C5896
Choline Oxidase from Alcaligenes sp.
lyophilized powder, ≥10 units/mg solid
Synonym(s):
Choline: oxygen 1-oxidoreductase
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About This Item
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form
lyophilized powder
specific activity
≥10 units/mg solid
mol wt
~95 kDa
technique(s)
cell based assay: suitable
suitability
suitable for assay of choline
suitable for molecular biology
storage temp.
−20°C
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Application
Choline oxidase from Alcaligenes sp. has been used:
- in a study to investigate a choline biosensor constructed with a chitinous membrane and its application in measuring cholinesterase inhibitory activities
- has also been used in spectroscopic studies on the photoreaction of choline oxidase with covalently bound flavin.
- as a component of phosphate buffer saline (PBS) buffer for measurement of choline acetyltransferase (ChAT) activity in brain tissue homogenates
- to measure the choline generated by autotaxin (ATX) by ATX activity assay and phospholipase-D (PLD) by PLD activity assay
Biochem/physiol Actions
Choline oxidase catalyzes the four-electron oxidation of choline to glycine-betaine, with betaine-aldehyde as the intermediate and molecular oxygen as th eprimary electron acceptor. The enzyme can also accept betaine-aldehyde as a substrate. This allows the study of the reaction mechanism for the conversion of choline to the aldehyde intermediate, & of betaine-aldehyde to glycine-betaine. The enzyme is a flavoprotein with a molecular weight of approx. 72,000 Da (gel filtration) or 66,000 Da (SDS gel electrophoresis).
Physical properties
Isoelectric point:4.1 ± 0.1
Michaelis constants:2.84 x 10¯3M (Choline), 5.33 x 10¯3M (Betaine aldehyde)
Structure:One mol of FAD is covalently bound to mol of the enzyme
Inhibitors:p-Chloromercuribenzoate, Cu++, Co++, Hg++, Ag+
Optimum pH:8.0 – 8.5
Optimum temp:40 – 45°C
pH Stability:pH 7.0 – 9.0 (30°C, 2hr)
Thermal stability:Below 37°C (pH 7.5, 10min)
Michaelis constants:2.84 x 10¯3M (Choline), 5.33 x 10¯3M (Betaine aldehyde)
Structure:One mol of FAD is covalently bound to mol of the enzyme
Inhibitors:p-Chloromercuribenzoate, Cu++, Co++, Hg++, Ag+
Optimum pH:8.0 – 8.5
Optimum temp:40 – 45°C
pH Stability:pH 7.0 – 9.0 (30°C, 2hr)
Thermal stability:Below 37°C (pH 7.5, 10min)
Unit Definition
One unit will form 1 μmole of H2O2 with oxidation of 1 μmole of choline to betaine aldehyde per min at pH 8.0 at 37 °C. Note: During the conversion of choline to betaine by choline oxidase, 2 μmoles of H2O2 are produced for every μmole of choline.
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Description
Pricing
Signal Word
Danger
Hazard Statements
Precautionary Statements
Hazard Classifications
Resp. Sens. 1
Storage Class Code
11 - Combustible Solids
WGK
WGK 3
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Personal Protective Equipment
dust mask type N95 (US), Eyeshields, Gloves
Certificates of Analysis (COA)
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Bo-Chuan Hsieh et al.
Journal of pharmaceutical and biomedical analysis, 45(4), 673-678 (2007-03-03)
An amperometric flow-injection choline biosensor was assembled utilizing natural chitinous membrane as the supporting material for biocatalyst immobilization, and the membrane was purified from Taiwanese soldier crab, Mictyris brevidactylus. The chitinous membrane (<50.0 microm in thickness) was covalently immobilized with
M Ohta-Fukuyama et al.
Journal of biochemistry, 88(1), 197-203 (1980-07-01)
Choline oxidase from Alcaligenes sp. catalyzed the oxidation of choline and betaine aldehyde to betaine with concomitant consumption of oxygen and production of hydrogen peroxide. The values of Km for choline and betaine aldehyde were 0.87 and 6.2 mM, respectively.
Shengyuan Deng et al.
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Giovanni Gadda
Biochimica et biophysica acta, 1646(1-2), 112-118 (2003-03-15)
Choline oxidase catalyzes the four-electron oxidation of choline to glycine-betaine, with betaine-aldehyde as intermediate and molecular oxygen as primary electron acceptor. The enzyme is capable of accepting betaine-aldehyde as a substrate, allowing the investigation of the reaction mechanism for both
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