In order to provide a new idea for molecular modification of xylanase, heterozygous xylanase was designed and synthetized in this study. Based on the xylanase XynZF-2 gene sequence of Aspergillus niger XZ-3S, 48 N-terminal amino acids were replaced by 34 N-terminal amino acids of EvXyn11. Heterozygous enzyme Xyn-ZL was designed by introducing aromatic amino acids P9Y and H14F, and disulfide bond Cys38-Cys191 at the C-terminal, hydrophobic amino acids K164M, G166A, N160I, and V111A at α-helix and G109A at cord area. After gene synthesis, the recombinant expression plasmid pPIC9K-ZL was constructed and transformed into P. pastoris GS115 to obtain the recombinant strain GS115-Xyn-ZL. It was found that the optimal fermentation condition was as follows: using potato medium as the medium, inoculated for 20 h before induction at 28 ℃ for 168 h at pH 6.5, and the optimal concentration of methanol for induction was 15 mL/L. The optimal reaction temperature and pH of the recombinant enzyme was 55 ℃ and pH 5.0, respectively. It was concluded that the XynZL expressed in P. pastoris exhibited specific properties and functions, which broadens the idea of modifying xylanase molecularly.
SHAO Tianci
,
HE You
,
ZHANG Fengkai
,
CAI Liutengzi
,
TIAN Yanjie
,
ZHOU Chenyan
. Design of heterozygous xylanase and the expression in Pichia pastoris[J]. Food and Fermentation Industries, 2019
, 45(19)
: 58
-62
.
DOI: 10.13995/j.cnki.11-1802/ts.021208
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