Zymography for Picogram Detection of Lipase and Esterase Activities
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 li...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-03-01
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| Series: | Molecules |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1420-3049/26/6/1542 |
| _version_ | 1797541713709891584 |
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| author | Andre Mong Jie Ng Hongfang Zhang Giang Kien Truc Nguyen |
| author_facet | Andre Mong Jie Ng Hongfang Zhang Giang Kien Truc Nguyen |
| author_sort | Andre Mong Jie Ng |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-10T13:20:39Z |
| format | Article |
| id | doaj.art-c7c262ada6b14a7789bdcdf05ee39aca |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-10T13:20:39Z |
| publishDate | 2021-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-c7c262ada6b14a7789bdcdf05ee39aca2023-11-21T10:06:36ZengMDPI AGMolecules1420-30492021-03-01266154210.3390/molecules26061542Zymography for Picogram Detection of Lipase and Esterase ActivitiesAndre Mong Jie Ng0Hongfang Zhang1Giang Kien Truc Nguyen2Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, 8 Medical Drive, Singapore 117596, SingaporeWIL@NUS Corporate Laboratory, Wilmar International Limited, Centre for Translational Medicine, 14 Medical Drive, Singapore 117599, SingaporeWIL@NUS Corporate Laboratory, Wilmar International Limited, Centre for Translational Medicine, 14 Medical Drive, Singapore 117599, SingaporeLipases 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.https://www.mdpi.com/1420-3049/26/6/1542esteraselipasepicogramzymogram |
| spellingShingle | Andre Mong Jie Ng Hongfang Zhang Giang Kien Truc Nguyen Zymography for Picogram Detection of Lipase and Esterase Activities Molecules esterase lipase picogram zymogram |
| title | Zymography for Picogram Detection of Lipase and Esterase Activities |
| title_full | Zymography for Picogram Detection of Lipase and Esterase Activities |
| title_fullStr | Zymography for Picogram Detection of Lipase and Esterase Activities |
| title_full_unstemmed | Zymography for Picogram Detection of Lipase and Esterase Activities |
| title_short | Zymography for Picogram Detection of Lipase and Esterase Activities |
| title_sort | zymography for picogram detection of lipase and esterase activities |
| topic | esterase lipase picogram zymogram |
| url | https://www.mdpi.com/1420-3049/26/6/1542 |
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