Production,optimization, purification,characterization, and anti-cancer application of extracellular L-glutaminase produced from the marine bacterial isolate |
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Authors: | Hanaa Orabi Eman Abdelkhalek Nagwa Sidkey |
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Institution: | 1. Protein Research Department, Genetic Engineering and Biotechnology Research Institute, City of Scientific Research and Technology Applications (SRTA-City), Alexandria, Egypt;2. Faculty of Science, Botany and Microbiology Department, Al-Azhar University, Cairo, Egypt;3. Faculty of Science, Botany and Microbiology Department, Al-Azhar University, Cairo, Egypt |
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Abstract: | AbstractL-glutaminase from bacterial sources has been proven to be effective and economical agents in cancer therapy, food industry and high-value chemicals like threonine. In the present study, a newly isolated bacterial strain was potentially producing extracellular L-glutaminase, it identified as Bacillus subtilis OHEM11 (MK389501) using the 16S rRNA gene. L-glutaminase production optimized and the optimum factors for production under submerged fermentation were at pH 6.5–7.0 and 35?°C after 28?hr using rhamnose and glutamine as carbon and nitrogen sources, respectively, while bagasse was the best inducer for the production under solid-state fermentation. Ethanol precipitation and ion-exchange chromatography using QFF are the purification steps. L-glutaminase was purified to 2-fold with specific activity 89.78?U/mg and its molecular weight about 54.8?kDa with the alkaline property of the enzyme makes it clear having carcinostatic property; maximum enzyme activity at pH 8.2 and 40?°C and retained about 90% activity for 1?hr. The cytotoxicity effect of L-glutaminase indicated a significant safety on Vero cells with high anticancer activity against NFS-60, HepG-2, and MCF-7 cancer cell lines. The outcomes demonstrated that L-glutaminase could be applied in many biotechnological applications such as pharmaceutical and food processing. |
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Keywords: | Apoptosis characterization L-glutaminase optimization purification |
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