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A method to type the potential angucycline producers in actinomycetes isolated from marine sponges

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Abstract

Angucyclines are aromatic polyketides with antimicrobial, antitumor, antiviral and enzyme inhibition activities. In this study, a new pair of degenerate primers targeting the cyclase genes that are involved in the aromatization of the first and/or second ring of angucycline, were designed and evaluated in a PCR protocol targeting the jadomycin cyclase gene of Streptomyces venezuelae ISP5230. The identity of the target amplicon was confirmed by sequencing. After validation, the primers were used to screen 49 actinomycete isolates from three different marine sponges to identify putative angucycline producers. Seven isolates were positively identified using this method. Sequence analysis of the positive amplicons confirmed their identity as putative angucycline cyclases with sequence highly similar to known angucycline cyclases. Phylogenetic analysis clustered these positives into the angucycline group of cyclases. Furthermore, amplifications of the seven isolates using ketosynthase-specific primers were positive, backing the results using the cyclase primers. Together these results provided strong support for the presence of angucycline biosynthetic genes in these isolates. The specific primer set targeting the cyclase can be used to identify putative angucycline producers among marine actinobacteria, and aid in the discovery of novel angucyclines.

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Acknowledgments

This research was partially funded by the State Principal and Basic Research and Development Program of the Ministry of Sciences and Technology of China (2010CB833801-3) and Provincial Collaborative Foundation Project of Guangdong (9351007002000001, 2008A030203004). We also thank the financial support of the China Ocean Mineral Resources R&D Association Project (DYXM115-02) and the Open Project Program (LMB091001) of the Key Laboratory of Marine Bio-resources Sustainable Utilization, SCSIO-CAS. Many thanks are given to Dr Ravi Kumar for critical reading of this manuscript and Chen W. Z. for identifying the marine sponge.

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Authors and Affiliations

  1. Key Laboratory of Marine Bio-resources Sustainable Utilization (LMB-CAS), Guangdong Key Laboratory of Marine Materia Medica (LMMM-GD), South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou, 510301, People’s Republic of China

    Yongchang Ouyang, Houbo Wu, Lianwu Xie, Guanghua Wang, Shikun Dai, Minjie Chen & Xiang Li

  2. Guangzhou Medical University, Guangzhou, 510182, People’s Republic of China

    Yongchang Ouyang

  3. Central South University of Forestry and Technology, Changsha, 410004, People’s Republic of China

    Lianwu Xie

  4. Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100080, People’s Republic of China

    Keqian Yang

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  1. Yongchang Ouyang
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  2. Houbo Wu
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  3. Lianwu Xie
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  4. Guanghua Wang
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Correspondence to Houbo Wu or Xiang Li.

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Ouyang, Y., Wu, H., Xie, L. et al. A method to type the potential angucycline producers in actinomycetes isolated from marine sponges. Antonie van Leeuwenhoek 99, 807–815 (2011). https://doi.org/10.1007/s10482-011-9554-5

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