Carboxyl-terminal domain characterization of polyene-specific P450 hydroxylase in Pseudonocardia autotrophica
- Authors
- Kim, Min-Kyung; Won, Hyung-Jin; Kim, Hye-Jin; Choi, Si-Sun; Lee, Heung-Shick; Kim, Pil; Kim, Eung-Soo
- Issue Date
- 11월-2016
- Publisher
- SPRINGER HEIDELBERG
- Keywords
- Polyene; P450 hydroxylase; Pseudonocardia; Substrate specificity
- Citation
- JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY, v.43, no.11, pp.1625 - 1630
- Indexed
- SCIE
SCOPUS
- Journal Title
- JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY
- Volume
- 43
- Number
- 11
- Start Page
- 1625
- End Page
- 1630
- URI
- https://scholar.korea.ac.kr/handle/2021.sw.korea/87121
- DOI
- 10.1007/s10295-016-1813-z
- ISSN
- 1367-5435
- Abstract
- A polyene compound NPP identified in Pseudonocardia autotrophica was shown to contain an aglycone identical to nystatin, but to harbor a unique disaccharide moiety that led to higher solubility and reduced hemolytic activity. Recently, it was revealed that the final step of NPP (nystatin-like polyene) biosynthesis is C10 regio-specific hydroxylation by the cytochrome P450 hydroxylase (CYP) NppL (Kim et al. [7]). Through mutation and cross-complementation, here we found that NppL preferred a polyene substrate containing a disaccharide moiety for C10 hydroxylation, while its orthologue NysL involved in nystatin biosynthesis showed no substrate preference toward mono- and disaccharide moieties, suggesting that two homologous polyene CYPs, NppL and NysL might possess a unique domain recognizing a sugar moiety. Two hybrid NppL constructs containing the C-terminal domain of NysL exhibited no substrate preference toward 10-deoxy NPP and 10-deoxy nystatin-like NysL, implying that the C-terminal domain plays a major role in differentiating the sugar moiety responsible for substrate specificity. Further C-terminal domain dissection of NppL revealed that the last fifty amino acids play a critical role in determining substrate specificity of polyene-specific hydroxylation, setting the stage for the biotechnological application of hydroxyl diversification for novel polyene biosynthesis in actinomycetes.
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