Jesuraj S Aravinth Vijay, Sarker Md Moklesur Rahman, Ming Long Chiau, Praya S Marylin Jeya, Ravikumar M, Wui Wong Tin
Centre for Pharmaceutical Sciences, JNT University, Kukatpally, Hyderabad, Telengana State, India.
Faculty of Pharmacy, Lincoln University College, Petaling Jaya, Selangor Darul Ehsan, Malaysia.
PLoS One. 2017 Aug 16;12(8):e0181745. doi: 10.1371/journal.pone.0181745. eCollection 2017.
Microbial anti-cancer enzymes have been proven to be effective and economical agents for cancer treatment. Aeromonas veronii has been identified as a microorganism with the potential to produce L-glutaminase, an anticancer agent effective against acute lymphocytic leukaemia. In this study, a selective medium of Aeromonas veronii was used to culture the microorganism. Strain improvement was done by adaptive and induced mutational techniques. A selective minimal agar media was incorporated for the growth of the strain which further supports adaptive mutation. Strains were also UV-irradiated and successively treated with N-methyl-N'-nitro-N-nitrosoguanidine to find a resilient strain capable of producing L-glutaminase efficiently. The Plackett-Burman design and central composite designs were used to screen and optimize additional carbon and nitrogen sources. Adaptive mutation resulted in promising yield improvements compared to native strain (P<0.001). The mean yield of 30 treated colonies from the induced mutation was significantly increased compared to the non-induced strain (P< 0.001). The economically feasible statistical designs were found to reinforce each other in order to maximize the yield of the enzyme. The interactions of nutrient factors were understood from the 3D response surface plots. The model was found to be a perfect fit in terms of maximizing enzyme yield, with the productivity improving at every stage to a fourfold output of enzyme (591.11 ±7.97 IU/mL) compared to the native strain (135±3.51 IU/mL).
微生物抗癌酶已被证明是治疗癌症的有效且经济的药物。维氏气单胞菌已被鉴定为一种有潜力产生L-谷氨酰胺酶的微生物,L-谷氨酰胺酶是一种对急性淋巴细胞白血病有效的抗癌剂。在本研究中,使用维氏气单胞菌的选择性培养基来培养该微生物。通过适应性和诱导突变技术进行菌株改良。加入选择性基本琼脂培养基以支持该菌株生长,这进一步支持适应性突变。菌株还经过紫外线照射,并依次用N-甲基-N'-硝基-N-亚硝基胍处理,以找到能够高效产生L-谷氨酰胺酶的抗性菌株。采用Plackett-Burman设计和中心复合设计来筛选和优化额外的碳源和氮源。与原始菌株相比,适应性突变导致产量有显著提高(P<0.001)。与未诱导的菌株相比,诱导突变产生的30个处理菌落的平均产量显著增加(P<0.001)。发现经济可行的统计设计相互加强,以最大化酶的产量。从三维响应面图中了解营养因子的相互作用。发现该模型在最大化酶产量方面非常合适,与原始菌株(135±3.51 IU/mL)相比,生产力在每个阶段都有所提高,酶产量提高到四倍(591.11±7.97 IU/mL)。