A solvent-free delipidation method for functional validation of lipases

Achintya Kumar Dolui, Panneerselvam Vijayaraj

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

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 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.

Original languageEnglish
Article number343
Journal3 Biotech
Volume10
Issue number8
DOIs
StatePublished - 1 Aug 2020
Externally publishedYes

Keywords

  • Activity-based protein profiling
  • Delipidation
  • Enzymes
  • Lipases
  • Lipids
  • Rice bran

ASJC Scopus subject areas

  • Biotechnology
  • Environmental Science (miscellaneous)
  • Agricultural and Biological Sciences (miscellaneous)

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