Effect of Medium Composition and Kinetic Studies on Extracellular and Intracellular Production of L-asparaginase from Pectobacterium carotovorum

Abstract

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.

Screening and characterization of extracelluar L-asparaginase producing Bacillus subtilis strain hswx88, isolated from Taptapani hotspring of Odisha, India

“…LAse also known commercially as Oncaspar, Colaspase, Crasnitin, Kidrolase, Erwinase and Elspar [5] . It is also used in food industry for the production of acrylamide (a potent carcinogen and a neurotoxic compound) free starchy fry food [6] . Preventase and acrylaway produced from Aspergillus niger and Aspergillus oryzae respectively are commercially available LAse currently used in food industry [7] .…”

“…The results of the morphological, biochemical and physiological tests were put into Bergey's Manual of Determinative Bacteriology [19] and bacterial identification software's like PIBwin version 2.0 and ABIS online for accurate identification. The isolate TPS-12 was exposed to temperature range of 40-100°C, pH range of [3][4][5][6][7][8][9][10][11][12] and sodium chloride concentrations range of 2%-10% for evaluation of physiological characteristics. The antibiogram pattern of the isolate TPS-12 was tested for sensitivity against different antibiotic discs (Himedia, Mumbai) viz., erythromycin (15 …”

“…The isolate TPS-12 when exposed to temperature range of 40-100 °C, showed growth in the temperature range of 150-70 °C, while the growth was restricted above and below the above specified range. The isolate TPS-12 when exposed to different pH range (3)(4)(5)(6)(7)(8)(9)(10)(11)(12) and tolerance to different concentrations of sodium chloride (2%-10%), exhibited growth at pH range of [5][6][7][8][9][10] and tolerated sodium chloride concentrations 2%-7% respectively. The isolate showed no growth at pH 3 The results of the antibiogram pattern revealed that the isolate TPS-12 was very sensitive to all the tested antibiotics.…”

Optimization of L-asparaginase activity of Actinobacteria isolated from Guaviare river sediments in Colombia

“…The best three carbon sources were determined among soluble starch, lactose, glycerol, glucose and sucrose [2,10,15,19] at 1 % (w/v) each, using the phenol red plate assay [15,20]. The best carbon source was then confirmed using nesslerization.…”

“…The best carbon source was then confirmed using nesslerization. The same tests were performed to determine the best nitrogen source among meat extract, yeast extract, potassium nitrate, peptone, malt extract, L-asparagine, and Lglutamine [6,20] at 1% (w/v), and two combinations of L-asparagine with malt extract or potassium nitrate (both at 0.05%).…”