Electron transfer engineering of artificially designed cell factory for complete biosynthesis of steroids

Abstract Biosynthesis of steroids by artificially designed cell factories often involves numerous nicotinamide adenine dinucleotide phosphate (NADPH)-dependent enzymes that mediate electron transfer reactions. However, the unclear mechanisms of electron transfer from regeneration to the final delive...

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Main Authors: Qihang Chen, Wenqian Wei, Zikai Chao, Rui Qi, Jianhong He, Huating Chen, Ke Wang, Xinglong Wang, Yijian Rao, Jingwen Zhou
Format: Article
Language:English
Published: Nature Portfolio 2025-04-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-58926-9
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author Qihang Chen
Wenqian Wei
Zikai Chao
Rui Qi
Jianhong He
Huating Chen
Ke Wang
Xinglong Wang
Yijian Rao
Jingwen Zhou
author_facet Qihang Chen
Wenqian Wei
Zikai Chao
Rui Qi
Jianhong He
Huating Chen
Ke Wang
Xinglong Wang
Yijian Rao
Jingwen Zhou
author_sort Qihang Chen
collection DOAJ
description Abstract Biosynthesis of steroids by artificially designed cell factories often involves numerous nicotinamide adenine dinucleotide phosphate (NADPH)-dependent enzymes that mediate electron transfer reactions. However, the unclear mechanisms of electron transfer from regeneration to the final delivery to the NADPH-dependent active centers limit systematically engineering electron transfer to improve steroids production. Here, we elucidate the electron transfer mechanisms of NADPH-dependent enzymes for systematically engineer electron transfer of Saccharomyces cerevisiae, including step-by-step engineering the electron transfer residues of 7-Dehydrocholesterol reductase (DHCR7) and P450 sterol side chain cleaving enzyme (P450scc), electron transfer components for directing carbon flux, and NADPH regeneration pathways, for high-level production of the cholesterol (1.78 g/L) and pregnenolone (0.83 g/L). The electron transfer engineering (ETE) process makes the electron transfer chains shorter and more stable which significantly accelerates deprotonation and proton coupled electron transfer process. This study underscores the significance of ETE strategies in steroids biosynthesis and expands synthetic biology approaches.
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issn 2041-1723
language English
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publisher Nature Portfolio
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series Nature Communications
spelling doaj-art-9f867aaca14747fab044e4b9d52516d32025-08-20T03:15:09ZengNature PortfolioNature Communications2041-17232025-04-0116111910.1038/s41467-025-58926-9Electron transfer engineering of artificially designed cell factory for complete biosynthesis of steroidsQihang Chen0Wenqian Wei1Zikai Chao2Rui Qi3Jianhong He4Huating Chen5Ke Wang6Xinglong Wang7Yijian Rao8Jingwen Zhou9Key Laboratory of Industrial Biotechnology, Ministry of Education and School of Biotechnology, Jiangnan University, 1800 Lihu RoadKey Laboratory of Industrial Biotechnology, Ministry of Education and School of Biotechnology, Jiangnan University, 1800 Lihu RoadKey Laboratory of Industrial Biotechnology, Ministry of Education and School of Biotechnology, Jiangnan University, 1800 Lihu RoadKey Laboratory of Industrial Biotechnology, Ministry of Education and School of Biotechnology, Jiangnan University, 1800 Lihu RoadKey Laboratory of Industrial Biotechnology, Ministry of Education and School of Biotechnology, Jiangnan University, 1800 Lihu RoadKey Laboratory of Industrial Biotechnology, Ministry of Education and School of Biotechnology, Jiangnan University, 1800 Lihu RoadKey Laboratory of Industrial Biotechnology, Ministry of Education and School of Biotechnology, Jiangnan University, 1800 Lihu RoadKey Laboratory of Industrial Biotechnology, Ministry of Education and School of Biotechnology, Jiangnan University, 1800 Lihu RoadKey Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education and School of Biotechnology, Jiangnan University, 1800 Lihu RoadKey Laboratory of Industrial Biotechnology, Ministry of Education and School of Biotechnology, Jiangnan University, 1800 Lihu RoadAbstract Biosynthesis of steroids by artificially designed cell factories often involves numerous nicotinamide adenine dinucleotide phosphate (NADPH)-dependent enzymes that mediate electron transfer reactions. However, the unclear mechanisms of electron transfer from regeneration to the final delivery to the NADPH-dependent active centers limit systematically engineering electron transfer to improve steroids production. Here, we elucidate the electron transfer mechanisms of NADPH-dependent enzymes for systematically engineer electron transfer of Saccharomyces cerevisiae, including step-by-step engineering the electron transfer residues of 7-Dehydrocholesterol reductase (DHCR7) and P450 sterol side chain cleaving enzyme (P450scc), electron transfer components for directing carbon flux, and NADPH regeneration pathways, for high-level production of the cholesterol (1.78 g/L) and pregnenolone (0.83 g/L). The electron transfer engineering (ETE) process makes the electron transfer chains shorter and more stable which significantly accelerates deprotonation and proton coupled electron transfer process. This study underscores the significance of ETE strategies in steroids biosynthesis and expands synthetic biology approaches.https://doi.org/10.1038/s41467-025-58926-9
spellingShingle Qihang Chen
Wenqian Wei
Zikai Chao
Rui Qi
Jianhong He
Huating Chen
Ke Wang
Xinglong Wang
Yijian Rao
Jingwen Zhou
Electron transfer engineering of artificially designed cell factory for complete biosynthesis of steroids
Nature Communications
title Electron transfer engineering of artificially designed cell factory for complete biosynthesis of steroids
title_full Electron transfer engineering of artificially designed cell factory for complete biosynthesis of steroids
title_fullStr Electron transfer engineering of artificially designed cell factory for complete biosynthesis of steroids
title_full_unstemmed Electron transfer engineering of artificially designed cell factory for complete biosynthesis of steroids
title_short Electron transfer engineering of artificially designed cell factory for complete biosynthesis of steroids
title_sort electron transfer engineering of artificially designed cell factory for complete biosynthesis of steroids
url https://doi.org/10.1038/s41467-025-58926-9
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