Preparation and Properties of a Cholesterol Oxidase from Nocardia sp. and Its Application to the Enzymatic Assay of Total Cholesterol in Serum
Получение и свойства холестеролоксидазы из Nocardia sp. и ее применение для ферментативного определения общего холестерина в сыворотке
1973-12-01
SCID: 54.1/d2yqenj8
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Nocardia sp.cholesterol oxidasehydrogen peroxide measurementserum total cholesterol assayxylenol orange
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Abstract (AI)
Abstract I describe the characterization, extraction, and purification of a cholesterol:oxygen oxidoreductase (EC 1.1.3.6) from Nocardia sp. This enzyme catalyzes oxidation of cholesterol to Δ4-choIestenone, with production of hydrogen peroxide. It is very stable, active over a wide pH range, and has a Km of 1.4 x 10-5 mol/ liter. It is highly specific for Δ4- or Δ5-3β-hydroxycholestanes, and may be applied to the assay of serum total cholesterol. In the procedure presented here, hydrogen peroxide is measured by reaction with quadrivalent titanium and xylenol orange. This constitutes a one-enzyme assay with stable reagents, which does not require protein precipitation and is not subject to interference from hemoglobin or bilirubin.
Key Findings
1
A cholesterol oxidase from Nocardia sp. was characterized, extracted, and purified for analytical use.
2
A one-enzyme serum total-cholesterol assay measures generated hydrogen peroxide using quadrivalent titanium and xylenol orange.
3
It is highly specific for Δ4- or Δ5-3β-hydroxycholestanes.
4
The assay uses stable reagents, requires no protein precipitation, and is not affected by hemoglobin or bilirubin interference.
5
The enzyme is highly stable, active across a wide pH range, and has a Km of 1.4 × 10^-5 mol/L.
6
The enzyme oxidizes cholesterol to Δ4-cholestenone while producing hydrogen peroxide.
Research Object
Cholesterol oxidase from Nocardia sp. and its application in serum total-cholesterol measurement
Research Subject
Enzyme characterization, catalytic specificity and stability, and the performance of a one-enzyme hydrogen-peroxide assay for serum total cholesterol
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1973-12-01
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