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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제20권 제4호
- 발행연도
- 2010.1
- 수록면
- 659 - 665 (7page)
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초록· 키워드
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A cis-epoxysuccinate hydrolase (CESH) from Bordetella sp. BK-52 was purified 51.4-fold with a yield of 27.1%using ammonium sulfate precipitation, ionic exchange,hydrophobic interaction, molecular sieve chromatography and an additional anion-exchange chromatography. The CESH was stable in a broad range of temperature (up to 50oC) and pH (4.0-10.0) with optima of 40oC and pH 6.5,respectively. It could be partially inhibited by EDTA-Na2,Ag+, SDS, and DTT, and slightly enhanced by Ba2+ and Ca2+. The enzyme exhibited high stereospecificity in D(-)-tartaric acid (enantiomeric excess value higher than 99%)with Km and Vmax values of 18.67 mM and 94.34 μM/min/mg for disodium cis-epoxysuccinate, respectively. The Bordetella sp. BK-52 CESH gene, which contained 885 nucleotides (open reading frame) encoding 294 amino acids with a molecular mass of about 32 kDa, was successfully overexpressed in Escherichia coli using a T7/lac promoter vector and the enzyme activity was increased 42-times compared with the original strain. It may be an industrial biocatalyst for the preparation of D(-)-tartaric acid.
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참고문헌 (20)
참고문헌 신청
Archelas, A / 1998 / Epoxide hydrolases: New tools for the synthesis of fine organic chemicals / Trends Biotechnol 16 : 108 ~ 116
Bradford, M. M / 1976 / A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding / Anal. Biochem 72
Chung, C. T / 1989 / One-step preparation of competent Escherichia coli: Transformation and storage of bacterial cells in the same solution / Proc. Natl. Acad. Sci. U.S.A : 2172 ~
Ghosh, A. K / 2001 / Tartaric acid and tartarates in the synthesis of bioactive molecules / Synthesis 9 : 1281 ~ 1301
Huang, T. H / 1990 / Production of L (+) tartaric acid / Gong Ye Wei Sheng Wu 6 : 14 ~ 17
Bradford, M. M / 1976 / A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding / Anal. Biochem 72
Chung, C. T / 1989 / One-step preparation of competent Escherichia coli: Transformation and storage of bacterial cells in the same solution / Proc. Natl. Acad. Sci. U.S.A : 2172 ~
Ghosh, A. K / 2001 / Tartaric acid and tartarates in the synthesis of bioactive molecules / Synthesis 9 : 1281 ~ 1301
Huang, T. H / 1990 / Production of L (+) tartaric acid / Gong Ye Wei Sheng Wu 6 : 14 ~ 17
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