S. Sharma et Mn. Gupta, Purification of phospholipase D from Dacus carota by three-phase partitioning and its characterization, PROT EX PUR, 21(2), 2001, pp. 310-316
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 w
as unstable above 30 degreesC. Kinetic studies showed a K-m value of 9.5 mM
and a V-max of 0.35 mL min-l. The enzyme purified by three phase partition
ing was found to resolve into two isoenzymes on a DEAE cellulose column. (C
) 2001 Academic Press.