Antonie van Leeuwenhoek

, Volume 99, Issue 4, pp 807–815

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

Original Paper

DOI: 10.1007/s10482-011-9554-5

Cite this article as:
Ouyang, Y., Wu, H., Xie, L. et al. Antonie van Leeuwenhoek (2011) 99: 807. doi:10.1007/s10482-011-9554-5

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.

Keywords

Actinomycete Angucycline Marine sponge Polyketide synthase 

Supplementary material

10482_2011_9554_MOESM1_ESM.docx (105 kb)
Supplementary material 1 (DOCX 105 kb)

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Yongchang Ouyang
    • 1
    • 2
  • Houbo Wu
    • 1
  • Lianwu Xie
    • 1
    • 3
  • Guanghua Wang
    • 1
  • Shikun Dai
    • 1
  • Minjie Chen
    • 1
  • Keqian Yang
    • 4
  • Xiang Li
    • 1
  1. 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 SciencesGuangzhouPeople’s Republic of China
  2. 2.Guangzhou Medical UniversityGuangzhouPeople’s Republic of China
  3. 3.Central South University of Forestry and TechnologyChangshaPeople’s Republic of China
  4. 4.Institute of Microbiology, Chinese Academy of SciencesBeijingPeople’s Republic of China