title: A method to type the potential angucycline producers in actinomycetes isolated from marine sponges.
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.
authors.affiliation: 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.
authors.forename: Yongchang
authors.initials: Y
authors.lastname: Ouyang
authors.forename: Houbo
authors.initials: H
authors.lastname: Wu
authors.forename: Lianwu
authors.initials: L
authors.lastname: Xie
authors.forename: Guanghua
authors.initials: G
authors.lastname: Wang
authors.forename: Shikun
authors.initials: S
authors.lastname: Dai
authors.forename: Minjie
authors.initials: M
authors.lastname: Chen
authors.forename: Keqian
authors.initials: K
authors.lastname: Yang
authors.forename: Xiang
authors.initials: X
authors.lastname: Li
mesh_terms: D039903:Actinobacteria / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D000818:Animals; D000900:Anti-Bacterial Agents / Q000096:biosynthesis*; D001551:Benz(a)Anthracenes / Q000378:metabolism*; D055808:Drug Discovery; D018942:Macrolides; D010802:Phylogeny; D048630:Polyketide Synthases / Q000096:biosynthesis; D016133:Polymerase Chain Reaction; D011161:Porifera / Q000382:microbiology*; D017422:Sequence Analysis, DNA; D013302:Streptomyces / Q000145:classification / Q000201:enzymology / Q000378:metabolism
chemical_list: D000900:Anti-Bacterial Agents; D001551:Benz(a)Anthracenes; D018942:Macrolides; D048630:Polyketide Synthases