Molecules (Mar 2021)

Zymography for Picogram Detection of Lipase and Esterase Activities

  • Andre Mong Jie Ng,
  • Hongfang Zhang,
  • Giang Kien Truc Nguyen

DOI
https://doi.org/10.3390/molecules26061542
Journal volume & issue
Vol. 26, no. 6
p. 1542

Abstract

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Lipases and esterases are important catalysts with wide varieties of industrial applications. Although many methods have been established for detecting their activities, a simple and sensitive approach for picogram detection of lipolytic enzyme quantity is still highly desirable. Here we report a lipase detection assay which is 1000-fold more sensitive than previously reported methods. Our assay enables the detection of as low as 5 pg and 180 pg of lipolytic activity by direct spotting and zymography, respectively. Furthermore, we demonstrated that the detection sensitivity was adjustable by varying the buffering capacity, which allows for screening of both high and low abundance lipolytic enzymes. Coupled with liquid chromatography-mass spectrometry, our method provides a useful tool for sensitive detection and identification of lipolytic enzymes.

Keywords