation of ovalbumin with tungstophosphoric decreases as the ionic strength of the buffer increases and is independent, of the protein concentration. Mixtures of ovalbumin and bovine serum albumin, though having close isoelectric points, can be separated by varying the concentration of the precipitant. The electropositive groups which combine with the tungstophosphoric acid are guanidino, ε-amino and imidazole. No precipitation is given by the α-amino groups. Filtrates of microbial fermentations containing lactase, glucose aerode-hydrogenase, alkaline protease, amyloglucosidase, and transglucosylase have been purified by precipitation with heteropolyacids.
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    Biotechnology and Bioengineering 12 (1970), S. 63-74 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A pilot-plant process has been developed for the continuous extraction and partial purification of prolyl-tRNA synthetase from mung bean. The bean slurry was wet ground in a hammer mill, clarified by two-stage centrifugation, and the protein in the effluent fractionated by precipitation at pH values of 5.2 and 4.2. The throughput was 13 kg dry bean/hr. The improved extraction process and reduced processing time resulted in an enzyme product with a specific activity 16 × that previously obtained in the batch process. The yield was also 50-60 times higher.
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    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The processing of fermentor-grown, edible yeast involves the removal of water. This can be accomplished through concentration followed by drum or spray drying. This study presents the essential physical properties of yeast solutions necessary for calculation of production economics. In addition, our initial studies of vacuum concentration show that some of the cell leakage necessary for good drying characteristics occurs. The residence time during concentration is also sufficient, to yield 1-2 log cycles of kill which are mandatory since the final product, should contain no viable cells.
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    Biotechnology and Bioengineering 12 (1970) 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
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  • 41
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    Biotechnology and Bioengineering 12 (1970), S. 273-290 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The concept of a “critical oxygen concentration” is conventionally considered to hold for the submerged aerobic fermentation of glucose to gluconic acid. Above the critical level the fermentation rate is supposedly independent of oxygen concentration. In this work it is shown that, at a given agitation rate, the fermentation is independent of dissolved oxygen when above the critical. However, an increase in the agitation rate results in an increase in the fermentation rate. This increase was shown to be accompanied by an increase in the gluconolactone concentration in the broth. Gluconolactone, an intermediate in the reaction pathway, is hydrolyzed nonenzymatically to gluconic acid. Evidence is presented to suggest that the increased gas-liquid interfacial area brought about by increased agitation causes an increased net rate of lactone formation. This in turn results in an increased rate of hydrolysis of the lactone to gluconic acid. A model is presented hypothesizing that negatively charged cells adsorb at the gas-liquid interface. These cells attract hydrogen ions, causing a lowering of the pH in the film around the bubbles. It is this lowered pH which is considered to bring about increased fermentation rates when the interfacial area is increased. Supporting evidence is presented.
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    Biotechnology and Bioengineering 12 (1970) 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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    Biotechnology and Bioengineering 12 (1970), S. 353-378 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Mathematical models of the interaction between predator and host populations have been expressed as systems of nonlinear ordinary differential equations. Solutions of such systems may be periodic or aperiodic. Periodic, oscillatory solutions may depend on the initial conditions of the system or may be limit cycles. Aperiodic solutions can, but do not necessarily, exhibit oscillatory behavior. Therefore, it is important to characterize predatory-prey models on the basis of the possible types of solutions they may possess. This characterization can be accomplished using some well-known methods of nonlinear analysis. Examination of the system singular points and inspection of phase plane portraits have proved to be useful techniques for evaluating the effect of various modifications of early predator-prey models. Of particular interest is the existence of limit cycle oscillations in a model in which predator growth rate is a function of the concentration of prey.
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    Biotechnology and Bioengineering 12 (1970), S. 399-407 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The enzyme L-amino acid oxidase of Crotalus adamanteus was covalently coupled to porous 96% silica glass particles. The insolubilized enzyme was active on several L-amino acids including: leucine, isoleucine, cysteine, phenylalanine, tryptophane, and methionine. No activity was observed with D-amino acids, L-asparagine, or L-proline. Maximum activity was observed at pH 7.8. Stability of the enzyme derivative was demonstrated by continuous operation of an enzyme column for 35 days, during which the bound enzyme oxidized over 5000 times its own weight of substrate.
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    Biotechnology and Bioengineering 12 (1970), S. 465-482 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A method for analyzing the reactor behavior of a continuous, multistage tower fermentor is described. A model consisting of a system of interconnected, ideal subreactors is set up on the basis of the fermentor's configuration and flow pattern. The residence time distribution curve is used to test the validity of the model and the relative quantities of flow streams and regions in the model are determined. A least-square fitting procedure between measured and calculated distribution curves is used to identify the proper model. The application of this method to real cultivation conditions is also discussed. Using this approach, the multistage tower fermentor is shown to be equivalent to a cascade of four perfectly mixed tanks with a backtracking stream between stages. The extent of backflow under various conditions has also been determined.
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    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
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    Notes: The microbiological extraction of zinc from a high-grade zinc sulfide concentrate has been investigated, using a pure strain of Thiobacillus ferrooxidans. Conditions such as temperature, pH, pulp density, nutrient, concentration, and specific surface of solids have been studied in terms of their effects on zinc extraction rate and in some instances on final zinc concentration in solution. Where appropriate, optimum conditions for leaching have been specified.
