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A Myo-Inositol-Inducible Expression System for Corynebacterium glutamicum and Its Application

Corynebacterium glutamicum is one of the important industrial microorganisms for production of amino acids and other value-added compounds. Most expression vectors used in C. glutamicum are based on inducible promoter (P( tac ) or P( trc )) activated by isopropyl-β-D-thiogalactopyranoside (IPTG). Ho...

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Autores principales: Lu, Nan, Zhang, Chenglin, Zhang, Wenjie, Xu, Haoran, Li, Yuhong, Wei, Minhua, Meng, Jing, Meng, Yan, Wang, Junzhe, Chen, Ning
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8634428/
https://www.ncbi.nlm.nih.gov/pubmed/34869258
http://dx.doi.org/10.3389/fbioe.2021.746322
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author Lu, Nan
Zhang, Chenglin
Zhang, Wenjie
Xu, Haoran
Li, Yuhong
Wei, Minhua
Meng, Jing
Meng, Yan
Wang, Junzhe
Chen, Ning
author_facet Lu, Nan
Zhang, Chenglin
Zhang, Wenjie
Xu, Haoran
Li, Yuhong
Wei, Minhua
Meng, Jing
Meng, Yan
Wang, Junzhe
Chen, Ning
author_sort Lu, Nan
collection PubMed
description Corynebacterium glutamicum is one of the important industrial microorganisms for production of amino acids and other value-added compounds. Most expression vectors used in C. glutamicum are based on inducible promoter (P( tac ) or P( trc )) activated by isopropyl-β-D-thiogalactopyranoside (IPTG). However, these vectors seem unsuitable for large-scale industrial production due to the high cost and toxicity of IPTG. Myo-inositol is an ideal inducer because of its non-toxicity and lower price. In this study, a myo-inositol-inducible expression vector pMI-4, derived from the expression vector pXMJ19, was constructed. Besides the original chloramphenicol resistance gene cat, multiple cloning sites, and rrnB terminator, the pMI-4 (6,643 bp) contains the iolR ( q ) cassette and the myo-inositol-inducible promoter P( iolT1 ). The pMI-4 could stably replicate in the C. glutamicum host. Meanwhile, the non-myo-inositol degradation host strain C. glutamicumΔiolGΔoxiCΔoxiDΔoxiE for maintaining the pMI-4 was developed. Overexpression of hemA ( M ) and hemL using pMI-4 resulted in a significant accumulation of 5-aminolevulinic acid, indicating its potential application in metabolic engineering and industrial fermentation.
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spelling pubmed-86344282021-12-02 A Myo-Inositol-Inducible Expression System for Corynebacterium glutamicum and Its Application Lu, Nan Zhang, Chenglin Zhang, Wenjie Xu, Haoran Li, Yuhong Wei, Minhua Meng, Jing Meng, Yan Wang, Junzhe Chen, Ning Front Bioeng Biotechnol Bioengineering and Biotechnology Corynebacterium glutamicum is one of the important industrial microorganisms for production of amino acids and other value-added compounds. Most expression vectors used in C. glutamicum are based on inducible promoter (P( tac ) or P( trc )) activated by isopropyl-β-D-thiogalactopyranoside (IPTG). However, these vectors seem unsuitable for large-scale industrial production due to the high cost and toxicity of IPTG. Myo-inositol is an ideal inducer because of its non-toxicity and lower price. In this study, a myo-inositol-inducible expression vector pMI-4, derived from the expression vector pXMJ19, was constructed. Besides the original chloramphenicol resistance gene cat, multiple cloning sites, and rrnB terminator, the pMI-4 (6,643 bp) contains the iolR ( q ) cassette and the myo-inositol-inducible promoter P( iolT1 ). The pMI-4 could stably replicate in the C. glutamicum host. Meanwhile, the non-myo-inositol degradation host strain C. glutamicumΔiolGΔoxiCΔoxiDΔoxiE for maintaining the pMI-4 was developed. Overexpression of hemA ( M ) and hemL using pMI-4 resulted in a significant accumulation of 5-aminolevulinic acid, indicating its potential application in metabolic engineering and industrial fermentation. Frontiers Media S.A. 2021-11-15 /pmc/articles/PMC8634428/ /pubmed/34869258 http://dx.doi.org/10.3389/fbioe.2021.746322 Text en Copyright © 2021 Lu, Zhang, Zhang, Xu, Li, Wei, Meng, Meng, Wang and Chen. https://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(s) 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 Bioengineering and Biotechnology
Lu, Nan
Zhang, Chenglin
Zhang, Wenjie
Xu, Haoran
Li, Yuhong
Wei, Minhua
Meng, Jing
Meng, Yan
Wang, Junzhe
Chen, Ning
A Myo-Inositol-Inducible Expression System for Corynebacterium glutamicum and Its Application
title A Myo-Inositol-Inducible Expression System for Corynebacterium glutamicum and Its Application
title_full A Myo-Inositol-Inducible Expression System for Corynebacterium glutamicum and Its Application
title_fullStr A Myo-Inositol-Inducible Expression System for Corynebacterium glutamicum and Its Application
title_full_unstemmed A Myo-Inositol-Inducible Expression System for Corynebacterium glutamicum and Its Application
title_short A Myo-Inositol-Inducible Expression System for Corynebacterium glutamicum and Its Application
title_sort myo-inositol-inducible expression system for corynebacterium glutamicum and its application
topic Bioengineering and Biotechnology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8634428/
https://www.ncbi.nlm.nih.gov/pubmed/34869258
http://dx.doi.org/10.3389/fbioe.2021.746322
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