title: Cloning and characterization of a novel CoA-ligase gene from Penicillium chrysogenum.
abstract: A novel phenylacetic acid (PAA)-induced CoA-ligase-encoding gene, designated as phlC, has been cloned from penicillin-producing fungus Penicillium chrysogenum. The open reading frame of phlC cDNA was 1671 bp and encoded a 556 amino acid residues protein with the consensus AMP binding site and a peroxisomal targeting signal 1 on its C terminus. The deduced amino acid sequence showed 37% and 38% identity with characterized P. chrysogenum Phl and PhlB protein, respectively. Functional recombinant PhlC protein was overexpressed in Escherichia coli. The purified recombinant enzyme was capable to convert PAA into its corresponding CoA ester with a specific activity of 129.5 ± 3.026 pmol/min per mg protein. Similar to Phl and PhlB, PhlC displayed broad substrate spectrum and showed higher activities to medium- and long-chain fatty acids. The catalytic properties of PhlC have been determined and compared to those of Phl and PhlB.
authors.affiliation: Department of Bioresources, New Drug R&D Center of North China Pharmaceutical Corporation and National Engineering Research Center for Microbial Medicine, 388 East Heping Road, Shijiazhuang, 050015, China.
authors.forename: Zhou-Liang
authors.initials: ZL
authors.lastname: Yu
authors.forename: Jing
authors.initials: J
authors.lastname: Liu
authors.forename: Fu-Qiang
authors.initials: FQ
authors.lastname: Wang
authors.forename: Meng
authors.initials: M
authors.lastname: Dai
authors.forename: Bao-Hua
authors.initials: BH
authors.lastname: Zhao
authors.forename: Jian-Gong
authors.initials: JG
authors.lastname: He
authors.forename: Hua
authors.initials: H
authors.lastname: Zhang
mesh_terms: D000105:Acetyl Coenzyme A / Q000096:biosynthesis*; D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D003066:Coenzyme A Ligases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D010408:Penicillium chrysogenum / Q000201:enzymology* / Q000235:genetics; D010648:Phenylacetates / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment
chemical_list: D010648:Phenylacetates; D011994:Recombinant Proteins; D000105:Acetyl Coenzyme A; C037816:phenylacetyl-coenzyme A; D003066:Coenzyme A Ligases; C023914:phenylacetate - CoA ligase; C025136:phenylacetic acid