Native Uricase from Microorganism

Products

Native Uricase from Microorganism

Product Specifications

Cat#	NATE-1017
Descriptions	Microbial Uricase selectively catalyzes the decomposition of serum uric acid into soluble allantoin, eliminating uric acid’s absorbance interference to realize direct colorimetric uric acid quantification. This enzyme is the core raw material for gout risk screening and renal function monitoring diagnostic kits, with wide linear ranges covering normal and hyperuricemia patient sample concentrations.
Applications	This product can be applied to perform enzymatic quantitative detection of uric acid in serum for clinical diagnostic tests.
Synonyms	Urate: oxygen oxidoreductase
EC Number	EC 1.7.3.3
Catalysis	Uric acid + O₂ + 2H₂O → Allantoin + H₂O₂+ CO₂
Activity	≥ 2.5 U/mg (Uricase activity)
Appearance	White to light yellowish brown powder, lyophilized
Additive	Sodium tetraborate decahydrate
Molecular Weight	120,000 (Gel filtration)
Isoelectric Point	5.4
Km	5.9 × 10⁻⁶ M (Uric acid)
Optimum pH	8.0
pH Stability	7.0-10.0 (25°C, 18 h)
Optimum Temperature	40°C
Thermal Stability	up to 50°C (pH 8.5, 10 min)
Unit Definition	1 unit is defined as the enzyme quantity which oxidizes 1 µ mole of uric acid per minute under the conditions described below.
Storage	36 months from the date of analysis when stored at -20°C or below in a dry place under sealed conditions.

General Properties

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Assay Principle

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Activity Assay

Unit Definition

1 unit is defined as the enzyme quantity which oxidizes 1 µ mole of uric acid per minute under the conditions described below.

Reagents

A. 50 mM Borate Buffer (pH 8.5): Weigh 1.55 g of boric acid and dissolve in 400 mL of deionized water. Adjust the pH to 8.5 with 4M NaOH and fill up to 500 mL with deionized water.
B. Uric acid stock solution: Weigh 21 mg of uric acid and dissolve in approx. 50 mL of 50 mM Borate Buffer (A). Fill up to 100 mL with 50 mM Borate Buffer (A). (Can be used for 1 week if kept refrigerated)
C. Substrate solution: Dilute 5 mL of Uric acid stock solution (B) with 50 mM Borate buffer (A) and fill up to 50 mL. Keep refrigerated. (Make a fresh solution for each use)
D. Diluent: Weigh 1.36 g of potassium dihydrogen phosphate and 0.5 g of bovine serum albumin, and dissolve in approx. 80 mL of deionized water. Adjust the pH to 7.5 with 4N NaOH and fill up to 100 mL with deionized water. Keep refrigerated.
E. Enzyme solution: Weigh out Uricase and dissolve in Diluent (D). Enzyme solution should be prepared so that the value of ΔOD/minute becomes in the range of 0.085±0.008.

Procedure

Pipette 3 mL of Substrate solution (C) into a quartz cell (d=10 mm) and keep at 25±0.5℃ for 10 minutes. Add 0.02 mL of Enzyme solution (E) into the quartz cell and mix well. Keep the reaction mixture at 25±0.5℃. Exactly at 2 minutes and 5 minutes after the addition of Enzyme Solution (E), measure the absorbance of the reaction mixture at 293 nm. (A2 and A5)

Calculation

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Safe Handling Avoid inhalation; in the event of direct skin or eye contact, immediately flush the affected area with abundant water, and consult the SDS for further information.