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1.
Kirsten Kottmeier Carsten Müller Robert Huber Jochen Büchs 《Applied microbiology and biotechnology》2010,86(1):93-101
By the use of directed limitations of secondary substrates, the metabolic flux should be deflected from biomass production
to product formation. In order to study the impact of directed limitations caused by various secondary substrates on the growth
and product formation of the methylotrophic yeast Hansenula polymorpha, the cultivation systems respiration activity monitoring system (RAMOS) and BioLector were used in parallel. While the RAMOS
device allows the online monitoring of the oxygen transfer rate in shake flasks, the BioLector enables in microtiter plates
the monitoring of scattered light and the fluorescence intensity of the green fluorescent protein (GFP). Secondary substrate
limitations of phosphate, potassium, and magnesium were analyzed in batch fermentations. The sole carbon source was either
10 g/L glucose or 10 g/L glycerol. The expression of the GFP gene is controlled by the FMD promoter (formate dehydrogenase). In batch cultures with glucose as carbon source, a directed limitation of phosphate increased
the GFP production 1.87-fold, compared to phosphate unlimited conditions. Under potassium-limited conditions with glycerol
as sole carbon source, the GFP production was 1.41-fold higher compared to unlimited conditions. A limitation of the substrate
magnesium resulted in a 1.22-fold increase GFP formation in the case of glycerol as carbon source. 相似文献
2.
Modulation of Acetone-Butanol-Ethanol Fermentation by Carbon Monoxide and Organic Acids 总被引:9,自引:5,他引:4
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Metabolic modulation of acetone-butanol-ethanol fermentation by Clostridium acetobutylicum with carbon monoxide (CO) and organic acids is described. CO, which is a known inhibitor of hydrogenase, was found to be effective in the concentration range of dissolved CO corresponding to a CO partial pressure of 0.1 to 0.2 atm. Metabolic modulation by CO was particularly effective when organic acids such as acetic and butyric acids were added to the fermentation as electron sinks. The uptake of organic acids was enhanced, and increases in butyric acid uptake by 50 to 200% over control were observed. Hydrogen production could be reduced by 50% and the ratio of solvents could be controlled by CO modulation and organic acid addition. Acetone production could be eliminated if desired. Butanol yield could be increased by 10 to 15%. Total solvent yield could be increased 1 to 3% and the electron efficiency to acetone-butanol-ethanol solvents could be increased from 73 to 78% for controls to 80 to 85% for CO- and organic acid-modulated fermentations. Based on these results, the dynamic nature of electron flow in this fermentation has been elucidated and mechanisms for metabolic control have been hypothesized. 相似文献
3.
Hyaluronic acid is routinely produced through fermentation of both Group A and C streptococci. Despite significant production costs associated with short fermentations and removal of contaminating proteins released during entry into stationary phase, hyaluronic acid is typically produced in batch rather than continuous culture. The main reason is that hyaluronic acid synthesis has been found to be unstable in continuous culture except at very low dilution rates. Here, we investigated the mechanisms underlying this instability and developed a stable, high dilution rate (0.4 h-1) chemostat process for both chemically defined and complex media operating for more than 150 h of production. In chemically defined medium, the product yield was 25% higher in chemostat cultures than in conventional batch culture when arginine or glucose was the limiting substrate. In contrast, glutamine limitation resulted in higher ATP requirements and a yield similar to that observed in batch culture. In complex, glucose-limited medium, ATP requirements were greatly reduced but biomass synthesis was favored over hyaluronic acid and no improvement in hyaluronic acid yield was observed. The successful establishment of continuous culture at high dilution rate enables both commercial production at reduced cost and a more rational characterization and optimization of hyaluronic acid production in streptococci. 相似文献
4.
N Zwicker U Theobald H Zähner H-P Fiedler 《Journal of industrial microbiology & biotechnology》1997,19(4):280-285
The production of EDDS (ethylene-diamine-disuccinic acid), a potential substitute for EDTA, has been optimized up to a product
concentration of 20 grams per litre in fermentations of Amycolatopsis orientalis. Decisive steps for the increase in productivity were variation of the synthetic medium composition, investigation of the
influence of metal ions on product formation, controlled feeding of carbon and nitrogen sources in fed-batch fermentations
and improvement of the downstream processing steps.
Received 05 May 1997/ Accepted in revised form 13 August 1997 相似文献
5.
