title: [Purification and some properties of D-hydantoinase produced by Pseudomonas 2262].
abstract: A D-hydantoinase produced by Pseudomonas 2262 was purified to electrophoretic homogeneity by the steps of thermal treatment, (NH4)2SO4 fractionation and column chromatography with Q-Sepharose fast flow, phenyl-Sepharose fast flow and Superose 12. Purification of about 60 fold was achieved with an overall yield of 16%. The relative molecular mass of the native enzyme is 109 kD and that of subunit is 53.7 kD by the analysis of Native and SDS-PAGE as well as gel filtration respectively. Some properties of the enzyme such as the sensitivity to thiol reagent and the effects of metal ions, for instance inhibited by Zn2+ and activited by Mn2+, Mg2+ are identical to dihydropyrimidinase. The optimum temperature and pH for enzymatic catalysis are 70 degrees C and 8.0 respectively. The enzyme activity is stable under 60 degrees C and in the pH range of 6-10. The N-terminal sequence for 10 amino acid residues is MDKLIKNGTI.
authors.affiliation: Biotechnol Center of Shanxi University, Taiyuan 030006, China.
authors.forename: Y
authors.initials: Y
authors.lastname: Shi
authors.forename: H
authors.initials: H
authors.lastname: Li
authors.forename: J
authors.initials: J
authors.lastname: Yuan
authors.forename: Y
authors.initials: Y
authors.lastname: Qi
authors.forename: J
authors.initials: J
authors.lastname: Li
mesh_terms: D000581:Amidohydrolases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D000595:Amino Acid Sequence; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D008970:Molecular Weight; D011549:Pseudomonas / Q000145:classification / Q000201:enzymology*; D017386:Sequence Homology, Amino Acid; D013696:Temperature; D015032:Zinc / Q000494:pharmacology
chemical_list: D008345:Manganese; D000581:Amidohydrolases; C020047:dihydropyrimidinase; D008274:Magnesium; D015032:Zinc