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A regulatory gene ( ECO‐orf4 ) required for ECO‐0501 biosynthesis in Amycolatopsis orientalis
Author(s) -
Shen Yang,
Huang He,
Zhu Li,
Luo Minyu,
Chen Daijie
Publication year - 2014
Publication title -
journal of basic microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.58
H-Index - 54
eISSN - 1521-4028
pISSN - 0233-111X
DOI - 10.1002/jobm.201200314
Subject(s) - gene cluster , gene , biology , biosynthesis , regulator gene , transcription factor , genetics , biochemistry , regulation of gene expression
ECO‐0501 is a novel linear polyene antibiotic, which was discovered from Amycolatopsis orientalis . Recent study of ECO‐0501 biosynthesis pathway revealed the presence of regulatory gene: ECO‐orf4 . The A. orientalis ECO‐orf4 gene from the ECO‐0501 biosynthesis cluster was analyzed, and its deduced protein (ECO‐orf4) was found to have amino acid sequence homology with large ATP‐binding regulators of the LuxR (LAL) family regulators. Database comparison revealed two hypothetical domains, a LuxR‐type helix‐turn‐helix (HTH) DNA binding motif near the C‐terminal and an N‐terminal nucleotide triphosphate (NTP) binding motif included. Deletion of the corresponding gene ( ECO‐orf4 ) resulted in complete loss of ECO‐0501 production. Complementation by one copy of intact ECO‐orf4 restored the polyene biosynthesis demonstrating that ECO‐orf4 is required for ECO‐0501 biosynthesis. The results of overexpression ECO‐orf4 on ECO‐0501 production indicated that it is a positive regulatory gene. Gene expression analysis by reverse transcription PCR of the ECO‐0501 gene cluster showed that the transcription of ECO‐orf4 correlates with that of genes involved in polyketide biosynthesis. These results demonstrated that ECO‐orf4 is a pathway‐specific positive regulatory gene that is essential for ECO‐0501 biosynthesis.