Abstract

BackgroundDue to broad range of clinical and industrial applications of cholesterol oxidase, isolation and screening of bacterial strains producing extracellular form of cholesterol oxidase is of great importance.ResultsOne hundred and thirty actinomycete isolates were screened for their cholesterol oxidase activity. Among them, a potential culture, strain NEAE-42 is displayed the highest extracellular cholesterol oxidase activity. It was selected and identified as Streptomyces cavourensis strain NEAE-42. The optimization of different process parameters for cholesterol oxidase production by Streptomyces cavourensis strain NEAE-42 using Plackett–Burman experimental design and response surface methodology was carried out. Fifteen variables were screened using Plackett–Burman experimental design. Cholesterol, initial pH and (NH4)2SO4 were the most significant positive independent variables affecting cholesterol oxidase production. Central composite design was chosen to elucidate the optimal concentrations of the selected process variables on cholesterol oxidase production. It was found that, cholesterol oxidase production by Streptomyces cavourensis strain NEAE-42 after optimization process was 20.521U/mL which is higher than result obtained from the basal medium before screening process using Plackett-Burman (3.31 U/mL) with a fold of increase 6.19.ConclusionsThe cholesterol oxidase level production obtained in this study (20.521U/mL) by the statistical method is higher than many of the reported values.

Highlights

  • Due to broad range of clinical and industrial applications of cholesterol oxidase, isolation and screening of bacterial strains producing extracellular form of cholesterol oxidase is of great importance

  • Cholesterol oxidase (3ß-hydroxysteroid oxidase, EC 1.1.3.6) is a flavin adenine dinucleotide (FAD)-dependent enzyme belongs to the family of oxidoreductases, those acting on the CH-OH group of donor with oxygen as acceptor that in most cases catalyzes the oxidation and isomerization of cholesterol using oxygen as an electron acceptor to form 4-cholesten-3one and hydrogen peroxide [1]

  • The total of one hundred and thirty morphologically different actinomycete strains were qualitative screened for their cholesterol oxidase activity using colony staining method, formation of pink zones around the colonies indicated the presence of cholesterol oxidase activity

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Summary

Introduction

Due to broad range of clinical and industrial applications of cholesterol oxidase, isolation and screening of bacterial strains producing extracellular form of cholesterol oxidase is of great importance. Cholesterol oxidase enzyme is simple, specific, and profoundly sensitive; utilized for the clinical determination of cholesterol levels in serum, HDL, or LDL for the evaluation of atherosclerotic illnesses and different lipid problems and for assessing the danger of thrombosis [2]. It has been proposed that bacterial degradation of cholesterol in cholesterol containing foods might be beneficial for human health [4]. Cholesterol oxidases can be used to produce a precursor for chemical synthesis of steroid hormones [1] and to degrade of dietary cholesterol in foods [4]

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