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- Biosynthesis of 2-amino-3-hydroxycyclopent-2-enone moiety of bafilomycin in Kitasatospora cheerisanensis KCTC2395
- Nguyen Phan Kieu Hanh , Jae Yoon Hwang , Hye Ryeung Oh , Geum Jin Kim , Hyukjae Choi , Doo Hyun Nam
- J. Microbiol. 2018;56(8):571-578. Published online July 25, 2018
- DOI: https://doi.org/10.1007/s12275-018-8267-0
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Abstract
- Bafilomycins produced by Kitasatospora cheerisanensis KCTC- 2395 belong to the 16-membered macrolactone family plecomacrolide antibiotics. Bafilomycin B1 contains 2-amino- 3-hydroxycyclopent-2-enone (C5N), a five membered ring, which gets condensed via an amide linkage to bafilomycin polyketide. To study the biosynthetic pathway of C5N during bafilomycin biosynthesis in K. cheerisanensis KCTC2395, we attempted the functional analysis of two putative genes, encoding 5-aminolevulinic acid synthase (ALAS) and acyl- CoA ligase (ACL). The amplified putative genes for ALAS and ACL were cloned into the E. coli expression vector pET- 32a(+) plasmid, following which the soluble recombinant ALAS and ACL proteins were purified through nickel-affinity column chromatography. Through HPLC analysis of the enzyme reaction mixture, we confirmed the products of putative ALAS and ACL reaction as 5-aminolevulinic acid (5- ALA) and 5-ALA-CoA, respectively. The optimal pH for the putative ALAS reaction was 7.5, and for putative ACL reaction was 7.0, as confirmed by the colorimetric assay. Furthermore, pyridoxal 5-phosphate (PLP) was found to be an essential cofactor in the putative ALAS reaction, and ATP was a cofactor for the putative ACL catalysis. Finally, we also confirmed that the simultaneous treatment of putative ACL and putative ALAS enzymes resulted in the production of C5N compound from 5-ALA.
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Citations
Citations to this article as recorded by
- The Secondary Metabolites from Genus Kitasatospora: A Promising Source for Drug Discovery
Yuanjuan Wei, Guiyang Wang, Yan Li, Maoluo Gan
Chemistry & Biodiversity.2024;[Epub] CrossRef - Elucidation of the Late Steps during Hexacosalactone A Biosynthesis in Streptomyces samsunensis OUCT16-12
He Duan, Fang Wang, Chuchu Zhang, Yujing Dong, Huayue Li, Fei Xiao, Wenli Li, Haruyuki Atomi
Applied and Environmental Microbiology.2023;[Epub] CrossRef - A Secondary Metabolic Enzyme Functioned as an Evolutionary Seed of a Primary Metabolic Enzyme
Jun Kawaguchi, Hikaru Mori, Noritaka Iwai, Masaaki Wachi, Miriam Barlow
Molecular Biology and Evolution.2022;[Epub] CrossRef - Biosynthesis of Methoxymalonyl-acyl Carrier Protein (ACP) as an Extender Unit for Bafilomycin Polyketide in Streptomyces griseus DSM 2608
Nguyen Phan Kieu Hanh, Jae Yoon Hwang, Doo Hyun Nam
Biotechnology and Bioprocess Engineering.2018; 23(6): 693. CrossRef
- The Secondary Metabolites from Genus Kitasatospora: A Promising Source for Drug Discovery
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