Identification of functionally clustered nystatin-like biosynthetic genes in a rare actinomycetes, Pseudonocardia autotrophica.
The polyene antibiotics, including nystatin, pimaricin, amphotericin, and candicidin, comprise a family of very valuable antifungal polyketide compounds, and they are typically produced by soil actinomycetes. Previously, using a polyene cytochrome P450 hydroxylase-specific genome screening strategy, Pseudonocardia autotrophica KCTC9441 was determined to contain genes potentially encoding polyene biosynthesis. Here, sequence information of an approximately 125.7-kb contiguous DNA region in five overlapping cosmids isolated from the P. autotrophica KCTC9441 genomic library revealed a total of 23 open reading frames, which are presumably involved in the biosynthesis of a nystatin-like compound tentatively named NPP. The deduced roles for six multi-modular polyketide synthase (PKS) catalytic domains were found to be highly homologous to those of previously identified nystatin biosynthetic genes. Low NPP productivity suggests that the functionally clustered NPP biosynthetic pathway genes are tightly regulated in P. autotrophica. Disruption of a NPP PKS gene completely abolished both NPP biosynthesis and antifungal activity against Candida albicans, suggesting that polyene-specific genome screening may constitute an efficient method for isolation of potentially valuable previously identified polyene genes and compounds from various rare actinomycetes widespread in nature.
Kim
Byung-Gyun
BG
Department of Biological Engineering, Inha University, Incheon, Korea.
Lee
Mi-Jin
MJ
Seo
Jiyoon
J
Hwang
Young-Bin
YB
Lee
Mi-Yeon
MY
Han
Kyuboen
K
Sherman
David H
DH
Kim
Eung-Soo
ES
D000192:Actinomycetales / Q000235:genetics*; D000935:Antifungal Agents / Q000378:metabolism*; D053898:Biosynthetic Pathways / Q000235:genetics; D002176:Candida albicans / Q000187:drug effects; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics; D023061:Gene Order; D005800:Genes, Fungal; D008826:Microbial Sensitivity Tests; D008954:Models, Biological; D008969:Molecular Sequence Data; D005810:Multigene Family; D016366:Open Reading Frames; D011090:Polyenes / Q000378:metabolism*; D048630:Polyketide Synthases / Q000235:genetics; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D000935:Antifungal Agents; D004271:DNA, Fungal; D005656:Fungal Proteins; D011090:Polyenes; D048630:Polyketide Synthases
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