Creatine amidinohydrolase (CR), CAS No.37340-58-2

CAS: 37340-58-2 Cat. No.: rp216175 Molecular Weight: ~100 kDa EC Number: 253-471-1

GRADE & PURITY Recombinant, BioReagent, ActiBioPure™, High Performance, EnzymoPure™, ≥90%(SDS-PAGE), ≥5.8 U/mg enzyme powder

Accession #

P38488

Bioactivity

≥5.8 U/mg enzyme powder

Overview

Molecular weight: 48 kDa 
Enzymology Committee Number: EC 3.5.3.3
Physical appearance: Lyophilized powder 

Hydrolyzed creatine, used in the development and mass formulation of enzymatic creatinine reagents.
Enzymatic properties 
Source: Microorganism
Enzymology Committee No. : EC 3.5.3.3
Molecular weight: 48 kDa (SDS-PAGE)
Isoelectric point: 4.6
Km value: 8.81×10-3M(Creatine)

Inhibitors: Ag⁺ , Cu²⁺, Hg²⁺
Optimal pH: 8.0                                                                 图1
Optimum temperature: 45℃                                           图2
pH stability: 4.5-10.0(25°C,16h)                                     图 3
Thermal stability: Stable below 50°C (pH7.5,30min)  图 4
Protective agents: EDTA and sugar

Assay method for activity
1. Principle

Urea (Urea) and DAB (rho - dimethylaminobenzaldenhyde) reaction to form the yellow dye amount shall be tested by spectrophotometer at 435 nm.
2. Definition of enzyme activity
Unit enzyme activity is defined as the amount of enzyme required to catalyze a reaction to produce 1 μmol of creatine per minute for complete conversion to sarcosine under the following conditions.
3. Reagent preparation
Reagent I: 0.1M creatine solution.
Reagent II: 2.0gDAB was dissolved in 100mL dimethyl sulfoxide and 15mL concentrated hydrochloric acid was added.
Enzyme diluent: 50mMPBS, pH7.5, enzyme diluent was used to dilute the sample to 2.0-3.0U/mL.

4. Operation procedure
1. Take 1mL reagent I and preheat it in 5mLEP tube at 37°C for 5min.
2. Add 0.1mL of enzyme liquid and mix well.
3. After 10min reaction in a water bath at 37°C, 2.0mL reagent II was added to terminate the reaction and placed at 25°C for 20min.
4. Use an ultraviolet spectrophotometer to determine the absorbance value of the reaction liquid at 435nm.
5. Enzyme diluent was used as blank control instead of enzyme liquid.

5. Vitality computing

3.1: total volume of reaction liquid (mL);
0.1: enzyme liquid volume (mL);
10: Reaction time (min);
1.0: optical path length (cm);
df: dilution ratio;

C: Enzyme concentration (mg/mL);
0.321: millimolar absorption coefficient of yellow dye at 435nm (cm²/μmol).

Specifications

Product Name

Creatine amidinohydrolase (CR), CAS No.37340-58-2

Grade

ActiBioPure™, BioReagent, High Performance, Recombinant, EnzymoPure™

Specifications & Purity

Recombinant, BioReagent, ActiBioPure™, High Performance, EnzymoPure™, ≥90%(SDS-PAGE), ≥5.8 U/mg enzyme powder

Bioactivity

≥5.8 U/mg enzyme powder

Accession #

P38488

CAS

37340-58-2

Enzyme Commission Number

EC 3.5.3.3

Molecule Type

Enzyme

Storage and Shipping

Concentration

≥5.8 U/mg enzyme powder

Reconstitution

Reconstitute in 50mM PBS, pH7.5.

Storage

Store at -20°C,Avoid repeated freezing and thawing

Shipped In

Ice chest + Ice pads

Stability And Storage

Store at -20℃ long term (18 months). Upon reconstitution, it is recommended to aliquot. Avoid freeze/thaw cycle.

Unit definition

The amount of enzyme that catalyzes the complete conversion of 1 μmol of creatine to sarcosine per minute under the specified conditions.

References

1. Yan Tao, Chongying Liao, Fangli He, Yixuan He, Sihui Yang, Liu-Pan Yang, Li-Li Wang.  (2025)  Smartphone-assisted colorimetric sensing platform based on Fe-LCDs nanozymes for detection of creatine in meat.  FOOD CHEMISTRY,      [PMID:40381557] [10.1016/j.foodchem.2025.144754]

2. Fangbing Wang, Qiwen Peng, Yongheng Zhang, Min Zhang, Guoyue Shi.  (2026)  Five-in-one colorimetric multiplexed point-of-care testing of blood biomarkers via Janus separation-sensing membrane.  CHEMICAL ENGINEERING JOURNAL,      [PMID:] [10.1016/j.cej.2026.174104]