中国生物工程杂志

Many bioactive peptides from neural and endocrine tissue are amidated at C-terminals, which is essential for their activities. The α-amide comes from post-translational modification that is catalyzed by α-AE (α-amidating enzyme) or PAM (pepdilylglycine α-amidating monooxygenase). In this paper, the gene encoding α-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 α-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 α-AE, which will benefit for studies of amidation at the C-terminals of peptides.