title: Expression and characterization of a novel deoxyribose 5-phosphate aldolase from Paenibacillus sp. EA001.
abstract: A novel deoC gene was identified from Paenibacillus sp. EA001 isolated from soil. The gene had an open reading frame (ORF) of 663 base pairs encoding 220 amino acids with a molecular mass of 24.5 kDa. The amino acid sequence was 79 % identical to that of deoxyribose 5-phosphate aldolase (DERA) from Geobacillus sp. Y412MC10. The deoC gene encoding DERA was cloned into expression vector and the protein was expressed in Escherichia coli. The recombinant DERA was purified by using Ni-NTA affinity chromatography and characterized. The optimum temperature and pH for DERA were 50 degrees C and 6.0, respectively. The specific activity for deoxyribose 5-phosphate (DR5P), substrate, was 62 micronmol/min/mg. The Km value for DR5P was determined to be 145 mM with the Kcat value of 3.2 times 10(2 /sec) from Lineweaver-Burk plots. The EA001 DERA showed stability toward a high concentration of acetaldehyde (100 mM).
authors.affiliation: Enzyme Fusion Technology Research Team, Molecular Bioprocess Research Center, Jeonbuk Branch Institute, Korea Research Institute of Bioscience and Biotechnology, Jeonbuk 580-185, Korea.
authors.forename: Yong-Mo
authors.initials: YM
authors.lastname: Kim
authors.forename: Nack-Shick
authors.initials: NS
authors.lastname: Choi
authors.forename: Yong Ook
authors.initials: YO
authors.lastname: Kim
authors.forename: Dong Ho
authors.initials: DH
authors.lastname: Son
authors.forename: Young-Hyo
authors.initials: YH
authors.lastname: Chang
authors.forename: Jae Jun
authors.initials: JJ
authors.lastname: Song
authors.forename: Joong Su
authors.initials: JS
authors.lastname: Kim
mesh_terms: D000446:Aldehyde-Lyases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D003001:Cloning, Molecular; D004795:Enzyme Stability; D015870:Gene Expression; D007700:Kinetics; D008969:Molecular Sequence Data; D056507:Paenibacillus / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D010802:Phylogeny; D017386:Sequence Homology, Amino Acid; D012988:Soil Microbiology
chemical_list: D001426:Bacterial Proteins; D000446:Aldehyde-Lyases; C020019:deoxyribose-phosphate aldolase