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. 2010 Dec 9;1(12):1093–1105. doi: 10.1007/s13238-010-0127-6

Characterization of the tunicamycin gene cluster unveiling unique steps involved in its biosynthesis

Wenqing Chen 1,3, Dongjing Qu 1, Lipeng Zhai 1, Meifeng Tao 1, Yemin Wang 1, Shuangjun Lin 1, Neil P J Price 2,✉, Zixin Deng 1,3,✉
PMCID: PMC4875072  PMID: 21153459

Abstract

Tunicamycin, a potent reversible translocase I inhibitor, is produced by several Actinomycetes species. The tunicamycin structure is highly unusual, and contains an 11-carbon dialdose sugar and an α, β-1″,11′-glycosidic linkage. Here we report the identification of a gene cluster essential for tunicamycin biosynthesis by high-throughput heterologous expression (HHE) strategy combined with a bioassay. Introduction of the genes into heterologous non-producing Streptomyces hosts results in production of tunicamycin by these strains, demonstrating the role of the genes for the biosynthesis of tunicamycins. Gene disruption experiments coupled with bioinformatic analysis revealed that the tunicamycin gene cluster is minimally composed of 12 genes (tunA-tunL). Amongst these is a putative radical SAM enzyme (Tun B) with a potentially unique role in biosynthetic carbon-carbon bond formation. Hence, a seven-step novel pathway is proposed for tunicamycin biosynthesis. Moreover, two gene clusters for the potential biosynthesis of tunicamycin-like antibiotics were also identified in Streptomyces clavuligerus ATCC 27064 and Actinosynnema mirums DSM 43827. These data provide clarification of the novel mechanisms for tunicamycin biosynthesis, and for the generation of new-designer tunicamycin analogs with selective/enhanced bioactivity via combinatorial biosynthesis strategies.

Electronic Supplementary Material

Supplementary material is available for this article at 10.1007/s13238-010-0127-6 and is accessible for authorized users.

Keywords: tunicamycin, biosynthetic gene cluster, high-throughput heterologous expression, bioassay, combinatorial biosynthesis

Electronic supplementary material

13238_2010_127_MOESM1_ESM.pdf (749.3KB, pdf)

Supplementary material, approximately 340 KB.

Footnotes

These authors contributed equally to the work.

Electronic Supplementary Material

Supplementary material is available for this article at 10.1007/s13238-010-0127-6 and is accessible for authorized users.

Contributor Information

Neil P. J. Price, Email: zxdeng@sjtu.edu.cn

Zixin Deng, Email: neil.price@ars.usda.gov.

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Supplementary Materials

13238_2010_127_MOESM1_ESM.pdf (749.3KB, pdf)

Supplementary material, approximately 340 KB.

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