European Journal of Biology and Medical Science Research (EJBMSR)

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PURIFICATION AND CHARACTERIZATION OF STENOTROPHOMONASSP.FZ L-ASPARAGINASE UNDER SOLID STATE FERMENTATION

Abstract

L-asparaginase has emerged as one of the most important clinically used enzymes as it exhibits chemotherapeutic potential in treatment of acute lymphoblastic leukimia and lymphosacroma. A novel bacterium L-asparaginaseproducerwas isolated from Lebanesesoil andwas identified asStenotrophomonas sp.FZ using 16srRNA. The enzyme was produced under solid state fermentation using the wheat bran as carbon and nitrogen source The enzyme was partially purified by acetone precipitation with 73.32 % yield and a purification factor of 6.32 fold. Further purification includes gel filtration chromatoghraphy on Sephadex G-75 and C-25 ion exchange chromatogrraphy with final purification factor of 329.061 fold and 54.312 yield%. The total protein was reduced by 99.83% and the specific activity was increased to be 1.6124IU/mgX1000. The maximum enzyme activity was recorded at pH 7 and 35°C with a linear relationship concerning the increase in enzyme concentration. The effect of substrate concentration showed a progressive increase in the enzyme activity in a concentration dependent manner till it reaches a plateau where saturation was reached. The kinetics parameters(Km and Vmax ) of Stenotrophomonassp L-asparaginase production were 96.71 mg/ml and 3333.33 umol/ml/min respectively. The theapeutic potential of this enzyme is well established.

Keywords: Characterization, L-Asparaginase., Purification, Stenotrophomonassp, Therapeutic Potential

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Email ID: editor.ejbmsr@ea-journals.org
Impact Factor: 7.77
Print ISSN: 2053-406X
Online ISSN: 2053-4078
DOI: https://doi.org/10.37745/ejbmsr.2013

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