?-galactosidase from Watermelon (Citrullus lanatus) Seedlings: Partial Purification and Properties. | Pakistan Journal of Biotechnology 2017

Author(s)

Asma Saeed Department of Biological Sciences, Gomal University, Dera Ismail Khan, Pakistan

Muhammad Salim Department of Biological Sciences, Gomal University, Dera Ismail Khan, Pakistan

Umber Zaman Department of Chemistry, Gomal University, Dera Ismail Khan, Pakistan

Rubina Naz Department of Chemistry, Gomal University, Dera Ismail Khan, Pakistan

Saleem Jan Department of Chemistry, University of Science and Information Technology, Bannu, Pakistan

Ahmad Saeed Department of Chemistry, Qurtuba University of Science and Information Technology, Dera Ismail Khan, Pakistan

Abstract

ß-Galactosidase from watermelon (Citrullus lanatus) seedlings was partially purified. Extract was fractionated in two steps with ammonium sulfate at 30 & 80% saturations. Ammonium sulfate precipitated enzyme yielded 60% recovery and two folds purification of the enzyme. Further, the enzyme was subjected to CM-Cellulose and Sephadex G-100 chromatography to obtain 24-folds purification. The specific activity was increased to from 1.5 U to 36 U per milligram of protein. The overall yield was 13.2%. The enzyme had optimum pH 4.8 and pH stability between pH 4.0 and 7.0 was observed. Optimum temperature was 50°C and temperature stability was also shown up to 50°C. Thereafter enzyme activity fell and inactivated completely at 70°C. The Km and Vmax were found to be 0.2 mM and 34Umin-1mg-1of protein, respectively. ß-mercaptoethanol exhibited very small activation at moderate concentration, but the inhibition was displayed at high concentrations. Heavy metal ions such as Al+3, Ag+1, Hg+2, etc. inhibited the enzyme strongly. The inhibition by heavy metals and SH-reacting reagents suggests that cystein was necessary for enzyme activity. On the addition of ß-mercaptoethanol to pre-incubated enzyme with metal ions, the inhibition was relieved. The ß-Galactosidase activity was increased in the presence of alcohols. This activation reflected glycosyltransferase activity.

Publication Details

Page(s) 271-278

DOI DOI not available

Published Journal: Pakistan Journal of Biotechnology, Volume: 14, Issue: 2, Year: 2017

Keywords

Keywords are not available for this article.

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