Native L-α-Glycerophosphate Oxidase from Microorganism

Products

Native L-α-Glycerophosphate Oxidase from Microorganism

Product Specifications

Cat#	NATE-1007
Descriptions	L-α-Glycerophosphate Oxidase catalyzes the oxidation of glycerol-3-phosphate to produce hydrogen peroxide for colorimetric signal readout, acting as the terminal signal-generating enzyme in standard triglyceride diagnostic kits. Our microbial-derived product features high catalytic turnover and low detection limits, fully compatible with automated analyzer workflows for large-volume serum lipid panel testing.
Applications	This product can be paired with lipoprotein lipase and glycerol kinase to conduct enzymatic quantitative testing of triglycerides in serum for clinical diagnostic use.
Synonyms	Sn-Glycerol-3-phosphate : oxygen 2-oxidoreductase
EC Number	EC 1.1.3.21
Catalysis	L-α-Glycerophosphate + O₂ → Dihydroxyacetone phosphate + H₂O₂
Activity	≥ 35 U/mg (L-α-Glycerophosphate oxidase activity)
Contaminant	≤ 3 × 10⁻³% (Lactase oxidase activity); ≤ 4 × 10⁻³% (Catalase activity); ≤ 6 × 10⁻³% (ATPase activity)
Appearance	Yellow powder, lyophilized
Additive	Not added
Molecular Weight	67,000 (SDS-PAGE)
Isoelectric Point	4.4
Km	1.18 × 10⁻⁴ M
Optimum pH	7.0
pH Stability	5.0-7.0 (37°C, 30 min)
Optimum Temperature	45°C
Thermal Stability	up to 40°C (pH 7.0, 10 min)
Inhibitors	Ag⁺, Hg²⁺
Stabilizers	FAD, (NH₄)₂SO₄
Unit Definition	1 unit is defined as the enzyme quantity which oxidizes 1 μ mole of L-α-glycerophosphate per minute under the conditions described below.
Storage	24 months from the date of analysis when stored at 5°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 L-α-glycerophosphate per minute under the conditions described below.

Reagents

A. 4-Aminoantipyrine solution (5 mg/mL deionized water)
B. Phenol solution (50 mg/mL deionized water)
C. 0.2 M Phosphate buffer (KH2PO4-NaOH, pH 7.0)
D. 0.02 M Phosphate buffer (KH2PO4-NaOH, pH 7.0)
E. Peroxidase solution: Weigh 1000 units of Peroxidase and dissolved in 5 mL of chilled 0.2 M Phosphate buffer (C).
F. Aminoantipyrine-phenol solution: Mix approx. 30 mL of chilled 0.2 M Phosphate buffer (C), 1 mL of Peroxidase solution (E), 6 mL of 4-Aminoantipyrine solution (A) and 0.5 mL of Phenol solution (B), then fill up to 50 mL with chilled 0.2 M Phosphate buffer (C). (Can be used for 14 days in light-shielded and refrigerated storage)
G. Substrate solution: Weigh 36.7 g of disodium (+/-)-1-glycerophosphate n-hydrate and dissolve in approx. 40 mL of deionized water. Adjust the pH to 7.0 with 4N HCI and fill up to 100 mL with deionized water. Then, filtrate the solution with 0.45μm membrane filter. (Can be used 14 days if kept refrigerated)
H. Diluent: Weigh 66.0 g of ammonium sulfate and dissolved in approx. 800 mL of 0.02 M Phosphate buffer (D). Add 1.0 g of bovine serum albumin and dissolve, then fill up to 1000 mL with 0.02 M Phosphate buffer (D).
I. Enzyme solution: Weigh out L-alpha-Glycerophosphate Oxidase and dissolve in diluent (H). Enzyme solution should be prepared so that the value of ΔOD/minute becomes in the range of 0.020 ± 0.005.

Procedure

Pipette 1.5 mL of Aminoantipyrine-phenol solution (F), 1.5 mL of Substrate Solution (G) respectively into a quartz cell (d=10 mm) and keep at 37±0.5°C for 10 minutes. Then, pipette 0.1 mL of Enzyme solution (I) into the quartz cell and mix well immediately. Keep the reaction mixture at 37±0.5°C. Exactly at 1 minute and 3 minutes after the addition of Enzyme Solution (I), measure the absorbances of the reaction mixture at 505 nm (A1 and A3). As a blank, pipette Diluent (H) into another quartz cell (d= 10 mm) instead of Enzyme solution (I) and take the same procedure described above (Ab1 and Ab3).

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.