Penicillin acylase is mainly used in the semi-synthetic industrial production of β-lactam antibiotics.This thesis, the pga gene was obtained from Providencia rettgeri, heterologously expressed in Komagataella pastoris GS115, and the enzyme production conditions were optimized for the recombinant enzyme PA.The results of the heterologous expression constructs showed that the enzyme activity of reorganization K. pastoris fermentation supernatant was 2 155.59 U/L, and the precipitated enzyme activity was 1 375.75 U/L. The optimal conditions for induction of enzyme production of recombinant enzyme were 29 ℃, 72 h, and methanol concentration of 1.5%, and the activity of recombinant enzyme PA was (5 003.72±50.623) U/L, which was 1.57-fold higher than the pre-optimization enzyme activity, and the purified The protein size was 72 kDa, the protein concentration was 1.58 mg/mL, the optimum reaction temperature of the purified recombinant enzyme was 60 ℃, the optimum pH was 7, and it was strongly inhibited by Mn2+ and activated by Al3+.The results of this study can provide guidance for the heterologous expression of penicillin acylase and further industrial applications.
JIA Yuhan
,
ZHANG Xiangjun
,
MA Cheng
,
ZHU Tengteng
,
WEI Xiaobo
,
CHEN Long
,
LIU Huiyan
,
FANG Haitian
. Heterologous expression of penicillin acylase in Komagataella pastoris and optimization of its enzyme production conditions[J]. Food and Fermentation Industries, 2025
, 51(9)
: 33
-39
.
DOI: 10.13995/j.cnki.11-1802/ts.039587
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