title: Molecular cloning and expression of perhydrolase genes from Pseudomonas aeruginosa and Burkholderia cepacia in Escherichia coli.
abstract: Two bacterial perhydrolase genes, perPA and perBC, were cloned from Pseudomonas aeruginosa and Burkholderia cepacia, respectively, using PCR amplification with primers designed to be specific for conserved amino acid sequences of the already-known perhydrolases. The amino acid sequence of PerPA was identical to a putative perhydrolase of P. aeruginosa PAO1 genome sequences, whereas PerBC of B. cepacia was a novel bacterial perhydrolase showing similarity of less than 80% with all other existing perhydrolases. Most importantly, the perPA gene was expressed as a soluble intracellular form to an extent of more than 50% of the total protein content in Escherichia coli. Two perhydrolase enzymes were confirmed to exhibit the halogenation activity towards Phenol Red and monochlorodimedone. These results suggested that we successfully obtained the newly identified members of the bacterial perhydrolase family, expanding the pool of available perhydrolases.
authors.affiliation: Chemical Biotechnology Research Center, Korea Research Institute of Chemical Technology, Daejeon, Korea. ajee@krict.re.kr
authors.forename: Jae Kwang
authors.initials: JK
authors.lastname: Song
authors.forename: Hyo Jeong
authors.initials: HJ
authors.lastname: Ahn
authors.forename: Hyun Suk
authors.initials: HS
authors.lastname: Kim
authors.forename: Bong Keun
authors.initials: BK
authors.lastname: Song
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D016956:Burkholderia cepacia / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression / Q000235:genetics*; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D010544:Peroxidases / Q000096:biosynthesis* / Q000235:genetics; D011550:Pseudomonas aeruginosa / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; D010544:Peroxidases