Cargando…

Enhancing poly-γ-glutamic acid production in Bacillus amyloliquefaciens by introducing the glutamate synthesis features from Corynebacterium glutamicum

BACKGROUND: Poly-γ-glutamic acid (γ-PGA) is a valuable polymer with glutamate as its sole precursor. Enhancement of the intracellular glutamate synthesis is a very important strategy for the improvement of γ-PGA production, especially for those glutamate-independent γ-PGA producing strains. Coryneba...

Descripción completa

Detalles Bibliográficos
Autores principales: Feng, Jun, Quan, Yufen, Gu, Yanyan, Liu, Fenghong, Huang, Xiaozhong, Shen, Haosheng, Dang, Yulei, Cao, Mingfeng, Gao, Weixia, Lu, Xiaoyun, Wang, Yi, Song, Cunjiang, Wang, Shufang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5440981/
https://www.ncbi.nlm.nih.gov/pubmed/28532451
http://dx.doi.org/10.1186/s12934-017-0704-y
_version_ 1783238171244363776
author Feng, Jun
Quan, Yufen
Gu, Yanyan
Liu, Fenghong
Huang, Xiaozhong
Shen, Haosheng
Dang, Yulei
Cao, Mingfeng
Gao, Weixia
Lu, Xiaoyun
Wang, Yi
Song, Cunjiang
Wang, Shufang
author_facet Feng, Jun
Quan, Yufen
Gu, Yanyan
Liu, Fenghong
Huang, Xiaozhong
Shen, Haosheng
Dang, Yulei
Cao, Mingfeng
Gao, Weixia
Lu, Xiaoyun
Wang, Yi
Song, Cunjiang
Wang, Shufang
author_sort Feng, Jun
collection PubMed
description BACKGROUND: Poly-γ-glutamic acid (γ-PGA) is a valuable polymer with glutamate as its sole precursor. Enhancement of the intracellular glutamate synthesis is a very important strategy for the improvement of γ-PGA production, especially for those glutamate-independent γ-PGA producing strains. Corynebacterium glutamicum has long been used for industrial glutamate production and it exhibits some unique features for glutamate synthesis; therefore introduction of these metabolic characters into the γ-PGA producing strain might lead to increased intracellular glutamate availability, and thus ultimate γ-PGA production. RESULTS: In this study, the unique glutamate synthesis features from C. glutamicum was introduced into the glutamate-independent γ-PGA producing Bacillus amyloliquefaciens NK-1 strain. After introducing the energy-saving NADPH-dependent glutamate dehydrogenase (NADPH-GDH) pathway, the NK-1 (pHT315-gdh) strain showed slightly increase (by 9.1%) in γ-PGA production. Moreover, an optimized metabolic toggle switch for controlling the expression of ɑ-oxoglutarate dehydrogenase complex (ODHC) was introduced into the NK-1 strain, because it was previously shown that the ODHC in C. glutamicum was completely inhibited when glutamate was actively produced. The obtained NK-PO1 (pHT01-xylR) strain showed 66.2% higher γ-PGA production than the NK-1 strain. However, the further combination of these two strategies (introducing both NADPH-GDH pathway and the metabolic toggle switch) did not lead to further increase of γ-PGA production but rather the resultant γ-PGA production was even lower than that in the NK-1 strain. CONCLUSIONS: We proposed new metabolic engineering strategies to improve the γ-PGA production in B. amyloliquefaciens. The NK-1 (pHT315-gdh) strain with the introduction of NADPH-GDH pathway showed 9.1% improvement in γ-PGA production. The NK-PO1 (pHT01-xylR) strain with the introduction of a metabolic toggle switch for controlling the expression of ODHC showed 66.2% higher γ-PGA production than the NK-1 strain. This work proposed a new strategy for improving the target product in microbial cell factories. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12934-017-0704-y) contains supplementary material, which is available to authorized users.
format Online
Article
Text
id pubmed-5440981
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-54409812017-05-24 Enhancing poly-γ-glutamic acid production in Bacillus amyloliquefaciens by introducing the glutamate synthesis features from Corynebacterium glutamicum Feng, Jun Quan, Yufen Gu, Yanyan Liu, Fenghong Huang, Xiaozhong Shen, Haosheng Dang, Yulei Cao, Mingfeng Gao, Weixia Lu, Xiaoyun Wang, Yi Song, Cunjiang Wang, Shufang Microb Cell Fact Research BACKGROUND: Poly-γ-glutamic acid (γ-PGA) is a valuable polymer with glutamate as its sole precursor. Enhancement of the intracellular glutamate synthesis is a very important strategy for the improvement of γ-PGA production, especially for those glutamate-independent γ-PGA producing strains. Corynebacterium glutamicum has long been used for industrial glutamate production and it exhibits some unique features for glutamate synthesis; therefore introduction of these metabolic characters into the γ-PGA producing strain might lead to increased intracellular glutamate availability, and thus ultimate γ-PGA production. RESULTS: In this study, the unique glutamate synthesis features from C. glutamicum was introduced into the glutamate-independent γ-PGA producing Bacillus amyloliquefaciens NK-1 strain. After introducing the energy-saving NADPH-dependent glutamate dehydrogenase (NADPH-GDH) pathway, the NK-1 (pHT315-gdh) strain