• Title/Summary/Keyword: catabolic repression

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Modulation of Phosphoenolpyruvate Metabolism of Anaerobiospirillum succiniciproducens ATCC 29305

  • Yoo, Jin Young;J. Gregory Zeikus
    • Journal of Microbiology and Biotechnology
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    • v.6 no.1
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    • pp.43-49
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    • 1996
  • Modulation of the catabolic PEP-pathway of Anaerobiospirillum succiniciproducens was tried using some enzymatic inhibitors such as gases and chemicals in order to enhance succinic acid production. 10$\%$ CO increased the succinic acid/acetic acid (S/A) ratio but inhibited growth as well as production of succinic and acetic acid. Hydrogen gas also increased the S/A ratio and inhibited the synthesis of pyruvate: ferredoxin oxidoreductase when used in mixture with $CO_2$, Catabolic repression by acetic, lactic and formic acid was not recognized and other modulators such as glyoxylate, pyruvate derivatives, arsenic salt, phosphate and sulfate were shown not to be effective. Magesium carbonate was shown effective for repressing acetate production. Palmitic acid, myristic acid and phenylalanine did not affect acetate production but carprylic acid completely inhibited growth.

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Effects of Glucose and Acetic Acid on the Growth of Recombinant E.coli and the Production of Pyruvate Dehydrogenase Complex-E2 Specific Human Monoclonal Antibody (유전자 재조합 대장균의 세포성장과 Pyruvate Dehydrogenase Complex-E2 특이성 인간 모노클론 항체 생산에 대한 포도당과 초산의 영향)

  • 이미숙;전주미;차상훈;정연호
    • KSBB Journal
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    • v.15 no.5
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    • pp.482-488
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    • 2000
  • The Fab fraction of PDC-E2 specific human monoclonal antibody was produced using recombinant E. coli, and the effects of glucose and acetate were investigated to develop an optimal strategy for recombinant human antibody production. Higher glucose concentration in the culture media resulted inn higher cell growth and glucose consumption rate, which in turn resulted in an increased acetate production rate. When glucose was depleted, cells began to consume acetate as an energy source, and this consumption rate depended on the glucose concentration. When the residual glucose concentration was high, the accumulation of acetate was accelerated due to an increase in the acetate production rate and a decrease in the acetate consumption rate. Futhermore, it was found that a high accumulation of acetate, accompanied by a high glucose concentration, inhibited human antibody formation; the critical acetate concentration was $0.6g/\ell$. During production, a high glucose concentration enhanced cell growth, but inhibited antibody formation due to catabolic repression. Therefore, it is important to keep the concentration of both glucose and acetate as low as possible to increase antibody production after induction. Accordingly, it is important to accurately control the concentration of glucose and acetate in the culture media to obtain high cell densities and high productivity levels of recombinant human antibody.

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Analysis of Trans-Acting Elements for Regulation of moc Operons of pTi15955 in Agrobacterium tumefaciens

  • Jung, Won-Hee;Baek, Chang-Ho;Lee, Jeong-Kug;Kim, Kun-Soo
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.637-645
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    • 1999
  • Two putative regulator genes, mocR and mocS, of the moc (mannityl opine catabolism) operons in pTi15955 of the octopine-/mannityl opine-type Agrobacterium tumefaciens strain 15955, were tested for their possible roles as repressors in the moc operons. The regions upstream of macC and mocD, the first structural genes in the two divergently oriented moc operons, were transcriptionally fused into the promoterless lacZ reporter gene. Each of the lacZ-fusions was introduced into Agrobacterium strain UIA5, a Ti plasmid-cured derivative, harboring either a mocR or a mocS clone. The resulting strains were grown in media containing various sugar sources, and the $\beta$-galactosidase activities were quantitatively measured. The results suggested that MocR repressed the expression of macC and macD. The expression of the fused $\beta$-galactosidase was not induced by mannopine (MOP) or possible catabolic intermediates of the opine, e.g. santhopine (SOP), glucose, mannose, or glutamine. However, the repression was significantly relieved by the supplementation of MOP and the concomitant introduction of the agcA gene encoding MOP cyclase that catalyzes the lactonization of MOP to agropine (AGR). These results suggested that AGR, rather than MOP or the other catabolic intermediates, is the inducer for the expression of the operon. On the contrary to previous report showing that the induction levels of macC and macD were lowered by the supplementation of inorganic nitrogen in media, the expression of these genes was not affected by the level of nitrogen in our reporter system. MocS did not strongly repress the expressions of macC and mocD. It is possible that MocS may be involved in the regulation of the operons present downstream of the moc operon, which are responsible for the utilization of mannopinic acid and agropinic acid.

