title: Purification of inactivated photoresponsive nitrile hydratase.
abstract: Photoresponsive nitrile hydratase from Rhodococcus sp. N-771 was purified in its inactivated form. The enzyme had a molecular weight of approximately 60 kDa and consisted of 2 subunits each having molecular weight of 27.5 and 28 kDa. The enzyme also contained 2 iron atoms/enzyme as a cofactor. The enzyme was more stable in its inactivated form, rather than the activated during storage in the dark. The enzyme was most stable in the temperature region of 0-35 degrees C, and lost its activity above 40 degrees C. The enzyme was most stable in the pH region of 6-8. The optimum temperature and pH for the enzyme activity was 30 degrees C and 7.8, respectively. The enzyme showed wide substrate specificity, and most of the metal ions did not affect enzyme activity significantly. The absorption spectrum revealed the presence of some cofactor which changed form after photoirradiation.
authors.affiliation: Frontier Research Program, RIKEN, Saitama, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Nagamune
authors.forename: H
authors.initials: H
authors.lastname: Kurata
authors.forename: M
authors.initials: M
authors.lastname: Hirata
authors.forename: J
authors.initials: J
authors.lastname: Honda
authors.forename: H
authors.initials: H
authors.lastname: Koike
authors.forename: M
authors.initials: M
authors.lastname: Ikeuchi
authors.forename: Y
authors.initials: Y
authors.lastname: Inoue
authors.forename: A
authors.initials: A
authors.lastname: Hirata
authors.forename: I
authors.initials: I
authors.lastname: Endo
mesh_terms: D004789:Enzyme Activation / Q000187:drug effects; D004795:Enzyme Stability; D006836:Hydro-Lyases / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D008670:Metals / Q000494:pharmacology; D012240:Rhodococcus / Q000187:drug effects / Q000201:enzymology*; D013056:Spectrophotometry, Ultraviolet; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D008670:Metals; D006836:Hydro-Lyases; C048831:nitrile hydratase