TY - JOUR
T1 - Structure-based design of agarase AgWH50C from Agarivorans gilvus WH0801 to enhance thermostability
AU - Zhang, Pujuan
AU - Zhang, Jinru
AU - Zhang, Lujia
AU - Sun, Jianan
AU - Li, Yuan
AU - Wu, Lian
AU - Zhou, Jiahai
AU - Xue, Changhu
AU - Mao, Xiangzhao
N1 - Publisher Copyright:
© 2018, Springer-Verlag GmbH Germany, part of Springer Nature.
PY - 2019/2/1
Y1 - 2019/2/1
N2 - 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.
AB - 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.
KW - Agarase
KW - Mutants
KW - Predicting softwares
KW - Structure
KW - Thermostability
UR - https://www.scopus.com/pages/publications/85058012606
U2 - 10.1007/s00253-018-9540-1
DO - 10.1007/s00253-018-9540-1
M3 - 文章
C2 - 30523371
AN - SCOPUS:85058012606
SN - 0175-7598
VL - 103
SP - 1289
EP - 1298
JO - Applied Microbiology and Biotechnology
JF - Applied Microbiology and Biotechnology
IS - 3
ER -