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Synthetic engineering of Corynebacterium crenatum to selectively produce acetoin or 2,3-butanediol by one step bioconversion method

BACKGROUND: Acetoin (AC) and 2,3-butanediol (2,3-BD) as highly promising bio-based platform chemicals have received more attentions due to their wide range of applications. However, the non-efficient substrate conversion and mutually transition between AC and 2,3-BD in their natural producing strain...

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Autores principales: Zhang, Xian, Han, Rumeng, Bao, Teng, Zhao, Xiaojing, Li, Xiangfei, Zhu, Manchi, Yang, Taowei, Xu, Meijuan, Shao, Minglong, Zhao, Youxi, Rao, Zhiming
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6683508/
https://www.ncbi.nlm.nih.gov/pubmed/31387595
http://dx.doi.org/10.1186/s12934-019-1183-0
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author Zhang, Xian
Han, Rumeng
Bao, Teng
Zhao, Xiaojing
Li, Xiangfei
Zhu, Manchi
Yang, Taowei
Xu, Meijuan
Shao, Minglong
Zhao, Youxi
Rao, Zhiming
author_facet Zhang, Xian
Han, Rumeng
Bao, Teng
Zhao, Xiaojing
Li, Xiangfei
Zhu, Manchi
Yang, Taowei
Xu, Meijuan
Shao, Minglong
Zhao, Youxi
Rao, Zhiming
author_sort Zhang, Xian
collection PubMed
description BACKGROUND: Acetoin (AC) and 2,3-butanediol (2,3-BD) as highly promising bio-based platform chemicals have received more attentions due to their wide range of applications. However, the non-efficient substrate conversion and mutually transition between AC and 2,3-BD in their natural producing strains not only led to a low selectivity but also increase the difficulty of downstream purification. Therefore, synthetic engineering of more suitable strains should be a reliable strategy to selectively produce AC and 2,3-BD, respectively. RESULTS: In this study, the respective AC (alsS and alsD) and 2,3-BD biosynthesis pathway genes (alsS, alsD, and bdhA) derived from Bacillus subtilis 168 were successfully expressed in non-natural AC and 2,3-BD producing Corynebacterium crenatum, and generated recombinant strains, C. crenatum SD and C. crenatum SDA, were proved to produce 9.86 g L(−1) of AC and 17.08 g L(−1) of 2,3-BD, respectively. To further increase AC and 2,3-BD selectivity, the AC reducing gene (butA) and lactic acid dehydrogenase gene (ldh) in C. crenatum were then deleted. Finally, C. crenatumΔbutAΔldh SD produced 76.93 g L(−1) AC in one-step biocatalysis with the yield of 0.67 mol mol(−1). Meanwhile, after eliminating the lactic acid production and enhancing 2,3-butanediol dehydrogenase activity, C. crenatumΔldh SDA synthesized 88.83 g L(−1) of 2,3-BD with the yield of 0.80 mol mol(−1). CONCLUSIONS: The synthetically engineered C. crenatumΔbutAΔldh SD and C. crenatumΔldh SDA in this study were proved as an efficient microbial cell factory for selective AC and 2,3-BD production. Based on the insights from this study, further synthetic engineering of C. crenatum for AC and 2,3-BD production is suggested. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12934-019-1183-0) contains supplementary material, which is available to authorized users.
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spelling pubmed-66835082019-08-09 Synthetic engineering of Corynebacterium crenatum to selectively produce acetoin or 2,3-butanediol by one step bioconversion method Zhang, Xian Han, Rumeng Bao, Teng Zhao, Xiaojing Li, Xiangfei Zhu, Manchi Yang, Taowei Xu, Meijuan Shao, Minglong Zhao, Youxi Rao, Zhiming Microb Cell Fact Research BACKGROUND: Acetoin (AC) and 2,3-butanediol (2,3-BD) as highly promising bio-based platform chemicals have received more attentions due to their wide range of applications. However, the non-efficient substrate conversion and mutually transition between AC and 2,3-BD in their natural producing strains not only led to a low selectivity but also increase the difficulty of downstream purification. Therefore, synthetic engineering of more suitable strains should be a reliable strategy to selectively produce AC and 2,3-BD, respectively. RESULTS: In this study, the respective AC (alsS and alsD) and 2,3-BD biosynthesis pathway genes (alsS, alsD, and bdhA) derived from Bacillus subtilis 168 were successfully expressed in non-natural AC and 2,3-BD producing Corynebacterium crenatum, and generated recombinant strains, C. crenatum SD and C. crenatum SDA, were proved to produce 9.86 g L(−1) of AC and 17.08 g L(−1) of 2,3-BD, respectively. To further increase AC and 2,3-BD selectivity, the AC reducing gene (butA) and lactic acid dehydrogenase gene (ldh) in C. crenatum were then deleted. Finally, C. crenatumΔbutAΔldh SD produced 76.93 g L(−1) AC in one-step biocatalysis with the yield of 0.67 mol mol(−1). Meanwhile, after eliminating the lactic acid production and enhancing 2,3-butanediol dehydrogenase activity, C. crenatumΔldh SDA synthesized 88.83 g L(−1) of 2,3-BD with the yield of 0.80 mol mol(−1). CONCLUSIONS: The synthetically engineered C. crenatumΔbutAΔldh SD and C. crenatumΔldh SDA in this study were proved as an efficient microbial cell factory for selective AC and 2,3-BD production. Based on the insights from this study, further synthetic engineering of C. crenatum for AC and 2,3-BD production is suggested. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12934-019-1183-0) contains supplementary material, which is available to authorized users. BioMed Central 2019-08-06 /pmc/articles/PMC6683508/ /pubmed/31387595 http://dx.doi.org/10.1186/s12934-019-1183-0 Text en © The Author(s) 2019 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Zhang, Xian
Han, Rumeng
Bao, Teng
Zhao, Xiaojing
Li, Xiangfei
Zhu, Manchi
Yang, Taowei
Xu, Meijuan
Shao, Minglong
Zhao, Youxi
Rao, Zhiming
Synthetic engineering of Corynebacterium crenatum to selectively produce acetoin or 2,3-butanediol by one step bioconversion method
title Synthetic engineering of Corynebacterium crenatum to selectively produce acetoin or 2,3-butanediol by one step bioconversion method
title_full Synthetic engineering of Corynebacterium crenatum to selectively produce acetoin or 2,3-butanediol by one step bioconversion method
title_fullStr Synthetic engineering of Corynebacterium crenatum to selectively produce acetoin or 2,3-butanediol by one step bioconversion method
title_full_unstemmed Synthetic engineering of Corynebacterium crenatum to selectively produce acetoin or 2,3-butanediol by one step bioconversion method
title_short Synthetic engineering of Corynebacterium crenatum to selectively produce acetoin or 2,3-butanediol by one step bioconversion method
title_sort synthetic engineering of corynebacterium crenatum to selectively produce acetoin or 2,3-butanediol by one step bioconversion method
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6683508/
https://www.ncbi.nlm.nih.gov/pubmed/31387595
http://dx.doi.org/10.1186/s12934-019-1183-0
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