Comparative Analysis of Universal Protein Extraction Methodologies for Screening of Lipase Activity from Agricultural Products

Catalysts

Jun-Young Park¹  Yoonseok Choi¹  Pahn-Shick Chang¹  Jisu Ha²  Kyung-Min Park² 

[1] Department of Agricultural Biotechnology, Seoul National University, Seoul 08826, Korea;Department of Food Science and Biotechnology, Wonkwang University, Iksan 54538, Korea;

关键词: lipase;    agricultural product;    protein extraction;    extraction yield;    specific activity;    spectrophotometric assay;   

DOI  :  10.3390/catal11070816

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown