A model for finding new L-asparaginase producing microorganisms using Taguchi design of experiments
Paper Details
A model for finding new L-asparaginase producing microorganisms using Taguchi design of experiments
Abstract
The current study suggests the screening of soil samples for L-asparaginase producing microorganisms and optimization of enzyme production by Taguchi Design of Experiments as a suitable model for finding alternate L-asparaginase enzyme to the ones used in current therapeutic methods. A bacterium designated as strain MH01 was isolated that could effectively breakdown L-asparagine. It was found to be specie of genus Bacillus based on morphological, biochemical analysis and 16S rRNA gene sequencing, resulting in 96% similarity with Bacillus marcorestinctum strain LQQ (GQ900516). Enzyme production was statistically optimized using Taguchi design of experiment. The maximum specific activity 0.8 IU/µg was obtained at 30°C temperature, 2:1 inoculum to substrate ratio and a pH 5.0. L-asparaginase produced under optimized conditions was partially purified by size exclusion column chromatography using sephadex G-100. The molecular weight was estimated to be 41.88 kDa. The enzyme was purified 1.79 folds with 31.98% yield and 1.586 IU/µg specific activity with an optimum temperature of 30°C and pH of 8.0-8.5. The enzyme had Km value of 0.389 mM, Vmax value of 30395 µML-1 min-1 and Kcat value of 429.36 s-1 which indicates its efficiency and potential for further study and proving the success of our model in finding an alternate L-asparaginase enzyme.
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