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    Biotechnology and Bioengineering 12 (1970), S. 577-589 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The conversion of naphthalene to salicylic acid by Pseudomonas fluorescens was studied as a model for dialysis fermentation. In a demonstration experiment, the continual removal of the product by dialysis and by intermittent replenishment of the dialysate reservoir caused cyclical changes in the concentrations of viable cells and product. The cumulative total amount of both cell mass and salicylate, however, continued to increase steadily until the experiment was terminated after 15 days. At this time the rate of product formation was highest and still increasing, although less than 10% of the cells were viable. The terminal amount of salicylate was about 20-fold greater than the maximum reached in the control fermentation, and was calculated to be 2.6-fold more productive even if the control were optimally recycled. Methods were projected to achieve still further improvements.
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    Biotechnology and Bioengineering 12 (1970), S. 633-634 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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  • 49
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    Biotechnology and Bioengineering 12 (1970), S. 635-639 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Of several candidate disinfectants for use in tissue culture work, especially suspension cultures, sodium hypochlorite solution was selected to test its effect on growing cells. Metabolizing cells reduce, sodium hypochlorite oxidizes ; therefore NaOCl leakage into such systems must be neutralized with no untoward effects on the cells. Dilutions of routine disinfectant-grade sodium hypochlorite were tested against cell cultures. Those exposed to 15.62 to 31.25 ppm of NaOCl grew with no apparent cell damage.
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    Biotechnology and Bioengineering 12 (1970), S. 679-712 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: High substrate concentrations inhibit growth and may distort the metabolism of microorganisms. Mechanisms causing substrate inhibition are discussed and used to derive several mathematical models representative of the entire concentration range, including stimulation of growth by low substrate concentrations. These kinetic models are tested with a variety of batch culture measurements of specific growth rate and respiration rate at widely-ranging substrate concentrations. Using one of the kinetic models, equations are developed for batch, continuous, and exponential-feed reactors. Comparison of results obtained in continuous culture with results from exponential-feed culture systems is shown to offer a novel experimental method for evaluating the effect of the cell age distribution on the properties and metabolic activity of a culture.
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    Biotechnology and Bioengineering 12 (1970), S. 803-830 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Two kinds of mathematical models have been developed for batch penicillin fermentations: (1) general models, based on averaged, nondimensionalized cell and penicillin synthesis curves from plant, scale fermentors and (2) particular models developed from specific sets of experimental data from two sources. Parameter-temperature functions used with the general models were assumed to have general shapes which could apply to many fermentations, i.e., they were based on the familiar temperature response of enzyme-catalyzed reactions. Parameter-temperature functions for the particular models were determined from experimental data for batch runs at various temperatures.
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  • 52
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    Biotechnology and Bioengineering 12 (1970), S. 913-920 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In view of the recent development that some petrochemical products are efficiently available as substrates for the fermentation industry, glycerol manufactured from propylene by chemical synthesis would also be hoped for the purpose. This paper describes some of the factors influencing mannitol production from glycerol by Torulopsis yeasts and a microbial conversion of glycerol to D-fructose via mannitol, in which two sequential steps of yeast and Acetobacter fermentation are involved. Torulopsis mannitofaciens CBS 5981 and Torulopsis vcrsatilis CBS 1752, exceptionally good mannitol producers, were selected for the study. High concentrations of nitrogen sources and KH2PO4 in the medium markedly decreased mannitol yield in spite of good utilization of the substrate. T. mannitofaciens produced mannitol in yield of 31% of the glycerol consumed at optimal condition. The fermentation by washed yeast cells gave much higher mannitol yield of more than 50%. A sequential fermentation process was carried out without isolation and purification of the intermediate and yielded.51.7%. D-fructose from the glycerol.
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  • 53
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    Biotechnology and Bioengineering 12 (1970), S. 961-974 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Growth and lactic acid production by L. delbrueckii was studied in a dialysis culture system and the inhibitory effect of lactate confirmed by removing lactate from the culture medium by dialysis. It has been shown that lactate inhibits growth after the log phase and that the maintenance of low lactate concentrations after this point permits higher specific growth rates and higher maximum cell concentrations. Acid production is also significantly higher in a dialysis culture system. Finally, a modification of the Luedeking-Piret model, incorporating the lactate inhibition effect, is proposed.
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  • 54
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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    Notes: Extended culture, a special type of semicontinuous culture, permits prolonged maintenance of a constant or programmed environment in a growing culture by a controlled addition of one or more substrates. Differences between extended culture and continuous culture data are a measure of differences in the properties of cell populations with different cell age distributions but identical steady-state environments. Both extended culture and continuous culture were used to study the growth kinetics of Candida utilis (ATCC 9226) under conditions of substrate inhibition at controlled concentrations of sodium acetate in a carbon-limited mineral salts medium supplemented with 0.01 g/1 yeast extract. Acetate concentrations ranged from 1.2 g/l to 10.8 g/l (expressed as acetic acid), while yeast concentrations varied from 0.3 to 7.8 (g dry cells)/1. Rate parameters such as growth yields (Y), specific growth rates (μ), and linear growth rates (K), were calculated by computer from the data and theory presented herein. Specific growth rates as high as 0.54/hr were observed, although extended culture growth was more nearly linear than exponential in these experiments. Growth yields usually varied between 0.2 and 0.4 (g dry cells)/(g acetate), although values were as high as 0.8 for a brief period during one experiment. Growth yields at a given acetate concentration were correlated by an equation of the form 1/Y = 1/YG + m/μ. A maintenance coefficient (m) of 0.17 (g acetate)/(g dry cell-hr) was observed at acetate concentrations of 4.5 and 10. g/1. A typical maximum growth yield (YG) of 0.51 (g dry cell)/(g acetate) was obtained at 4.5 g/1 acetate, but an unusually high YG of 1.33 was found at 10. g/1 acetate. Oxygen uptake measurements are compared with these cell yield measurements. Linear growth rates in expended culture were correlated by the equation K = 0.89-0.70 (S/S0) where K has units of (g dry cell)/(l-hr), S is the instantaneous acetate concentration, and S0 is the initial acetate concentration. The extended culture kinetic data are shown to be substantially different from continuous culture kinetic data. Reason for these differences are discussed in light of diffrences in the cell age distributions, as well as possible differences in experimental conditions.
