Purification and characterization of recombinant protease with thrombolytic activity of Perinereis aibuhitensis Grube
- VernacularTitle:重组沙蚕溶栓活性蛋白酶的纯化及其性质研究
- Author:
Ronggui LI
;
Feng ZHAO
;
Hong YANG
;
Guicai DU
;
Jmgchao WANG
;
Bin ANG
- Publication Type:Journal Article
- Keywords:
P.aibuhitensis;
protease with thrombolytic activity;
purification;
characterization
- From:
Chinese Journal of Marine Drugs
1994;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To express and purify recombinant protease with thrombolysis activity of Perinereis aibuhitensis Grube and study on its characterization.Methods pMAL-PPA was in- troduced into E.coli DH5?to construct engineering bacteria and overexpression of the prote- ase of fused with maltose binding protein(MBP-PPA)was achieved with IPTG induction. The fusion protein was purified by affinity chromatography on amylose-resin column fol- lowed by chromatography on a DEAE-sepharose FF column.PPA cut with Factor Xa was assayed using casein plates supplied with plasminogen.Results Engineering bacteria express- ing maltose binding protein-thrombolytic protease of P.aibuhitensis were constructed and overexpression of MBP-PPA was achieved with IPTG induction.A recombinant fusion pro- tein of 51kD was purified,and PPA cut down from the fusion protein had a plasminogen-acti- rating activity.The protease showed a good thermal stability with an optimal pH of 8.0. This enzyme was also relatively stable in a pH range of 6.0～9.0 and still active after stored in organic solvents for 20d.Conclusion PPA was verified as a plasminogen activator,and might be a new thrombolytic medicine in the future.
