Structure-based design of agarase AgWH50C from Agarivorans gilvus WH0801 to enhance thermostability
| Autor: | Lujia Zhang, Changhu Xue, Jiahai Zhou, Lian Wu, Jianan Sun, Yuan Li, Xiangzhao Mao, Pujuan Zhang, Jinru Zhang |
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| Rok vydání: | 2018 |
| Předmět: |
0106 biological sciences
0301 basic medicine Hot Temperature Glycoside Hydrolases Mutant Crystallography X-Ray Disaccharides 01 natural sciences Applied Microbiology and Biotechnology Protein Structure Secondary 03 medical and health sciences chemistry.chemical_compound Hydrolysis Bacterial Proteins 010608 biotechnology Hydrolase Glycoside hydrolase Thermostability biology Alteromonadaceae Agarase Rational design General Medicine 030104 developmental biology chemistry Biochemistry Mutagenesis Mutation biology.protein Agarose Biotechnology |
| Zdroj: | Applied microbiology and biotechnology. 103(3) |
| ISSN: | 1432-0614 |
| Popis: | AgWH50C, an exo-β-agarase of GH50 isolated from Agarivorans gilvus WH0801, plays a key role in the enzymatic production of neoagarobiose, which has great application prospect in the cosmetics and pharmaceutical industry. In contrast, the poor thermostability becomes the main obstructive factor of glycoside hydrolase (GH) family 50 agarases, including AgWH50C. Herein, based on the AgWH50C crystal structure, we designed several mutants by a multiple cross-linked rational design protocol used thermostability predicting softwares ETSS, PoPMuSiC, and HotMuSiC. To our surprise, the mutant K621F increased its relative activity by as much as 45% and the optimal temperature increased to 38 °C compared to that of wild-type, AgWH50C (30 °C). The thermostability of K621F also exhibited a substantial improvement. Considering that the gelling temperature of the agarose is higher than 35 °C, K621F can be used to hydrolyze agarose for neoagarobiose production. |
| Databáze: | OpenAIRE |
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