A model for finding new L-asparaginase producing microorganisms using Taguchi design of experiments

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Research Paper 01/03/2021
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A model for finding new L-asparaginase producing microorganisms using Taguchi design of experiments

Hassan Mustafa Arif, Zulfiqar Ali Malik, Muhammad Irfan, Fakhra Hanif, Fariha Hasan, Samiullah Khan, Malik Badshah, Aamer Ali Shah
Int. J. Biosci.18( 3), 276-289, March 2021.
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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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