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Substitution of His260 residue alters the thermostability of Pseudoalteromonas carrageenovora arylsulfatase

Authors :
Hui Ni
Hebin Li
Lijun Li
Xiaoqian Yin
Yanbing Zhu
Han Liu
Yanhong Chen
Anfeng Xiao
Source :
Acta Oceanologica Sinica. 38:75-82
Publication Year :
2019
Publisher :
Springer Science and Business Media LLC, 2019.

Abstract

This study aimed to improve the thermostability of arylsulfatase from Pseudoalteromonas carrageenovora. A library of P. carrageenovora arylsulfatase mutants was constructed by introducing random mutagenesis using error-prone PCR. After screening, two mutants of H260L and D84A/H260L showed enhanced thermal stability than the wild-type predecessor (WT). Site-directed mutagenesis demonstrated that only amino acid residue at Position 260 plays an important role in the thermostability of P. carrageenovora arylsulfatase. Thermal inactivation analysis showed that the half-life (t1/2) values at 55°C for H260L, H260I, H260Q, H260F and H260R were 40.6, 48.4, 30.9, 29.1 and 34.5 min, respectively, while that of WT was 9.1 min. Structure modeling demonstrated that the additional hydrogen bonds and/or optimization of surface charge-charge interactions could be responsible for the increased thermostability imparted by H260L, H260I, H260Q, H260F and H260R.

Details

ISSN :
18691099 and 0253505X
Volume :
38
Database :
OpenAIRE
Journal :
Acta Oceanologica Sinica
Accession number :
edsair.doi...........b51680df575c923c966f62e02af18c7d
Full Text :
https://doi.org/10.1007/s13131-019-1356-z