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Moderate expression of the transcriptional regulator ALsR enhances acetoin production by Bacillus subtilis

  • Metabolic Engineering and Synthetic Biology
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Acetoin, a major extracellular catabolic product of Bacillus subtilis cultured on glucose, is widely used to add flavor to food and also serves as a precursor for chemical synthesis. The biosynthesis of acetoin from pyruvate requires the enzymes α-acetolactate synthase (ALS) and α-acetolactate decarboxylase (ALDC), both of which are encoded by the alsSD operon. The transcriptional regulator ALsR is essential for the expression of alsSD. Here we focused on enhancing the production of acetoin by B. subtilis using different promoters to express ALsR. The expression of reporter genes was much higher under the control of the HpaII promoter than under control of the P bdhA promoter. Although the HpaII promoter highly enhanced transcription of the alsSD operon through overexpression of ALsR, the production of acetoin was not significantly increased. In contrast, moderate enhancement of ALsR expression using the P bdhA promoter significantly improved acetoin production. Compared with the wild-type, the enzyme activities of ALS and ALDC in B. subtilis harboring P bdhA were increased by approximately twofold, and the molar yield of acetoin from glucose was improved by 62.9 % in shake flask fermentation. In a 5-L fermentor, the engineered B. subtilis ultimately yielded 41.5 g/L of acetoin. Based on these results, we conclude that enhanced expression of ALDC and ALS by moderately elevated expression of the transcriptional regulator ALsR could increase acetoin production in recombinant B. subtilis.

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Abbreviations

ALDC:

α-Acetolactate decarboxylase

ALS:

α-Acetolactate synthase

AlsR:

Transcriptional regulator

AR/BDH:

Acetoin reductase/2,3-butanediol dehydrogenase

CAT:

Chloramphenicol acetyltransferase

GFP:

Green fluorescent protein

GRAS:

Generally recognized as safe

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Acknowledgments

This work was supported by the Program for New Century Excellent Talents in University (NCET-10-0459), the National Basic Research Program of China (973 Program) (2012CB725202), the National Natural Science Foundation of China (30970056, 21276110), the Research Fund for the Doctoral Program of Higher Education of China (20110093120001), the Fundamental Research Funds for the Central Universities (JUSRP51306A and JUSRP21121), the 111 Project (111-2-06), and a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institution.

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Author notes

  1. Zhiming Rao

    Present address: School of Biotechnology, Jiangnan University, 1800 Lihu Avenue, Wuxi, Jiangsu, 214122, People’s Republic of China

Authors and Affiliations

  1. The Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, 214122, People’s Republic of China

    Xian Zhang, Rongzhen Zhang, Teng Bao, Taowei Yang, Meijuan Xu, Huazhong Li & Zhiming Rao

  2. School of Medicine and Pharmaceuticals, Jiangnan University, Wuxi, Jiangsu, 214122, People’s Republic of China

    Zhenghong Xu

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  1. Xian Zhang
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Correspondence to Rongzhen Zhang or Zhiming Rao.

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Zhang, X., Zhang, R., Bao, T. et al. Moderate expression of the transcriptional regulator ALsR enhances acetoin production by Bacillus subtilis . J Ind Microbiol Biotechnol 40, 1067–1076 (2013). https://doi.org/10.1007/s10295-013-1303-5

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