showed slightly increase (by 9.1%) in γ-PGA production. Moreover, an optimized metabolic toggle switch for controlling the expression of ɑ-oxoglutarate dehydrogenase complex (ODHC) was introduced into the NK-1 strain, because it was previously shown that the ODHC in C. glutamicum was completely inhibited when glutamate was actively produced. The obtained NK-PO1 (pHT01-xylR) strain showed 66.2% higher γ-PGA production than the NK-1 strain. However, the further combination of these two strategies (introducing both NADPH-GDH pathway and the metabolic toggle switch) did not lead to further increase of γ-PGA production but rather the resultant γ-PGA production was even lower than that in the NK-1 strain. CONCLUSIONS: We proposed new metabolic engineering strategies to improve the γ-PGA production in B. amyloliquefaciens. The NK-1 (pHT315-gdh) strain with the introduction of NADPH-GDH pathway showed 9.1% improvement in γ-PGA production. The NK-PO1 (pHT01-xylR) strain with the introduction of a metabolic toggle switch for controlling the expression of ODHC showed 66.2% higher γ-PGA production than the NK-1 strain. This work proposed a new strategy for improving the target product in microbial cell factories. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12934-017-0704-y) contains supplementary material, which is available to authorized users. BioMed Central 2017-05-22 /pmc/articles/PMC5440981/ /pubmed/28532451 http://dx.doi.org/10.1186/s12934-017-0704-y Text en © The Author(s) 2017 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
Feng, Jun
Quan, Yufen
Gu, Yanyan
Liu, Fenghong
Huang, Xiaozhong
Shen, Haosheng
Dang, Yulei
Cao, Mingfeng
Gao, Weixia
Lu, Xiaoyun
Wang, Yi
Song, Cunjiang
Wang, Shufang
Enhancing poly-γ-glutamic acid production in Bacillus amyloliquefaciens by introducing the glutamate synthesis features from Corynebacterium glutamicum
title Enhancing poly-γ-glutamic acid production in Bacillus amyloliquefaciens by introducing the glutamate synthesis features from Corynebacterium glutamicum
title_full Enhancing poly-γ-glutamic acid production in Bacillus amyloliquefaciens by introducing the glutamate synthesis features from Corynebacterium glutamicum
title_fullStr Enhancing poly-γ-glutamic acid production in Bacillus amyloliquefaciens by introducing the glutamate synthesis features from Corynebacterium glutamicum
title_full_unstemmed Enhancing poly-γ-glutamic acid production in Bacillus amyloliquefaciens by introducing the glutamate synthesis features from Corynebacterium glutamicum
title_short Enhancing poly-γ-glutamic acid production in Bacillus amyloliquefaciens by introducing the glutamate synthesis features from Corynebacterium glutamicum
title_sort enhancing poly-γ-glutamic acid production in bacillus amyloliquefaciens by introducing the glutamate synthesis features from corynebacterium glutamicum
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5440981/
https://www.ncbi.nlm.nih.gov/pubmed/28532451
http://dx.doi.org/10.1186/s12934-017-0704-y
work_keys_str_mv AT fengjun enhancingpolygglutamicacidproductioninbacillusamyloliquefaciensbyintroducingtheglutamatesynthesisfeaturesfromcorynebacteriumglutamicum
AT quanyufen enhancingpolygglutamicacidproductioninbacillusamyloliquefaciensbyintroducingtheglutamatesynthesisfeaturesfromcorynebacteriumglutamicum
AT guyanyan enhancingpolygglutamicacidproductioninbacillusamyloliquefaciensbyintroducingtheglutamatesynthesisfeaturesfromcorynebacteriumglutamicum
AT liufenghong enhancingpolygglutamicacidproductioninbacillusamyloliquefaciensbyintroducingtheglutamatesynthesisfeaturesfromcorynebacteriumglutamicum
AT huangxiaozhong enhancingpolygglutamicacidproductioninbacillusamyloliquefaciensbyintroducingtheglutamatesynthesisfeaturesfromcorynebacteriumglutamicum
AT shenhaosheng enhancingpolygglutamicacidproductioninbacillusamyloliquefaciensbyintroducingtheglutamatesynthesisfeaturesfromcorynebacteriumglutamicum
AT dangyulei enhancingpolygglutamicacidproductioninbacillusamyloliquefaciensbyintroducingtheglutamatesynthesisfeaturesfromcorynebacteriumglutamicum
AT caomingfeng enhancingpolygglutamicacidproductioninbacillusamyloliquefaciensbyintroducingtheglutamatesynthesisfeaturesfromcorynebacteriumglutamicum
AT gaoweixia enhancingpolygglutamicacidproductioninbacillusamyloliquefaciensbyintroducingtheglutamatesynthesisfeaturesfromcorynebacteriumglutamicum
AT luxiaoyun enhancingpolygglutamicacidproductioninbacillusamyloliquefaciensbyintroducingtheglutamatesynthesisfeaturesfromcorynebacteriumglutamicum
AT wangyi enhancingpolygglutamicacidproductioninbacillusamyloliquefaciensbyintroducingtheglutamatesynthesisfeaturesfromcorynebacteriumglutamicum
AT songcunjiang enhancingpolygglutamicacidproductioninbacillusamyloliquefaciensbyintroducingtheglutamatesynthesisfeaturesfromcorynebacteriumglutamicum
AT wangshufang enhancingpolygglutamicacidproductioninbacillusamyloliquefaciensbyintroducingtheglutamatesynthesisfeaturesfromcorynebacteriumglutamicum