RRC ID 40708
著者 Ross AC, Gulland LE, Dorrestein PC, Moore BS.
タイトル Targeted capture and heterologous expression of the Pseudoalteromonas alterochromide gene cluster in Escherichia coli represents a promising natural product exploratory platform.
ジャーナル ACS Synth Biol
Abstract Marine pseudoalteromonads represent a very promising source of biologically important natural product molecules. To access and exploit the full chemical capacity of these cosmopolitan Gram-(-) bacteria, we sought to apply universal synthetic biology tools to capture, refactor, and express biosynthetic gene clusters for the production of complex organic compounds in reliable host organisms. Here, we report a platform for the capture of proteobacterial gene clusters using a transformation-associated recombination (TAR) strategy coupled with direct pathway manipulation and expression in Escherichia coli. The ~34 kb pathway for production of alterochromide lipopeptides by Pseudoalteromonas piscicida JCM 20779 was captured and heterologously expressed in E. coli utilizing native and E. coli-based T7 promoter sequences. Our approach enabled both facile production of the alterochromides and in vivo interrogation of gene function associated with alterochromide's unusual brominated lipid side chain. This platform represents a simple but effective strategy for the discovery and biosynthetic characterization of natural products from marine proteobacteria.
巻・号 4(4)
ページ 414-20
公開日 2015-4-17
DOI 10.1021/sb500280q
PMID 25140825
PMC PMC4410906
MeSH Bacterial Proteins* / biosynthesis Bacterial Proteins* / genetics Escherichia coli* / genetics Escherichia coli* / metabolism Gene Expression* Genes, Bacterial Lipopeptides* / biosynthesis Lipopeptides* / genetics Multigene Family* Pseudoalteromonas / genetics* Recombinant Proteins / biosynthesis Recombinant Proteins / genetics Transformation, Bacterial*
IF 4.411
引用数 50
WOS 分野 BIOCHEMICAL RESEARCH METHODS
リソース情報
一般微生物 JCM 20779