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Engineered EryF hydroxylase improving heterologous polyketide erythronolide B production in Escherichia coli

In the last two decades, the production of complex polyketides such as erythromycin and its precursor 6‐deoxyerythronolide B (6‐dEB) was demonstrated feasible in Escherichia coli. Although the heterologous production of polyketide skeleton 6‐dEB has reached 210 mg l(−1) in E. coli, the yield of its...

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Autores principales: Liu, Zhifeng, Xu, Jianlin, Liu, Haili, Wang, Yong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9049603/
https://www.ncbi.nlm.nih.gov/pubmed/35174640
http://dx.doi.org/10.1111/1751-7915.14000
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author Liu, Zhifeng
Xu, Jianlin
Liu, Haili
Wang, Yong
author_facet Liu, Zhifeng
Xu, Jianlin
Liu, Haili
Wang, Yong
author_sort Liu, Zhifeng
collection PubMed
description In the last two decades, the production of complex polyketides such as erythromycin and its precursor 6‐deoxyerythronolide B (6‐dEB) was demonstrated feasible in Escherichia coli. Although the heterologous production of polyketide skeleton 6‐dEB has reached 210 mg l(−1) in E. coli, the yield of its post‐modification products erythromycins remains to be improved. Cytochrome P450EryF catalyses the C6 hydroxylation of 6‐dEB to form erythronolide B (EB), which is the initial rate‐limiting modification in a multi‐step pathway to convert 6‐dEB into erythromycin. Here, we engineered hydroxylase EryF to improve the production of heterologous polyketide EB in E. coli. By comparative analysis of various versions of P450EryFs, we confirmed the optimal SaEryF for the biosynthesis of EB. Further mutation of SaEryF based on the crystal structure of SaEryF and homology modelling of AcEryF and AeEryF afforded the enhancement of EB production. The designed mutant of SaEryF, I379V, achieved the yield of 131 mg l(−1) EB, which was fourfold to that produced by wild‐type SaEryF. Moreover, the combined mutagenesis of multiple residues led to further boost the EB concentration by another 41%, which laid the foundation for efficient heterologous biosynthesis of erythromycin or other complex polyketides.
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spelling pubmed-90496032022-05-02 Engineered EryF hydroxylase improving heterologous polyketide erythronolide B production in Escherichia coli Liu, Zhifeng Xu, Jianlin Liu, Haili Wang, Yong Microb Biotechnol Engineering Biology and Synthetic Biology In the last two decades, the production of complex polyketides such as erythromycin and its precursor 6‐deoxyerythronolide B (6‐dEB) was demonstrated feasible in Escherichia coli. Although the heterologous production of polyketide skeleton 6‐dEB has reached 210 mg l(−1) in E. coli, the yield of its post‐modification products erythromycins remains to be improved. Cytochrome P450EryF catalyses the C6 hydroxylation of 6‐dEB to form erythronolide B (EB), which is the initial rate‐limiting modification in a multi‐step pathway to convert 6‐dEB into erythromycin. Here, we engineered hydroxylase EryF to improve the production of heterologous polyketide EB in E. coli. By comparative analysis of various versions of P450EryFs, we confirmed the optimal SaEryF for the biosynthesis of EB. Further mutation of SaEryF based on the crystal structure of SaEryF and homology modelling of AcEryF and AeEryF afforded the enhancement of EB production. The designed mutant of SaEryF, I379V, achieved the yield of 131 mg l(−1) EB, which was fourfold to that produced by wild‐type SaEryF. Moreover, the combined mutagenesis of multiple residues led to further boost the EB concentration by another 41%, which laid the foundation for efficient heterologous biosynthesis of erythromycin or other complex polyketides. John Wiley and Sons Inc. 2022-02-17 /pmc/articles/PMC9049603/ /pubmed/35174640 http://dx.doi.org/10.1111/1751-7915.14000 Text en © 2022 The Authors. Microbial Biotechnology published by Society for Applied Microbiology and John Wiley & Sons Ltd. https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Engineering Biology and Synthetic Biology
Liu, Zhifeng
Xu, Jianlin
Liu, Haili
Wang, Yong
Engineered EryF hydroxylase improving heterologous polyketide erythronolide B production in Escherichia coli
title Engineered EryF hydroxylase improving heterologous polyketide erythronolide B production in Escherichia coli
title_full Engineered EryF hydroxylase improving heterologous polyketide erythronolide B production in Escherichia coli
title_fullStr Engineered EryF hydroxylase improving heterologous polyketide erythronolide B production in Escherichia coli
title_full_unstemmed Engineered EryF hydroxylase improving heterologous polyketide erythronolide B production in Escherichia coli
title_short Engineered EryF hydroxylase improving heterologous polyketide erythronolide B production in Escherichia coli
title_sort engineered eryf hydroxylase improving heterologous polyketide erythronolide b production in escherichia coli
topic Engineering Biology and Synthetic Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9049603/
https://www.ncbi.nlm.nih.gov/pubmed/35174640
http://dx.doi.org/10.1111/1751-7915.14000
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