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C5896

Sigma-Aldrich

Choline Oxidase from Alcaligenes sp.

lyophilized powder, ≥10 units/mg solid

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Synonym(s):
Choline: oxygen 1-oxidoreductase
CAS Number:
Enzyme Commission number:
EC Number:
MDL number:
NACRES:
NA.54

form

lyophilized powder

Quality Level

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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This Item
C4405C8868C1235
vibrant-m

C1235

Cholesterol Oxidase microbial

specific activity

≥10 units/mg solid

specific activity

8-20 units/mg solid

specific activity

≥50 units/mg protein

specific activity

≥10 units/mg protein

suitability

suitable for assay of choline, suitable for molecular biology

suitability

-

suitability

-

suitability

-

form

lyophilized powder

form

lyophilized powder

form

lyophilized powder

form

lyophilized powder

mol wt

~95 kDa

mol wt

71 kDa by SDS-PAGE, 83 kDa by gel filtration

mol wt

64 kDa

mol wt

55 kDa

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

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)

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.

pictograms

Health hazard

signalword

Danger

hcodes

Hazard Classifications

Resp. Sens. 1

Storage Class

11 - Combustible Solids

wgk_germany

WGK 3

flash_point_f

Not applicable

flash_point_c

Not applicable

ppe

Eyeshields, Gloves, type N95 (US)


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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.
S Ikuta et al.
Journal of biochemistry, 82(6), 1741-1749 (1977-12-01)
Choline oxidase was purified from the cells of Arthrobacter globiformis by fractionations with acetone and ammonium sulfate, and column chromatographies on DEAE-cellulose and on Sephadex G-200. The purified enzyme preparation appeared homogeneous on disc gel electrophoresis. The enzyme was a
S Pundir et al.
Analytical biochemistry, 427(1), 26-32 (2012-05-09)
A bienzymatic choline biosensor was constructed by coimmobilizing acetylcholinesterase (AChE) and choline oxidase (ChO) onto nanocomposite of carboxylated multiwalled carbon nanotubes (c-MWCNTs) and zirconium oxide nanoparticles (ZrO(2)NPs) electrodeposited on the surface of a glassy carbon electrode (GCE) and using it
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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