Identification of biosynthetic gene clusters from metagenomic libraries using PPTase complementation in a Streptomyces host.
作者： J. Kipchirchir Bitok , Christophe Lemetre , Melinda A. Ternei , Sean F. Brady
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摘要: The majority of environmental bacteria are not readily cultured in the lab, leaving natural products they make inaccessible using culture-dependent discovery methods. Cloning and heterologous expression DNA extracted from samples (environmental DNA, eDNA) provides a means circumventing this bottleneck. To facilitate identification clones containing biosynthetic gene clusters, we developed model reporter strain Streptomyces albus::bpsA ΔPPTase. This carries 4΄-phosphopantetheinyl transferase (PPTase)-dependent blue pigment synthase A gene, bpsA, PPTase deletion background. eDNA that express functional restore production pigment, indigoidine. As genes often occur clusters (BGCs), indigoidine can be used to identify BGCs. We screened soil library hosted S. ΔPPTase identified non-ribosomal peptide synthetase (NRPS), polyketide (PKS) mixed NRPS/PKS clusters. One NRPS cluster was shown confer myxochelin
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