• Title/Summary/Keyword: metabolic flux shift

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Inter-scale Observation and Process Optimization for Guanosine Fermentation

  • Chu, Ju;Zhang, Si-Liang;Zhuang, Ying-Ping
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2005.06a
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    • pp.233-244
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    • 2005
  • Guanosine fermentation process can be well predicted and analyzed by the proposed state equations describing the dynamic change of a bioreactor. Pyruvate and alanine were found to be characteristically accumulated along with the decline of the guanosine formation rate during the mid-late phase of the process. The enzymological study of the main pathways in glucose catabolism and the quantitative stoichiometric calculation of metabolic flux distribution revealed that it was entirely attributed to the shift of metabolic flux from hexose monophosphate (HMP) pathway to glycolysis pathway. The process optimization by focusing on the restore of the shift of metabolic flux was conducted and the overcoming the decrease of oxygen uptake rate (OUR) was taken as the relevant factor of the trans-scale operation. As a result, the production of guanosinewas increased from 17 g/L to over 34 g/I.

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Metabolic Flux Shift of Weissella kimchii sk10 Grown Under Aerobic Conditions

  • Park, Sun-Mi;Kang, Hye-Sun;Park, Doo-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.14 no.5
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    • pp.919-923
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    • 2004
  • The sk10 isolated from kimchi was identified as W. kimchii on the basis of l6s-rDNA sequencing. Studies were made to analyze the metabolic flux shift of the sk10 on glucose under aerobic growth conditions. The sk10 produced 38.2 mM acetate, 16.3 mM ethanol, and 33.2 mM lactate under aerobic conditions, but 2.4 mM acetate, 48.0 mM ethanol, and 44.1 mM lactate under anaerobic conditions. The NADH peroxidase (NADH-dependent hydrogen peroxidase) activity of sk10 grown under aerobic conditions was 11 times higher than that under anaerobic conditions. Under the low ratio of $NADH/NAD^+$, the metabolic flux toward lactate and ethanol was shifted to the flux through acetate kinase without NADH oxidation. The kinds of enzymes and metabolites of sk10 were close to those in the pathway of Leuconostoc sp., but the metabolites produced under aerobic growth conditions were different from those of Leuconostoc sp. The stoichiometric balance calculated using the concentrations of metabolites and substrate was about 97%, coincident with the theoretical values under both aerobic and anaerobic conditions. From these results, it was concluded that the metabolic flux of W. kimchii sk10 was partially shifted from lactate and ethanol to acetate under aerobic conditions only.

pH-dependent Metabolic Flux Shift in Novel Hydrogen-Producing Bacterium Enterobacter sp. SNU-1453 (새로운 수소 생산 균주인 Enterobacter sp. SNU-1453의 pH에 따른 Metabolic Flux 변화)

  • Shin, Jong-Hwan;Yoon, Jong-Hyun;Ahn, Eun-Kyoung;Sim, Sang-Jun;Kim, Mi-Sun;Park, Tai-Hyun
    • KSBB Journal
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    • v.20 no.6
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    • pp.464-469
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    • 2005
  • For the biological production of hydrogen, a new fermentative hydrogen-producing bacterium, Enterobacter sp. SNU-1453, was isolated from a domestic landfill. During the culture of this bacterium, pH significantly decreased with the accumulation of various organic acids, and consequently this inhibited the production of hydrogen. It was found that the metabolic flux in this bacterium depended on the pH and affected the hydrogen production. A butanediol pathway was dominant during the fermentation when pH was not controlled. By controlling the pH at 7 this pathway can be shifted to a mixed acid pathway, which is favorable to the production of hydrogen.