In batch fermentations of C. acetobutylicum, with 5 g/L yeast extract and 50mM glucose, the ratio of ammonium to glucose affected solvent production when the pH was left to vary uncontrolled from 4.5 to 3.65. High solvent production was observed for a low ratio. When the pH was controlled at 4.5, only acids were produced for all ratio values. At a low ammonium-to-glucose ratio, solvents were produced when the pH was controlled at 3.7. Acids only were produced for a low ratio value at pH 4.0 or for a high ratio value at pH 3.7. In continuous cultures, mostly acids were produced under glucose limitation, but solvents were produced under nitrogen limitation. It was concluded that the nitrogen availability controls solvent production and that the pH affects the availability of organic nitrogen. Biomass autolysis at the stationary phase of batch cultures was reversibly inhibited at pH values less than 3.8. In batch fermentations, the overall molar growth yields on ATP (Y(ATP)) varied from 5.5 to 9.0 and the transient yields from 5.5 to 15.5. In continuous cultures, the Y(ATP) values varied from 5.5 to 14.7 under glucose limitation, and from 6.1 to 9.3 under nitrogen limitation. The Y(ATP) depended on the ammonium to glucose ratio and the culture pH, but did not show the usual dependence on the specific growth rate in batch cultures. The experiments seem to confirm the hypothesis that solvent production is controlled by the demand and availability of ATP. 相似文献
6.
Batch, fed-batch, and continuous A-B-E fermentations were conducted and compared with pH controlled at 4.5, the optimal range for solvent production. While the batch mode provides the highest solvent yield, the continuous mode was preferred in terms of butanol yield and productivity. The highest butanol yield and productivity found in the continuous fermentation at dilution rate of 0.1 h−1 were 0.21 g-butanol/g-glucose and 0.81 g/L/h, respectively. In the continuous and fed-batch fermentation, the time needed for passing acidogenesis to solventogenesis was an intrinsic hindrance to higher butanol productivity. Therefore, a low dilution rate is suggested for the continuous A-B-E fermentation, while the fed-batch mode is not suggested for solvent production. While 3:6:1 ratio of acetone, butanol, and ethanol is commonly observed from A-B-E batch fermentation by Clostridium acetobutylicum when the pH is uncontrolled, up to 94% of the produced solvent was butanol in the chemostat with pH controlled at 4.5. 相似文献
7.
Stephan Lane Yanfei Zhang Eun Ju Yun Leah Ziolkowski Guochang Zhang Yong-Su Jin José L. Avalos 《Biotechnology and bioengineering》2020,117(2):372-381
Bioconversion of xylose—the second most abundant sugar in nature—into high-value fuels and chemicals by engineered Saccharomyces cerevisiae has been a long-term goal of the metabolic engineering community. Although most efforts have heavily focused on the production of ethanol by engineered S. cerevisiae, yields and productivities of ethanol produced from xylose have remained inferior as compared with ethanol produced from glucose. However, this entrenched focus on ethanol has concealed the fact that many aspects of xylose metabolism favor the production of nonethanol products. Through reduced overall metabolic flux, a more respiratory nature of consumption, and evading glucose signaling pathways, the bioconversion of xylose can be more amenable to redirecting flux away from ethanol towards the desired target product. In this report, we show that coupling xylose consumption via the oxidoreductive pathway with a mitochondrially-targeted isobutanol biosynthesis pathway leads to enhanced product yields and titers as compared to cultures utilizing glucose or galactose as a carbon source. Through the optimization of culture conditions, we achieve 2.6 g/L of isobutanol in the fed-batch flask and bioreactor fermentations. These results suggest that there may be synergistic benefits of coupling xylose assimilation with the production of nonethanol value-added products. 相似文献
8.
Summary The effect of product gases, H2 and CO2, on solvent production was studied using a continuous culture of alginate-immobilized Clostridium acetobutylicum. Initially, in order to find the optimum dilution rate for aceton--butanol production in this system, fermentations were carried out at various dilution rates. With 10% H2 and 10% CO2 in the sparging gas, a dilution rate of 0.07 h–1 was found to maximize volumetric productivity (0.58 g·l–1·h–1), while the maximum specific productivity of 0.27 g–·h–1 occured at 0.12 h–1. Continuous cultures with vigorous sparging of N2 produced only acids. It was concluded that in the case of continuous fermentation H2 is essential for good solvent production, although good solvent production is possible in an H2-absent environment in the case of batch fermentations. When the fermentation was carried out at atmospheric pressure under H2-enriched conditions, the presence of CO2 in the sparging gas did not slow down glucose metabolism; rather it changed the direction of the phosphoroclastic reaction and as a result increased the butanol/acetone ratio. 相似文献
9.
