title: Metabolic engineering of Nocardia sp. CS682 for enhanced production of nargenicin A₁.
abstract: A number of secondary metabolites having therapeutic importance have been reported from the genus Nocardia. One of the polyketide antibiotic compounds isolated from Nocardia is nargenicin A(1). Recently, nargenicin A(1) has been isolated from Nocardia sp. CS682, a new Nocardia strain isolated from soil in Jeonnam, South Korea. It possesses strong antibacterial activity against methicillin-resistant Staphylococcus aureus. In this study, we applied a metabolic engineering approach based on recombinant DNA technology in order to boost the production of nargenicin A(1) from Nocardia sp. CS682. Initially, we optimized the transformation of this new strain by electroporation method. Heterologous expression of S-adenosylmethionine synthetase (MetK1-sp) in Nocardia sp. CS682 enhanced the production of nargenicin A(1) by about 2.8 times due to transcriptional activation of biosynthetic genes as revealed by reverse transcription polymerase chain reaction analysis. Similarly, expression of acetyl-CoA carboxylase genes improved nargenicin A(1) production by about 3.8 times in Nocardia sp. ACC18 compared to that in Nocardia sp. CS682 and Nocardia sp. NV18 by increasing precursor pool. Thus, enhanced production of nargenicin A(1) from Nocardia sp. CS682 can be achieved by expression of transcriptional activator genes and precursor genes from Streptomyces strains.
authors.affiliation: Institute of Biomolecule Reconstruction, Sun Moon University, 100, Kalsan-ri, Tangjeonmyun, Asansi, Chungnam 336-708, Republic of Korea.
authors.forename: Sushila
authors.initials: S
authors.lastname: Maharjan
authors.forename: Dinesh
authors.initials: D
authors.lastname: Koju
authors.forename: Hei Chan
authors.initials: HC
authors.lastname: Lee
authors.forename: Jin Cheol
authors.initials: JC
authors.lastname: Yoo
authors.forename: Jae Kyung
authors.initials: JK
authors.lastname: Sohng
mesh_terms: D000103:Acetyl-CoA Carboxylase / Q000235:genetics / Q000378:metabolism*; D000900:Anti-Bacterial Agents / Q000096:biosynthesis / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D003470:Culture Media; D018274:Electroporation; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D007783:Lactones / Q000378:metabolism / Q000494:pharmacology; D060847:Metabolic Engineering / Q000379:methods; D055624:Methicillin-Resistant Staphylococcus aureus / Q000187:drug effects; D008716:Methionine Adenosyltransferase / Q000235:genetics / Q000378:metabolism*; D009615:Nocardia / Q000201:enzymology / Q000235:genetics*; D010957:Plasmids; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012988:Soil Microbiology; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics; D015533:Transcriptional Activation; D014169:Transformation, Bacterial
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D003470:Culture Media; D007783:Lactones; C034624:nargenicin A1; D008716:Methionine Adenosyltransferase; D000103:Acetyl-CoA Carboxylase