Merck, C3172-1KU, Creatininase from microorganisms

General description
Isoelectric point:4.7
Michaelis constants:3.2 x 10‾²M (Creatinine), 5.7 x 10‾²M (Creatine)
Structure:6 subunits per mol of enzyme (One mol of zinc is bound to each subunit)
Inhibitors:Ag⁺, Hg⁺⁺, N-bromosuccinimide, EDTA
Optimum pH:6.5 − 7.5
Optimum temp:70°C
pH Stability:pH 7.5 − 9.0 (5°C, 16hr)
Thermal stability:Below 70°C (pH 7.5, 30 min)
Application
Creatininase from microorganisms may be used in the preparation of amperometric biosensor by co-immobilization with other enzymes for the determination of creatinine.
This enzyme is useful for enzymatic determination of creatinine when coupled with creatine amidinohydrolase, sarcosine dehydrogenase or sarcosine oxidase and formaldehyde dehydrogenase in clinical analysis.
Biochem/physiol Actions
Creatininase from Pseudomonas sp. is a homohexameric enzyme with a molecular mass of 28.4 kDa per subunit. It is a cyclic amidohydrolase catalysing the reversible conversion of creatinine to creatine. Each monomer contains a binuclear zinc centre near the C termini of the β-strands and the N termini of the main α-helices. These zinc ions indicate the location of the active site.
Physical form
Lyophilized powder containing sucrose and BSA as stabilizers
Analysis Note
Protein determined by biuret.
Other Notes
One unit will hydrolyze 1.0 μmole of creatinine to creatine per min at pH 8.0 and 25 °C