Mahmoud, S., Ghaly, M., Abd-El Razik, M. (2023). Extraction, Purification and Characterization of Cholesterol Oxidase Enzyme Biosynthesized by Probiotic Lactiplantibacillus plantarum MF1. Suez Canal Veterinary Medical Journal. SCVMJ, (), -. doi: 10.21608/scvmj.2023.176182.1110
samar Mahmoud; Mohamed Farouk Ghaly; Mohamed Abd-El Razik. "Extraction, Purification and Characterization of Cholesterol Oxidase Enzyme Biosynthesized by Probiotic Lactiplantibacillus plantarum MF1". Suez Canal Veterinary Medical Journal. SCVMJ, , , 2023, -. doi: 10.21608/scvmj.2023.176182.1110
Mahmoud, S., Ghaly, M., Abd-El Razik, M. (2023). 'Extraction, Purification and Characterization of Cholesterol Oxidase Enzyme Biosynthesized by Probiotic Lactiplantibacillus plantarum MF1', Suez Canal Veterinary Medical Journal. SCVMJ, (), pp. -. doi: 10.21608/scvmj.2023.176182.1110
Mahmoud, S., Ghaly, M., Abd-El Razik, M. Extraction, Purification and Characterization of Cholesterol Oxidase Enzyme Biosynthesized by Probiotic Lactiplantibacillus plantarum MF1. Suez Canal Veterinary Medical Journal. SCVMJ, 2023; (): -. doi: 10.21608/scvmj.2023.176182.1110
Extraction, Purification and Characterization of Cholesterol Oxidase Enzyme Biosynthesized by Probiotic Lactiplantibacillus plantarum MF1
Articles in Press, Accepted Manuscript, Available Online from 09 February 2023
1Department of Botany and Microbiology, Faculty of science, Suez Canal University, Egypt
2Botany and Microbiology Department ,Faculty of Science,Suez Canal University
Abstract
This work concerned with purification and characterization of cholesterol oxidase enzyme produced by probiotic bacteria isolated from raw cow’s milk. The bacterial isolate was genotyped as Lactiplantibacillus plantarum by 16S rRNA and deposited with GenBank under accession number MW242720. The L. plantarum MF1 cholesterol oxidase enzyme was purified using acetone and sephadex G 100 column chromatography, yielding enzyme recovery and purification folds of 19.3% and 4.2, respectively. The purified cholesterol oxidase appeared as a single protein band at 51 kDa using SDS-PAGE technique. For maximum activity of the purified CHO enzyme the optimal conditions were a 30 minutes incubation period at 30°C, a substrate concentration of 150 μL in the reaction mixture, and an enzyme concentration of 100 μL in the reaction mixture. The purified cholesterol oxidase was activated by metal ions (CuSO4), whereas it was inhibited by CaCl2, MgSO4, ZnSO4, and MnCl2. After storage at -20°C for 4 days, the purified enzyme retained 57% of its initial enzyme activity. The purified enzyme demonstrated an active effect on serum human cholesterol samples, resulting in a 29% reduction.
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