Laccase is an important green biocatalyst with wide application prospects in the field of environmental remediation. However, owing to the low tolerance to environmental factors, e.g. unsuitable pH and temperature, could lead to the disruption of the structure, and the rapid loss of catalytic activity, while might limit the application of laccase. In this paper, the laccase BILac (GenBank: AAU23292.2) from Bacillus licheniformis was recombinantly expressed and modified through homologous modeling and molecular docking. Two mutagenesis libraries including the shift of binding energy and the designing of disulfide bond were built, and the multiple mutations between the two libraries were also constructed. Finally, the mutant C189A-S251C with promoted activity and thermal stability was screened. The catalytic efficiency against substrate ABTS was up to 6.3 times as that of the native one, the Tm was enhance from 54 ℃ to 57 ℃,while the half-life at 40 ℃ was promoted from 2.5 h to 5.5 h. The investigation of optimal pH and temperature indicated that the tolerance to acidic environments and temperature were all increased, the optimum temperature also increased from 37 ℃ to 50 ℃. This substitution will bring great potential of laccase BlLac in environmental application. At the same time, the design strategy of this study provides a feasible solution for the modification of other enzymes.
HUANG Aimin
,
GUO Zhongpeng
,
LI Moying
,
GUO Zitao
,
GU Zhenghua
,
ZHANG Liang
,
XIN Yu
. Activity and thermal stability modification of laccase BlLac derived from Bacillus licheniformis[J]. Food and Fermentation Industries, 2023
, 49(21)
: 30
-38
.
DOI: 10.13995/j.cnki.11-1802/ts.034552
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