Peng Zhu, Yanling Zheng, Yurong You, Xiaojun Yan, Jianzhong Shao
College of Life Sciences, Zhejiang University, Hangzhou 310058, China.
Canadian journal of microbiology 2009 MarIn the present study, we sought to confirm a putative non-ribosomal peptide synthase (NRPS) - polyketide synthase (PKS) gene cluster in marine sponge-associated bacterium with cytotoxic activity and elucidate the gene's structural information. The genomic library of the marine sponge Hymeniacidon perleve-associated bacterium Pseudoalteromonas sp. strain NJ631 was constructed using a pCC1FOS fosmid. Positive clones that covered the whole gene cluster region of hybrid NRPS-PKS were selected for shotgun sequencing. The results obtained from BlastX and open reading frame (ORF) analysis indicated that there are 3 big ORFs, NJA1, NJA2, and NJA3, that encoded proteins with similarities to amino acid adenylation, beta-ketoacyl synthase, and non-ribosomal peptide synthase, respectively, from different organisms. The results gave us a clue that there could be PKS or NRPS modules in the 3 ORFs. Further analysis demonstrated 3 ORFs encoding 2 NRPS modules, 1 PKS module, and 3 NRPS modules. Using the specificity-conferring selection rule, the substrate specificity of 4 adenylation (A) domains (A2, A3, A4, and A5) were successfully predicted, and the amino acids of the substrate specificity were glutamic acid - glutamine, serine, D-serine, and Aeo (2-amino-9,10-epoxy-8-oxodecanoic acid), respectively.
Peng Zhu, Yanling Zheng, Yurong You, Xiaojun Yan, Jianzhong Shao. Sequencing and modular analysis of the hybrid non-ribosomal peptide synthase - polyketide synthase gene cluster from the marine sponge Hymeniacidon perleve-associated bacterium Pseudoalteromonas sp. strain NJ631. Canadian journal of microbiology. 2009 Mar;55(3):219-27
PMID: 19370064
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