| Attributs | Valeurs |
|---|
| type
| |
| Is Part Of
| |
| Subject
| |
| Title
| - Effect of Glycosylation Modification (N-Q-108I → N-Q-108T) onthe Freezing Stability of Recombinant Chicken CystatinOverexpressed in Pichiapastoris X-33
|
| has manifestation of work
| |
| related by
| |
| Author
| |
| Abstract
| - The cDNAs encoding chicken cystatin and its N-glycosylation-modified mutant (Asn106−Ile108→Asn106−Thr108) were cloned into the pGAPZαC expression vector, using the GAP as promoter and Zeocin asresistant agent, and transformed into Pichiapastoris X-33 expression host. The effect of N-glycosylation on the stability of recombinant chicken cystatin was investigated. A large quantity ofrecombinant chicken cystatin and the Asn106-glycosylated cystatins were expressed and secretedinto broth using α-factor preprosequence. The Ki of the recombinant chicken cystatin (0.08 nM) wassimilar to that of wild-type chicken cystatin (0.05 nM). They acted as a competitive inhibition reactionagainst papain. According to the Ki, the inhibition ability of Asn106-glycosylated mutant cystatin (Ki =9.5 nM) was weaker than that of the wild-type one. However, N-glycosylation at Asn106 substantiallyenhanced the freezing stability of recombinant chicken cystatin overexpressed in P. pastoris. Keywords: P. pastoris; glycosylation modification; recombinant cystatin; overexpression
|
| article type
| |
| is part of this journal
| |
