Performance characteristics of cholesterol oxidase for kinetic determination of total cholesterol

Performance characteristics of cholesterol oxidase for kinetic determination of total cholesterol.

The enzymatic method for cholesterol determination can use either an endpoint or a kinetic method. Not much is known concerning the properties (K(m) and V(max)) of the commercial enzyme for the kinetic method. We measured the K(m) and V(max) of Brevibacterium, Streptomyces, Pseudomonas fluorescens, and Cellulomonas cholesterol oxidase. Brevibacterium gave the highest K(m) value (230.3 x 10(-4) ...

Cholesterol oxidase-based determination, by continuous-flow analysis, of total and free cholesterol in serum.

We describe a continuous-flow, automated determination of total cholesterol in serum, which is based on enzymatic hydrolysis of cholesterol esters, oxidation of cholesterol by cholesterol oxidase, and colorimetric measurement of liberated perioxide with 4-aminoantipyrine, phenol, and peroxidase. Free cholesterol is determined with the same AutoAnalyzer II manifold and reagents, except that chol...

A simple method for the determination of serum cholesterol

STREPTOMYCES FRADIAE, A POTENT PRODUCER OF CHOLESTEROL OXIDASE

Thirteen species of Streptomycetes, Arthrobacters and Mycobacterium were studied for the determination of a potential cholesterol oxidase producer. Only Streptomyces fradiae and Streptomyces rishiriences were found to be capable of producing this enzyme. Streptomyces fradiae was selected for the production of cholesterol oxidase and cultured in different media to find optimum media and con...

Enzymatic determination of total serum cholesterol.

An enzymatic method is described for determination of total serum cholesterol by use of a single aqueous reagent. The method requires no prior treatment of sample and the Calibration curve is linear to 600 mg/dl. Cholesterol esters are hydrolyzed to free cholesterol by cholesterol ester hydrolase (EC 3.1.1.13). The free cholesterol produced is oxidized by cholesterol oxidase to cholest-4-en-3-o...