The production of pharmaceuticals in a ‘green’ way is a concern in the recent years. In this work, we have examined an oxidation reaction in a continuous flow supercritical CO2 system using individual CLEA (Cross Linked Enzyme Aggregate) of cholesterol oxidase and combi-CLEA of cholesterol oxidase and catalase. Besides the activity studies, the research was aimed to be extended to kinetic studi...

High-pressure EPR spectroscopy indicates that cholesterol forms aggregates in supercritical carbon dioxide. In pure carbon dioxide, changes in cholesterol aggregate size or packing structure are observed with changing pressure. Near the critical point of carbon dioxide, cholesterol solubility is too low to permit significant aggregation, and monomeric cholesterol is observed. Addition of small ...

The NADPH oxidase Nox2, a multi-subunit enzyme complex comprising membrane and cytosolic proteins, catalyzes a very intense production of superoxide ions O2(•-), which are transformed into other reactive oxygen species (ROS). In vitro, it has to be activated by addition of amphiphiles like arachidonic acid (AA). It has been shown that the membrane part of phagocyte NADPH oxidase is present in l...

Site-directed mutagenesis was used to identify key amino acid residues of the cholesterol oxidase from Streptomyces sp., which catalyzes the oxidation of cholesterol and the isomerization of 5-cholesten-3-one. Eight mutant enzymes were constructed and the following amino acid substitutions were identified: N318A, N318H, E356A, E356D, H441A, H441N, N480A and N480Q. Mutants N318A and N318H retain...

Cholesterol oxidase (EC1.1.3.6; CHO) is an enzyme, which catalyzes the oxidation of cholesterol and converts 5cholesten-3β-ol into 4cholesten-3-one. The objective of this study is to isolate extracellular cholesterol oxidase (CHO) producing microorganisms to obtain an abundant source of cholesterol oxidase (CHO) for industrial and medicinal needs. Cholesterol oxidase producing bacteria were iso...

We use bilirubin oxidase (EC 1.3.3.5) to remove interference by bilirubin in the assay of cholesterol concentration in bile by standard enzymatic methods. Samples are treated for 10 min with nonlimiting amounts of bilirubin oxidase to form biliverdin from bilirubin before the reagent for cholesterol is added. The relatively small interference by biliverdin is easily eliminated by use of sample ...

Recent studies have indicated the importance of cholesterol-rich membrane lipid rafts (LRs) in oxidative stress-induced signal transduction. Reduced nicotinamide-adenine dinucleotide phosphate (NADPH) oxidases, the major sources of reactive oxygen species, are implicated in cardiovascular diseases, including hypertension. We tested the hypothesis that NADPH oxidase subunits and activity are reg...

Cholesterol oxidase (COX), a bi-functional FAD-containing microbial enzyme belongs to the family oxidoreductases. COX catalyses the oxidation of cholesterol into 4-cholesten-3-one. In recent time, cholesterol oxidase has received great attention due to its wider use in clinical (determination of serum cholesterol) laboratories practice and in the biocatalysis for the production of a number of s...