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Title: Effect of Glycosylation Modification (N-Q-108I f N-Q-108T) on the Freezing Stability of Recombinant Chicken Cystatin Overexpressed in Pichia pastoris X-33
Authors: Shann-Tzong Jiang
Gen-Hung Chen
Shye-Jye Tang
Ching-San Chen
Contributors: 國立臺灣海洋大學:食品科學系
Keywords: glycosylation modification
P. pastoris
recombinant cystatin
Date: 2002
Issue Date: 2018-05-23T01:35:13Z
Publisher: Journal of Agricultural and Food Chemistry
Abstract: 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 as resistant agent, and transformed into Pichia pastoris X-33 expression host. The effect of N-glycosylation on the stability of recombinant chicken cystatin was investigated. A large quantity of recombinant chicken cystatin and the Asn106-glycosylated cystatins were expressed and secreted into broth using α-factor preprosequence. The Ki of the recombinant chicken cystatin (0.08 nM) was similar to that of wild-type chicken cystatin (0.05 nM). They acted as a competitive inhibition reaction against 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 substantially enhanced the freezing stability of recombinant chicken cystatin overexpressed in P. pastoris.
Relation: 50(19) pp.5313–5317
URI: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/46536
Appears in Collections:[食品科學系] 期刊論文

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