New Platform for Cytochrome P450 Reaction Combining in Situ Immobilization on Biopolymer
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- 作者：Jae Hyung Lee ; Dong Heon Nam ; Sahng Ha Lee ; Jong Hyun Park ; Si Jae Park ; Seung Hwan Lee ; Chan Beum Park ; Ki Jun Jeong
- 刊名：Bioconjugate Chemistry
- 出版年：2014
- 出版时间：December 17, 2014
- 年：2014
- 卷：25
- 期：12
- 页码：2101-2104
- 全文大小：306K
- ISSN：1520-4812
文摘
We describe an efficienct chemical conversion platform with in situ immobilization of P450-BM3 on poly(3-hydroxybutyrate) granules. Through fusion with phasin, P450-BM3 is easily immobilized on poly(3-hydroxybutyrate) granules in Escherichia coli. In our work, the immobilized P450 exhibited higher stability and catalytic activity compared to free P450 against changes of pH, temperature, and concentrations of urea and ions. Through quick recovery of immobilized enzyme, the P450鈥揚(3HB) complex successfully catalyzed an O-dealkylation reaction several times with maintained activity. Using the robust P450鈥揚(3HB) complex, we performed a P450-catalyzed reaction on a preparative reactor scale (100 mL) and high-level production (12.3 渭M) of 7-hydroxycoumarine from 7-ethoxycoumarin could be achieved.