His260残基的替代改变食鹿角菜假交替单胞菌芳香基硫酸酯酶的热稳定性

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 H260 L and D84 A/H260 L 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(t_(1/2)) values at 55°C for H260 L, H260 I, H260 Q, H260 F and H260 R 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 H260 L, H260 I, H260 Q, H260 F and H260 R.