title: Chaperone-aided expression of LipA and LplA followed by the increase in alpha-lipoic acid production.
abstract: Alpha-lipoic acid (LA), a naturally occurring cofactor reported to be present in a diverse group of microorganisms, plants, and animal tissues, has been widely and successfully used as a therapy for a variety of diseases, including diabetes and heart disease. However, to date, recombinant DNA technology has not been applied for higher LA production due mainly to difficulties in the functional expression of key enzymes involved in LA production. Here, we report a study for higher LA production with the aid of chaperone plasmids, DnaKJE and trigger factor (Tf). The lipA and lplA genes encoding lipoate synthase and lipoate protein ligase in Pseudomonas fluorescens, respectively, were cloned and transformed into Escherichia coli K12. When they were overexpressed in E. coli, both LipA and LplA were expressed as inclusion bodies leading to no increase in LA production. However, when chaperone plasmids DnaKJE and Tf were coexpressed with lipA and lplA, the resulting recombinant E. coli strains showed higher LA production than the wild-type E. coli by 32-111%, respectively.
authors.affiliation: Department of Bioscience and Biotechnology, Konkuk University, 1 Hwayang-Dong Gwangjin-Gu, Seoul, 143-701, South Korea.
authors.forename: Hee-Jung
authors.initials: HJ
authors.lastname: Moon
authors.forename: Marimuthu
authors.initials: M
authors.lastname: Jeya
authors.forename: In-Sik
authors.initials: IS
authors.lastname: Yu
authors.forename: Jung-Hwan
authors.initials: JH
authors.lastname: Ji
authors.forename: Deok-Kun
authors.initials: DK
authors.lastname: Oh
authors.forename: Jung-Kul
authors.initials: JK
authors.lastname: Lee
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism*; D010453:Peptide Synthases / Q000235:genetics / Q000378:metabolism*; D011551:Pseudomonas fluorescens / Q000235:genetics; D008063:Thioctic Acid / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; C079999:LipA protein, Bacteria; D018832:Molecular Chaperones; D008063:Thioctic Acid; D010453:Peptide Synthases; C072322:lipoate-protein ligase