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    Biotechnology and Bioengineering 12 (1970), S. 75-84 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Xanthan biopolymer has been produced by single-stage continuous fermentation with Xanthomonas campestris NRRL B-1459 in a medium of glucose, minerals, distillers' solubles, and urea for as long as 20 days. At the highest dilution rate studied (D = 0.0285 hr-1), the steady state rate of xanthan production was 0.36 g/kg/hr and the steady state yield, basis glucose consumed, was 68%. Observations indicate that xanthan production rate is a function of pH and D.
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    Biotechnology and Bioengineering 12 (1970), S. 483-500 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Aerobiology studies at the Naval Biological Laboratory require use of a vacuum system to provide safe disposal of air contaminated with pathogenic, microorganisms. A system for thermal decontamination of this process air has been installed and tested. The system uses a natural gas burner to heat approximately 550 cfm of air to temperatures exceeding 750°F. Tests showed a reduction in number of acrosolized viable hardy spores (Bacillus subtilis var. niger) of more than 8 logs at design flow rates. The kill rate (D values) measured in this system is somewhat higher than those reported by other workers. The annual owning and operating cost of the system is approximately $9000.
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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    Biotechnology and Bioengineering 12 (1970), S. 1001-1017 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A simple model is proposed for continuous stirred microbial culture system to explain observed deviations from the behavior predicted on the basis that the fluid is completely mixed, which arises from the use of radial flow impellers in such apparatus.
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  • 59
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    Notes: The effects of mixing on the critical mean holding time for washout and the steady state performance of growth processes in continuous flow reactors are investigated. Macromixing, micromixing, and cell recycle arc considered. The tanks-in-series model composed of N completely mixed flow reactors, the dispersion model, the plug flow model, and a combined model composed of a plug flow reactor and a continuous stirred tank flow reactor connected in series arc used to represent the macro-mixing or residence time distribution. The extreme cases of micromixing, namely, complete segregation and maximum mixedness, as well as intermediate states of micromixing are investigated to determine their effects on washout and the occurence of multiple steady states. A technique for predicting the maximum mixedness washout condition from a knowledge of the residence time distribution is presented and used to determine the washout condition for the dispersion model under maximum mixedness conditions.
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  • 60
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    Notes: Cellulase of Trichoderma viride was concentrated in various molecular cutoff membranes, and flux rates and retention of activity were studied under ultra-filtration conditions. Little or no Cellulase was discharged through the membranes tested. The concentrated (5-8-fold) enzymes were used to saccharify finely ground substrate (Solka Floe) in stirred tank (STR) and membrane reactors (MR). A pressure filtration vessel provided with a membrane for simultaneous removal of low molecular weight products (glucose) from the reacting system (Cellulose-Cellulase) is designated as a membrane reactor. Continuous digestion of dense cellulose suspension in the membrane reactor was achieved. Using PM-30 (Amicon) membrane reasonably high mass flux values (9.7-23.3 gals/ft2 - day) were obtained in separating glucose from a digest of 30% cellulose suspension. Abcor membrane (HFA 300) was equally effective and necessitated less care in handling. Nearly 14% glucose concentration has been achieved in less than 50 hrs in STR by digesting a 30% cellulose suspension. Based on experimental data a model system is proposed for the continuous steady state Saccharification of ground substrate in which there is continuous removal of concentrated glucose syrup, and a feedback of enzyme.
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    Biotechnology and Bioengineering 12 (1970), S. 1069-1079 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The effect of the rate of ‘natural death’ of bacteria on the steady state behaviour of continuous culture has been studied. A model which has no real biochemical basis but which gives good experimental correlation has been proposed. Populations of Acrobacter Acrogenes harvested from the chemostat at dilution rates greater than 0.1 HR-1 were found to be over 95% viable. The effect of the rate of death on the steady state yields becomes significant only at very low dilution rates. The experimental work of other workers has also been simulated to test the validity of the model. Theoretically it is also shown that an ‘apparent’ lag in batch cultures will be observed if the innoculum is of very low viability.
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    Notes: Bacteriophages are widely distributed in nature and may be important factors in regulating populations of their hosts. Model continuous culture systems of a single bacterial species and a temperate parasitic phage have been studied. Steady state cultures of lysogenic Escherichia coli 159T- (λcts) produced a small quantity of free λ cts phage. Temperature shocking such a culture resulted in a sharp increase in phage concentration with a concomitant fall in cell population. With time the system returned to a steady state condition.
