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Metabolic Regulation of Organic Acid Biosynthesis in Actinobacillus succinogenes

Actinobacillus succinogenes is one of the most promising strains for succinic acid production; however, the lack of efficient genetic tools for strain modification development hinders its further application. In this study, a markerless knockout method for A. succinogenes using in-frame deletion was...

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Autores principales: Zhang, Wenming, Yang, Qiao, Wu, Min, Liu, Haojie, Zhou, Jie, Dong, Weiliang, Ma, Jiangfeng, Jiang, Min, Xin, Fengxue
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6759810/
https://www.ncbi.nlm.nih.gov/pubmed/31620431
http://dx.doi.org/10.3389/fbioe.2019.00216
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author Zhang, Wenming
Yang, Qiao
Wu, Min
Liu, Haojie
Zhou, Jie
Dong, Weiliang
Ma, Jiangfeng
Jiang, Min
Xin, Fengxue
author_facet Zhang, Wenming
Yang, Qiao
Wu, Min
Liu, Haojie
Zhou, Jie
Dong, Weiliang
Ma, Jiangfeng
Jiang, Min
Xin, Fengxue
author_sort Zhang, Wenming
collection PubMed
description Actinobacillus succinogenes is one of the most promising strains for succinic acid production; however, the lack of efficient genetic tools for strain modification development hinders its further application. In this study, a markerless knockout method for A. succinogenes using in-frame deletion was first developed. The resulting ΔpflA (encode pyruvate formate lyase 1-activating protein) strain displayed distinctive organic acid synthesis capacity under different cultivation modes. Additional acetate accumulation was observed in the ΔpflA strain relative to that of the wild type under aerobic conditions, indicating that acetate biosynthetic pathway was activated. Importantly, pyruvate was completely converted to lactate under anaerobic fermentation. The transcription analysis and enzyme assay revealed that the expression level and specific activity of lactate dehydrogenase (LDH) were significantly increased. In addition, the mRNA expression level of ldh was nearly increased 85-fold compared to that of the wild-type strain during aerobic–anaerobic dual-phase fermentation, resulting in 43.05 g/L lactate. These results demonstrate that pflA plays an important role in the regulation of C3 flux distribution. The deletion of pflA leads to the improvement of acetic acid production under aerobic conditions and activates lactic acid biosynthesis under anaerobic conditions. This study will help elaborate the mechanism governing organic acid biosynthesis in A. succinogenes.
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spelling pubmed-67598102019-10-16 Metabolic Regulation of Organic Acid Biosynthesis in Actinobacillus succinogenes Zhang, Wenming Yang, Qiao Wu, Min Liu, Haojie Zhou, Jie Dong, Weiliang Ma, Jiangfeng Jiang, Min Xin, Fengxue Front Bioeng Biotechnol Bioengineering and Biotechnology Actinobacillus succinogenes is one of the most promising strains for succinic acid production; however, the lack of efficient genetic tools for strain modification development hinders its further application. In this study, a markerless knockout method for A. succinogenes using in-frame deletion was first developed. The resulting ΔpflA (encode pyruvate formate lyase 1-activating protein) strain displayed distinctive organic acid synthesis capacity under different cultivation modes. Additional acetate accumulation was observed in the ΔpflA strain relative to that of the wild type under aerobic conditions, indicating that acetate biosynthetic pathway was activated. Importantly, pyruvate was completely converted to lactate under anaerobic fermentation. The transcription analysis and enzyme assay revealed that the expression level and specific activity of lactate dehydrogenase (LDH) were significantly increased. In addition, the mRNA expression level of ldh was nearly increased 85-fold compared to that of the wild-type strain during aerobic–anaerobic dual-phase fermentation, resulting in 43.05 g/L lactate. These results demonstrate that pflA plays an important role in the regulation of C3 flux distribution. The deletion of pflA leads to the improvement of acetic acid production under aerobic conditions and activates lactic acid biosynthesis under anaerobic conditions. This study will help elaborate the mechanism governing organic acid biosynthesis in A. succinogenes. Frontiers Media S.A. 2019-09-18 /pmc/articles/PMC6759810/ /pubmed/31620431 http://dx.doi.org/10.3389/fbioe.2019.00216 Text en Copyright © 2019 Zhang, Yang, Wu, Liu, Zhou, Dong, Ma, Jiang and Xin. 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(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
Zhang, Wenming
Yang, Qiao
Wu, Min
Liu, Haojie
Zhou, Jie
Dong, Weiliang
Ma, Jiangfeng
Jiang, Min
Xin, Fengxue
Metabolic Regulation of Organic Acid Biosynthesis in Actinobacillus succinogenes
title Metabolic Regulation of Organic Acid Biosynthesis in Actinobacillus succinogenes
title_full Metabolic Regulation of Organic Acid Biosynthesis in Actinobacillus succinogenes
title_fullStr Metabolic Regulation of Organic Acid Biosynthesis in Actinobacillus succinogenes
title_full_unstemmed Metabolic Regulation of Organic Acid Biosynthesis in Actinobacillus succinogenes
title_short Metabolic Regulation of Organic Acid Biosynthesis in Actinobacillus succinogenes
title_sort metabolic regulation of organic acid biosynthesis in actinobacillus succinogenes
topic Bioengineering and Biotechnology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6759810/
https://www.ncbi.nlm.nih.gov/pubmed/31620431
http://dx.doi.org/10.3389/fbioe.2019.00216
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