Purification and Characterization of ?-AE Protein Expressed in E.coli
- VernacularTitle:大鼠?-酰胺化酶在大肠杆菌中的表达、纯化及活性研究
- Author:
Lian-Hong GUO
;
Xiao-Qiang QI
;
Rong JIANG
;
Chen YAO
;
Yuan LI
;
- Publication Type:Journal Article
- Keywords:
?-AE Cloning and expression Affinity chromatography Amidating Bioactive peptide
- From:
China Biotechnology
2006;0(11):-
- CountryChina
- Language:Chinese
-
Abstract:
Many bioactive peptides from neural and endocrine tissue are amidated at C-terminals,which is essential for their activities.The ?-amide comes from post-translational modification that is catalyzed by ?-AE (?-amidating enzyme) or PAM (pepdilylglycine ?-amidating monooxygenase).The gene encoding ?-AE was amplified with PCR and cloned into the plasmid pET-30a.After the recombinant plasmid pET-A was transformed into E.coli BL21,the ?-AE was expressed and purified by the Ni2+affinity chromatography,which has the ability catalyzing Dns-Tyr-Val-Gly to Dns-Tyr-Val-NH2.It identified that the recombinant protein producing by E.coli BL21 is ?-AE,which will benefit for studies of amidation at the C-terminals of peptides.