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    Biotechnology and Bioengineering 12 (1970), S. 409-417 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Earlier observations revealed that incubation of media and the attendant changes in oxidation-reduction potential (ORP) were related to improved cell production. This is a report, of work done to show that the higher levels and increased rates of growth of cells grown in incubated medium are associated with the ORP level of the medium before inoculation of the medium with cells. Work was done using 250-ml centrifuge spinner bottles as the culture vessels. Further work is needed to establish the desirability of deliberate poising of media prior to use for studies in small vessels and flasks.
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    Biotechnology and Bioengineering 12 (1970), S. 1103-1109 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Enzymes subjected to shearing in a viscometer are partially inactivated. It is possible with viscometry to calculate the degree of inactivation that occura when an enzyme solution flows through a capillary tube. When shear rate × exposure time is less than 104, there is little or no inactivation.The masa average shear-rate × time or shear, for laminar flow in a cylindrical tube is simply 16L/3D. It is surprising that for a single pass through a tube, the masa average shear is independent of flow rate and shear rate.
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    Biotechnology and Bioengineering 12 (1970), S. 141-143 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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    Biotechnology and Bioengineering 12 (1970), S. 157-158 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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    Biotechnology and Bioengineering 12 (1970), S. 251-271 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The amylase of Bacillus subtilis NRRL B3411 has been purified and partially characterized. The specific activity can be increased from 300,000 units/g to 6,000,000 units/g with a 60% recovery of total units. The purified material consists of one major and one trace anodic component as determined by disc gel electrophoresis. The molecular weight was 48,000 as determined by bio-gel filtration; the molecular weight was 44,900 ± 2400 as determined by sedimentation equilibrium methods. This purified enzyme is stable at, 70°C in the presence of 0.01 M Ca++ and 0.1 M NaCl over a broad pH range from 5.5-9.5. The pH activity profile indicates optimum activity at pH 6.0. This amylase exhibits maximum activity at 60°C. The enzyme is a liquefying α-amylase as determined by analysis of hydrolysis products and immunological studies.
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    Biotechnology and Bioengineering 12 (1970), S. 341-346 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The ecological significance of bacterial capsules when virulent bacteriophages are present was explored by exposing continuous cultures of Escherichia coli ATCC 11303, in various stages of capsulation and clumping, to a virulent coliphage, T2. Only partial protection was provided by capsulation, but this could be a factor affecting survival in complex mixed cultures.
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    Biotechnology and Bioengineering 12 (1970), S. 379-397 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The main purpose of the work reported here was to establish the effectiveness of aeration and agitation, and to determine the best conditions of aeration for the growth and production of glucose oxidase of Aspergillus niger, on a semi-industrial scale. Concentration of dissolved O2, O2 consumption and CO2 production were measured. It was found that the rate of growth and the activity of glucose oxidase per gram mycelium increased with the increase of speed of agitation. The concentration of dissolved oxygen of the fermentation broth, as well as the rate of respiration (O2 consumption and CO2 production) increased in direct proportion to the increase of speed of agitation, while assimilation of sugars was accelerated. The values of the respiratory ratio showed a fluctuation according to the presence or absence of sugar in the medium.
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    Biotechnology and Bioengineering 12 (1970) 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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    Biotechnology and Bioengineering 12 (1970), S. 519-539 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The mechanisms and kinetic course of BOD exertion were compared in both open and closed systems. Two open reactors, a simulated stream device, and an open stirred reactor were employed, and the closed systems consisted of standard BOD bottles and 2.4-liter vessels. In the closed systems, both quiescent and stirred conditions of incubation were examined. Biological solids concentration, bacteria and protozoa concentration, substrate analysis, and chemical oxygen demand as well as biochemical oxygen utilization were employed to assess the performance of these systems.Oxygen uptake rate constants were observed to increase with increasing concentration o carbon source, thus militating against irect use of the usual dilution technique for predicting rate of deoxygenation in receiving streams. The relationship between specific O2 uptake rate and substrate concentration approximated a hyperbolic function similar to the Mono relationship for specific growth rate and substrate concentration. A technique using an open stirred reactor than the standard BOD bottle dilution technique is recommended.
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    Biotechnology and Bioengineering 12 (1970), S. 591-601 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A kinetic analysis was made of the relationship between salicylate production from naphthalene and growth of Pseudomonas fluorescens in semicontinuous dialysis culture. The specific rates both of product formation and growth initially were increased by the diffusional withdrawal of salicylate, but subsequently were reduced to low levels despite continued salicylate removal. Productivity and growth were correlated by the Luedeking-Piret equation in an initial nondialysis period and in the early stages of dialysis fermentation, when specific growth rates exceeded. 005 hr-1. Below this level of growth at later stages of dialysis fermentation, the specific production rate was correlated only with total cell mass by a proportionality constant of .035 hr-1, which was attributed to maintenance metabolism. Maintenance accounted for about 84% of the total salicylate produced, while growth-associated metabolism accounted for the remainder.
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    Biotechnology and Bioengineering 12 (1970) 
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    Biotechnology and Bioengineering 12 (1970), S. 713-746 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In this work, a mathematical model which can be used to describe butch growth in fermentations with two liquid phases present is developed for systems in which the growth limiting substrate is dissolved in the dispersed phase. The model takes into account the drop size distribution, the rate of adsorption of cells on the drop surface, the rate of desorption of cells from the drop surface, substrate transport between phases, phase equilibrium, and growth kinetics. The model also considers the effect, of coalescence and redispersion of oil drops in the system. It is assumed that the composition of the dispersed phase is such that substrate utilization from it causes little or no change in the interfacial area. A discrete uniform distribution and a discrete normal distribution which is obtained from an experimental distribution curve are used as drop size distributions. Simulation results are obtained for a wide range of parameter values using the IBM S/360 Continuous System Modeling Program.
