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Alternative Carbon Source (alternative + carbon_source)
Selected AbstractsUtilization of an Alternative Carbon Source for Efficient Production of Human ,1 -Antitrypsin by Genetically Engineered Rice Cell CultureBIOTECHNOLOGY PROGRESS, Issue 3 2001Masaaki Terashima Human ,1 -antitrypsin was produced by genetically engineered rice cells using promoter and signal peptide of a rice ,-amylase isozyme. Batch and continuous cultures were employed to investigate the effects of alternative carbon sources on the ,1 -antitrypsin production. While this expression system is inducible by sugar depletion, we have found that the productivity of ,1 -antitrypsin increased 2.4- to 3.4-fold, compared with the control medium without carbon source, in medium containing an alternative carbon source, such as pyruvic acid and glyoxylic acid. The accumulated ,1 -antitrypsin in the medium containing pyruvic acid reached 18.2,24.2 mg/g-dry cell in 50,70 h by batch culture. [source] Selection and identification of bacterial strains with methyl- tert -butyl ether, ethyl- tert -butyl ether, and tert -amyl methyl ether degrading capacitiesENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 11 2008Jessica Purswani Abstract Nine bacterial strains isolated from two hydrocarbon-contaminated soils were selected because of their capacity for growth in culture media amended with 200 mg/L of one of the following gasoline oxygenates: Methyl- tert -butyl ether (MTBE), ethyl- tert -butyl ether (ETBE), and tert -amyl methyl ether (TAME). These strains were identified by amplification of their 16S rRNA gene, using fD1 and rD1 primers, and were tested for their capacity to grow and biotransform these oxygenates in both mineral and cometabolic media. The isolates were classified as Bacillus simplex, Bacillus drentensis, Arthrobacter sp., Acinetobacter calcoaceticus, Acinetobacter sp., Gordonia amicalis (two strains), Nocardioides sp., and Rhodococcus ruber. Arthrobacter sp. (strain MG) and A. calcoaceticus (strain M10) consumed 100 (cometabolic medium) and 82 mg/L (mineral medium) of oxygenate TAME in 21 d, respectively, under aerobic conditions. Rhodococcus ruber (strain E10) was observed to use MTBE and ETBE as the sole carbon and energy source, whereas G. amicalis (strain T3) used TAME as the sole carbon and energy source for growth. All the bacterial strains transformed oxygenates better in the presence of an alternative carbon source (ethanol) with the exception of A. calcoaceticus (strain M10). The capacity of the selected strains to remove MTBE, ETBE, and TAME looks promising for application in bioremediation technologies. [source] Physiological adaptation of Corynebacterium glutamicum to benzoate as alternative carbon source , a membrane proteome-centric viewPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 14 2009Ute Haußmann Abstract The ability of microorganisms to assimilate aromatic substances as alternative carbon sources is the basis of biodegradation of natural as well as industrial aromatic compounds. In this study, Corynebacterium glutamicum was grown on benzoate as sole carbon and energy source. To extend the scarce knowledge about physiological adaptation processes occurring in this cell compartment, the membrane proteome was investigated under quantitative and qualitative aspects by applying shotgun proteomics to reach a comprehensive survey. Membrane proteins were relatively quantified using an internal standard metabolically labeled with 15N. Altogether, 40 proteins were found to change their abundance during growth on benzoate in comparison to glucose. A global adaptation was observed in the membrane of benzoate-grown cells, characterized by increased abundance of proteins of the respiratory chain, by a starvation response, and by changes in sulfur metabolism involving the regulator McbR. Additional to the relative quantification, stable isotope-labeled synthetic peptides were used for the absolute quantification of the two benzoate transporters of C. glutamicum, BenK and BenE. It was found that both transporters were expressed during growth on benzoate, suggesting that both contribute substantially to benzoate uptake. [source] Effect of carbon source addition on toluene biodegradation by an Escherichia coli DH5, transconjugant harboring the TOL plasmidBIOTECHNOLOGY & BIOENGINEERING, Issue 2 2010Kaoru Ikuma Abstract Horizontal gene transfer (HGT) of plasmids is a naturally occurring phenomenon which could be manipulated for bioremediation applications. Specifically, HGT may prove useful to enhance bioremediation through genetic bioaugmentation. However, because the transfer of a plasmid between donor and recipient cells does not always result in useful functional phenotypes, the conditions under which HGT events result in enhanced degradative capabilities must first be elucidated. The objective of this study was to determine if the addition of alternate carbon substrates could improve toluene degradation in Escherichia coli DH5, transconjugants. The addition of glucose (0.5,5,g/L) and Luria,Bertani (LB) broth (10,100%) resulted in enhanced toluene degradation. On average, the toluene degradation rate increased 14.1 (±2.1)-fold in the presence of glucose while the maximum increase was 18.4 (±1.7)-fold in the presence of 25% LB broth. Gene expression of xyl genes was upregulated in the presence of glucose but not LB broth, which implies different inducing mechanisms by the two types of alternate carbon source. The increased toluene degradation by the addition of glucose or LB broth was persistent over the short-term, suggesting the pulse amendment of an alternative carbon source may be helpful in bioremediation. While the effects of recipient genome GC content and other conditions must still be examined, our results suggest that changes in environmental conditions such as alternate substrate availability may significantly improve the functionality of the transferred phenotypes in HGT and therefore may be an important parameter for genetic bioaugmentation optimization. Biotechnol. Bioeng. 