| RRC ID |
71286
|
| 著者 |
Wang S, Meng K, Su X, Hakulinen N, Wang Y, Zhang J, Luo H, Yao B, Huang H, Tu T.
|
| タイトル |
Cysteine Engineering of an Endo-polygalacturonase from Talaromyces leycettanus JCM 12802 to Improve Its Thermostability.
|
| ジャーナル |
J Agric Food Chem
|
| Abstract |
Thermostable enzymes have many advantages for industrial applications. Therefore, in this study, computer-aided design technology was used to improve the thermostability of a highly active endo-polygalacturonase from Talaromyces leycettanus JCM12802 at an optimal temperature of 70 °C. The melting temperature and specific activity of the obtained mutant T316C/G344C were increased by 10 °C and 36.5%, respectively, compared with the wild-type enzyme. The crystal structure of the T316C/G344C mutant showed no formation of a disulfide bond between the introduced cysteines, indicating a different mechanism than the conventional mechanism underlying improved enzyme thermostability. The cysteine substitutions directly formed a new alkyl hydrophobic interaction and caused conformational changes in the side chains of the adjacent residues Asn315 and Thr343, which in turn caused a local reconstruction of hydrogen bonds. This method greatly improved the thermostability of the enzyme without affecting its activity; thus, our findings are of great significance for both theoretical research and practical applications.
|
| 巻・号 |
69(22)
|
| ページ |
6351-6359
|
| 公開日 |
2021-6-9
|
| DOI |
10.1021/acs.jafc.1c01618
|
| PMID |
34043362
|
| MeSH |
Cysteine
Enzyme Stability
Polygalacturonase* / genetics
Polygalacturonase* / metabolism
Talaromyces* / genetics
Talaromyces* / metabolism
Temperature
|
| リソース情報 |
| 一般微生物 |
JCM12802 |
