期刊
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
卷 153, 期 -, 页码 441-450出版社
ELSEVIER
DOI: 10.1016/j.ijbiomac.2020.02.319
关键词
Esterase; Functional metagenomics; Protein purification; Nudeic acid aptamers
资金
- Project on the Integration of Industry, Education and Research of Guangdong Province [2013B090500094]
- National Key Research and Development Program of China [2016YFE0117100]
- Science and Technology Program of Guangdong Province [2016B090918019]
- GDAS Special Project of Science and Technology Development [2017GDASCX-0107]
A new esterase gene est906 was identified from paper mill wastewater sediments via a function-based metagenomic approach. The gene encoded a protein of 331 amino acids, that shared 86% homology with known esterases. Based on the results of multiple sequence alignment and phylogenetic analysis, it was confirmed that Est906 contained a characteristic hexapeptide motif (G-F-S-M-G-G), which classified it as a lipolytic enzyme family V protein. Est906 displayed the highest hydrolysis activity to p-nitrophenyl caproate (C6), and its optimal temperature and pH were 54 degrees C and 93, respectively. Additionally, this enzyme had good stability under strong alkaline conditions (pH 10.0-11.0) in addition to moderate heat resistance and good tolerance against several metal ions and organic solvents. Furthermore, a specific nucleic acid aptamer (Apt1) bound to Est906 was obtained after five rounds of magnetic bead SELEX screening. Apt1 displayed high specific recognition and capture ability to Est906. In conclusion, this study not only identified a new esterase of family V with potential industrial application by metagenomic technology but also provided a new method to purify recombinant esterases via nucleic acid aptamers, which will facilitate the isolation and purification of target proteins in the future. (C) 2020 Elsevier B.V. All rights reserved.
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