SEQUENTIAL PRECIPITATION WITH REVERSIBLY SOLUBLE INSOLUBLE POLYMERS AS A BIOSEPARATION STRATEGY - PURIFICATION OF BETA-GLUCOSIDASE FROM TRICHODERMA-LONGIBRACHIATUM
R. Agarwal et Mn. Gupta, SEQUENTIAL PRECIPITATION WITH REVERSIBLY SOLUBLE INSOLUBLE POLYMERS AS A BIOSEPARATION STRATEGY - PURIFICATION OF BETA-GLUCOSIDASE FROM TRICHODERMA-LONGIBRACHIATUM, Protein expression and purification, 7(3), 1996, pp. 294-298
Precipitation of proteins/enzymes with reversibly soluble-insoluble po
lymers is a simple approach which can be easily scaled up. it is shown
here that; considerable purification may be achieved if more than one
polymer (chosen judiciously) is used in sequence for purification. Th
e principle is illustrated with purification of beta-glucosidase from
a commercial preparation of Trichoderma longibrachiatum. Precipitation
with chitosan removed cellulase activity from the preparation with 4-
fold purification and 99% recovery of beta-glucosidase activity in the
supernatant. The treatment of the supernatant with Eudragit S-100 pre
cipitated 98% of the enzyme activity. The two different elution proced
ures gave 82% recovery (with 14-fold purification) and 86% recovery (w
ith 12-fold purification) of the enzyme activity from the precipitate.
A low molecular weight protein still present in these preparations co
uld be removed by gel filtration. This finally yielded an electrophore
tically homogeneous enzyme with 74% recovery and 29-fold purification.
(C) 1996 Academic Press, Inc.