Constructing Protein-Scaffolded Multienzyme Assembly Enhances the Coupling Efficiency of the P450 System for Efficient Daidzein Biosynthesis from (2S)-Naringenin

• DOI: 10.1021/acs.jafc.3c09854
• Publication Date: 2024-03-12T05:27:50Z
• Authors: Zhe Wang; Yiqiang Dai; Fidelis Azi; Zhongjiang Wang; Weimin Xu; Daoying Wang; Mingsheng Dong; Xiudong Xia

Daidzein is a major isoflavone compound with an immense pharmaceutical value. This study applied novel P450 CYP82D26 which can biosynthesize daidzein from (2S)-naringenin. However, the recombinant systems often suffer low coupling efficiency, leading to electron transfer efficiency decrease and harmful reactive oxygen species release, thereby compromising their stability catalytic efficiency. To address these challenges, SH3-GBD-PDZ (SGP) protein scaffold was assemble multienzyme system comprising CYP82D26, reductase, NADP+-dependent aldehyde reductase in desired stoichiometric ratios. Results showed that of significantly increased, primarily attributed channeling effect NADPH resulting proximity tethered enzymes electrostatic interactions between SGP. Assembling this SGP-scaffolded assembly Escherichia coli yielded titer 240.5 mg/L 86% (2S)-naringenin conversion rate, 9-fold increase over free system. These results underscore potential application enhancing