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C3172

Sigma-Aldrich

Creatininase from microorganisms

lyophilized powder, 100-300 units/mg protein

Synonym(s):

Creatinine Amidohydrolase

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About This Item

CAS Number:
Enzyme Commission number:
EC Number:
MDL number:
UNSPSC Code:
12352204
NACRES:
NA.54

form

lyophilized powder

Quality Level

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

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 μmole 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.

Pictograms

Health hazard

Signal Word

Danger

Hazard Statements

Precautionary Statements

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

dust mask type N95 (US), Eyeshields, Gloves

Certificates of Analysis (COA)

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Tadashi Yoshimoto et al.
Journal of molecular biology, 337(2), 399-416 (2004-03-09)
Creatininase from Pseudomonas putida is a member of the urease-related amidohydrolase superfamily. The crystal structure of the Mn-activated enzyme has been solved by the single isomorphous replacement method at 1.8A resolution. The structures of the native creatininase and the Mn-activated
Manoj Nahar et al.
Nanomedicine : nanotechnology, biology, and medicine, 4(3), 252-261 (2008-05-27)
Our aim in the present investigation was to develop a nanoparticulate carrier of amphotericin B (AmB) for controlled delivery as well as reduced toxicity. Nanoparticles of different gelatins (GNPs) (type A or B) were prepared by two-step desolvation method and
Chhanda Mallick et al.
Pancreas, 38(3), 322-329 (2009-01-27)
The study focused on the ability of the extracts of Musa paradisiaca and Coccinia indica on protein metabolic disorders in streptozotocin (STZ)-induced diabetes. Wistar strain rats were divided into 6 groups as control, control + composite extract treated, STZ-induced diabetes
Jason A Berberich et al.
Acta biomaterialia, 1(2), 193-199 (2006-05-17)
We describe the development of an extended use amperometric three-enzyme creatinine biosensor and the successful chemical modification and immobilization of the enzyme creatinine amidohydrolase using polyurethane prepolymers. Creatinine amidohydrolase is significantly stabilized by immobilization in polyurethane polymers. The half-life increases
Anne-Annette P Raso et al.
International journal of rheumatic diseases, 12(3), 243-249 (2010-04-09)
To determine the relationship of steroid use with tophus formation and other comorbid conditions among male gout patients. Review of medical records of Filipino gout patients under the care of rheumatologists was conducted. Univariate analysis (chi-square, Student's t-test) and multiple

Protocols

Assay Procedure for Creatininase

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