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KMID : 0545120090190121612
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Journal of Microbiology and Biotechnology
2009 Volume.19 No. 12 p.1612 ~ p.1616
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Production of Genistein from Naringenin using Escherichia coli Containing Isoflavone Synthase-Cytochrome P450 Reductase Fusion Protein
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Kim Dae-Hwan
Kim Bong-Gyu
Jung Na-Ri
Ahn Joong-Hoon
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Abstract
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Isoflavonoid is known as phytoestrogen. Isoflavnone synthase (IFS) is responsible to convert naringenin to genistein. IFS is one of cytochrome P450 and requires cytochrome P450 reductase (CPR) for its activity. In addition, most of cytochrome P450s contain membrane binding domain and thus it is difficult to express in Escherichia coli. To overcome these problems, we made an in-frame fusion of the IFS from red clover (RCIFS) and CPR gene from rice (RCPR) after removal of the membrane binding domain from IFS and CPR reductase. The resulting fusion gene, RCIFS-RCPR, was expressed in E. coli. Conversion of naringenin into genistein was verified using this E. coli transformant. After optimization of medium and cell density for biotransformation, 60 μM of genistein from 100 μM of naringenin was able to produce. This fusion protein approach might be applicable to express other P450s in E. coli.
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KEYWORD
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cytochrome P450, cytochrome P450 reductase, genistein, naringenin
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