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    Biotechnology and Bioengineering 12 (1970), S. 651-677 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The utilization of an exogenous substrate by enzyme inside a bacterial cell can be limited by diffusion up to the cell, penetration of the cell, diffusion within the cell, and/or attack by internal enzyme. For small molecular weight substrates such as galactosides, and for bacteria such as Escherichia coli the diffusion steps are not rate limiting even with the permeases fully induced and the external concentration of substrate low. In permeaseless organisms with more than about 20 enzyme molecules per cell, permeation of O-nitrophenyl-β-D-galactoside through the membrane is limiting. Thus, a single initiation of transcription of a lactose message suffices to yield enough enzyme molecules to switch an uninduced cell from enzyme limitation to permeability limitation. Subsequent initiations change the cellular activity very little. This transition can be followed by assaying enzyme activity of both intact and lysed cell suspensions. In this way the induction response amongst cells in growing populations at high inducer concentrations has been found to be uniform. It was found that nearly all of the cells from balanced growing culture are immediately inducible even with doubling times as short as 7.6 hrs. At 24 hrs about 1/3 of the cells are inert at any time, but all cells synthesize enzyme within a 3-hour period. At low inducer concentration or in the present of catabolite repressor the rate of initiation is greatly decreased; this leads to a non uniform distribution of enzyme within the cells, which is readily detected by the experimental technique. In addition to developing the kinetics for enzyme contained in cells, the distribution of the enzyme among uninduced bacteria is presented.
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    Biotechnology and Bioengineering 12 (1970), S. 845-847 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
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    Biotechnology and Bioengineering 12 (1970), S. 849-871 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
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    Notes: Two types of steam-sterilizable dissolved-oxygen probs were evaluated for use in fermentations. A galvanic-cell probe was selected over a polarographic probe because of its demonstrated ruggedness and dependability. Various methods for determining kLa in fermentors were compared and the oxygen balance method selected for use in viscous streptomycete fermentations. Rheological data are presented to identify a range of mash viscosity where many kLa measurement methods are not applicable. Oxygen transfer data are presented for streptomycete fermentations pilot fermentors.
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    Notes: One important economical method for producing singlecell protein is to spray dry the cultured cells. This study presents some preliminary data on the effects of spray drying on cell viability. Under conditions similar to those for the production of spray-dried milk, 4-5 log cycles destruction occurred. The results indicate that, the activation energy for thermal destruction of yeast was reduced from the normal heat treatment value of 84 kcal/°K mole to about 38 kcal/°K mole.
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    Biotechnology and Bioengineering 12 (1970), S. 159-166 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
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    Notes: Streptomyces mannosidase, like the enzyme from other sources, is shown to require a divalent cation for enzyme activity. N-Ethylmaleimide pretreatment of enzyme-containing cells eliminated the requirement of aeration for enzyme activity. Methyl-α-D-mannoside was found to be a strong inhibitor of the hydrolysis of both p-nitrophenyl-α-D-mannoside and mannosidost reptomycin. The enzyme is bound at or near the surface of the cell and is inactivated by sonic oscillation. Small participate matter containing most of the activity can be released from the cells into water, such release being inhibited by phosphate, Tris, or sodium chloride.
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    Biotechnology and Bioengineering 12 (1970), S. 213-249 
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    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The alkaline proteases of B. subtilis NRRL B3411, B. pumilis, and B. licheniformis have been isolated by fractionation followed by ion exchange chromatography and their homogeneity demonstrated. General enzyme properties of the B. sublitis NRRL B3411 alkaline protease have been studied and attempts made to differentiate a group of alkaline proteases. It is clear that the alkaline proteases known as Subtilisins or Subtilopeptidases are not, exclusive to B. subtilis but are common to many Bacilli and therefore the generic name Bacillopeptidases has been proposed. It is clear too that on the basis of the effect of pH on activity, amino acid composition, esterase activity, and immunological cross-reactions the Bacillopeptidases can be divided into two groups or types: (a) Bacillopcptidase A (Subtilisin A or Subtilopeptidase A) which includes Subtilisin Carlsberg, B. licheniformis, and B. pumilis alkaline proteases; (b) Bacillopeptidase B (Subtilisin B or Subtilopeptidase B) which includes B subtilis NRRL B3411, Subtilisin Novo, Subtilisin BPN' (Nagarse), alkaline protease Daiwa Kasei, and (probably) B. subtilis var. amylosacchariticus. At present, no further differentiation is possible and whether or not the enzymes within group A or B are identical remains an open question. Methods for examination of crude enzyme mixtures or fermentation beers are described and from the examination of a number of crude enzymes and fermentation beers it appears that organisms producing Bacillopeptidase A do not produce neutral protease or amylase, while organisms producing Bacillopeptidase B produce a neutral protease and amylase as well.
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  • 81
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    Biotechnology and Bioengineering 12 (1970), S. 321-331 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Through the use of pilot plant equipment, transaldolase types I, II, and III (from Candida utilis) have been separated and purified. The procedure includes a time sensitive solvent fractionation below 0°C, ion exchange chromatography, and crystalization. The enzyme yield represents a 41% recovery of crystalline type III and partially purified types I and II.
