Characterization Of Crmm As A O-Methyltransferase In Caerulomycin A Biosynthesis

Actinomycetes are the most important type of bacteria in Marine sediments, anddistributed widely. Marine actinomycetes lived in special marine environment with complexunique metabolic pathways and its metabolites are very special in such as structure andbiological activity, which make actinomyces the most important medicinal microbial population.Caerulomycin A is a kind of secondary metabolite, isolated from marine actinomyceteActinoalloteichus cyanogriseus. Caerulomycin A was found to have strong antifungal, antitumoractivity and moderately antibacterial activity. Tiacumicin B is one of macrolides antibiotics with18member ring, has obvious antibacterial activity against Clostridium difficile,produced byTerrestrial actinomycetes Dactylosporangium aurantiacum subsp. Hamdenensis NRRL18085.The biosynthesis of natural products can be broadly divided into three parts: forming theprecursor, skeleton Construction and modification. The representative modification enzymesinclude methyl transferase enzyme, P450oxidase, acylase, glycosyltransferase, a halogenenzymes aminotransferase and phosphorylase, and so on. The post-modification reactionsubstantially increases the species and structural diversity of the natural products is acombination of biological research tools. Integrating the biosynthesis of Caerulomycin andTiacumicin, we explore the post-modification and its functioning enzymes, and lay thefoundation of Combination of biotechnology, expecting to design and manufacture the betterproducts.In this study, we attempt to evaluate the function of CrmM, TiaP1and TiaP2. We clonedCrmM, tiaP1and tiaP2, respectively constructed this three genes in vectors with histidine tagand GST tag, and successfully expression and purification the related proteins in E.coli BL21,and use the coarse enzyme reaction, pure enzyme reaction, as well as biological transformationstudy CrmM, TiaP1and TiaP2enzyme activity in vitro. TiaP2may play a role in hydroxylationthrough biotransformation verification.