Screening for pancreatic lipase inhibitors: evaluating assay conditions using p-nitrophenyl palmitate as substrate
Current in vitro pancreatic lipase inhibitor screenings are based on previous spectrophotometric lipase assays. Nevertheless, they are with little evaluation of assay conditions. This study focuses on the impacts of experimental factors on enzyme activity in the assay with p-nitrophenyl palmitate as...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Taylor & Francis Group
2022-12-01
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| Series: | All Life |
| Subjects: | |
| Online Access: | http://dx.doi.org/10.1080/26895293.2021.2019131 |
| _version_ | 1797236929578663936 |
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| author | Cam-Van T. Vo Nhan V. H. Luu Thoai T. H. Nguyen Truc T. Nguyen Bach Q. Ho Thuong H. Nguyen Thanh-Dao Tran Quoc-Thai Nguyen |
| author_facet | Cam-Van T. Vo Nhan V. H. Luu Thoai T. H. Nguyen Truc T. Nguyen Bach Q. Ho Thuong H. Nguyen Thanh-Dao Tran Quoc-Thai Nguyen |
| author_sort | Cam-Van T. Vo |
| collection | DOAJ |
| description | Current in vitro pancreatic lipase inhibitor screenings are based on previous spectrophotometric lipase assays. Nevertheless, they are with little evaluation of assay conditions. This study focuses on the impacts of experimental factors on enzyme activity in the assay with p-nitrophenyl palmitate as substrate by monitoring their effects on the hydrolysis rates. On the results, experimental conditions for lipase inhibitory assay were proposed. Notably, 5 mM sodium deoxycholate as emulsifier not only maintains the assay homogeneity but also enhances lipase activity. Organic co-solvents to dissolve organic inhibitors including DMSO, EtOH, MeOH, IPA, AcCN [0–30% (v/v)] was found well tolerated by the enzyme. With 10% (v/v) glycerol, lipase solutions can be stored at –20°C for up to one month without significant loss of activity. The results reported here provide researchers the assay condition sets in which most inhibitors can be dissolved, and lipase activity is not severely affected. This could accelerate the rational development of novel lipase inhibitors. |
| first_indexed | 2024-03-08T16:58:52Z |
| format | Article |
| id | doaj.art-6c0e7390ffdd4895909f9967a699dc7d |
| institution | Directory Open Access Journal |
| issn | 2689-5307 |
| language | English |
| last_indexed | 2024-04-24T17:11:40Z |
| publishDate | 2022-12-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | All Life |
| spelling | doaj.art-6c0e7390ffdd4895909f9967a699dc7d2024-03-28T09:48:50ZengTaylor & Francis GroupAll Life2689-53072022-12-01151132210.1080/26895293.2021.20191312019131Screening for pancreatic lipase inhibitors: evaluating assay conditions using p-nitrophenyl palmitate as substrateCam-Van T. Vo0Nhan V. H. Luu1Thoai T. H. Nguyen2Truc T. Nguyen3Bach Q. Ho4Thuong H. Nguyen5Thanh-Dao Tran6Quoc-Thai Nguyen7University of Medicine and Pharmacy at Ho Chi Minh CityUniversity of Medicine and Pharmacy at Ho Chi Minh CityUniversity of Medicine and Pharmacy at Ho Chi Minh CityUniversity of Medicine and Pharmacy at Ho Chi Minh CityUniversity of Medicine and Pharmacy at Ho Chi Minh CityUniversity of Medicine and Pharmacy at Ho Chi Minh CityUniversity of Medicine and Pharmacy at Ho Chi Minh CityUniversity of Medicine and Pharmacy at Ho Chi Minh CityCurrent in vitro pancreatic lipase inhibitor screenings are based on previous spectrophotometric lipase assays. Nevertheless, they are with little evaluation of assay conditions. This study focuses on the impacts of experimental factors on enzyme activity in the assay with p-nitrophenyl palmitate as substrate by monitoring their effects on the hydrolysis rates. On the results, experimental conditions for lipase inhibitory assay were proposed. Notably, 5 mM sodium deoxycholate as emulsifier not only maintains the assay homogeneity but also enhances lipase activity. Organic co-solvents to dissolve organic inhibitors including DMSO, EtOH, MeOH, IPA, AcCN [0–30% (v/v)] was found well tolerated by the enzyme. With 10% (v/v) glycerol, lipase solutions can be stored at –20°C for up to one month without significant loss of activity. The results reported here provide researchers the assay condition sets in which most inhibitors can be dissolved, and lipase activity is not severely affected. This could accelerate the rational development of novel lipase inhibitors.http://dx.doi.org/10.1080/26895293.2021.2019131lipase assaypancreatic lipase inhibitorsp-nitrophenyl esters |
| spellingShingle | Cam-Van T. Vo Nhan V. H. Luu Thoai T. H. Nguyen Truc T. Nguyen Bach Q. Ho Thuong H. Nguyen Thanh-Dao Tran Quoc-Thai Nguyen Screening for pancreatic lipase inhibitors: evaluating assay conditions using p-nitrophenyl palmitate as substrate All Life lipase assay pancreatic lipase inhibitors p-nitrophenyl esters |
| title | Screening for pancreatic lipase inhibitors: evaluating assay conditions using p-nitrophenyl palmitate as substrate |
| title_full | Screening for pancreatic lipase inhibitors: evaluating assay conditions using p-nitrophenyl palmitate as substrate |
| title_fullStr | Screening for pancreatic lipase inhibitors: evaluating assay conditions using p-nitrophenyl palmitate as substrate |
| title_full_unstemmed | Screening for pancreatic lipase inhibitors: evaluating assay conditions using p-nitrophenyl palmitate as substrate |
| title_short | Screening for pancreatic lipase inhibitors: evaluating assay conditions using p-nitrophenyl palmitate as substrate |
| title_sort | screening for pancreatic lipase inhibitors evaluating assay conditions using p nitrophenyl palmitate as substrate |
| topic | lipase assay pancreatic lipase inhibitors p-nitrophenyl esters |
| url | http://dx.doi.org/10.1080/26895293.2021.2019131 |
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