• Title/Summary/Keyword: Biological Synthesis

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Selection and Directed Evolution of New Microbial Biocatalysts and Their Application to Organic Synthesis

  • Asano, Yasuhisa
    • Journal of Applied Biological Chemistry
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    • v.43 no.4
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    • pp.207-210
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    • 2000
  • As a typical example of the screening for a microbial biocatalyst from nature, isolation of nitrilesynthesizing microorganisms, characterization of a new enzyme aldoxime dehydratase, and its function in the aldoxime-nitrile pathway are introduced. Catalytic properties of some of our enzymes were improved through a direct evolutionary approach.

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Polyamine Stimulation of arcA Expression in Escherichia coli

  • Rhee, Mun-Su;Kim, Young-Sik;Park, Seon-Young;Park, Myung-Hun;Kim, Bo-Min;Kang, Seong-Uk;Lee, Kui-Joo;Lee, Jong-Ho
    • Journal of Microbiology
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    • v.40 no.4
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    • pp.305-312
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    • 2002
  • The effects of two natural polyamines (putrescine and spermidine) on the synthesis of ArcA, a response regulator of the Arc two-component signal transduction system, were studied using an E. coli mutant deficient in polyamine biosynthesis. Endogenous polyamine deficiency of the mutant resulted in marked reduction in the ArcA level determined by Western blot analysis. Putrescine supplement to the growth medium effectively increased the ArcA level of the mutant in a concentration-dependent manner. Spermidine also stimulated the ArcA level in the mutant to a greater degree than putrescine. Expression of arcA'::lacZ operon fusion in the mutant was stimulated 6-fold and 10-fold by putrescine and spermidine at a 1mM concentration, respectively, indicating that the stimulatory effect of the polyamines on ArcA synthesis is due to transcriptional induction, and that spermidine is a more potent arcA inducer than putrescine. The polyamine-dependent arcA'::lacZ induction was growth-phase-dependent and independent of either arcA or fnr which are two regulators involved in anaerobic stimulation of the Arch level. These results suggested that putrescine and spermidine polyamines may be potential intracellular signal molecules in the control of arcA expression, and thereby may play an important role in cellular metabolism.

Response Surface Methodological Approach for Optimization of Enzymatic Synthesis of Sorbitan Methacrylate

  • Jeong, Gwi-Taek;Lee, Kyoung-Min;Kim, Hae-Sung;Lee, Woo-Tai;Sunwoo, Chang-Shin;Park, Don-Hee
    • 한국생물공학회:학술대회논문집
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    • 2005.10a
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    • pp.511-516
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    • 2005
  • Sorbitan methacrylate was synthesized from sorbitan dehydrated from D-sorbitol using an immobilized lipase. To optimize the enzymatic synthesis of sorbitan methacrylate, response surface methodology was applied to determine the effects of five-level-four-factors and their reciprocal interactions on sorbitan methacrylate biosynthesis. A total of 30 individual experiments were performed, which were designed to study reaction temperature, reaction time, enzyme amount and substrate molar ratio. A statistical model predicted that the highest conversion yield of sorbitan methacrylate was 100%, at the following optimized reaction conditions: a reaction temperature of 43.06 $^{\circ}C$, a reaction time of 164.25 mins., an enzyme amount of 7.47%, and a substrate molar ratio of 3.98:1. Using these optimal factor values under experimental conditions in four independent replicates, the average conversion yield reached 98.7%${\pm}$1.2% and was well within the value predicted by the model.

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Effect of Expression of Genes in the Sphingolipid Synthesis Pathway on the Biosynthesis of Ceramide in Saccharomyces cerevisiae

  • Kim, Se-Kyung;Noh, Yong-Ho;Koo, Ja-Ryong;Yun, Hyun-Shik
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.356-362
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    • 2010
  • Ceramide is important not only for the maintenance of the barrier function of the skin but also for the water-binding capacity of the stratum corneum. Although the exact role of ceramide in the human skin is not fully understood, ceramide has become a widely used ingredient in cosmetic and pharmaceutical industries. Compared with other microorganisms, yeast is more suitable for the production of ceramide because yeast grows fast and is non-toxic. However, production of ceramide from yeast has not been widely studied and most work in this area has been carried out using Saccharomyces cerevisiae. Regulating the genes that are involved in sphingolipid synthesis is necessary to increase ceramide production. In this study, we investigated the effect of the genes involved in the synthesis of ceramide, lcb1, lcb2, tsc10, lac1, lag1, and sur2, on ceramide production levels. The genes were cloned into pYES2 high copy number vectors. S. cerevisiae was cultivated on YPDG medium at $30^{\circ}C$. Ceramide was purified from the cell extracts by solvent extraction and the ceramide content was analyzed by HPLC using ELSD. The maximum production of ceramide (9.8 mg ceramide/g cell) was obtained when the tsc10 gene was amplified by the pYES2 vector. Real-time RT-PCR analysis showed that the increase in ceramide content was proportional to the increase in the tsc10 gene expression level, which was 4.56 times higher than that of the control strain.