4.6 Article

Zymography for Picogram Detection of Lipase and Esterase Activities

期刊

MOLECULES
卷 26, 期 6, 页码 -

出版社

MDPI
DOI: 10.3390/molecules26061542

关键词

esterase; lipase; picogram; zymogram

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  1. Wilmar International Limited
  2. WIL@NUS Corporate Laboratory

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The study introduces a new method for detecting lipolytic enzyme activity which is 1000-fold more sensitive than previous methods, allowing for detection of as low as 5pg and 180pg of lipolytic activity. By adjusting buffering capacity, the detection sensitivity can be controlled, facilitating screening of both high and low abundance lipolytic enzymes. Coupled with liquid chromatography-mass spectrometry, this method provides a useful tool for sensitive detection and identification of lipolytic enzymes.
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.

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