Improvement of chloramphenicol production in Streptomyces venezuelae ATCC 10712 by overexpression of the aroB and aroK genes catalysing steps in the shikimate pathway

Streptomyces venezuelae ATCC 10712 produces chloramphenicol in small amounts. To enhance chloramphenicol production, two genes, aroB and aroK, encoding rate-limiting enzymes of the shikimate pathway were overexpressed using the expression vector pIJ86 under the control of the strong constitutive erm...

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Bibliographic Details
Published in:	Antonie van Leeuwenhoek Vol. 109; no. 3; pp. 379 - 388
Main Authors:	Vitayakritsirikul, Vipawan, Jaemsaeng, Ratchaniwan, Lohmaneeratana, Karan, Thanapipatsiri, Anyarat, Daduang, Ratama, Chuawong, Pitak, Thamchaipenet, Arinthip
Format:	Journal Article
Language:	English
Published:	Cham Springer International Publishing 01-03-2016 Springer Nature B.V
Subjects:	Bacteria Biomedical and Life Sciences Biosynthesis carbon Catalysis chloramphenicol Chloramphenicol - biosynthesis chorismic acid Enzymes Gene Expression Gene Expression Regulation, Enzymologic Gene Expression Regulation, Fungal gene overexpression genes Life Sciences Medical Microbiology Metabolic Networks and Pathways Metabolites Microbiology Original Paper Phosphotransferases (Alcohol Group Acceptor) - genetics Plant Sciences shikimate pathway Shikimic Acid - metabolism Soil Science & Conservation Streptomyces - genetics Streptomyces - metabolism Streptomyces venezuelae transcription (genetics) Transcription factors Transcription, Genetic Chloramphenicol Overexpression aroK aroB Streptomyces venezuelae
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Summary:	Streptomyces venezuelae ATCC 10712 produces chloramphenicol in small amounts. To enhance chloramphenicol production, two genes, aroB and aroK, encoding rate-limiting enzymes of the shikimate pathway were overexpressed using the expression vector pIJ86 under the control of the strong constitutive ermE* promoter. The recombinant strains, S. venezuelae/pIJ86-aroB and S. venezuelae/pIJ86-aroK, produced 2.5- and 4.3-fold greater amounts respectively of chloramphenicol than wild type at early stationary phase of growth. High transcriptional levels of aroB and aroK genes were detected at the early exponential growth of both recombinant strains and consistent with the enhanced expression of pabB gene encoding an early enzyme in chloramphenicol biosynthesis. The results suggested that the increment of carbon flux was directed towards intermediates in the shikimate pathway required for the production of chorismic acid, and consequently resulted in the enhancement of chloramphenicol production. This work is the first report of a convenient genetic approach to manipulate primary metabolite genes in S. venezuelae in order to increase chloramphenicol production.
Bibliography:	http://dx.doi.org/10.1007/s10482-015-0640-y ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0003-6072 1572-9699
DOI:	10.1007/s10482-015-0640-y