CREATININASE FROM PSEUDOMONAS SP.

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 properties

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)

Unit Definition

One unit will hydrolyze 1.0 mmole of creatinine to creatine per min at pH 8.0 and 25 °C

Physical form

Lyophilized powder containing sucrose and BSA as stabilizers

Analysis Note

Protein determined by biuret.

form: lyophilized powder. Quality Level: 200. specific activity: 100-300 . units/mg protein. mol wt: ~175 . kDa. composition: Protein, 65-85% . foreign activity: Creatinase and urease ≤. 1%, Hexokinase and ATPase ≤. 0.1%. storage temp.: 2-8°C. Pictograms: GHS08. Signal Word: Danger. Hazard Statements: H334. Precautionary Statements: P261 - P284 - P501. Hazard Classifications: Resp. Sens. 1. Storage Class Code: 11 - Combustible Solids. WGK: WGK 3. Flash Point(F): Not applicable. Flash Point(C): Not applicable. Personal Protective Equipment: Eyeshields, Gloves, multi-purpose combination respirator cartridge (US).