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Production of DagA, a ${\beta}$-Agarase, by Streptomyces lividans in Glucose Medium or Mixed-Sugar Medium Simulating Microalgae Hydrolysate

  • Park, Juyi;Hong, Soon-Kwang;Chang, Yong Keun
    • Journal of Microbiology and Biotechnology
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    • v.24 no.12
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    • pp.1622-1628
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    • 2014
  • DagA, a ${\beta}$-agarase, was produced by cultivating a recombinant Streptomyces lividans in a glucose medium or a mixed-sugar medium simulating microalgae hydrolysate. The optimum composition of the glucose medium was identified as 25 g/l glucose, 10 g/l yeast extract, and $5g/l\;MgCl_2{\cdot}6H_2O$. With this, a DagA activity of 7.26 U/ml could be obtained. When a mixed-sugar medium containing 25 g/l of sugars was used, a DagA activity of 4.81 U/ml was obtained with very low substrate utilization efficiency owing to the catabolic repression of glucose against the other sugars. When glucose and galactose were removed from the medium, an unexpectedly high DagA activity of about 8.7 U/ml was obtained, even though a smaller amount of sugars was used. It is recommended for better substrate utilization and process economics that glucose and galactose be eliminated from the medium, by being consumed by some other useful applications, before the production of DagA.

Isolation and Characterization of Cryptococcus sp. CS-2 Secreting Polygalacturonase from Soil (토양으로부터 Cryptococcus sp. CS-2의 분리 및 균주가 분비하는 Polygalacturonase의 특성에 관한 연구)

  • 강희경;문명님;임채영;양영기
    • Korean Journal of Microbiology
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    • v.35 no.2
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    • pp.158-163
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    • 1999
  • A ploygalacturonase-produchg yeast was isolated from Cheju soil by selective eivichment media. One strain which has the highesl activity of polygalacturonase was selected. The characle~ishcs of the strain CS-2 were as follows: CS-2 utilized xylose. sucrose, maltose, u.ehalose, cellobiose. melibiose, lactose, raffinose, inosiiol, dulicilol, and dextrose, but did not utilized galactose, nitrate. nit~te, and lysine. Growth of CS-2 was inhibited by cyclohexamide, 1% acetic acid, and high concenaation (over 50%) of glucose. It grew at $30^{\circ}C$ but did 'IIOL $35^{\circ}C$. The cell size ofthe strain CS-2 was 2.9 p ~ n in length and 1.3 $\mu$ in diameter. Vegetable reproductmn was multiple budding and ascospre was present I to 4. Pseudomycelia or true myceliua formation were not observed In any of the cullureq. These results suggest that strain CS-2 is most likely a strain related Cryptococcus spp. (Cryptococcu spp. CS-2). When polygalacturonase or ihe yeast was induced by addition of polygalactoronic acid, polygalacturonase activity was detected in culture supernatent. There was a peak of specific activity a1 he mid-stationary phase(3 days culture) of growth. Polygalacturonase specific activity of Crylmcoccus sp. CS-2 was 2.96 unitsling. The molecular weighl ol'polygalacturonase was showed to be 46 KDa by both SDS-PAGE and activity stailling.

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