The effect of the dilution rate on biomass and product synthesis in fermentations of glucose, fructose and a commercial mixture of fructooligosaccharides (FOS) by Bifidobacterium longum ATCC 15707 was studied. Kinetic parameters (maximum specific growth rate, Monod constant, maintenance, and yield coefficients) in the mathematical model of the fermentation were estimated from experimental data. In the FOS mixture fermentations, approximately 12% of the total reducing sugars (mainly fructose) in the feed were not metabolized by the bacterium. In fermentations of fructose and the FOS mixture, biomass concentration increased as the dilution rate increased and, once maximum values were reached [3.90 (D=0.20 h–1) and 2.54 g l–1 (D=0.15 h–1), respectively], decreased rapidly as the culture was washed out. Formic acid was detected at low dilution rates in glucose and fructose fermentations. The main products in fermentations of the three carbon sources were lactic and acetic acids. Average values of the molar ratio between acetic and lactic acids of 1.18, 1.21 and 0.83 mol mol–1 were obtained in glucose, fructose and FOS mixture fermentations, respectively. In batch fermentations carried out without pH control this molar ratio was lower than 1.5 only when fructose was used as the carbon source. 相似文献
10.
Proteomic Profiling of Recombinant Escherichia coli in High-Cell- Density Fermentations for Improved Production of an Antibody Fragment Biopharmaceutical
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Ilana S. Aldor Denise C. Krawitz William Forrest Christina Chen Julie C. Nishihara John C. Joly Kathleen M. Champion 《Applied microbiology》2005,71(4):1717-1728
By using two-dimensional polyacrylamide gel electrophoresis, a proteomic analysis over time was conducted with high-cell-density, industrial, phosphate-limited Escherichia coli fermentations at the 10-liter scale. During production, a recombinant, humanized antibody fragment was secreted and assembled in a soluble form in the periplasm. E. coli protein changes associated with culture conditions were distinguished from protein changes associated with heterologous protein expression. Protein spots were monitored quantitatively and qualitatively. Differentially expressed proteins were quantitatively assessed by using a t-test method with a 1% false discovery rate as a significance criterion. As determined by this criterion, 81 protein spots changed significantly between 14 and 72 h (final time) of the control fermentations (vector only). Qualitative (on-off) comparisons indicated that 20 more protein spots were present only at 14 or 72 h in the control fermentations. These changes reflected physiological responses to the culture conditions. In control and production fermentations at 72 h, 25 protein spots were significantly differentially expressed. In addition, 19 protein spots were present only in control or production fermentations at this time. The quantitative and qualitative changes were attributable to overexpression of recombinant protein. The physiological changes observed during the fermentations included the up-regulation of phosphate starvation proteins and the down-regulation of ribosomal proteins and nucleotide biosynthesis proteins. Synthesis of the stress protein phage shock protein A (PspA) was strongly correlated with synthesis of a recombinant product. This suggested that manipulation of PspA levels might improve the soluble recombinant protein yield in the periplasm for this bioprocess. Indeed, controlled coexpression of PspA during production led to a moderate, but statistically significant, improvement in the yield. 相似文献
11.
The effects of nitrogen and phosphate in batch and continuous AEB fermentations were tested. Both nitrogen- and phosphate-limited fermentations favored acid formation but not solvent production. A coupled two-stage continuous fermentation was performed for 30 days with a nitrogen-limited first stage fermentation for enhanced acid production. The bacteria from the acidogenic phase (first stage) fermentation were continuously pumped into a 14-l second stage fermentor with supplemental glucose and nitrogen for solvent production. The second stage fermentor had a maximum butanol productivity of 0.4 g l−1 h−1 (total solvent production was 0.6 g l−1 h−1) at a dilution rate of 0.06 h−1. 相似文献
12.
Cells of Zymomonas mobilis ATCC 10988 were immobilized in 1.5% calcium alginate and packed in a column bioreactor for a series of fermentations utilizing 10.0% glucose media with the addition of one of the following amino acids or keto acids: L-leucine, L-isoleucine, L-valine, α-ketoisocaproic acid, α-ketobutyric acid, or α-ketoisovaleric acid. This was done in order to study the rates of production of higher alcohols during ethanolic fermentations at varying dilution rates while under the influence of amino acids or keto acids. Results indicate that the EHRLICH mechanism is operative in Zymomonas sp. α-Ketobutyrate enhanced the production of n-propanol and act-amyl alcohol. α-Ketoisocaproic acid stimulated the production of isoamyl alcohol. α-Ketoisovaleric acid increased the levels of isobutanol. The amino acids also gave rise to their corresponding alcohols but to a far lesser degree than did the keto acids. During high glucose utilization, ethanol yields ranged from 87% to 94% of theoretical with productivity ranging from 60.08 g/l/h in one fermentation (at a dilution rate of 1.35 h?1) to 70.42 g/l/h in another (at a dilution rate of 1.58 h?1). At dilution rates of 1.58 h?1, higher alcohol productivity rose to as high as 4,313 mg/l/h in the presence of α-ketoisocaproic acid, 1,734.49 mg/l/h using α-ketoisovaleric acid, and 1,618.05 mg/l/h in α-ketobutyric acid. The concomitant production of ethanol and higher alcohols in all of the fermentations indicates that glucose is required for the production of the higher alcohols from their corresponding amino acids or keto acids. 相似文献
13.
