Screening and production of bacterial L-glutaminase as an anticancer agent from different soil environments in Egypt | ||
| Middle East Journal of Medical Genetics | ||
| Volume 12, Issue 1, January 2023, Pages 1-12 PDF (1.27 M) | ||
| Document Type: Original Article | ||
| DOI: 10.21608/mxe.2024.228814.1006 | ||
| Author | ||
| Mohammed M Kassab* | ||
| Instructor of microbiology,immunology,faculty of pharmacy,Cairo university,Egypt Specialist of pharmacology, toxicology and clinical pharmacy. | ||
| Abstract | ||
| Background: Breast cancer and other neoplasms like acute lymphocytic leukaemia pose a serious threat to public health worldwide. Treatment for malignancies including acute lymphocytic leukaemia and breast cancer requires enzymes that break down L-glutamine. An amidohydrolase called L-glutaminase can be an effective chemotherapeutic tool for treating a variety of cancers. Aim of Study: This is a screening experimental representation was conducted to study and industry of bacterial L-glutaminase as an anticancer substance from several Egyptian soil regions. Patients and Methods: On mineral L-glutamine agar selective medium (MGA), a few bacterial isolates were examined in the current investigation to see if they produced L-glutaminase. The most effective positive bacterial isolates generating L-glutaminase, however, were identified using morphological and biochemical testing. Molecular detection using the direct Southern blotting approach was also used to identify the main positive isolate that produces L-glutaminase. L-glutaminase synthesis by bacteria was evaluated for its characteristics. Using an MTT test, the anticancer activity was evaluated. Results: Exclusive bacterial isolates that used L-glutamine as their sole source of metabolic nitrogen exhibited favorable growth on MGA. PH 7.4 and a temperature of 37oC were the ideal environmental and physiological conditions for developing positive bacterial isolates. From the soil samples taken from various soil conditions in Egypt, the morphological and biochemical analyses showed that Bacillus cereus 14579 was the main positive bacterial generating L-glutaminase isolate. The produced L-glutaminase has shown excellent bioavailability and effectiveness as an anticancer therapy. The yield [productivity] was 5.2 U/ml during the original manufacture from MGA, but it increased to 42.96 U/ml by bacterial recombinant DNA manufacturing. L-glutaminase was purified, yielding final enzyme recovery of 55 1.23%, total activity 12,990±19.76 (U), specific activity 384.66±8.92 (U/mg of protein), and purification fold 2±2.99. Furthermore, the L-glutaminase activity was raised by 19%, 23%, 15%, and 9%, respectively, by the enzymatic activators Mn2+, K+, Na+, and Fe3+. It demonstrated strong DPPH (2, 2-diphenyl-2-picryl-hydroxyl) scavenging activity (IC50= 61 μg/ml) and anticancer activity (IC50= 40.72, 9.7, 7.39, 20.61, 51.28, and 11.55 μg/ml, respectively) against human liver (HepG-2), colon (HCT-116), breast (MCF-7), lung (A-549), lymphocytic (CCL-120), and cervical (Hela) cancer cell lines. A stronger affinity for its substrate was shown by the kinetic parameters of Km and Vmax, which were 13.2 10-5 M and 119.86 μmol/ml/min, respectively. Conclusion: Acute lymphocytic leukemia and breast cancer are examples of auxotrophic malignancies for which L-glutamine serves as the only metabolic source, and L-glutaminase generated by Bacillus cereus was an appropriate enzyme in the therapy of these diseases. | ||
| Keywords | ||
| Acute lymphocytic leukaemia; Auxotrophic; Breast cancer; L-glutaminase | ||
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