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  • 82
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    Biotechnology and Bioengineering 12 (1970), S. 615-631 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In most enzymatic reactions, batch or continuous, separation of the enzyme for reuse is difficult if not impossible. A process will be presented in which an Ultrafiltration membrane serves to separate the reaction products from the enzyme and the substrate. In this manner the enzyme may be retained and re-used. Furthermore, under these conditions, the enzyme need only be present in catalytic amounts regardless of the amount of product produced.Under proper operating conditions and proper ultrafiltration membrane selection, a pure solution of α-amylase from Bacillus subtilis may be retained with no loss in enzyme activity over a test period of 30 hr after steadystate has been achieved. In the presence of substrate, the membrane support and ultrafiltration cell serve as the reaction vessel for the hydrolysis of starch. The substrate is continuously pumped into the cell under constant ultrafiltration pressure. The di-, oligo-, and polysaccharides formed from the enzyme reaction then either pass through the membrane as products or are retained. The molecular weight distribution of the products is dependent on the nominal molecular weight cut-off of the membrane, absolute ultrafiltration pressure, enzyme-to-substrate ratio, temperature, and residence time of the substrate in the reactor. In addition to the partial hydrolysis of starch by α-amylase, some preliminary findings on the complete hydrolysis of starch by glucoamylase will also be presented. In these latter studies, the substrate may be completely hydrolyzed to glucose units.
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  • 83
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 84
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    Biotechnology and Bioengineering 12 (1970), S. 747-769 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A number of experimental studies on the dynamic, behavior of the chemostat have shown that the specific growth rate does not, instantaneously adjust to changes in the concentration of limiting substrate in the chemostat following disturbances in the steady state input limiting substrate concentration or in the steady state dilution rate. Instead of an instantaneous response, as would be predicted by the Monod equation, experimental studies have shown that the specific growth rate experiences a dynamic lag in responding to the changes in the concentration of limiting substrate in the culture vessel. The observed dynamic lag has been recognized by researchers in such terms as an inertial phenomenon and as a hysteresis effect, but as yet a systems engineering approach has not been applied to the observed data. The present paper criticizes the use of the Monod equation as a dynamic relationship and offers as an alternative a dynamic equation relating specific growth rate to the limiting substrate concentration in the chemostat. Following the development of equations, experimental methods of evaluating parameters are discussed. Dynamic responses of analog simulations (incorporating the newly derived equations) are compared with the dynamic responses predicted by the Monod equation and with the dynamic responses of experimental chemostats.
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  • 85
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: An enzyme kinetic model has been proposed to describe fermentation processes. This type of model was chosen because it is biologically sound, can incorporate all of the important engineering control variables, and can draw upon, in its development, the extensive kinetic literature. An intial qualitative test for this model was made on the gluconic acid fermentation. A necessary check of the model was that Monod's empirical cell growth and yield equations were derived as a special case. The model also offered an explanation for the hysteresis behavior of the gluconic acid production rate as a function of gluconolactone.
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  • 86
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Gram-sized quantities of purified arginine, formylmethionine, glutamic acid, and phenylalanine-2 tRNAs have been prepared from pools of E. coli K-12 MO7 mixed tRNAs by reversed-phase chromatography after preliminary fractionation on DEAE-cellulose. Purified formylmethionine tRNA and partially purified arginine tRNA and glutamie acid tRNA were obtained from large-scale RPC-3 runs (4 × 36 in. column). The arginine tRNA was further purified by rechromatography on RPC-4 columns, and the gluatmic acid tRNA by rechromatography on an RPC-3 column. Two phenylalanine tRNAs were resolved on large-scale (2 × 96 in. column) RPC-3 runs; only the second phenylalanine tRNA reached a satisfactory degree of activity. About 0.88 g of arginine tRNA, 70% activity; 3.32 g of formylmethionine tRNA, 97% activity; 0.80 g of glutamic acid tRNA, 83% activity and O.92 g of phenylalanine-2 tRNA, 78% activity, were produced. The processing steps employed are reliable and reproducible and the procedure is amenable to routine production of these tRNAs.
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  • 87
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    Biotechnology and Bioengineering 12 (1970), S. 29-50 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The kinetics of the batch-wise liquid-phase oxidation of ferrous sulfate by the organism Thiobacillus ferrooxidans has been studied over a range of temperatures from 20°C to 31°C and in the presence of an abundant supply of oxygen, carbon dioxide, and other nutrients.The rate of oxidation was found to be accurately described by the equation \documentclass{article}\pagestyle{empty}\begin{document}$$ \frac{{dS}}{{dt}} = \frac{{\mu _m SX}}{{Y(K + S)}} $$\end{document} where t = time hr, S = concentration of ferrous ions g Fe++/1., μm = maximum specific growth rate of bacteria, hr-1. Y = mass of bacteria produced per gram of iron oxidized g/g, K = saturation constant, g Fe++/l., and X = concentration of bacteria g/1.The value for the maximum specific growth rate, μm, was found to vary from 0.12 hr-1 at 20°C to 0.20 hr-1 at 31°C, while the value for the saturation constant K varied randomly between 1 and 2 g/1.A method has also been described which permitted evaluation of the relevant rate constants μm and K without direct knowledge of the bacterial population. This method was found to yield values of μm and K which agreed with values determined accurately by a statistical regression analysis of the experimental data.
