Cloning, Characterization and Heterologous Expression of the Indolocarbazole Biosynthetic Gene Cluster from Marine-Derived Streptomyces sanyensis FMA.docVIP

Mar. Drugs 2013, 11, 466-488; doi:10.3390/m OPEN ACCESS Marine Drugs ISSN 1660-3397 /journal/marinedrugs Article Cloning, Characterization and Heterologous Expression of the Indolocarbazole Biosynthetic Gene Cluster from Marine-Derived Streptomyces sanyensis FMA 1 1 2 2 1 3, Tong Li , Yuanyuan Du , Qiu Cui , Jingtao Zhang , Weiming Zhu , Kui Hong * and Wenli Li 1,* 1 Key Laboratory of Marine Drugs, Ministry of Education of China, School of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, China; E-Mails: oaixlittle@163.com (T.L.); awetobean@126.com (Y.D.); weimingzhu@ (W.Z.) 2 Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, No. 189 Songling Road, Qingdao 266101, China; E-Mails: cuiqiu@ (Q.C.); zhang_jt@ (J.Z.) 3 Ministry of Education of China, School of Pharmaceutical Sciences, Wuhan University, Wuhan 430071, China * Authors to whom correspondence should be addressed; E-Mails: kuihong31@ (K.H.); liwenli@ (W.L.); Tel./Fax: +86-27(K.H.); Tel./Fax: +86-532(W.L.). Received: 21 November 2012; in revised form: 24 December 2012 / Accepted: 21 January 2013 / Published: 6 February 2013 Abstract: The indolocarbazole (ICZ) alkaloids have attracted much attention due to their unique structures and potential therapeutic applications. A series of ICZs were recently isolated and identified from a marine-derived actinomycete strain, Streptomyces sanyensis FMA. To elucidate the biosynthetic machinery associated with ICZs production in S. sanyensis FMA, PCR using degenerate primers was carried out to clone the FAD-dependent monooxygenase gene fragment for ICZ ring formation, which was used as a probe to isolate the 34.6-kb DNA region containing the spc gene cluster. Sequence analysis revealed genes for ICZ ring formation (spcO, D, P, C), sugar unit formation (spcA, B, E, K, J, I), glycosylation (spcN, G), me