2010;107: 269,277. © 2010 Wiley Periodicals, Inc. [source] Effects of glucose and insulin on HepG2-C3A cell metabolismBIOTECHNOLOGY & BIOENGINEERING, Issue 2 2010Vidya V. Iyer Abstract HepG2, hepatocellular carcinoma cells, are used in drug toxicity studies and have also been explored for bioartificial livers. For these applications, the cells are under variable levels of nutrients and hormones, the effects of which on metabolism are poorly understood. In this study, HepG2-C3A cells were cultured under varying levels of glucose (high, low, and glucose-free) and insulin (without and with physiological levels of insulin) for 5 days. Cell growth was found to be comparable between high and low glucose media and lowest for glucose-free medium. Several features of central metabolism were affected profoundly by the medium glucose levels. Glucose consumption was greater for low glucose medium compared to high glucose medium, consistent with known glucose feedback regulation mechanisms. Urea productivity was highest in glucose-free medium. Further, it was seen that lactate acted as an alternative carbon source in the absence of glucose, whereas it acted as a sink for the high and low glucose media. Using a metabolic network flexibility analysis (MNFA) framework with stoichiometric and thermodynamic constraints, intracellular fluxes under varying levels of glucose and insulin were evaluated. The analysis indicates that urea production in HepG2-C3A cells arises via the arginase II pathway rather than from ammonia detoxification. Further, involvement of the putrescine metabolism with glutamine metabolism caused higher urea production in glucose-free medium consistent with higher glutamine uptake. MNFA indicated that in high and low glucose media, glycolysis, glutaminolysis, and oxidative phosphorylation were the main sources of energy (NADH, NADPH, and ATP). In the glucose-free medium, due to very low glycolytic flux, higher malate to pyruvate glutaminolytic flux and TCA cycle contributed more significantly to energy metabolism. The presence of insulin lowered glycerol uptake and corresponding fluxes involved in lipid metabolism for all glucose levels but otherwise exerted negligible effect on metabolism. HepG2-C3A cells thus show distinct differences from primary hepatocytes in terms of energy metabolism and urea production. This knowledge can be used to design media supplements and metabolically engineer cells to restore necessary hepatic functions to HepG2-C3A cells for a range of applications. Biotechnol. Bioeng. 2010;107: 347,356. © 2010 Wiley Periodicals, Inc. [source] Utilization of an Alternative Carbon Source for Efficient Production of Human ,1 -Antitrypsin by Genetically Engineered Rice Cell CultureBIOTECHNOLOGY PROGRESS, Issue 3 2001Masaaki Terashima Human ,1 -antitrypsin was produced by genetically engineered rice cells using promoter and signal peptide of a rice ,-amylase isozyme. Batch and continuous cultures were employed to investigate the effects of alternative carbon sources on the ,1 -antitrypsin production. While this expression system is inducible by sugar depletion, we have found that the productivity of ,1 -antitrypsin increased 2.4- to 3.4-fold, compared with the control medium without carbon source, in medium containing an alternative carbon source, such as pyruvic acid and glyoxylic acid. The accumulated ,1 -antitrypsin in the medium containing pyruvic acid reached 18.2,24.2 mg/g-dry cell in 50,70 h by batch culture. [source] Physiological adaptation of Corynebacterium glutamicum to benzoate as alternative carbon source , a membrane proteome-centric viewPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 14 2009Ute Haußmann Abstract The ability of microorganisms to assimilate aromatic substances as alternative carbon sources is the basis of biodegradation of natural as well as industrial aromatic compounds. In this study, Corynebacterium glutamicum was grown on benzoate as sole carbon and energy source. To extend the scarce knowledge about physiological adaptation processes occurring in this cell compartment, the membrane proteome was investigated under quantitative and qualitative aspects by applying shotgun proteomics to reach a comprehensive survey. Membrane proteins were relatively quantified using an internal standard metabolically labeled with 15N. Altogether, 40 proteins were found to change their abundance during growth on benzoate in comparison to glucose. A global adaptation was observed in the membrane of benzoate-grown cells, characterized by increased abundance of proteins of the respiratory chain, by a starvation response, and by changes in sulfur metabolism involving the regulator McbR. Additional to the relative quantification, stable isotope-labeled synthetic peptides were