C1235
Cholesterol Oxidase microbial
recombinant, lyophilized powder, ≥10 units/mg protein
Synonym(s):
Cholesterol: oxygen oxidoreductase
Quality Level
form
lyophilized powder
specific activity
≥10 units/mg protein
mol wt
55 kDa
solubility
50 mM potassium phosphate buffer, pH 7.0: soluble
storage temp.
−20°C
Application
Cholesterol oxidase is used to determine serum cholesterol. The enzyme also finds application in the microanalysis of steroids in food samples and in distinguishing 3-ketosteroids from 3b-hydroxysteroids. Transgenic plants expressing cholesterol oxidase are being investigated in the fight against the cotton boll weevil. CHOD has also been used as a molecular probe to elucidate cellular membrane structures.
Biochem/physiol Actions
Cholesterol oxidase (CHOD) is a monomeric flavoprotein containing FAD that catalyzes the first step in cholesterol catabolism. This bifunctional enzyme oxidizes cholesterol to cholest-5-en-3-one in an FAD-requiring step. This is subsequently isomerized to cholest-4-en-3-one with the release of H2O2. Optimum pH of the enzyme is 7.0. Hg2+, Ag+, ionic detergents inhibit the enzyme activity.
Preparation Note
Dissolves in cold 50 mM potassium phosphate buffer, pH 7.0. Solution is to be prepared just before use.
Other Notes
One unit will convert 1.0 μmol of cholesterol to 4-cholesten-3-one per minute at 37 °C and pH 7.0 in a peroxidase linked system.
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
Regulatory Information
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Bruno M Castro et al.
The Journal of biological chemistry, 284(34), 22978-22987 (2009-06-13)
A uniquely sensitive method for ceramide domain detection allowed us to study in detail cholesterol-ceramide interactions in lipid bilayers with low (physiological) ceramide concentrations, ranging from low or no cholesterol (a situation similar to intracellular membranes, such as endoplasmic reticulum)
Christophe A Marquette et al.
Analytical and bioanalytical chemistry, 390(1), 155-168 (2007-10-03)
The present review draws a general picture of the bioanalytical applications of electro-chemiluminescent reactions (ECL). Only the two main ECL reactions-i.e. the luminol-based and Ru(bpy)(3)(2+)-based reactions-are considered for application in the fields of enzyme biosensors, immunochemical biosensors, DNA biosensors, and
Porntip H Lolekha et al.
Clinica chimica acta; international journal of clinical chemistry, 339(1-2), 135-145 (2003-12-23)
Cholesterol oxidase is used for the determination of serum cholesterol. It can be derived from Streptomyces, Pseudomonas fluorescens, Cellulomonas, and Brevibacterium. This study compared the performance characteristics of four enzymes in the endpoint cholesterol determination. Using the Mega analyzer, we
Kwang-wook Ahn et al.
Biochemistry, 43(3), 827-836 (2004-01-21)
We investigated the dependence of cholesterol oxidase catalytic activity and membrane affinity on lipid structure in model membrane bilayers. The binding affinities of cholesterol oxidase to 100-nm unilamellar vesicles composed of mixtures of DOPC or DPPC and cholesterol are not
Marco P C Marques et al.
New biotechnology, 29(2), 227-234 (2011-10-20)
The use of microchannel reactor based technologies within the scope of bioprocesses as process intensification and production platforms is gaining momentum. Such trend can be ascribed a particular set of characteristics of microchannel reactors, namely the enhanced mass and heat
Protocols
Enzymatic activity assay for cholesterol oxidase, excluding specific product numbers, providing guidelines for accurate cholesterol oxidase assays.
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