Comparative Analysis of Universal Protein Extraction Methodologies for Screening of Lipase Activity from Agricultural Products
Protein extraction techniques are absolutely required for the research of biological catalysts. The present study compared four universal protein extraction methodologies (ammonium sulfate precipitation, TCA/acetone precipitation, and two commercial kits) to provide practical information on protein...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-07-01
|
| Series: | Catalysts |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2073-4344/11/7/816 |
| _version_ | 1797527487676153856 |
|---|---|
| author | Jisu Ha Jun-Young Park Yoonseok Choi Pahn-Shick Chang Kyung-Min Park |
| author_facet | Jisu Ha Jun-Young Park Yoonseok Choi Pahn-Shick Chang Kyung-Min Park |
| author_sort | Jisu Ha |
| collection | DOAJ |
| description | Protein extraction techniques are absolutely required for the research of biological catalysts. The present study compared four universal protein extraction methodologies (ammonium sulfate precipitation, TCA/acetone precipitation, and two commercial kits) to provide practical information on protein extraction in order to discover a novel lipase in agricultural products. Yields of protein extraction from 24 domestic agricultural products and their specific activities were evaluated and compared with each other. TCA/acetone precipitation showed a relatively higher extraction yield (on average, 3.41 ± 1.08 mg protein/0.1 g sample) in crude protein extraction, whereas the Pierce™ Plant Total Protein Extraction Kit showed the highest specific lipase activity on average in both spectrophotometric (266.61 ± 235.78 μU/mg protein) and fluorometric (41.52 ± 32.63 μU/mg protein) assays. Our results suggest that commercial kits for the rapid extraction of soluble functional proteins would be a better choice than conventional precipitation techniques to perform the high-throughput screening of enzyme activity from plant sources. Finally, several agricultural products such as cordyceps, pepper, bracken, and hemp, all of which exhibited an excellent specific lipase activity, were proposed as promising candidates for a source of novel lipases. |
| first_indexed | 2024-03-10T09:43:30Z |
| format | Article |
| id | doaj.art-d110a51044f948e8ba6b72d5a364f925 |
| institution | Directory Open Access Journal |
| issn | 2073-4344 |
| language | English |
| last_indexed | 2024-03-10T09:43:30Z |
| publishDate | 2021-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Catalysts |
| spelling | doaj.art-d110a51044f948e8ba6b72d5a364f9252023-11-22T03:26:56ZengMDPI AGCatalysts2073-43442021-07-0111781610.3390/catal11070816Comparative Analysis of Universal Protein Extraction Methodologies for Screening of Lipase Activity from Agricultural ProductsJisu Ha0Jun-Young Park1Yoonseok Choi2Pahn-Shick Chang3Kyung-Min Park4Department of Food Science and Biotechnology, Wonkwang University, Iksan 54538, KoreaDepartment of Agricultural Biotechnology, Seoul National University, Seoul 08826, KoreaDepartment of Agricultural Biotechnology, Seoul National University, Seoul 08826, KoreaDepartment of Agricultural Biotechnology, Seoul National University, Seoul 08826, KoreaDepartment of Food Science and Biotechnology, Wonkwang University, Iksan 54538, KoreaProtein extraction techniques are absolutely required for the research of biological catalysts. The present study compared four universal protein extraction methodologies (ammonium sulfate precipitation, TCA/acetone precipitation, and two commercial kits) to provide practical information on protein extraction in order to discover a novel lipase in agricultural products. Yields of protein extraction from 24 domestic agricultural products and their specific activities were evaluated and compared with each other. TCA/acetone precipitation showed a relatively higher extraction yield (on average, 3.41 ± 1.08 mg protein/0.1 g sample) in crude protein extraction, whereas the Pierce™ Plant Total Protein Extraction Kit showed the highest specific lipase activity on average in both spectrophotometric (266.61 ± 235.78 μU/mg protein) and fluorometric (41.52 ± 32.63 μU/mg protein) assays. Our results suggest that commercial kits for the rapid extraction of soluble functional proteins would be a better choice than conventional precipitation techniques to perform the high-throughput screening of enzyme activity from plant sources. Finally, several agricultural products such as cordyceps, pepper, bracken, and hemp, all of which exhibited an excellent specific lipase activity, were proposed as promising candidates for a source of novel lipases.https://www.mdpi.com/2073-4344/11/7/816lipaseagricultural productprotein extractionextraction yieldspecific activityspectrophotometric assay |
| spellingShingle | Jisu Ha Jun-Young Park Yoonseok Choi Pahn-Shick Chang Kyung-Min Park Comparative Analysis of Universal Protein Extraction Methodologies for Screening of Lipase Activity from Agricultural Products Catalysts lipase agricultural product protein extraction extraction yield specific activity spectrophotometric assay |
| title | Comparative Analysis of Universal Protein Extraction Methodologies for Screening of Lipase Activity from Agricultural Products |
| title_full | Comparative Analysis of Universal Protein Extraction Methodologies for Screening of Lipase Activity from Agricultural Products |
| title_fullStr | Comparative Analysis of Universal Protein Extraction Methodologies for Screening of Lipase Activity from Agricultural Products |
| title_full_unstemmed | Comparative Analysis of Universal Protein Extraction Methodologies for Screening of Lipase Activity from Agricultural Products |
| title_short | Comparative Analysis of Universal Protein Extraction Methodologies for Screening of Lipase Activity from Agricultural Products |
| title_sort | comparative analysis of universal protein extraction methodologies for screening of lipase activity from agricultural products |
| topic | lipase agricultural product protein extraction extraction yield specific activity spectrophotometric assay |
| url | https://www.mdpi.com/2073-4344/11/7/816 |
| work_keys_str_mv | AT jisuha comparativeanalysisofuniversalproteinextractionmethodologiesforscreeningoflipaseactivityfromagriculturalproducts AT junyoungpark comparativeanalysisofuniversalproteinextractionmethodologiesforscreeningoflipaseactivityfromagriculturalproducts AT yoonseokchoi comparativeanalysisofuniversalproteinextractionmethodologiesforscreeningoflipaseactivityfromagriculturalproducts AT pahnshickchang comparativeanalysisofuniversalproteinextractionmethodologiesforscreeningoflipaseactivityfromagriculturalproducts AT kyungminpark comparativeanalysisofuniversalproteinextractionmethodologiesforscreeningoflipaseactivityfromagriculturalproducts |