title: Improvement of chloramphenicol production in Streptomyces venezuelae ATCC 10712 by overexpression of the aroB and aroK genes catalysing steps in the shikimate pathway.
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 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.
authors.affiliation: Department of Genetics, Faculty of Science, Kasetsart University, 50 Ngamwongwan Road, Chatuchak, Bangkok, 10900, Thailand.|Bioscience Program, Faculty of Science, Kasetsart University, Bangkok, 10900, Thailand.
authors.forename: Vipawan
authors.initials: V
authors.lastname: Vitayakritsirikul
authors.affiliation: Department of Genetics, Faculty of Science, Kasetsart University, 50 Ngamwongwan Road, Chatuchak, Bangkok, 10900, Thailand.|Center for Advanced Studies in Tropical Natural Resources, NRU-KU, Kasetsart University, Bangkok, 10900, Thailand.
authors.forename: Ratchaniwan
authors.initials: R
authors.lastname: Jaemsaeng
authors.affiliation: Department of Genetics, Faculty of Science, Kasetsart University, 50 Ngamwongwan Road, Chatuchak, Bangkok, 10900, Thailand.
authors.forename: Karan
authors.initials: K
authors.lastname: Lohmaneeratana
authors.affiliation: Department of Genetics, Faculty of Science, Kasetsart University, 50 Ngamwongwan Road, Chatuchak, Bangkok, 10900, Thailand.
authors.forename: Anyarat
authors.initials: A
authors.lastname: Thanapipatsiri
authors.affiliation: Department of Genetics, Faculty of Science, Kasetsart University, 50 Ngamwongwan Road, Chatuchak, Bangkok, 10900, Thailand.
authors.forename: Ratama
authors.initials: R
authors.lastname: Daduang
authors.affiliation: Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, and Special Research Unit for Advanced Magnetic Resonance (AMR), Kasetsart University, Bangkok, 10900, Thailand.
authors.forename: Pitak
authors.initials: P
authors.lastname: Chuawong
authors.affiliation: Department of Genetics, Faculty of Science, Kasetsart University, 50 Ngamwongwan Road, Chatuchak, Bangkok, 10900, Thailand. arinthip.t@ku.ac.th.|Center for Advanced Studies in Tropical Natural Resources, NRU-KU, Kasetsart University, Bangkok, 10900, Thailand. arinthip.t@ku.ac.th.
authors.forename: Arinthip
authors.initials: A
authors.lastname: Thamchaipenet
mesh_terms: D002384:Catalysis; D002701:Chloramphenicol / Q000096:biosynthesis*; D015870:Gene Expression; D015971:Gene Expression Regulation, Enzymologic; D015966:Gene Expression Regulation, Fungal; D053858:Metabolic Networks and Pathways; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics*; D012765:Shikimic Acid / Q000378:metabolism*; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism*; D014158:Transcription, Genetic
chemical_list: D012765:Shikimic Acid; D002701:Chloramphenicol; D017853:Phosphotransferases (Alcohol Group Acceptor)
keywords: Chloramphenicol; Overexpression; Streptomyces venezuelae; aroB; aroK