Ketoamine oxidase (KAOD)

Cat. No.: rp216165

GRADE & PURITY BioReagent, EnzymoPure™, ≥90%(SDS-PAGE), ≥ 4 U/mg

Overview

Application
It is used in the research and development of enzymatic glycosylated albumin reagents and mass formulation.
Enzymatic properties
Source: Microorganism
Enzymology Committee Number: EC1.5.3
Molecular weight: 55kDa (SDS-PAGE)
Isoelectric point: 6
Km value: 5.0×10 ⁻⁴ M (Fructosyl-Ala)
Inhibitors: Hg ²⁺ , Pb ²⁺
Optimal pH: 7.7 Figure 1
Optimum temperature: 42℃ Figure 2
pH stability: 5.0-9.5 (25℃, 16h) Figure 3
Thermal stability: Stable below 40℃ (pH8.0, 30min) Figure 4
Stability: -25 ~ -15℃ standing store for 12 months
More than 90% activity Figure 5
Protective agent: glycerin, trehalose

Assay method for activity

1. Principle

 
2. Definition of enzyme activityUnit enzyme activity is defined as the amount of enzyme required to catalyze the production of 1μmol H2O2 per minute under the following conditions.
3. Reagent preparation
Reagent I: 1M potassium phosphate buffer, pH8.0
Reagent II: 1kU/mL POD
Reagent III: 50mM TOOS solution
Reagent IV: 50mM 4-AA solution
Reagent V: 200mM Glycated alanine
Enzyme diluent: 20mM Tris-HCl, pH8.0
Prepare the reaction mixture as follows:
Reagent I: 10mL
Reagent II: 0.1mL
Reagent III: 1mL
Reagent IV: 1mL
Reagent V: 10mL
Double steam water set volume to 100mL
4. Operation procedure
4.1 Add 980μL reaction mixture into 1mL colorimetric dish.
4.2 Incubate at 37°C for 5 minutes.
4.3 Add 20 μL of enzyme solution to be tested to the reaction mixture.
4.4 Reaction at 37°C, the absorbance change (∆As) of the sample within 1min is detected by spectrophotometer at 555nm.
* Blank control measurement method: Use 20 μL of enzyme dilution solution instead of the enzyme solution to be tested, and measure the absorbance change (ΔAb) of the sample within 1 minute.
∆A=∆As-∆Ab
5. Vitality computing

Vt: Total volume of reaction liquid (1.0 mL);
Vs: Enzyme liquid volume (0.02 mL);
t: Reaction time (1 min);
df: Dilution ratio;
C: Enzyme concentration (mg/mL);
1.0: Optical path length (cm);
1/2: 1 mole hydrogen peroxide to generate 1/2 mole quinone imide dye;
39.2: Under standard reaction conditions, the millimolar absorption coefficient of the color group at 555nm (cm² /μmol).

Specifications

Product Name

Ketoamine oxidase (KAOD)

Synonyms

Ketoamine oxidase | KAOD

Grade

BioReagent, EnzymoPure™

Specifications & Purity

EnzymoPure™, BioReagent, ≥90%(SDS-PAGE), ≥ 4 U/mg

Molecule Type

Enzyme

Storage and Shipping

Concentration

≥ 4 U/mg

Storage

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

Shipped In

Ice chest + Ice pads

Stability And Storage

Store at -20℃ long term (12 months). Upon receipt, it is recommended to aliquot. Avoid freeze/thaw cycle.