Purification of phospholipase D from Dacus carota by three-phase partitioning and its characterization

Protein Expression and Purification
S Sharma, M N Gupta

Abstract

Phospholipase D from Dacus carota (carrot) was purified by subjecting it to three-phase partitioning. The single step of three phase partitioning led to 13-fold purification with an activity recovery of 72%. SDS-PAGE analysis showed a single band with minimum molecular weight corresponding to nearly 60 kDa. The purified enzyme had a pH optimum in the range of 6.0--6.5 and was unstable above 30 degrees C. Kinetic studies showed a K(m) value of 9.5 mM and a V(max) of 0.35 mL min(-1). The enzyme purified by three-phase partitioning was found to resolve into two isoenzymes on a DEAE-cellulose column.

Citations

Dec 15, 2012·PloS One·Gulam Mohmad RatherMunishwar N Gupta
Apr 25, 2007·The FEBS Journal·Susanne StumpeRenate Ulbrich-Hofmann
Jul 6, 2015·International Journal of Biological Macromolecules·M ShanmugaprakashD Amala Reddy
Sep 5, 2002·Biotechnology and Bioengineering·Aparna Sharma, M N Gupta
Jun 14, 2017·Critical Reviews in Food Science and Nutrition·Jing-Kun YanJian-Yong Wu
Nov 16, 2001·Phytochemical Analysis : PCA

Related Concepts

Ammonium Sulfate
Calcium
Catalysis
Chromatography, DEAE-Cellulose
Enzyme Stability
Hydrogen-Ion Concentration
Alloenzymes
Phosphatidylcholines
Phospholipase D
Tensides

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