title: [Purification and properties of pyrocatechase II from Pseudomonas putida strain 87].
abstract: Induction of modified ortho-pathway enzymes (catechol 1.2-dioxygenase II, muconate cycloisomerase II, dienelactone hydrolase, and maleylacetate reductase) was found in Pseudomonas putida 87, when 3-chlorobenzoic acid was used as a sole carbon and energy source. Catechol 1.2-dioxygenase II, the key chlorocatechol cleaving enzyme, was purified and characterized. The enzyme molecular mass as determined by gel filtration was 65,000 Da; the minimum molecular mass upon SDS electrophoresis was 33,000 Da. The pH and temperature optima for the enzyme were 7.2-7.8 and 35 degrees C, respectively. The highest stability of catechol 1.2-dioxygenase II upon storage was observed in 50 mM Tris-HCl buffer pH 7.8 at 4 degrees C. The relative values of Vmax for catechol 1.2-dioxygenase II with 3-chloro-, 4-chloro-, and 3.5-dichlorocatechols were 28%, 50%, and 41% of those for catechol. The enzyme affinity for chlorocatechols was 3-9 times higher than for methylcatechols and 10-20 times higher than for unsubstituted catechol.
authors.forename: I P
authors.initials: IP
authors.lastname: Solianikova
authors.forename: O V
authors.initials: OV
authors.lastname: Mal'tseva
authors.forename: L A
authors.initials: LA
authors.lastname: Golovleva
mesh_terms: D002384:Catalysis; D050548:Catechol 1,2-Dioxygenase; D002474:Cell-Free System; D002852:Chromatography, Ion Exchange; D049308:Dioxygenases; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D010105:Oxygenases / Q000302:isolation & purification* / Q000378:metabolism; D016958:Pseudomonas putida / Q000201:enzymology*; D013696:Temperature
chemical_list: D010105:Oxygenases; D049308:Dioxygenases; D050548:Catechol 1,2-Dioxygenase
vernacular_title: Ochistka i svoĭstva pirokatekhazy II iz shtamma Pseudomonas putida 87.