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A solvent‑free delipidation method for functional validation of lipases.

IR@CSIR-CFTRI

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Relation http://ir.cftri.com/14465/
https://doi.org/10.1007/s13205-020-02338-7
 
Title A solvent‑free delipidation method for functional validation of lipases.
 
Creator Achintya Kumar, Dolui
Vijayaraj, P.
 
Subject 05 Enzymes
07 Waste utilization
01 Rice
 
Description 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.
 
Date 2020
 
Type Article
PeerReviewed
 
Format pdf
 
Language en
 
Identifier http://ir.cftri.com/14465/1/3%20Biotech%20%282020%29%2010343.pdf
Achintya Kumar, Dolui and Vijayaraj, P. (2020) A solvent‑free delipidation method for functional validation of lipases. 3 Biotech, 10 (343). pp. 1-8.