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Differential partitioning of beta-galactosidase and beta-glucosidase using aqueous two phase extraction

Hemavathi, A. B. and Raghavarao, K. S. M. S. (2011) Differential partitioning of beta-galactosidase and beta-glucosidase using aqueous two phase extraction. Process Biochemistry, 46. pp. 649-655.

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Aqueous two phase extraction (ATPE) is used for the first time for simultaneous separation and purification of�-galactosidase and�-glucosidase from barley (Hordeum vulgare). The influence of various process parameters such as polymer molecular weight and its concentration, salt type and its concentration, system pH, tie line length, phase volume ratio and neutral salt addition on separation and purification of these two enzymes was evaluated. The �-galactosidase and �-glucosidase were selectively partitioned to top and bottom phases, respectively. Suitable conditions for purification were found in aqueous two phase system, having 14% (w/w) polyethylene glycol 1500/13% (w/w) ammonium sulphate, at tie line length of 19.65% (w/w). Single stage of ATPE resulted in an activity recovery of 98.26% with purification of 2.1 fold of�-galactosidase and an activity recovery of 92.58% with purification of 3.3 fold of�-glucosidase. Second stage of ATPE with respective new phases increased the purification of �-galactosidase and �- glucosidase to 2.4 and 4.1 fold, respectively. During ATPE, conditions which enabled a balance between yield and purification of both the enzymes were selected. Further ultrafiltration in diafiltration mode increased the purification of enzymes (�-galactosidase 6.8 fold and �-glucosidase 7.7 fold) besides the removal of the phase components.

Item Type: Article
Uncontrolled Keywords: ATPE, Differential partitioning, Barley, bet-Galactosidase beta-Glucosidase, PEG 1500
Subjects: 600 Technology > 08 Food technology > 21 Cereals > 02 Barley
600 Technology > 08 Food technology > 05 Processing and Engineering
Divisions: Food Engineering
Depositing User: Food Sci. & Technol. Information Services
Date Deposited: 17 Jun 2011 05:55
Last Modified: 01 Oct 2018 08:24
URI: http://ir.cftri.res.in/id/eprint/10128

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