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To Construct an Engineered (S)-Equol Resistant E. coli for in Vitro (S)-Equol Production

(S)-equol is one of the major metabolites of daidzein that is produced by human and animal gut bacteria. Most of the physiological functions of soybean isoflavones, such as anti-oxidative activity, anti-cancer activity, and cardiovascular protection have been ascribed to (S)-equol. However, only 30–...

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Autores principales: Li, Hailiang, Mao, Shaoming, Chen, Huahai, Zhu, Liying, Liu, Wei, Wang, Xin, Yin, Yeshi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5994542/
https://www.ncbi.nlm.nih.gov/pubmed/29915570
http://dx.doi.org/10.3389/fmicb.2018.01182
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author Li, Hailiang
Mao, Shaoming
Chen, Huahai
Zhu, Liying
Liu, Wei
Wang, Xin
Yin, Yeshi
author_facet Li, Hailiang
Mao, Shaoming
Chen, Huahai
Zhu, Liying
Liu, Wei
Wang, Xin
Yin, Yeshi
author_sort Li, Hailiang
collection PubMed
description (S)-equol is one of the major metabolites of daidzein that is produced by human and animal gut bacteria. Most of the physiological functions of soybean isoflavones, such as anti-oxidative activity, anti-cancer activity, and cardiovascular protection have been ascribed to (S)-equol. However, only 30–50% people contain this kind of equol-producing bacteria, and therefore are able to convert daidzein to (S)-equol. Administration of (S)-equol may be more beneficial than soybean isoflavones. The aim of this study was to construct an engineered (S)-equol resistant Escherichia coli to enhance (S)-equol production in vitro. First, transposon mutagenesis libraries were constructed and screened to isolate the (S)-equol resistant mutant E. coli strain BL21 (ydiS) in order to overcome the inhibitory effects of (S)-equol on bacterial growth. Bacterial full genome scan sequencing and in vitro overexpression results revealed that the ydiS gene was responsible for this resistance. Second, the (S)-equol-producing genes L-dznr, L-ddrc, L-dhdr, and L-thdr of Lactococcus strain 20–92 were synthesized and cloned into compatible vectors, pETDuet-1 and pCDFDuet-1. These plasmids were subsequently transformed into BL21 (DE3) and its mutant BL21 (ydiS). Both engineered BL21 (DE3) and BL21 (ydiS) could use daidzein as substrate to produce (S)-equol under both anaerobic and aerobic conditions. As expected, engineered BL21 (ydiS) had faster growth rates than BL21 (DE3) when supplemented with high concentrations of (S)-equol. The yield and the daidzein utilization ratio were higher for engineered BL21 (ydiS). Interestingly, engineered BL21 (ydiS) was able to convert daidzein to (S)-equol efficiently under aerobic conditions, providing a convenient method for (S)-equol production in vitro. In addition, a two-step method was developed to produce (S)-equol using daidzin as substrate.
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spelling pubmed-59945422018-06-18 To Construct an Engineered (S)-Equol Resistant E. coli for in Vitro (S)-Equol Production Li, Hailiang Mao, Shaoming Chen, Huahai Zhu, Liying Liu, Wei Wang, Xin Yin, Yeshi Front Microbiol Microbiology (S)-equol is one of the major metabolites of daidzein that is produced by human and animal gut bacteria. Most of the physiological functions of soybean isoflavones, such as anti-oxidative activity, anti-cancer activity, and cardiovascular protection have been ascribed to (S)-equol. However, only 30–50% people contain this kind of equol-producing bacteria, and therefore are able to convert daidzein to (S)-equol. Administration of (S)-equol may be more beneficial than soybean isoflavones. The aim of this study was to construct an engineered (S)-equol resistant Escherichia coli to enhance (S)-equol production in vitro. First, transposon mutagenesis libraries were constructed and screened to isolate the (S)-equol resistant mutant E. coli strain BL21 (ydiS) in order to overcome the inhibitory effects of (S)-equol on bacterial growth. Bacterial full genome scan sequencing and in vitro overexpression results revealed that the ydiS gene was responsible for this resistance. Second, the (S)-equol-producing genes L-dznr, L-ddrc, L-dhdr, and L-thdr of Lactococcus strain 20–92 were synthesized and cloned into compatible vectors, pETDuet-1 and pCDFDuet-1. These plasmids were subsequently transformed into BL21 (DE3) and its mutant BL21 (ydiS). Both engineered BL21 (DE3) and BL21 (ydiS) could use daidzein as substrate to produce (S)-equol under both anaerobic and aerobic conditions. As expected, engineered BL21 (ydiS) had faster growth rates than BL21 (DE3) when supplemented with high concentrations of (S)-equol. The yield and the daidzein utilization ratio were higher for engineered BL21 (ydiS). Interestingly, engineered BL21 (ydiS) was able to convert daidzein to (S)-equol efficiently under aerobic conditions, providing a convenient method for (S)-equol production in vitro. In addition, a two-step method was developed to produce (S)-equol using daidzin as substrate. Frontiers Media S.A. 2018-06-04 /pmc/articles/PMC5994542/ /pubmed/29915570 http://dx.doi.org/10.3389/fmicb.2018.01182 Text en Copyright © 2018 Li, Mao, Chen, Zhu, Liu, Wang and Yin. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Li, Hailiang
Mao, Shaoming
Chen, Huahai
Zhu, Liying
Liu, Wei
Wang, Xin
Yin, Yeshi
To Construct an Engineered (S)-Equol Resistant E. coli for in Vitro (S)-Equol Production
title To Construct an Engineered (S)-Equol Resistant E. coli for in Vitro (S)-Equol Production
title_full To Construct an Engineered (S)-Equol Resistant E. coli for in Vitro (S)-Equol Production
title_fullStr To Construct an Engineered (S)-Equol Resistant E. coli for in Vitro (S)-Equol Production
title_full_unstemmed To Construct an Engineered (S)-Equol Resistant E. coli for in Vitro (S)-Equol Production
title_short To Construct an Engineered (S)-Equol Resistant E. coli for in Vitro (S)-Equol Production
title_sort to construct an engineered (s)-equol resistant e. coli for in vitro (s)-equol production
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5994542/
https://www.ncbi.nlm.nih.gov/pubmed/29915570
http://dx.doi.org/10.3389/fmicb.2018.01182
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