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  • 88
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    Biotechnology and Bioengineering 12 (1970), S. 93-121 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Samples of oriented DNA prepared by wet spinning have been found to be very useful for physicochemical and biochemical studies with various techniques. The results obtained yield information on such fundamental properties of DNA as its hydration, electrical conductivity, and its interaction with irradiation and mutagenic and carcinogenic substances. Against this background a detailed description is given of the wet, spinning apparatus and of the techniques developed to produce bigger samples from spun films of oriented DNA. Photographic illustrations are used to give a clear picture of the various details. Extensions of the wet spinning method are discussed.
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  • 89
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    Biotechnology and Bioengineering 12 (1970), S. 145-154 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
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  • 90
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    Biotechnology and Bioengineering 12 (1970), S. 167-178 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The possibility to use microorganisms as human food is limited by several factors. The intact cell is resistant to digestion, the cell wall is unbalanced in essential amino acids, and the nucleic acids are said to be harmful. For using single cell protein as food it may thus be necessary to disrupt the cell wall and separate the protein from nucleic acid. This paper is concerned with the production and properties of extracellular enzymes able to lyse cell walls of microorganisms. Soil bacteria and actinomycetes have been cultivated and lytic enzymes from these organisms have been used to lyse living cells of the yeast like organism E. ashbyii. Efforts were also made to use these enzymes for lysing cell of a Methanomonas sp.
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  • 91
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    Biotechnology and Bioengineering 12 (1970), S. 179-212 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: B. subtilis NRRL B3411 neutral protease has been extensively purified by solvent, and salt fractional ion, pigment removal with DEAE-cellulose followed by chromatography on hydroxylapatite, and a final passage through a Sephadex G-100 column. The neutral protease was shown to be homogeneous by disc gel and cellulose acetate electrophoresis, gel filtration chromatography, and ultra-centrifugation. The molecular weight was determined by osmometry and ultracentrifugation to be about 38-42,000 and the amino acid composition and zinc content determined. The general properties of the enzyme, pH-activity relationship, stability, effect of inhibitors, and specificity are discussed. Comparative studies were carried out on the B. subtilis NRRL B3411 and B. subtilis var. amylosacchariticus neutral proteases and these enzymes were found to be indistinguishable by the methods used, but quite distinct from the thermostable enzyme thermolysin from B. thermoprotcolyticus.
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  • 92
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    Biotechnology and Bioengineering 12 (1970), S. 291-311 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: This paper is concerned with the study of an enzymatic system in a repeated batch process where the enzyme is subject to deactivation. The particular system studied was the enzymatic hydrolysis of Penicillin G to 6-aminopenicillanic acid. Utilizing standard optimization techniques, pH and temperature control policies were determined that would maximize the product yield.
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  • 93
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    Biotechnology and Bioengineering 12 (1970), S. 313-319 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A comparative study of the Bacillus subtilis neutral protease and Bacillus thermoproteolyticus thermolysin calalyzed hydrolysis of a few dipeptide sustrates including furylacryloylglycyl-L-leucine amide is reported. While differences in the kcat/Km were observed between the two enzymes toward substrates in which phenylalanine or leucine donated the amino group of the peptide bond, secondary effects of substituents on the carbonyl donating amino acid and pH profiles were quite similar. Differences were also observed toward protein substrates as compared to dipeptides.
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  • 94
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    Biotechnology and Bioengineering 12 (1970), S. 333-340 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A microbial ecosystem represents a delicately balanced population of microorganisms each interacting with and influencing the other members of the population. An understanding of the nature and effects of these interactions is essential to improving the performance of these ecologies, which are important, in such diverse processes as biological waste treatment procedures, water pollution abatement, industrial fermentations, human or animal digestives processes and in soil. There are several types of mocrobial interactions, such as commensalism, inhibition, food competition, predation, parasitism, and synergism, which either singly or in combination may influence the functioning of the microbial ecology.To understand interactions, it is necessary to perform a detailed study of the physiology of the individual predominating microorganisms to establish their requirements with respect to such environmental factors as nutrients, temperature, pH, oxidation-reduction potential, removal of waste products, or toxic materials which may be involved in control processes and to determine how these factors affect their capabilities. The sum total of this information will indicate the possible interactions between the microorganisms and will form the basis for conducting experiments either in the laboratory or with mathematical models. Such experiments will lead to an understanding of microbial activities and to the formulation of control measures, often using an alteration of the environmental factors for regulation of the microbial ecologies. Extensive research remains to be done on the microbial interact inns in obtain the desired, precise control of these ecological processes.
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  • 95
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    Biotechnology and Bioengineering 12 (1970), S. 419-428 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Previous studies of oxidation-reduction potential (ORP) variation in monolayer (Roux bottle) cultures pointed out the need for data on pH and ORP patterns in simple spinner cultures. This information was desirable for optimizing conditions of growth in small 1-L and New Brunswick fermentors. Results of experiments in 250-ml centrifuge spinner vessels are presented showing that incubation of media prior to inoculation induces desirable qualities reflected in better growth. The importance of initial ORP values of the medium is discussed. The relationship of ORP levels to yield and longevity of cell growth is also considered. The ORP level of the medium at inoculation is shown to be effected by previous incubation.
