Purification, crystallization, and some properties of creatine amidinohydrolase from Pseudomonas putida.
A method was developed for purification and crystallization of creatinase [creatine amidinohydrolase, EC 3.5.3.3] from Pseudomonas putida var. naraensis C-83. The purified preparation appeared homogeneous on disc electrophoresis and ultracentrifugation and had a molecular weight of 94,000. It was most active at pH 8 and stable between pH 6 and 8 for 24 hr at 37 degrees. SDS-polyacrylamide gel electrophoresis indicated that the native enzyme was made up of two subunit monomers, the molecular weights of which were estimated to be 47,000. Inhibition experiments suggested that a sulfhydryl group is located in or near the active site of the enzyme.
D001665:Binding Sites; D003401:Creatine / Q000378:metabolism; D003460:Crystallization; D003545:Cysteine / Q000032:analysis; D004355:Drug Stability; D006863:Hydrogen-Ion Concentration; D006867:Hydrolases / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D046911:Macromolecular Substances; D008970:Molecular Weight; D011549:Pseudomonas / Q000201:enzymology*
D046911:Macromolecular Substances; D006867:Hydrolases; D003545:Cysteine; D003401:Creatine