Mapping of Carbon Flow Distribution in the Central Metabolic Pathways of Clostridium cellulolyticum: Direct Comparison of Bacterial Metabolism with a Soluble versus an Insoluble Carbon Source

  • DESVAUX, MICKAEL,
    • Journal of Microbiology and Biotechnology
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    • v.14 no.6
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    • pp.1200-1210
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    • 2004
  • Metabolic flux analysis was established by adapting previous stoichiometric model developed during growth with cellulose to cell grown with cellobiose for further direct comparison of the bacterial metabolism. In carbon limitation with cellobiose, a shift from acetate-ethanol fermentation to ethanol-lactate fermentation is observed and the pyruvate overflow is much higher than with cellulose. In nitrogen limitation with cellobiose, the cellodextrin and exopolysaccharide overflows are much higher than on cellulose. In carbon and nitrogen saturation with cellobiose, the cellodextrin, exopolysaccharide, and free amino acids overflows reach the highest levels observed but all remain limited on cellulose. By completely shunting the cellulosome, the use of cellobiose allows to reach much higher carbon consumption rates which, in return, highlights the metabolic limitation of C. cellulolyticum. Therefore, the physical nature of the carbon source has a profound impact on the metabolism of C. cellulolyticum and most probably of other cellulolytic bacteria. For cellulolytic bacteria, the use of soluble carbon substrate must carefully be taken into consideration for the interpretation of results. Direct comparison of metabolic flux analysis from cellobiose and cellulose revealed the importance of cellulosome, phosphoglucomutase and pyruvate-ferredoxin oxidoreductase in the distribution of carbon flow in the central metabolism. In the light of these findings, future directions for improvement of cellulose catabolism by this bacterium are discussed.

Anaerobic Respiration of Superoxide Dismutase-Deficient Saccharomyces cerevisiae under Oxidative Stress

  • Lee, Sun-Mi;Nam, Doo-Hyun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.3 no.1
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    • pp.15-18
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    • 1998
  • The entanol productivity of superoxide dismutase (SOD)-deficient mutants of Saccharo-Myces cerevisiae was examined under the oxidative stress by Paraquat. It was observed that MnSOD-deficient mutant of S. cerevisiae had higher ethanol productivity than wild type or CuZnSOD-deficient yeast both in aerobic and in anaerobic culture condition. Pyruvated dehydrogenase activity decreased by 35% and alcohol dehydrogenase activity increased by 32% were observed in MnSOD-deficient yeast grown aerobically. When generating oxygen radicals by Paraquat, the ehanol productivity was increased by 40% in CuZnSOD-deficient or wild strain, resulting from increased activity of alcohol dehydrogenase and decreased a activity of pyruvate dehydrogenase. However, the addition of ascorbic acid with Paraquat returned the enzyme activities at the level of control. These results imply that SOD-deficiency in yeast strains may cause the metabolic flux to shift into anaerobic ethanol fermentation in order to avoid their oxidative damages by Paraquat.

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Electrochemical Control of Metabolic Flux of Weissella kimchii sk10: Neutral Red Immobilized in Cytoplasmic Membrane as Electron Channel

  • PARK, SUN-MI;KANG, HYE-SUN;PARK, DAE-WON;PARK, DOO-HYUN
    • Journal of Microbiology and Biotechnology
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    • v.15 no.1
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    • pp.80-85
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    • 2005
  • Electrochemical control of the metabolic flux of W. kimchii sk10 on glucose and pyruvate was studied. The growing cell of W. kimchii sk10 produced 87.4 mM lactate, 69.3 mM ethanol, and 4.9mM lactate from 83.1mM glucose under oxidation condition of the anode compartment, but 98.9 mM lactate, 84.3mM ethanol, and 0.2 mM acetate were produced from 90.8 mM glucose under reduction condition of the cathode compartment for 24 h, respectively. The resting cell of W. kimchii sk10 produced 15.9 mM lactate and 15.2 mM acetate from 32.1 mM pyruvate under oxidation condition of the anode compartment, and 71.3 mM lactate and 3.8 mM acetate from 79.8mM pyruvate under reduction condition of the cathode compartment. The redox balance (NADH/$NAD^+$) of metabolites electrochemically produced from pyruvate was 1.05 and 18.76 under oxidation and reduction conditions, respectively. On the basis of these results, we suggest that the neutral red (NR) immobilized in bacterial membrane can function as an electron channel for the electron transfer between electrode and cytoplasm without dissipation of membrane potential, and that the bacterial fermentation of W. kimchii sk10 can be shifted to oxidized or reduced pathways by the electrochemical oxidation or reduction, respectively.