Li Jia-Da, Wang Ke-Yi
Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, the Chinese Academy of Sciences, Shanghai 200031, China.
Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai). 2001;33(5):513-518.
A phage-displayed peptide library was biopanned with Aspergillus niger glucoamylase. Three specifically binding phage clones (GB-1, GB-2 and GB-3) were identified after three cycles of screening and amplification. All three peptides contained a disulfide bridge, and the impairment of disulfide bond greatly lowered the binding activity. One peptide, cyclic KCHFEECLAY, representing the N-terminal 10 amino acid residues from GB-1, competitively inhibited A. niger glucoamylase (K(I) 0.2 mmol/L) as well as rat intestinal alpha-glucosidase (KI 1.4 mmol/L).
用黑曲霉葡糖淀粉酶对噬菌体展示肽库进行生物淘选。经过三轮筛选和扩增后,鉴定出三个特异性结合的噬菌体克隆(GB-1、GB-2和GB-3)。所有三个肽都含有一个二硫键,二硫键的破坏大大降低了结合活性。一种代表GB-1 N端10个氨基酸残基的环肽KCHFEECLAY,竞争性抑制黑曲霉葡糖淀粉酶(抑制常数K(I)为0.2 mmol/L)以及大鼠肠道α-葡糖苷酶(抑制常数KI为1.4 mmol/L)。