| Uses |
Cholesterol oxidases are part of a unique class of enzymes that are soluble proteins although they interact with highly insoluble substrates. The natural substrate, cholesterol, is an important membrane component exhibiting low solubility in the aqueous medium of the cell, hence the enzyme must interact with the lipid bilayer in order to for the substrate to partition out of the membrane and undergo oxidation at the enzyme active site. In this regard cholesterol oxidase is an interfacial enzyme as it binds transiently to the membrane surface during catalysis and can only access the substrate from the membrane phase. Other examples of interfacial enzymes include phospholipases (Berg et al., 2001; Gelb et al., 1995; Jain et al., 1995).
Cholesterol oxidase has a number of important commercial applications. Initial studies on the enzyme focused on its use for the determination of cholesterol in serum, HDL or LDL (see review, Smith and Brooks, 1976). Serum cholesterol levels are determined using a three-enzyme assay, including cholesterol esterase, cholesterol oxidase and peroxidase (Richmond, 1973; Allain et al., 1974; Richmond, 1976). More recently electrochemical biosensors with immobilized cholesterol oxidase have been employed to determine cholesterol levels in serum or food (Arya et al., 2007; Basu et al., 2007; Vidal et al., 2004). Determining the serum cholesterol levels is critical for the assessment of a variety of diseases including hypercholesterolemia, coronary heart disease and lipid disorders for estimating the risk of thrombosis and myocardial infarction.
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