PURIFICATION OF ARGINASE FROM EMBRYONIC AXES OF SOYBEAN (Glycine max L.) Merr.
Abstract
- In this research, arginase was extracted from embryonic axes of soybean by using MnCl2 salt solution. Arginase (L-arginine amidinohydrolase, (EC 3.5.3.1) is a binuclear manganese metalloenzyme that catalyzes the hydrolysis of arginine to ornithine and urea. The arginase activity was determined spectrophotometrically by measuring the urea liberated. In diacetylmonoxime method for urea determination, the wavelength of maximum absorption of the colored product was found at 480 nm. The arginase activity was defined as micromole of urea liberated per minute per milileter of enzyme solution. The variation of arginase activity with growth day of embryonic axes of soybean was studied. The maximum arginase activity was found after five day of germination. In the determination of protein content is Biuret method, the wavelength of maximum absorption of copper protein complex was found at 550 nm. After chromatographic separation with Sephadex G-200 gel, the specific activity the relative purity of the enzyme increased about (15) folds from crude to final purification step. The homogeneity of the purified arginase was confirmed by non SDS-PAGE as a single band. In this research effect of metal ions ( K+, Ca2+, Mn2+, Co2+, Fe2+, Cu2+, Zn2+) on arginase activities were studied. The Mn2+ ion showed the largest activating effect on arginase activities. The presence of manganese ion is essential to get the stable structure of arginase enzyme proteins.
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Year
- 2019
Author
-
Nan Yin Yin Htwe
Subject
- Chemistry
Publisher
- Myanmar Academy of Arts and Science (MAAS)