Enhancement of the production of L-glutaminase, an anticancer enzyme, from Aeromonas veronii by adaptive and induced mutation techniques

Abstract

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).

Novel halo- and thermo-tolerant Cohnella sp. A01 L-glutaminase: heterologous expression and biochemical characterization

“…L-glutaminase with its proven abilities to use in the food industry and cancer treatment has become more important in the last few decades 3–5 . The other L-glutaminase applications are in culture medium as glutamine biosensor 6,7 , in GABA 8 and theanine synthesis 9 and also to measure the reaction rate of threonine synthesis 10 . L-glutaminase by releasing L-glutamate as a flavor enhancer amino acid from L-glutamine can be used in the food industry 10,11 .…”

Studies on optimization of L-glutaminase production under submerged fermentation from marine Bacillus subtilis JK-79

“…Supplementation of sucrose and yeast extract as carbon and nitrogen source improved Lglutaminase production by Zygosaccharomyces rouxii while higher L-glutaminase production was noticed in Providencia species with glucose and urea as carbon and nitrogen source, respectively. Recently, there are several reports which confirm the fact that the carbon and nitrogen sources significantly affect the production of Lglutaminase by microorganisms (Sathish and Prakasham, 2013;Krishnakumar et al, 2011;Pandian et al, 2014;Jesuraj et al, 2017).…”

“…RSM was used for optimization and the bacterium grown in the optimized medium [arabinose (2%), skim milk (4%) and sodium chloride (2%)] yielded L-glutaminase activity of 1.34 IU/mg. Jesuraj et al (2017) has reported statistical optimization of L-glutaminase production by mutated strain Aeromonas veronii by PBD and CCD. 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).…”

Anticancer Glutaminase Production and Optimization Using Halotolerant Aspergillus Flavus Czcu-9, F1h

“…Also, Vibrio azureus JK-79 also was good producer of glutaminase [7]. Aeromonas veronii has been identified as potential producer of lglutaminase [8].…”