Enzymatic assay for cholesterol ester hydrolase activity.

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Enzymatic assay for cholesterol ester hydrolase activity.

A rapid and accurate method is described for the assay of cholesterol ester hydrolase (CEH) activity. Aliquots of the enzyme-substrate incubation mixture are extracted into isopropanol. The free cholesterol concentration in each extract is determined enzymatically using a single aqueous reagent containing cholesterol oxidase and peroxidase. The free cholesterol remaining after the cholesterol e...

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Studies on the cholesterol ester hydrolase of Trogoderma (Coleoptera).

The enzyme cholesterol ester hydrolase (EC 3.1.1.13) was detected in the larvae of the khapra beetle, Trogoderma granarium (Everts). The pH and temperature optima for the enzyme were 6.6 degrees and 37 degrees C respectively. The mol.wt. of the enzyme was 76000-80000. The enzyme was equally effective in hydrolysing cholesteryl acetate, stearate and oleate. Cholesterol derivatives, namely the ch...

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The hippurate hydrolase enzyme of Campylobacter jejuni was expressed in Escherichia coli as a six-histidine-tagged fusion protein. The purified recombinant enzyme was characterized to gain an understanding of the structure and activity of the hippurate hydrolase. The recombinant enzyme had a native molecular mass of 193+/- 11 kDa a reduced molecular mass of 42.4+/- 0.8 kDa, and possessed 1.98+/...

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Simple enzymatic assay for determining cholesterol concentrations in bile.

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 ...

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Hepatic overexpression of cholesteryl ester hydrolase enhances cholesterol elimination and in vivo reverse cholesterol transport.

Neutral cholesteryl ester hydrolase (CEH)-mediated hydrolysis of cellular cholesteryl esters (CEs) is required not only to generate free cholesterol (FC) for efflux from macrophages but also to release FC from lipoprotein-delivered CE in the liver for bile acid synthesis or direct secretion into the bile. We hypothesized that hepatic expression of CEH would regulate the hydrolysis of lipoprotei...

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ژورنال

عنوان ژورنال: Journal of Lipid Research

سال: 1978

ISSN: 0022-2275

DOI: 10.1016/s0022-2275(20)40706-0