The biosynthesis of the polyether antibiotic nanchangmycin is controlled by two pathway-specific transcriptional activators
- 675 Downloads
The nanchangmycin (NAN) produced by Streptomyces nanchangensis NS3226 is a polyether antibiotic resembling monensin in their gene clusters and the chemical structures. They can inhibit gram-positive bacteria and be a growth promoter for ruminants. Within the nanchangmycin gene cluster (nan), we identified that two SARP-family regulatory genes, nanR1 and nanR2, were both required to activate the transcription of all nan polyketide genes. Overexpression of NanR1 and NanR2 in wild-type increase NAN yields by at least three folds. Bioinformatic analysis of the immediate upstream DNA sequence of each nan gene and quantitative real-time RT-PCR analysis of the nan operons identified five putative SARP binding sites. Moreover, deletion of an AraC-family repressor gene nanR4 increased expression of NanR1 and R2 and led to a threefold increase in NAN production.
KeywordsSARP Nanchangmycin Streptomyces Regulation
Authors gave their thanks to Tobias Kieser for his helpful comment and manuscript editing. This work was supported by grants from the Ministry of Science and Technology (973 and 863 Programs), the National Science Foundation of China, the Ministry of Education, and the Science and Technology Commission of Shanghai Municipality.
- Bibb M (1996) 1995 Colworth Prize Lecture. The regulation of antibiotic production in Streptomyces coelicolor A3(2). Microbiology 142(Pt 6):1335–1344Google Scholar
- Champness WC, Chater KF (1994) Regulation and integration of antibiotic production and morphological differentiation in Streptomyces spp. In: Patrick Piggot CM, Youngman P (eds) Regulation of bacterial differentiation. American Society for Microbiology, Washington, D.C., pp 61–93Google Scholar
- Hopwood DA (2007) Streptomyces in nature and medicine: the antibiotic makers. Oxford University Press, New YorkGoogle Scholar
- Kieser T, Bibb MJ, Buttner MJ, Chater KF, Hopwood DA (2000) Practical streptomyces genetics. A laboratory manual. John Innes Foundation, NorwichGoogle Scholar
- Leadlay PF, Staunton J, Oliynyk M, Bisang C, Cortes J, Frost E, Hughes-Thomas ZA, Jones MA, Kendrew SG, Lester JB, Long PF, McArthur HA, McCormick EL, Oliynyk Z, Stark CB, Wilkinson CJ (2001) Engineering of complex polyketide biosynthesis—insights from sequencing of the monensin biosynthetic gene cluster. J Ind Microbiol Biotechnol 27:360–367PubMedCrossRefGoogle Scholar
- Novakova R, Rehakova A, Kutas P, Feckova L, Kormanec J (2011) The role of two SARP-family transcriptional regulators in regulation of the auricin gene cluster in Streptomyces aureofaciens CCM 3239. Microbiology 157:1629–1639Google Scholar
- Oliynyk M, Stark CB, Bhatt A, Jones MA, Hughes-Thomas ZA, Wilkinson C, Oliynyk Z, Demydchuk Y, Staunton J, Leadlay PF (2003) Analysis of the biosynthetic gene cluster for the polyether antibiotic monensin in Streptomyces cinnamonensis and evidence for the role of monB and monC genes in oxidative cyclization. Mol Microbiol 49:1179–1190PubMedCrossRefGoogle Scholar
- Ouyang L, Wan S, Tu G, Chen X, Zhen Y, Gao Y, Xie X (1984) A new species of Streptomyces producing insecticidal antibiotics. Chin J Microbiol 24:195–199Google Scholar
- Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring HarborGoogle Scholar
- Takano E, Gramajo HC, Strauch E, Andres N, White J, Bibb MJ (1992) Transcriptional regulation of the redD transcriptional activator gene accounts for growth-phase-dependent production of the antibiotic undecylprodigiosin in Streptomyces coelicolor A3(2). Mol Microbiol 6:2797–2804PubMedCrossRefGoogle Scholar