title: Identification and molecular characterization of a novel DyP-type peroxidase from Pseudomonas aeruginosa PKE117.
abstract: A new DyP-type peroxidase from Pseudomonas aeruginosa PKE117 was identified and characterized. The dypPa was first identified via sequence analysis and then cloned in Escherichia coli. Subsequently, the recombinant protein DyPPa was expressed and purified. Its DNA sequence analysis revealed an open reading frame of 897 bp, encoding a protein monomer of 299 amino acid residues with isoelectric point 4.62. According to SDS-PAGE analysis and FPLC result, DyPPa mainly existed as homodimer (64 kDa). DyPPa displayed typical heme absorbance of Soret band, with an Rz value of 1.18. Inductively coupled plasma-atomic absorption spectrum data also indicated DyPPa contained iron. Multiple amino acid sequence alignment of DyPPa with other members of the DyP-type peroxidases family showed the presence of conserved D139, H210, and R227 amino acids and GXXDG motifs, which were commonly shared by the DyP-type peroxidase family. Although the primary structure homology between DyPPa and other family members was very low, their secondary and tertiary structure displayed high homology, which explained the high decolorizing activity of DyPPa. Specifically, DyPPa displayed a good thermal stability and maximal activity on Reactive blue 5 under pH 3.5. Therefore, it was proposed that DyPPa, with a wide range of substrate specificity, was a novel member of the DyP-type peroxidases family.
authors.affiliation: State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100193, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.forename: Chen
authors.initials: C
authors.lastname: Liu
authors.forename: Baozhen
authors.initials: B
authors.lastname: Li
authors.forename: Hongli
authors.initials: H
authors.lastname: Yuan
authors.forename: Jinshui
authors.initials: J
authors.lastname: Yang
authors.forename: Beiwen
authors.initials: B
authors.lastname: Zheng
mesh_terms: D000595:Amino Acid Sequence; D000880:Anthraquinones; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D004396:Coloring Agents; D017124:Conserved Sequence; D004795:Enzyme Stability; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010544:Peroxidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010802:Phylogeny; D055503:Protein Multimerization; D011550:Pseudomonas aeruginosa / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
chemical_list: D000880:Anthraquinones; D001426:Bacterial Proteins; D004396:Coloring Agents; C544286:Reactive Blue 5; D011994:Recombinant Proteins; D010544:Peroxidases