title: Codon optimization enhances protein expression of human peptide deformylase in E. coli.
abstract: Human peptide deformylase (hPDF), located in the mitochondria, has recently become a promising target for anti-cancer therapy. However, the expression of the hPDF gene in Escherichia coli is not efficient likely due to extremely high levels of GC content as well as the presence of rare codons. We performed codon optimization of the hPDF gene in order to reduce GC content and to eliminate rare codons. Putative stable secondary structures of the optimized gene were also reduced. Codon optimization increased the expression of hPDF protein (residues 63-243) presumably by reducing the GC content. A large amount of soluble hPDF was obtained upon its fusion with thioredoxin (Trx-hPDF), although an insoluble fraction was still dominant. We confirmed that Co(2+) is an optimal metal for increasing the activity of purified Trx-hPDF, and that actinonin acts as an efficient inhibitor. Therefore, a large amount of purified hPDF protein would provide many benefits for the screening of various drug candidates.
authors.affiliation: Division of Magnetic Resonance, Korea Basic Science Institute Ochang Campus, Cheongwon-Gun, Ochang-Eup, Yangcheong-Ri 804-1, Chungcheongbuk-Do 363-883, Republic of Korea.
authors.forename: Ji-Hoon
authors.initials: JH
authors.lastname: Han
authors.forename: Yun-Seok
authors.initials: YS
authors.lastname: Choi
authors.forename: Won-Je
authors.initials: WJ
authors.lastname: Kim
authors.forename: Young Ho
authors.initials: YH
authors.lastname: Jeon
authors.forename: Seung Kyu
authors.initials: SK
authors.lastname: Lee
authors.forename: Bong-Jin
authors.initials: BJ
authors.lastname: Lee
authors.forename: Kyoung-Seok
authors.initials: KS
authors.lastname: Ryu
mesh_terms: D000581:Amidohydrolases / Q000096:biosynthesis* / Q000235:genetics; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D003062:Codon / Q000235:genetics*; D004926:Escherichia coli / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*
chemical_list: D003062:Codon; D011993:Recombinant Fusion Proteins; D000581:Amidohydrolases; C084507:peptide deformylase