Enhanced Butanol Production by Clostridium beijerinckii BA101 Grown in Semidefined P2 Medium Containing 6 Percent Maltodextrin or Glucose
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Dramatically elevated levels of butanol and acetone resulted in higher butanol and total solvent yields for hyperamylolytic Clostridium beijerinckii BA101 relative to the NCIMB 8052 parent strain grown in semidefined P2 medium containing either 6% glucose or STAR-DRI 5 maltodextrin. C. beijerinckii BA101 consistently produced on the order of 19 g of butanol per liter in 20-liter batch fermentations. This represents a greater than 100% increase in butanol concentration by the BA101 strain compared to the parent NCIMB 8052 strain. The kinetics of butanol production over time also indicate a more rapid rate of butanol production by BA101 in semidefined P2 medium containing glucose or maltodextrin. The lower levels of butyric and acetic acids produced over the course of the fermentation carried out by BA101 are consistent with an enhanced capacity for uptake and recycling of these acids. C. beijerinckii BA101 appears to more completely utilize carbohydrate compared to the 8052 strain. Carbon balance following fermentation by C. beijerinckii 8052 and BA101 indicates that sufficient carbon is available for the twofold increase in butanol concentration observed during BA101 fermentations. C. beijerinckii BA101 also has superior solvent production capacity during continuous culture fermentation in P2 medium containing 6% glucose. Volumetric solvent yields of 0.78 and 1.74 g/liter/h for BA101 and 0.34 and 1.17 g/liter/h for NCIMB 8052 were obtained at dilution rates of 0.05 and 0.20 h(sup-1), respectively. No drift towards acid synthesis (strain degeneration) was observed for up to 200 h (d = 0.05 h(sup-1)) and 100 h (d = 0.20 h(sup-1)). 相似文献
14.
Summary The stability of continuous fermentations of Clostridium acetobutylicum can be limited by cell flocculation under conditions of high solvent production. The addition of chloride salts to a synthetic medium prevents the flocculation of the bacteria, and thus results in stable continuous cultures. 相似文献
15.
Aldor IS Krawitz DC Forrest W Chen C Nishihara JC Joly JC Champion KM 《Applied and environmental microbiology》2005,71(4):1717-1728
By using two-dimensional polyacrylamide gel electrophoresis, a proteomic analysis over time was conducted with high-cell-density, industrial, phosphate-limited Escherichia coli fermentations at the 10-liter scale. During production, a recombinant, humanized antibody fragment was secreted and assembled in a soluble form in the periplasm. E. coli protein changes associated with culture conditions were distinguished from protein changes associated with heterologous protein expression. Protein spots were monitored quantitatively and qualitatively. Differentially expressed proteins were quantitatively assessed by using a t-test method with a 1% false discovery rate as a significance criterion. As determined by this criterion, 81 protein spots changed significantly between 14 and 72 h (final time) of the control fermentations (vector only). Qualitative (on-off) comparisons indicated that 20 more protein spots were present only at 14 or 72 h in the control fermentations. These changes reflected physiological responses to the culture conditions. In control and production fermentations at 72 h, 25 protein spots were significantly differentially expressed. In addition, 19 protein spots were present only in control or production fermentations at this time. The quantitative and qualitative changes were attributable to overexpression of recombinant protein. The physiological changes observed during the fermentations included the up-regulation of phosphate starvation proteins and the down-regulation of ribosomal proteins and nucleotide biosynthesis proteins. Synthesis of the stress protein phage shock protein A (PspA) was strongly correlated with synthesis of a recombinant product. This suggested that manipulation of PspA levels might improve the soluble recombinant protein yield in the periplasm for this bioprocess. Indeed, controlled coexpression of PspA during production led to a moderate, but statistically significant, improvement in the yield. 相似文献
16.
