Strategy for enhancing adenosine production under the guidance of transcriptional and metabolite pool analysis

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• 作者：Chenglin Zhang ; Shanshan Du ; Yue Liu ; Xixian Xie ; Qingyang Xu…
• 关键词：Adenosine ; Bacillus subtilis ; Metabolite pool ; 5 ; Phosphoribosyl 1 ; pyrophosphate ; Transcriptional analysis
• 刊名：Biotechnology Letters
• 出版年：2015
• 出版时间：July 2015
• 年：2015
• 卷：37
• 期：7
• 页码：1361-1369
• 全文大小：552 KB
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• 作者单位：Chenglin Zhang (1) (2) (3)
  Shanshan Du (1)
  Yue Liu (1)
  Xixian Xie (1) (2) (3)
  Qingyang Xu (1) (2) (3)
  Ning Chen (1) (2) (3)
  
  1. College of Biotechnology, Tianjin University of Science and Technology, Tianjin, 300457, China
  2. National and Local United Engineering Lab of Metabolic Control Fermentation Technology, Tianjin, 300457, China
  3. Tianjin Engineering Lab of Efficient and Green Amino Acid Manufacture, Tianjin, 300457, China
  
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Microbiology
  Biotechnology
  Applied Microbiology
  Biochemistry
  
• 出版者：Springer Netherlands
• ISSN：1573-6776

文摘

Objectives To rationally identify targets for enhancing adenosine production, transcription level of genes involved in adenosine synthesis of Bacillus subtilis XGL was detected during the fermentation process, complemented with metabolite pool analysis. Results PurR-regulated genes (pur operon and purA) and prs were down-regulated and 5-phosphoribosyl 1-pyrophosphate (PRPP) decreased considerably after 24?h when adenosine significantly accumulated. Since PRPP could strongly antagonize the binding of PurR to its targets, it was inferred that down-regulation of pur operon and purA might be due to a low PRPP pool, which was confirmed by metabolite analysis. So desensitized prs responsible for PRPP synthesis was overexpressed, resulting in increased PRPP concentration and pur operon transcription. To further enhance the adenosine production, desensitized purF and prs were co-overexpressed with integrating additional copy of purA to B. subtilis XGL genome, resulting in 24.3?% (1.29?g/g DCW) higher adenosine production than that by B. subtilis XG. Conclusions Overexpression of prs, purF and purA under the guidance of transcriptional and metabolite pool analysis significantly increased adenosine production. Strategies used in this study have potential applications for rational modification of industrial microorganisms.