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- Title
A regulatory gene ( ECO-orf4) required for ECO-0501 biosynthesis in Amycolatopsis orientalis.
- Authors
Shen, Yang; Huang, He; Zhu, Li; Luo, Minyu; Chen, Daijie
- Abstract
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.
- Subjects
BIOCHEMICAL templates; ORGANIC synthesis; POLYENE antibiotics; GENE expression; AMYCOLATOPSIS
- Publication
Journal of Basic Microbiology, 2014, Vol 54, Issue 2, p104
- ISSN
0233-111X
- Publication type
Article
- DOI
10.1002/jobm.201200314