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A solvent‑free delipidation method for functional validation of lipases.
Achintya Kumar, Dolui and Vijayaraj, P. (2020) A solvent‑free delipidation method for functional validation of lipases. 3 Biotech, 10 (343). pp. 1-8.
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Abstract
Extracting protein in its active form is critical for its functional characterization, and lipid removal is an essential step in the protein extraction process for further downstream applications. In the present study, we revisited the delipidation protocol and developed a rapid, solvent-free delipidation method using activated silica. The delipidated samples showed improved optical clarity and a signifcant reduction of endogenous lipids. The functional integrity of the lipases present in the delipidated sample was validated by in vitro enzyme assay using physiological substrate which includes neutral lipid as well as phospholipid. The accessibility of active site of the extracted enzymes was demonstrated by activity-based protein profling (ABPP), a functional chemoproteomic approach. Detection of serine hydrolases using ABPP probe labeling was enhanced upon delipidation. Further, the total polyphenol content was signifcantly reduced, which helps to enhance the protein enrichment and small-molecule inhibitor screening by ABPP. Collectively, these results suggest that the present solvent-free delipidation approach is efcient and highly compatible with the functional characterization of the enzymes, particularly lipid hydrolases.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | Delipidation · Enzymes · Lipases · Rice bran · Lipids · Activity-based protein profling |
| Subjects: | 600 Technology > 08 Food technology > 16 Nutritive value > 05 Enzymes 600 Technology > 08 Food technology > 16 Nutritive value > 07 Waste utilization 600 Technology > 08 Food technology > 21 Cereals > 01 Rice |
| Divisions: | Dept. of Lipid Science |
| Depositing User: | Food Sci. & Technol. Information Services |
| Date Deposited: | 30 Sep 2020 09:17 |
| Last Modified: | 30 Sep 2020 09:17 |
| URI: | http://ir.cftri.res.in/id/eprint/14465 |
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