Purification and characterisation of the extracellular cholesterol oxidase enzyme from Enterococcus hirae

BMC Microbiology

Shimaa M. Ibraheim¹  Wesam A. Hassanein¹  Hany M. Yehia² 

[1]Department of Botany (Microbiology), Faculty of Science, Zagazig University, Zagazig, Egypt

[2]Food Science and Nutrition, Faculty of Home Economics, Helwan University, Helwan, Egypt

关键词: ChoX molecular weight;    Food samples;    Decomposition;    Enterococcus hirae;    Cholesterol oxidase enzyme (ChoX);   

Others  :  1227560
DOI  :  10.1186/s12866-015-0517-2

 received in 2015-02-21, accepted in 2015-09-10,  发布年份 2015

【 摘 要 】

Background

Recently many efforts are being carried out to reduce cholesterol in foods. Out of the 50 selected isolates that were tested using the agar well diffusion method to assess their ability to decompose cholesterol, 24 bacterial isolates were screened based on their cholesterol-decomposition ability in liquid media.

Results

The bacterial isolate that displayed the highest cholesterol oxidase activity was identified as Enterococcus hirae. The maximal growth and cholesterol decomposition were achieved with a 1-day incubation under static conditions at 37 °C in cholesterol basal medium adjusted to pH 7 supplemented with 1 g/l cholesterol as the substrate, no additional carbon or nitrogen sources and 0.5 % CaSO ₄ . The cholesterol oxidase enzyme (ChoX) produced by E. hirae was extracted at an (NH ₄ ) ₂ SO ₄saturation level of 80 % and purified with 79 % yield, resulting in 2.3-fold purification. The molecular weight of (ChoX) was 60 kDa. The optimal conditions required for the maximal activity of the purified COD enzyme produced by E. hirae were 30 min, 40 °C, pH 7.8, substrate concentration of 1 g/l and 200 ppm of MgCl ₂ . The enzyme maintained approximately 36 % and 58.5 % of its activity after 18 days of storage at 4–8 °C. Also, the enzyme loss its activity by gradual thermal treatment, but it maintained 58.5 % of its activity at 95 °C for 2 hr.

Conclusions

E. hirae Mil-31 isolated from milk had a great capacity to decompose cholesterol in basal medium supplemented with cholesterol under its optimal growth conditions. Decomposition process of cholesterol by this strain results from its production of cholesterol oxidase enzyme (ChoX). The highest specific enzyme activity and highest purification fold of purified enzyme were achieved after using Sephadex G-100.

【 授权许可】

   
2015 Yehia et al.

【 图 表 】

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