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

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Synthetic and Systems Biotechnology 2023年第4期8卷 741-748页

作者： Chixiang Sun Baodong Hu Yanchun Li Zhimeng Wu Jingwen Zhou Jianghua Li Jian Chen Guocheng Du Xinrui Zhao Science Center for Future Foods Jiangnan University1800 Lihu RoadWuxiJiangsu214122China Key Laboratory of Industrial Biotechnology Ministry of EducationSchool of BiotechnologyJiangnan University1800 Lihu RoadWuxiJiangsu214122China Jiangsu Province Engineering Research Center of Food Synthetic Biotechnology Jiangnan University1800 Lihu RoadWuxiJiangsu214122China Engineering Research Center of Ministry of Education on Food Synthetic Biotechnology Jiangnan University1800 Lihu RoadWuxiJiangsu214122China Key Laboratory of Carbohydrate Chemistry and Biotechnology Ministry of EducationJiangnan University1800 Lihu RoadWuxiJiangsu214122China

Deoxycholic acid(DCA)has been authorized by the Federal Drug Agency for cosmetic reduction of redundant submental *** hydroxylated product(6β-OH DCA)was developed to improve the solubility and pharmaceutic properties of DCA for further ***,a combinatorial catalytic strategy was applied to construct a powerful Cytochrome P450 biocatalyst(CYP107D1,OleP)to convert DCA to 6β-OH ***,the weak expression of OleP was significantly improved using pRSFDuet-1 plasmid in the *** C41(DE3)***,the supply of heme was enhanced by the moderate overexpression of crucial genes in the heme biosynthetic *** addition,a new biosensor was developed to select the appropriate redox ***,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

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Functional expression and regulation of eukaryotic cytochrome P450 enzymes in surrogate microbial cell factories

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Engineering Microbiology 2022年第1期2卷 17-34页

作者： Pradeepraj Durairaj Shengying Li State Key Laboratory of Microbial Technology Shandong UniversityQingdao 266237ShandongChina Laboratory for Marine Biology and Biotechnology Qingdao National Laboratory for Marine Science and TechnologyQingdao 266237ShandongChina

Cytochrome P450(CYP)enzymes play crucial roles during the evolution and diversification of ancestral monocel-lular eukaryotes into multicellular eukaryotic organisms due to their essential functionalities including catalysis of housekeeping biochemical reactions,synthesis of diverse metabolites,detoxification of xenobiotics,and con-tribution to environmental *** CYPs with versatile functionalities are undeniably regarded as promising biocatalysts with great potential for biotechnological,pharmaceutical and chemical industry ***,the modes of action and the challenges associated with these membrane-bound proteins have hampered the effective utilization of eukaryotic CYPs in a broader *** review is focused on comprehen-sive and consolidated approaches to address the core challenges in heterologous expression of membrane-bound eukaryotic CYPs in different surrogate microbial cell factories,aiming to provide key insights for better studies and applications of diverse eukaryotic CYPs in the *** also highlight the functional significance of the previously underrated cytochrome P450 reductases(CPRs)and provide a rational justification on the progression of CPR from auxiliary redox partner to function modulator in CYP catalysis.

关键词： Cytochrome P450 enzymes Cytochrome P450 reductase Membrane-bound proteins N-terminal transmembrane domain Heterologous expression Microbial cell factories redox partners Electron transfer

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