Effect of medium composition and kinetic studies on extracellular and intracellular production of L-asparaginase from Pectobacterium carotovorum.

Microbial L-asparaginase occupies a prominent place among biocatalysts owing to their ability to catalyze the reaction that hydrolyze the asparagine molecule. Effect of various medium components on the production of L-asparaginase in submerged fermentation by Pectobacterium carotovorum was studied for optimal nutrient requirements. Six different media compositions were tested for the L-asparaginase production keeping fermentation conditions constant at temperature 30 °C, initial pH 7.0 and agitation speed of 120 rpm. Maximum intracellular and extracellular L-asparaginase activity was obtained in the medium containing tryptone, yeast extract, monosodium glutamate, K₂HPO₄ and L-asparagine. These medium components were further optimized by central composite experimental design using response surface methodology. Maximum intracellular and extracellular L-asparaginase activity of 2.282 U/mL and 0.587 U/mL were obtained respectively at the late logarithmic phase in optimized media. Unstructured kinetic models were used to describe the cell growth and product formation kinetics. The unstructured models predicted the cell growth and product formation profile accurately with high coefficient of determination.