Substrate specificity of two cytochrome P450 monooxygenases involved in lankamycin biosynthesis
| Autor: | Haruyasu Kinashi, Yuya Misaki, Aiko Teshima, Yosi Nindita, Kenji Arakawa, Yuji Konaka, Hisashi Kondo, Yasuhiro Itakura, Yu Tanaka, Tsugumi Tonokawa |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Stereochemistry Mutant 01 natural sciences Applied Microbiology and Biotechnology Biochemistry Streptomyces Analytical Chemistry Substrate Specificity Hydroxylation 03 medical and health sciences chemistry.chemical_compound Gene Knockout Techniques Biosynthesis Cytochrome P-450 Enzyme System Molecular Biology Biotransformation biology 010405 organic chemistry Organic Chemistry Glycosyltransferase Gene Substrate (chemistry) Cytochrome P450 General Medicine Monooxygenase biology.organism_classification Recombinant Proteins 0104 chemical sciences Erythromycin 030104 developmental biology chemistry biology.protein Biotechnology |
| Zdroj: | Bioscience, biotechnology, and biochemistry. 85(1) |
| ISSN: | 1347-6947 |
| Popis: | To elucidate the gross lankamycin biosynthetic pathway including two cytochrome P450 monooxygenases, LkmK and LkmF, we constructed two double mutants of P450 genes in combination with glycosyltransferase genes, lkmL and lkmI. An aglycon 8,15-dideoxylankanolide, a possible substrate for LkmK, was prepared from an lkmK–lkmL double mutant, while a monoglycoside 3-O-l-arcanosyl-8-deoxylankanolide, a substrate for LkmF, was from an lkmF–lkmI double mutant. Bioconversion of lankamycin derivatives was performed in the Escherichia coli recombinant for LkmK and the Streptomyces lividans recombinant for LkmF, respectively. LkmK catalyzes the C-15 hydroxylation on all 15-deoxy derivatives, including 8,15-dideoxylankanolide (a possible substrate), 8,15-dideoxylankamycin, and 15-deoxylankamycin, suggesting the relaxed substrate specificity of LkmK. On the other hand, LkmF hydroxylates the C-8 methine of 3-O-l-anosyl-8-deoxylankanolide. Other 8-deoxy lankamycin/lankanolide derivatives were not oxidized, suggesting the importance of a C-3 l-arcanosyl moiety for substrate recognition by LkmF in lankamycin biosynthesis. Thus, LkmF has a strict substrate specificity in lankamycin biosynthesis. |
| Databáze: | OpenAIRE |
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