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A6941

Sigma-Aldrich

Alcohol Oxidase from Candida boidinii

lyophilized powder, 5-15 units/mg protein

Synonym(s):

AOD1, AOX, Alcohol:oxygen oxidoreductase

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About This Item

CAS Number:
9073-63-6
Enzyme Commission number:
1.1.3.13 (BRENDA, IUBMB)
EC Number:
232-971-3
MDL number:
MFCD00130452
UNSPSC Code:
12352204
NACRES:
NA.54

biological source

fungus (Candida boidinii)

Quality Level

200

form

lyophilized powder

specific activity

5-15 units/mg protein

mol wt

octomer 600 kDa by sedimentation equilibrium

solubility

100 mM potassium phosphate, pH 7.5: soluble 1.0 mg/mL at 25 °C (Cold)

storage temp.

−20°C

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General description

Research area: Cell Signaling

Alcohol Oxidase (AOX) is a homo-octamer composed of eight flavin adenine dinucleotide (FAD) cofactors and belongs to the glucose-methanol-choline (GMC) family of oxidoreductases. The AOX1 and AOX2 genes are responsible for encoding AOX. Alcohol oxidase is primarily localized in the peroxisome but is also found in the cytoplasm. Alcohol oxidase is a 600 kDa homooctomeric flavoprotein with eight equal 74 kDa subunits; each containing a flavin adenine dinucleotide (FAD) molecule.

Application

Alcohol oxidase has been used:
  • to catalyze the oxidation of short-chain, primary, aliphatic alcohols to their respective aldehydes .
  • to study methanol metabolism in yeasts, such as Candida, Pichia, and Hansenula.
  • to study protein translocation into peroxisomes.
  • for the determination of ethanol concentration in alcoholic drinks using enzymatic assay.
  • in development of enzyme electrode for the determination of alcohols.

Biochem/physiol Actions

Alcohol oxidase has the highest affinity for methanol. The affinity decreases with increasing chain length of the alkyl (R) group. The enzyme shows little activity toward secondary, tertiary, or aromatic alcohols; or aliphatic alcohols with a chain length of more than 5 carbons. The pH range for activity of this product is 6.5-8.5, with the optimum pH being 7.5. Alcohol oxidases facilitate the conversion of alcohol into carbonyl compounds while producing hydrogen peroxide. It is also the first enzyme in the yeast methanol utilization pathway.

Unit Definition

One unit will oxidize 1.0 μmole of methanol to formaldehyde per min at pH 7.5 at 25 °C.

Physical form

Contains potassium phosphate buffer salts, DTE, and stabilizer

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)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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H R Waterham et al.
The Journal of cell biology, 139(6), 1419-1431 (1998-02-12)
Alcohol oxidase (AOX), the first enzyme in the yeast methanol utilization pathway is a homooctameric peroxisomal matrix protein. In peroxisome biogenesis-defective (pex) mutants of the yeast Pichia pastoris, AOX fails to assemble into active octamers and instead forms inactive cytoplasmic
Erol Akyilmaz et al.
Talanta, 61(2), 113-118 (2008-10-31)
An amperometric biosensor based on catalase enzyme for alcohol determination was developed. To construct the biosensor catalase was immobilized by using gelatin and glutaraldehyde on a Clark type dissolved oxygen (DO) probe covered with a teflon membrane which is sensitive
Alcohol oxidase from Candida boidinii.
H Sahm et al.
Methods in enzymology, 89 Pt D, 424-428 (1982-01-01)
B Vinet
Clinical chemistry, 33(12), 2204-2208 (1987-12-01)
This method for the specific determination of methanol in serum is based on the following two reactions: (formula; see text) Alcohol oxidase is not specific: it converts all lower alcohols to their corresponding aldehydes; however, formaldehyde dehydrogenase is specific and
H Gülce et al.
Biosensors & bioelectronics, 17(6-7), 517-521 (2002-04-18)
A new enzyme electrode for the determination of alcohols was developed by immobilizing alcohol oxidase in polvinylferrocenium matrix coated on a Pt electrode surface. The amperometric response due to the electrooxidation of enzymatically generated H(2)O(2) was measured at a constant
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Protocols

Enzymatic Assay of Alcohol Oxidase (EC 1.1.3.13)

To measure alcohol oxidase activity, this assay uses 2,2′-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) and a continuous spectrophotometric rate determination at 405 nm.

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