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C3172 Sigma-Aldrich

Creatininase from microorganisms

lyophilized powder, 100-300 units/mg protein

Synonym: Creatinine Amidohydrolase

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Properties

Related Categories 3.5.x.x Acting on C-N other than peptides, 3.x.x.x Hydrolases, Application Index, Biochemicals and Reagents, Cell Biology,
Quality Level   200
form   lyophilized powder
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

Description

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.

Packaging

1000 units in glass bottle

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.

Safety & Documentation

Safety Information

Symbol 
GHS08  GHS08
Signal word 
Danger
Hazard statements 
Precautionary statements 
RIDADR 
NONH for all modes of transport
WGK Germany 
WGK 3
Flash Point(F) 
Not applicable
Flash Point(C) 
Not applicable

Documents

Certificate of Analysis (COA)

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Certificate of Origin (COO)

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Protocols & Articles

Protocols

Assay Procedure for Creatininase

The appearance of creatinine-picrate (orange dye based on Jaffe's reaction) is measured at 520nm by spectrophotometry.
Keywords: Extinction coefficient, Sample preparations

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Peer-Reviewed Papers
15

References

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