• 제목/요약/키워드: Enzymatic Synthesis

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Enzymatic Synthesis of Cephaloglycin

  • Doo-Hyun Nam;Heon-Soo Sohn;Dewey D. Y. Ryu
    • Bulletin of the Korean Chemical Society
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    • 제4권2호
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    • pp.72-76
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    • 1983
  • Cephaloglycin was synthesized directly from D-${\alpha}$ -phenylglycine methyl ester and 7-aminocephalosporanic acid using whole cell enzyme of Xanthomonas citri (IFO 3835). Some optimal conditions for cephaloglycin synthesis were investigated, and yield improvements for its production by several methods were attempted. Using the whole cell enzyme system, the reaction kinetic model for cephaloglycin synthesis is proposed, and the kinetic constants for D-${\alpha}$ -phenylglycine methyl ester hydrolysis, cephaloglycin synthesis, and cephaloglycin hydrolysis were determined. The $K_m$ values of D-${\alpha}$-phenylglycine methyl ester, 7-aminocephalosporanic acid, and cephaloglycin were 11 mM, 24 mM, and 167 mM, and $K_i$ value of D-${\alpha}$-phenylglycine was 15 mM, respectively. The pattern of product inhibition was found to be competitive one.

N-Benzoylaspartame의 효소적 합성을 위한 용매계의 선정 (Development of Solvent System for Enzymatic Synthesis of N-Benzoylaspartame)

  • 한민수;김우정
    • 한국식품과학회지
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    • 제24권5호
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    • pp.504-510
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    • 1992
  • 고정화 thermolysin에 의한 아스파탐 전구체의 하나인 N-Benzoylaspertame(BzAPM)의 합성을 효율적으로 할 수 있는 수용성 유기용매가 함유된 용매계를 산정하고자 하였다. BzAPM 및 L-phenylalanine(Phe)의 용해도는 methanol 45%가 함유된 용매계에서 각각 1.84 및 1.79%로 가장 높았으며, dimethyl sulfoxlilde(DMSO) 25%와 polyethylene glycol(PEG) 200 20%가 함유된 혼합 용매계도 비교적 높은 용해도를 보였다. BzAPM의 용해도는 ethylene glycol류의 분자량이 커질수록 용해도가 증가하였으나, Phe의 경우에는 이러한 경향을 나타내지 않았다. 고정화 thermolysin에 의한 BzAPM의 합성역가는 methanol 45% 및 DMSO 45%의 난일 유기용매계와 DMSO 25% 및 PEG 200 20%가 함유된 혼합 용매계에서 높게 나타났으며, 초기 합성속도도 빠른 것으로 나타났다. $40^{\circ}C$에서 42일간 고정화 효소를 보관하였을 때, thermolysin은 DMSO 25%와 PEG 200 20%가 함유된 용매계에서 가장 안정하였다. L-phenyalanine methyl ester의 비효소적 가수분해 속도는 methanol함유 용매계에서 가장 낮았고, DMSO 25%와 PEG 200이 함유된 용매계에서는 이들의 중간 정도인 것으로 확인되었다. 그리하여 고정화 thermolysin에 의한 BzAPM의 합성에 적합한 용매계로서 DMSO 25% 및 PEG 200 20%가 함유된 용매계를 선정하였다.

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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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    • 한국생물공학회 2005년도 생물공학의 동향(XVII)
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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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효소공정에 의한 트립토판 생산 (Production of L-Tryptophan by Enzymatic Processes)

  • 이인영;안경섭;김의환;이선복
    • 한국미생물·생명공학회지
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    • 제20권1호
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    • pp.73-78
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    • 1992
  • Tryptophanase를 이용하여 트립토판 합성시 인돌의 효소활성 저해를 억제하기 위하여 유가식 조업, 유기용매 이상계의 사용, 그리고 cyclodextrin의 첨가등에 대하여 연구하였다. 효소 농도가 0.5mg/ml일때 인돌 농도 0.4mM 부근에서 트립토판 생성이 가장 빨랐으며 그 이상에서는 효소활성이 심한 저해를 받았다. 초기 인돌 농도가 20mM일 때는 27시간 반응후 인돌의 전환율이 20인데 비하여 반응기내 인돌 농도를 5mM 이하로 유가식 조업을 하였을 때 전환율이 80%로 향상되었다.

