title: Characterization of dephosphocoenzyme A kinase from Streptomyces peucetius ATCC27952, and its application for doxorubicin overproduction.
abstract: Dephosphocoenzyme A (CoaE) catalyzes the last step in the biosynthesis of the cofactor coenzyme A. In this study, we report the identification and application of CoaE from Stretomyces peucetius ATCC27952. After expression of coaE, the protein was found to have a molecular mass of 28.6 kDa. Purification of the His-tagged fused CoaE protein was done by immobilized metal-affinity chromatography, and then in vitro enzymatic coupling assay was performed. The increasing NADH consumption with time shed light on the phosphorylating activity of CoaE. Furthermore, the overexpression of coaA and coaE independently under the ermE(*) promoter in the doxorubicin -producing wild type strain, resulted in 1.4- and 1.5-fold enhancements in doxorubicin production, respectively. In addition, the overexpression of both genes together showed a 2.1-fold increase in doxorubicin production. These results established a positive role for secondary metabolite production from Streptomyces peucetius.
authors.affiliation: Department of Pharmaceutical Engineering, SunMoon University, Asansi 336-708, Republic of Korea.
authors.forename: Na-Rae
authors.initials: NR
authors.lastname: Lee
authors.forename: Hemraj
authors.initials: H
authors.lastname: Rimal
authors.forename: Joo-Ho
authors.initials: JH
authors.lastname: Lee
authors.forename: Tae-Jin
authors.initials: TJ
authors.lastname: Oh
mesh_terms: D001709:Biotechnology; D004317:Doxorubicin / Q000032:analysis / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D053858:Metabolic Networks and Pathways; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics*
chemical_list: D011993:Recombinant Fusion Proteins; D004317:Doxorubicin; D017853:Phosphotransferases (Alcohol Group Acceptor); C020021:dephospho-CoA kinase