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Enhanced recovery of L-asparaginase from isolated Penicillium sp. through modified cultural and nutritional amendments under submerged culture conditions

Author Affiliations

  • 1Regional Plant Resource Centre, Bhubaneswar 751015 Odisha, India
  • 2Regional Plant Resource Centre, Bhubaneswar 751015 Odisha, India

Int. Res. J. Biological Sci., Volume 11, Issue (1), Pages 24-32, February,10 (2022)

Abstract

Culture conditions were optimized for enhancing the amount of L-asparaginase in submerged culture medium of Penicillium sp. Optimized parameters involve different C and N ratio, culture conditions and additional compounds followed by extraction, purification and characterization of the enzyme. The fungus required 8 days and a pH of 4.5 for the optimum growth and enzyme production. The maximum L-asparaginase activity was observed with fructose, ammonium sulphate and L-arginine. The enzyme production was concentration dependant and maximum activity was recorded at fructose 35g/litre with addition of 5g/litre each of L-asparagine, ammonium sulphate and L arginine. The purification of enzyme from mass culture was developed in a modified media by following salt precipitation, Sephadex G-100 filtration and ion exchange chromatography by DEAE cellulose columns. Result revealed purified protein and enzyme preparations that was matched with standard (66kDa) and exhibited approximately 97k Dawith 0901x10-3MKm value. Enzyme purified through gel filtration was active within the fractions 4 to 7.2 and at higher temperature. The impact of nutritional and cultural conditions towards enhanced enzyme production is clearly evident in the present study. The thermal stability of the purified enzyme also ensures its suitability for long term preservation and the basic modification of the basal medium in order to achieve enhanced enzyme production may be a good support for the large scale fermentation and production of this enzyme.

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