SUMO-fusion expressions and catalytic efficiency of an organophosphate hydrolase mutant
10.7644/j.issn.1674-9960.2025.08.006
- VernacularTitle:有机磷水解酶突变体的SUMO融合表达及活性评价
- Author:
Zhuang LIU
1
;
Yanan ZHAI
;
Shunye WANG
;
Ming MA
;
Ziyang WANG
;
Yanqin LIU
;
Xiang GAO
;
Jing GAO
Author Information
1. 军事科学院军事医学研究院,北京 100850
- Keywords:
organophosphate hydrolase;
SUMO-Tag;
acetylcholinesterase;
ethyl paraoxon;
soman;
kinetic parameters
- From:
Military Medical Sciences
2025;49(8):598-603
- CountryChina
- Language:Chinese
-
Abstract:
Objective To heterologously express and purify the small ubiquitin-like modifier(SUMO)tag fused organo-phosphorus hydrolase mutant H257Y/L303T(YT),namely SUMO-YT,and evaluate its hydrolytic capacity against ethyl paraoxon and soman.Methods The SUMO tag encoding gene was constructed at the N-terminus of the YT encoding gene with a linker sequence via enzyme digestion and ligation before SUMO-YT was expressed in Escherichia coli.SUMO-YT and YT were purified through ammonium sulfate precipitation and affinity chromatography to obtain high-purity target proteins.The activity and kinetic parameters of the recombinant enzymes were examined using ethyl paraoxon and soman as substrates.Results The system for expression and purification of recombinant enzymes was established,yielding SUMO-YT and YT,and the former exhibited more significantly enhanced hydrolytic efficiency than the latter,with catalytic rates 11-fold higher for paraoxon and 4.4-fold higher for soman.At 37 ℃ and pH 7.2,SUMO-YT reduced the inhibition rate of acetylcholinesterase(AChE)by soman from 100％to 45.7％within 3 minutes,whereas YT reduced it to no more than 80％.Conclusion The high-activity recombinant SUMO-YT is prepared.SUMO tag fusion can significantly enhance the hydrolytic capacity of YT against ethyl paraoxon and soman.