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Phospholipase D에 의한 비천연 인지방질의 합성: I. 에멀젼계 내에서의 합성 (Biosynthesis of Unnatural Phospholipids by Phospholipase D: I. Synthesis in A Emulsion System)

  • 정의호;이해익이상영
    • KSBB Journal
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    • 제6권3호
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    • pp.271-279
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    • 1991
  • Phosphatidylglycerol(PG) and two unnatural phospholipids, phosphatidylethyleneglycol (PEG) and phosphatidylpropyleneglycol(PPG), were synthesized from ovolecithin using cabbage phospholipase D(PLD) in a emulsion system. Optimum pH and temperature for the enzymatic synthesis of PG, PEG and PPG in the emulsion system was 5.0-5.6 and 37$^{\circ}C$, respectively. The maximum activity for transphosphatidylation was obtained with 30-80 mM Ca++. Addition of 25% glycerol was required to convert completely ovolecithin to PG, whereas 16% glycerol was sufficient to attain the highest rate of conversion for both PEG and PPG syntheses, the highest conversion rate was obtained with addition of either 10% ethyleneglycol or propyleneglycol. However, the concentration of alcoholic acceptor should be increased up to 20% to improve selectivity up to 100% for PEG or PPG synthesis. Identification of PEG and PPG was made by analyzing the polyvalent alcohols released after their hydrolysis by HCl or PLD.

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Enzymatic Synthesis of 2-Phenoxyethanol Galactoside by Whole Cells of ${\beta}$-Galactosidase-Containing Escherichia coli

  • Lee, Hyang-Yeol;Jung, Kyung-Hwan
    • Journal of Microbiology and Biotechnology
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    • 제24권9호
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    • pp.1254-1259
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    • 2014
  • We investigated whether ${\beta}$-galactosidase (${\beta}$-gal)-containing Escherichia coli cells could transfer a galactose to 2-phenoxyethanol, resulting in 2-phenoxyethanol galactoside (PE-Gal). PE-Gal was confirmed by liquid chromatography-mass spectrometry. In addition, we also confirmed that a galactose molecule was covalently bonded with PE during thin-layer chromatography analysis of the ${\beta}$-gal hydrolysate of PE-Gal. The yield for PE-Gal synthesis was about 37.5% (weight basis), which was about 7-8 times greater than that of a previous report. In addition, the concentration of ${\beta}$-gal (0.96 U/ml) used in this PE-Gal synthesis was about 20 times less than that in a previous report.

One-pot Enzymatic Synthesis of UDP-D-glucose from UMP and Glucose-1-phosphate Using an ATP Regeneration System

  • Lee, Hei-Chan;Lee, Seung-Don;Sohng, Jae-Kyung;Liou, Kwang-Kyoung
    • BMB Reports
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    • 제37권4호
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    • pp.503-506
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    • 2004
  • Glucose-1-phosphate uridylyltransferase from E. coli K12 was used to convert uridine-5'-triphosphate and glucose-1-phosphate to UDP-D-glucose. The conversion was efficient and completed within 5 minutes under the employed conditions. In addition, thymidine-5'-monophosphate kinase and acetate kinase were proven to be non-specific, converting udridine-5'-monophosphate to uridine-5'-triphosphate with 55% conversion after 6 h, which was much slower than the production of TTP under the same conditions (complete conversion within one hour). Since these two reactions could proceed under the same conditions, a one-pot synthesis of UDP-D-glucose with ATP regeneration was designed from easily available starting materials, and conversion up to 40% by HPLC peak integration was achieved given a reaction time of 4 h.

Optimizing the Synthesis of Citronellyl Valerate Using Lipase from Rhizopus sp

  • De Melo, Lauro Luis M. M.;Pastore, Gbiucia M.;Macedo, Gabriela A.
    • Food Science and Biotechnology
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    • 제14권3호
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    • pp.368-370
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    • 2005
  • Citronellyl valerate was synthesized by a lipase from a Rhizopus sp strain isolated and the lipase produced, at UNICAMP, Brazil. Direct esterification was performed in a solvent-free medium to produce the flavor ester. Response surface methodology was used to optimize the process with respect to the substrate molar ratio and lipase concentration. The results show that the synthesis of citronellyl valerate can be carried out in a solvent-free medium, the maximum ester conversion rate achieved being 91.5% after 48 hours of reaction time.