Extraction and Characterization of L-asparaginase from Spinacea oleraceae

摘要

L-asparaginase (E.C. 3.5.1.1) is an enzyme that catalyzes the hydrolysis of L-asparagine into L-aspartate and ammonia. It is identified as an effective antitumor agent in human clinical trials and is now recognized as one of the important component of antitumor therapy. The purpose of the present study was to screen for the production of L-asparaginase enzyme in Spinacea oleracea. The presence of the enzyme was confirmed by the formation of ammonia which was detected using UV-Visible Spectrophotometer at 460nm. The enzyme activity was found to be 1.9IU and specific activity was 6.8 -moles/mg/min. The enzyme kinetics revealed that the optimum pH of the enzyme L-asparaginase was 8.0, optimum temperature was found to be 370C and optimum time was 15 minutes. The Km and V-max values of Spinacia oleracea was found to be 5 and 2.5 respectively by means of the double-reciprocal Lineweaver-Burk plot.