期刊
BIOTECHNOLOGY AND BIOENGINEERING
卷 116, 期 11, 页码 2852-2863出版社
WILEY
DOI: 10.1002/bit.27133
关键词
Baeyer-Villiger monooxygenase; biocatalysis; dehydrogenase; enzymatic cascade; neutral redox system
资金
- French Agence Nationale de la Recherche (NaturaDyRe) [ANR-10-CD2I-014]
- French Ministere de l'Enseignement Superieur et de la Recherche (Sidiky Menil fellowships)
- COST Action on Systems Biocatalysis [CM1303]
The efficiency of a versatile in vivo cascade involving a promiscuous alcohol dehydrogenase, obtained from a biodiversity search, and a Baeyer-Villiger monooxygenase was enhanced by the independent control of the production level of each enzyme to produce epsilon-caprolactone and 3,4-dihydrocoumarin. This goal was achieved by adjusting the copy number per cell of Escherichia coli plasmids. We started from the observation that this number generally correlates with the amount of produced enzyme and demonstrated that an in vivo multi-enzymatic system can be improved by the judicious choice of plasmid, the lower activity of the enzyme that drives the limiting step being counter-balanced by a higher concentration. Using a preconception-free approach to the choice of the plasmid type, we observed positive and negative synergetic effects, sometimes unexpected and depending on the enzyme and plasmid combinations. Experimental optimization of the culture conditions allowed us to obtain the complete conversion of cyclohexanol (16 mM) and 1-indanol (7.5 mM) at a 0.5-L scale. The yield for the conversion of cyclohexanol was 80% (0.7 g epsilon-caprolactone, for the productivity of 244 mg center dot L-1 center dot h(-1)) and that for 1-indanol 60% (0.3 g 3,4-dihydrocoumarin, for the productivity of 140 mg center dot L-1 center dot h(-1)).
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据