Olivier Fond G. Matta-Ammouri H. Petitdemange J. M. Engasser 《Applied microbiology and biotechnology》1985,22(3):195-200
Summary The production of solvent by Clostridium acetobutylicum was studied, using fed-batch fermentations. Different specific rates of carbohydrate utilisation were obtained by variations in feeding rates of sugar. At slow catabolic rates of sugar, addition of acetic acid or butyric acid, alone or together, increased the rate of the metabolic transition by a factor 10 to 20, the amount of solvents by a factor 6 and the percentage of fermented glucose to solvents by a factor 3. The same results were obtained with both glucose and xylose fermentations. Depending on the rates of growth, butanol production began at acid levels of 3–4 g·l-1 for fast metabolism and at acid levels of 8–10 g·l-1 for slow metabolism. Associated with slow metabolism, reassimilation of acids required values as high as 6.5 g·l-1 of acetic acid and 7.5 g·l-1 of butyric acid. At a high rate of metabolism, acetic and butyric acids were reassimilated at concentrations of 4.5 g·l-1. 相似文献
17.
Traugott C. Schüz Hans-Peter Fiedler Hans Zähner 《Applied microbiology and biotechnology》1993,39(4-5):433-437
We performed fed-batch and continuous fermentations to extend the time of maximal nikkomycin production by Streptomyces tendae Tü 901/S 2566. This was achieved by the fed-batch culture technique. Furthermore, high productivity was obtained at slow growth rates in a continuous fermentation process. Different dilution rates with and without carbon limitation were done and the results were compared.
Correspondence to : T. Schüz 相似文献
18.
Weak organic acids are well-known metabolic effectors in yeast and other micro-organisms. High concentrations of lactic acid due to infection of lactic acid bacteria often occurs in combination with growth under nutrient-limiting conditions in industrial yeast fermentations. The effects of lactic acid on growth and product formation of Saccharomyces cerevisiae were studied, with cells growing under carbon- or nitrogen-limiting conditions in anaerobic chemostat cultures (D=0.1 h−1) at pH values 3.25 and 5. It was shown that lactic acid in industrially relevant concentrations had a rather limited effect on the metabolism of S. cerevisiae. However, there was an effect on the energetic status of the cells, i.e. lactic acid addition provoked a reduction in the adenosine triphosphate (ATP) content of the cells. The decrease in ATP was not accompanied by a significant increase in the adenosine monophosphate levels. 相似文献
19.
Fumaric acid production by fermentation 总被引:3,自引:0,他引:3
Roa Engel CA Straathof AJ Zijlmans TW van Gulik WM van der Wielen LA 《Applied microbiology and biotechnology》2008,78(3):379-389
The potential of fumaric acid as a raw material in the polymer industry and the increment of cost of petroleum-based fumaric
acid raises interest in fermentation processes for production of this compound from renewable resources. Although the chemical
process yields 112% w/w fumaric acid from maleic anhydride and the fermentation process yields only 85% w/w from glucose, the latter raw material is three times cheaper. Besides, the fermentation fixes CO2. Production of fumaric acid by Rhizopus species and the involved metabolic pathways are reviewed. Submerged fermentation systems coupled with product recovery techniques
seem to have achieved economically attractive yields and productivities. Future prospects for improvement of fumaric acid
production include metabolic engineering approaches to achieve low pH fermentations. 相似文献
20.
Microbial contamination is a pervasive problem in any ethanol fermentation system. These infections can at minimum affect the efficiency of the fermentation and at their worse lead to stuck fermentations causing plants to shut down for cleaning before beginning anew. These delays can result in costly loss of time as well as lead to an increased cost of the final product. Lactic acid bacteria (LAB) are the most common bacterial contaminants found in ethanol production facilities and have been linked to decreased ethanol production during fermentation. Lactobacillus sp. generally predominant as these bacteria are well adapted for survival under high ethanol, low pH and low oxygen conditions found during fermentation. It has been generally accepted that lactobacilli cause inhibition of Saccharomyces sp. and limit ethanol production through two basic methods; either production of lactic and acetic acids or through competition for nutrients. However, a number of researchers have demonstrated that these mechanisms may not completely account for the amount of loss observed and have suggested other means by which bacteria can inhibit yeast growth and ethanol production. While LAB are the primary contaminates of concern in industrial ethanol fermentations, wild yeast may also affect the productivity of these fermentations. Though many yeast species have the ability to thrive in a fermentation environment, Dekkera bruxellensis has been repeatedly targeted and cited as one of the main contaminant yeasts in ethanol production. Though widely studied for its detrimental effects on wine, the specific species–species interactions between D. bruxellensis and S. cerevisiae are still poorly understood. 相似文献