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  • 96
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: In replicated 30 to 40-ml suspension cultures of rapidly proliferating monkey kidney cells of a comparatively fragile strain, the rates of glucose utilization and lactic acid accumulation averaged about 400 micrograms and 110 micrograms per 106 cells per day respectively, with average molar La/Gl ratios of 0.48. These two rates of glucose utilization and lactic acid accumulation were about 4 × and 10 × as high as the corresponding rates in comparable cultures of the hardier strain 2071-L mouse fibroblasts under the same conditions, with average molar La/Gl ratios of 0.16. In comparable but nonproliferating suspension cultures of the same strain of monkey kidney cells, during about 3 weeks the rates were extremely high, with about 710 micrograms glucose utilized and 445 micrograms lactic acid accumulated per 106 cells per day, with average molar La/Gl ratios of 1.37. The rates of glucose uptake and lactic acid accumulation were higher in the nonproliferating cultures aerated with 5% CO2 in air than in those aerated with 10% CO2 in air. This difference was associated with pH, which was higher in the former group.It was concluded that with this fragile strain of monkey Kidney cells(1) in nonproliferating cultures the cells were metabolizing actively but with a marked tendency to higher La/Gl ratios, (2) in the proliferating cultures the high rates of glucose utilization and lactic acid accumulation were definitely not directly correlated with the rate of growth, and (3) in none of the cultures was the amount of glucose remaining in the fluid at fluid changes so low as to have been a limiting factor.Information in the literature concering glucose utilization and lactic acid production by cells vitro is voluminous and in some respects contradictory. In the present study the rates were unexpectedly high for the monkey kidney cells, particularly those in the otherwise apparently inactive nonproliferating cultures. The data seem to be unique, in that an established strain of cells in chemically defined medium in suspension cultures has been characterized for these metabolic parameters in both proliferating cultures and in equivalent nonproliferating cultures under directly comparable conditions.The concept was developed that since these monkey kidney cells are obviously more fragile than the other cells examined, the complex physical stresses imposed upon these cells in agitated cultures can be modified and lessened in order to permit growth. Lessening of such mechanical stress waa brought about in several ways, of which only the smaller flask size seemed to be at least partly effective. Increasing either the concentration or the viscosity type of Methocel waa not effective.
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  • 97
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    Biotechnology and Bioengineering 12 (1970), S. 561-575 
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The stability characteristics of a continuous culture system were studied following the addition of the natural product inhibitor, ethanol. For a steady state culture of Klebsiella (Aerobacter) aerogenes there was a linear dependence of growth rate on ethanol concentration. Following impulse and step addition of the inhibitor, response patterns of the growth rate (μ) and overall metabolism (Qo2, QCo2, QAC) were observed. A mathematical model of the transient behavior of a product-limited system is proposed, and analog computer solutions fitted to the experimental data. The transient response of the growth rate could best be described by second or higher order equations, e.g., \documentclass{article}\pagestyle{empty}\begin{document}$$ T_2^2 \frac{{d^2 \mu }}{{dt^2 }} + 2T_2^2 \xi \frac{{d\mu }}{{dt}} + \mu = \mu _m (1.0 - q.P_t) $$\end{document} with values of the second order time constant (T2) = 5 min, and damping coefficient (ξ) = 0.4.
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  • 98
    ISSN: 0006-3592
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Fungal cells from Curvularia lunata were entrapped in a crosslinked polyacrylamide gel. The gel-cells obtained as granules were applied in the microbial transformation of Reichstein compound S leading to cortisol through an 11-β-hydroxylation step. Some kinetic studies of this conversion using gel-cells were carried out. In addition, it was shown that gel-cell granules which had lost part of their 11-β-hydroxylase activity on storage could be reactivated yielding preparations with increased activity.From Corynebacterium simplex a steroid dehydrogenase catalyzing the Δ1- dehydrogenation of cortisol leading to prednisolone was isolated and partially purified. The preparation was entrapped in a crosslinked polyacrylamide gel and the gel-enzyme granules obtained used in steroid dehydrogenation processes.
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  • 99
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    Weinheim : Wiley-Blackwell
    Liebigs Annalen 1999 (1999), S. 57-60 
    ISSN: 1434-193X
    Keywords: Benzyne ; Dehydroanthracene ; Matrix isolation ; Photochemistry ; Bergman reaction ; Chemistry ; General Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: -9,10-Didehydroanthracene (1) is an interesting derivative of p-benzyne that has been subject of several studies. In contrast to an earlier report, the photochemical decarbonylation of 9,10-dicarbonyl-9,10-dihydroanthracene (2) does not lead to 1 but rather to the ring-opened ene-diyne 4. The key intermediate for this reaction is keto carbene 7 which is formed by monodecarbonylation of 2. Carbene 7 is labile towards visible-light irradiation and easily looses the second CO molecule to give 4. Carbene 7 and diyne 4 are characterized by IR and UV/Vis spectra, the IR spectra are compared to calculations at the B3LYP/6-31G(d,p) level of theory.
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  • 100
    ISSN: 1434-193X
    Keywords: Nucleophilic addition ; Solvent effect ; Reversal of diastereoselectivity ; Temperature effect ; Imines ; Chemistry ; General Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: -Commonly observed, but rarely explored, is the possibility of modifying the diastereomeric excess (de%) by means of temperature. A complete reversal in the diastereofacial selectivity could be obtained whenever the diastereoisomers concerned are differentially favored by enthalpy and entropy. The enthalpic or entropic dominance of a diastereoisomer depends greatly on the reaction solvent used.
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