Lactate oxidase (LOX), CAS No.9028-72-2

CAS: 9028-72-2 Cat. No.: rp216190 EC Number: 232-841-6

GRADE & PURITY Suitable for molecular biology, BioReagent, EnzymoPure™, ≥90%(SDS-PAGE), ≥ 45 U/mg

Overview

Application
For the detection of lactic acid content.
Enzymatic properties
Source: Microorganism
Enzymology Committee Number: EC 1.1.3.2
Molecular weight: 42 kDa (SDS-PAGE)
Isoelectric point: pH 4.6
Km value: 7.5 × 10-4 M (L-Lactate)
Inhibitors:  Hg²⁺ , Pb²⁺ 
Optimum pH: 6.0-7.0                                                   Figure 1 
Optimum temperature: 50℃                                        Figure 2
pH stability: 6.0-8.5 (25℃,16h)                                    Figure 3 
Thermal stability: Stable below 50℃ (pH7.0, 30min)  Figure 4
Stability: -25 ~ -15℃ standing store for 12 months
More than 90% activity                                                Figure 5


Enzyme activity definition
Unit enzyme activity is defined as the amount of enzyme required to catalyze the production of 1μmol H2O2 per minute under the following conditions.
Assay method for activity
1. Principle

The amount of Quinoneimine dye produced by the reaction can be measured by spectrophotometer at 555 nm.
2. Definition of enzyme activity
Unit enzyme activity is defined as the amount of enzyme required to catalyze the production of 1 μmol H2O2 per minute under the following conditions.
3. Reagent preparation
Reagent I: 0.2 MpH 6.5 potassium phosphate buffer.
Reagent II: 1kU/mL peroxidase (POD) solution.
Reagent III: 50 mM4-AA solution.
Reagent IV: 0.5 MDL- lactic acid solution, pH6.5.
Reagent V: 50 mMTOOS solution.
Enzyme diluent: 10 mMpH7.0 potassium phosphate buffer containing 10 μM FAD.
Sample: Dilute the enzyme with enzyme diluent to 0.05-0.2U/mL.
Prepare the reaction mixture as follows:
Reagent I is 10 ml
Reagent II 0.25 mL
Reagent III 1.5 mL
Reagent IV is 5 mL
Reagent V 1.5 mL
Double steam water to 50 ml
4. Operation procedure
4.1 Add 1mL reaction mixture into 1mL colorimetric dish.
4.2 Preheat the reaction mixture at 37 °C for 5min.
4.3 Add 20μL of enzyme liquid to be measured and mix well.
4.4 The reaction is measured at 37 °C at 555 nm and the absorbance change (∆As) within 1min is recorded.
* Replace enzyme liquid with enzyme diluent, other steps are the same, the absorbance of the resulting solution is blank absorbance (∆Ab)
∆A=∆As-∆Ab
5. Vitality computing

1.020: total volume of reaction liquid (mL);
0.020: enzyme liquid volume (mL);
1: optical path length (cm);
1/2:1 mole of hydrogen peroxide to generate 1/2 mole of quinone imide dye;
df: dilution ratio;
C: Enzyme concentration (mg/mL);
39.2: Under standard reaction conditions, the millimolar absorption coefficient of the color group at 555 nm (cm2/μmol).

Specifications

Product Name

Lactate oxidase (LOX), CAS No.9028-72-2

Grade

BioReagent, Suitable for molecular biology, EnzymoPure™

Specifications & Purity

BioReagent, Suitable for molecular biology, EnzymoPure™, ≥90%(SDS-PAGE), ≥ 45 U/mg

CAS

9028-72-2

Molecule Type

Enzyme

Storage and Shipping

Concentration

≥ 45 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.

References

1. Yuanting Xie, Jinhua He, Wenqing He, Tayyaba Iftikhar, Chuangjie Zhang, Lei Su, Xueji Zhang.  (2024)  Enhanced Interstitial Fluid Extraction and Rapid Analysis via Vacuum Tube-Integrated Microneedle Array Device.  Advanced Science,      [PMID:38502884] [10.1002/advs.202308716]

2. Xiangling Li, Shantao Zheng, Mengyi He, Xinshuo Huang, Cheng Yang, Jingshan Mo, Jingbo Yang, Chengduan Yang, Huijiuan Chen, Xi Xie.  (2025)  Self-calibrating multiplexed microneedle electrode array for continuous mapping of subcutaneous multi-analytes in diabetes.  The Innovation,      [PMID:39991476] [10.1016/j.xinn.2024.100781]

3. Xiaotian Liu, Xiaoyun Chen, Wenjie Tao, Jiaxing Wang, Haotian Guo, Wenji Li, Zhankui Zhang, Xiaoguang Qiao.  (2025)  Meso-Hybridization of enzymes with silk fibroin and marked enhancement of flexible enzymic electrochemical sensing performance.  COLLOIDS AND SURFACES A-PHYSICOCHEMICAL AND ENGINEERING ASPECTS,      [PMID:] [10.1016/j.colsurfa.2025.139058]