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

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Web End = Antonie van Leeuwenhoek (2016) 109:379388 DOI 10.1007/s10482-015-0640-y

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Web End = Improvement of chloramphenicol productionin Streptomyces venezuelae ATCC 10712 by overexpression of the aroB and aroK genes catalysing steps in the shikimate pathway

Vipawan Vitayakritsirikul . Ratchaniwan Jaemsaeng .

Karan Lohmaneeratana . Anyarat Thanapipatsiri .

Ratama Daduang . Pitak Chuawong . Arinthip Thamchaipenet

Received: 24 November 2015 / Accepted: 18 December 2015 / Published online: 29 December 2015 Springer International Publishing Switzerland 2015

Abstract 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 biosyn-thesis. The results suggested that the increment of carbon ux 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 rst report of a convenient genetic approach to manipulate primary metabolite genes in S. venezuelae in order to increase chloramphenicol production.

Keywords Chloramphenicol aroB aroK

Overexpression Streptomyces venezuelae

Introduction

Chloramphenicol (Cm) is produced by actinomycetes including Streptomyces venezuelae ATCC 10712 (Vining and Stuttard 1995). It is a bacteriostatic antibiotic for the treatment of a number of bacterial infections including meningitis, typhoid, plague and cholera by specic binding to the 50S ribosomal subunit and thus inhibiting protein synthesis. The biosynthesis of Cm includes parts of glycolysis, the pentose phosphate pathway, and the shikimate pathway. Erythrose-4-phosphate (E4P) and phosphoenol pyruvate (PEP) are converted by seven sequential

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