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

Abstract:

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.

journal_name

Biotechnol Lett

journal_title

Biotechnology letters

authors

Zhang C,Du S,Liu Y,Xie X,Xu Q,Chen N

doi

10.1007/s10529-015-1801-9

subject

Has Abstract

pub_date

2015-07-01 00:00:00

pages

1361-9

issue

7

eissn

0141-5492

issn

1573-6776

journal_volume

37

pub_type

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