used for the absolute quantification of the two benzoate transporters of C. glutamicum, BenK and BenE. It was found that both transporters were expressed during growth on benzoate, suggesting that both contribute substantially to benzoate uptake. [source] Rhamnolipid Surfactants: An Update on the General Aspects of These Remarkable BiomoleculesBIOTECHNOLOGY PROGRESS, Issue 6 2005Marcia Nitschke Pseudomonas strains are able to biosynthesize rhamnose-containing surfactants also known as rhamnolipids. These surface-active compounds are reviewed with respect to chemical structure, properties, biosynthesis, and physiological role, focusing on their production and the use of low-cost substrates such as wastes from food industries as alternative carbon sources. The use of inexpensive raw materials such as agroindustrial wastes is an attractive strategy to reduce the production costs associated with biosurfactant production and, at same time, contribute to the reduction of environmental impact generated by the discard of residues, and the treatment costs. Carbohydrate-rich substrates generated low rhamnolipid levels, whereas oils and lipid-rich wastes have shown excellent potential as alternative carbon sources. [source] Global Gene Expression Differences Associated with Changes in Glycolytic Flux and Growth Rate in Escherichia coli during the Fermentation of Glucose and XyloseBIOTECHNOLOGY PROGRESS, Issue 1 2002Ramon Gonzalez The simplicity of the fermentation process (anaerobic with pH, temperature, and agitation control) in ethanologenic Escherichia coli KO11 and LY01 makes this an attractive system to investigate the utility of gene arrays for biotechnology applications. By using this system, gene expression, glycolytic flux, and growth rate have been compared in glucose-grown and xylose-grown cells. Although the initial metabolic steps differ, ethanol yields from both sugars were essentially identical on a weight basis, and little carbon was diverted to biosynthesis. Expression of only 27 genes changed by more than 2-fold in both strains. These included induction of xylose-specific operons ( xylE, xylFGHR, and xylAB) regulated by XylR and the cyclic AMP,CRP system and repression of Mlc-regulated genes encoding glucose uptake ( ptsHIcrr, ptsG) and mannose uptake ( manXYZ) during growth on xylose. However, expression of genes encoding central carbon metabolism and biosynthesis differed by less than 2-fold. Simple statistical methods were used to investigate these more subtle changes. The reproducibility (coefficient of variation of 12%) of expression measurements (mRNA as cDNA) was found to be similar to that typically observed for in vitro measurements of enzyme activities. Using Studentapos;s t test, many smaller but significant sugar-dependent changes were identified ( p < 0.05 in both strains). A total of 276 genes were more highly expressed during growth on xylose; 307 genes were more highly expressed with glucose. Slower growth (lower ATP yield) on xylose was accompanied by decreased expression of 62 genes concerned with the biosynthesis of small molecules (amino acids, nucleotides, cofactors, and lipids), transcription, and translation; 5 such genes were expressed at a higher level. In xylose-grown cells, 90 genes associated with the transport, catabolism, and regulation of pathways for alternative carbon sources were expressed at higher levels than in glucose-grown cells, consistent with a relaxation of control by the cyclic AMP,CRP regulatory system. Changes in expression of genes encoding the Embden,Meyerhof,Parnas (EMP) pathway were in excellent agreement with calculated changes in flux for individual metabolites. Flux through all but one step, pyruvate kinase, was predicted to be higher during glucose fermentation. Expression levels (glucose/xylose) were higher in glucose-grown cells for all EMP genes except the isoenzymes encoding pyruvate kinase ( pykA and pykF). Expression of both isoenzymes was generally higher during xylose fermentation but statistically higher in both strains only for pykF encoding the isoenzyme activated by fructose-6-phosphate, a key metabolite connecting pentose metabolism to the EMP pathway. The coordinated changes in expression of genes encoding the EMP pathway suggest the presence of a common regulatory system and that flux control within the EMP pathway may be broadly distributed. In contrast, expression levels for genes encoding the Pentose,Phosphate pathway did not differ significantly between glucose-grown and xylose-grown cells. [source] Utilization of an Alternative Carbon Source for Efficient Production of Human ,1 -Antitrypsin by Genetically Engineered Rice Cell CultureBIOTECHNOLOGY PROGRESS, Issue 3 2001Masaaki Terashima Human ,1 -antitrypsin was produced by genetically engineered rice cells using promoter and signal peptide of a rice ,-amylase isozyme. Batch and continuous cultures were employed to investigate the effects of alternative carbon sources on the ,1 -antitrypsin production. While this expression system is inducible by sugar depletion, we have found that the productivity of ,1 -antitrypsin increased 2.4- to 3.4-fold, compared with the control medium without carbon source, in medium containing an alternative carbon source, such as pyruvic acid and glyoxylic acid. The accumulated ,1 -antitrypsin in the medium containing pyruvic acid reached 18.2,24.2 mg/g-dry cell in 50,70 h by batch culture. [source] | ||||||||||