Efficient stereoselective hydroxylation of deoxycholic acid by the robust whole-cell cytochrome P450 CYP107D1 biocatalyst  

作　　者：Chixiang Sun Baodong Hu Yanchun Li Zhimeng Wu Jingwen Zhou Jianghua Li Jian Chen Guocheng Du Xinrui Zhao 

机构地区：[1]Science Center for Future Foods,Jiangnan University,1800 Lihu Road,Wuxi,Jiangsu,214122,China [2]Key Laboratory of Industrial Biotechnology,Ministry of Education,School of Biotechnology,Jiangnan University,1800 Lihu Road,Wuxi,Jiangsu,214122,China [3]Jiangsu Province Engineering Research Center of Food Synthetic Biotechnology,Jiangnan University,1800 Lihu Road,Wuxi,Jiangsu,214122,China [4]Engineering Research Center of Ministry of Education on Food Synthetic Biotechnology,Jiangnan University,1800 Lihu Road,Wuxi,Jiangsu,214122,China [5]Key Laboratory of Carbohydrate Chemistry and Biotechnology,Ministry of Education,Jiangnan University,1800 Lihu Road,Wuxi,Jiangsu,214122,China

出　　处：《Synthetic and Systems Biotechnology》2023年第4期741-748,共8页合成和系统生物技术（英文）

基　　金：supported by the National Key Research and Development Program of China(2019YFA0906400);the National First-class Discipline Program of Light Industry Technology and Engineering(LITE2018-08);Postgraduate Research&Practice Innovation Program of Jiangsu Province(KYCX23_2486);We thank Prof.Shengying Li(Shandong University,China)for providing plasmids pET28a-SelFdx1499 and pET28a-SelFdR0978.

摘　　要：Deoxycholic acid(DCA)has been authorized by the Federal Drug Agency for cosmetic reduction of redundant submental fat.The hydroxylated product(6β-OH DCA)was developed to improve the solubility and pharmaceutic properties of DCA for further applications.Herein,a combinatorial catalytic strategy was applied to construct a powerful Cytochrome P450 biocatalyst(CYP107D1,OleP)to convert DCA to 6β-OH DCA.Firstly,the weak expression of OleP was significantly improved using pRSFDuet-1 plasmid in the E.coli C41(DE3)strain.Next,the supply of heme was enhanced by the moderate overexpression of crucial genes in the heme biosynthetic pathway.In addition,a new biosensor was developed to select the appropriate redox partner.Furthermore,a cost-effective whole-cell catalytic system was constructed,resulting in the highest reported conversion rate of 6β-OH DCA(from 4.8%to 99.1%).The combinatorial catalytic strategies applied in this study provide an efficient method to synthesize high-value-added hydroxylated compounds by P450s.

关 键 词：OleP Deoxycholic acid HYDROXYLATION Redox partners Whole-cell catalysis 

分 类 号：Q81[生物学—生物工程]