BETA-GLUCOSIDASE AND A PROCESS FOR EXTRACTION THEREOF

Abstract

The present invention provides a novel beta glucosidase and a process for extraction of a beta-glucosidase from Rauvolfia serpentine useful for the cleaving of beta-1,4 linkage of PNPG and to convert other gluco-conjugates such as strictosidine and raucaffricine into their corresponding aglycon such as vomilenine, commercially through immobilizing the enzyme. The β glucosidase enzyme has shown maximum activity in the acid pH range, with high optimum temperature using PNPG as substrate. The crude enzyme, when stored at 4° C., was quite stable for 6 days with 50% loss of activity. The enzyme was activated in presence of FeSO4 in the assay mixture.

Description

Claims

1. A β glucosidase enzyme useful for the cleavage of β 1,4 linkage of p-nitrophenyl β-D-glucopyranoside (PNPG).

2. A β glucosidase enzyme as claimed in claim 1, wherein the said enzyme have following characteristics: a) it is stable for more than 7 days at 4 degree C.;b) it is active in acidic pH range;c) optimum activity of this enzyme is at about 60 degree C.;d) it is strong active in the presence of FeSO4;

3. A β glucosidase enzyme as claimed in claim 1, wherein the said enzyme is obtained from the extract of plant tissues of Rauvolfia serpentina.

4. A β glucosidase enzyme as claimed in claim 1, wherein the plant tissue used is a flower of Rauvolfia serpentina.

5. A process for extraction of a β-glucosidase from plant tissues of Rauvolfia serpentina wherein the said process comprises: a) homogenizing the plant tissue in a cold extraction medium in the ratio ranging from 1:1 to 1:3 (w/v)b) centrifuging the homogenized solution obtained from step (a) at 10,000 rpm to 12,000 rpm for 20-30 min at a temperature in the range 4 degree C. to 10 degree C. to obtain the clear supernatant containing desired product.

6. A process as claimed in claim 5, wherein the used plant tissue from Rauvolfia serpentina is selected from the group consisting of roots, stem, old, mature and young leaves, flower stem (petiole), flowers and fruits etc.

7. A process as claimed in claim 5, wherein the extraction medium used is comprises a disulphide bond reducing agent and a chelating agent in phosphate buffer at pH of about 6.0 in the ratio ranging from 1:5 to 2:5.

8. A process as claimed in claim 5, wherein the reducing agent used is beta mercaptoethanol.

9. A process as claimed in claim 5, wherein the chelating agent used is EDTA.