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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제24권 제12호
- 발행연도
- 2014.1
- 수록면
- 1,636 - 1,643 (8page)
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초록· 키워드
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3-Hydroxybutyryl-CoA dehydrogenase is an enzyme that catalyzes the second step in the biosynthesis of n-butanol from acetyl-CoA, in which acetoacetyl-CoA is reduced to 3-hydroxybutyryl-CoA. To understand the molecular mechanisms of n-butanol biosynthesis, we determined the crystal structure of 3-hydroxybutyryl-CoA dehydrogenase from Clostridium butyricum (CbHBD). The monomer structure of CbHBD exhibits a two-domain topology, with N- and C-terminal domains, and the dimerization of the enzyme was mostly constituted at the C-terminal domain. The mode of cofactor binding to CbHBD was elucidated by determining the crystal structure of the enzyme in complex with NAD+. We also determined the enzyme’s structure in complex with its acetoacetyl-CoA substrate, revealing that the adenosine diphosphate moiety was not highly stabilized compared with the remainder of the acetoacetyl-CoA molecule. Using this structural information, we performed a series of sitedirected mutagenesis experiments on the enzyme, such as changing residues located near the substrate-binding site, and finally developed a highly efficient CbHBD K50A/K54A/L232Y triple mutant enzyme that exhibited approximately 5-fold higher enzyme activity than did the wild type. The increased enzyme activity of the mutant was confirmed by enzyme kinetic measurements. The highly efficient mutant enzyme should be useful for increasing the production rate of n-butanol.
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참고문헌 (24)
참고문헌 신청
Atsumi S / 2008 / Metabolic engineering of Escherichia coli for 1-butanol production / Metab. Eng 10 : 305 ~ 311
Atsumi S / 2008 / Non-fermentative pathways for synthesis of branched-chain higher alcohols as biofuels / Nature 451 : 86 ~ 89
Barycki JJ / 1999 / Biochemical characterization and crystal structure determination of human heart short chain L-3-hydroxyacyl-CoA dehydrogenase provide insights into cataly
Barycki JJ / 2000 / Sequestration of the active site by interdomain shifting. Crystallographic and spectroscopic evidence for distinct conformations of L-3-hydroxyacyl-CoA de
Boynton ZL / 1996 / Cloning, sequencing, and expression of clustered genes encoding betahydroxybutyryl-coenzyme A (CoA) dehydrogenase, crotonase and butyryl-CoA dehydrogenase
Atsumi S / 2008 / Non-fermentative pathways for synthesis of branched-chain higher alcohols as biofuels / Nature 451 : 86 ~ 89
Barycki JJ / 1999 / Biochemical characterization and crystal structure determination of human heart short chain L-3-hydroxyacyl-CoA dehydrogenase provide insights into cataly
Barycki JJ / 2000 / Sequestration of the active site by interdomain shifting. Crystallographic and spectroscopic evidence for distinct conformations of L-3-hydroxyacyl-CoA de
Boynton ZL / 1996 / Cloning, sequencing, and expression of clustered genes encoding betahydroxybutyryl-coenzyme A (CoA) dehydrogenase, crotonase and butyryl-CoA dehydrogenase
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