A solvent-free delipidation method for functional validation of lipases

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 delipi...

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Bibliographic Details
Published in:3 Biotech 2020-08, Vol.10 (8), p.343-343, Article 343
Main Authors: Dolui, Achintya Kumar, Vijayaraj, Panneerselvam
Format: Article
Language:English
Subjects:
Agriculture
Bioinformatics
Biomaterials
Biotechnology
Cancer Research
Chemistry
Chemistry and Materials Science
Enzymes
Lipids
Phospholipids
Proteins
Protocols and Methods
Serine
Silicon dioxide
Solvents
Stem Cells
Substrates
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Summary: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 significant 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 profiling (ABPP), a functional chemoproteomic approach. Detection of serine hydrolases using ABPP probe labeling was enhanced upon delipidation. Further, the total polyphenol content was significantly 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 efficient and highly compatible with the functional characterization of the enzymes, particularly lipid hydrolases.
ISSN:2190-572X
2190-5738
DOI:10.1007/s13205-020-02338-7