Implementation of cellulomonas cholesterol oxidase for total serum cholesterol determination by the endpoint method

A simple method for the determination of serum cholesterol

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...

Endpoint colorimetric method for assaying total cholesterol in serum with cholesterol dehydrogenase.

BACKGROUND Various methods are available to measure serum cholesterol concentrations. Of these, the cholesterol ester hydrolase (CEH)-cholesterol oxidase-peroxidase chromogenic method is widely used. However, this method has the disadvantage of interference by reducing substances. We developed and evaluated an endpoint assay for serum cholesterol, based on a CEH-cholesterol dehydrogenase (CDH)-...

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...

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) ...