A novel approach for purification of recombinant proteins using the dextran‐binding domain

FEBS Letters

Kodama, Takao²  Fukui, Kazuhiro¹  Kaseda, Kuniyoshi³  Hirose, Keiko¹ 

[1] Department of Microbiology, Okayama University Dental School, Okayama 700-8525, Japan;Laboratory of Molecular Enzymology, Department of Biochemical Science and Engineering, Kyushu Institute of Technology, Ihzuka, Fukuoka 820-8502, Japan;Gene Discovery Research Center, National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1 Higashi, Tsukuba, Ibaraki 305-8562, Japan

关键词: Dextran;    Dextran-binding domain;    Glucosyltransferase;    Purification;    Recombinant protein;    Tag;    DBD;    dextran-binding domain;    GFP;    green fluorescent protein;    GTF;    glucosyltransferase;    RFP;    red fluorescent protein;    Triton X-100;    polyoxyethylene (10) octylphenyl ester;    Tween 20;    polyoxyethylenesorbitan monolaurate;   

DOI  :  10.1016/S0014-5793(01)02607-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Using the dextran-binding domain (DBD) of a type of glucosyltransferase (GTF) from Streptococcus sobrinus, we have developed a novel method for purifying recombinant proteins. DBD-tagged green and red fluorescent proteins as well as the parent GTF and DBD moiety were adsorbed well to commercially available cross-linked dextran (such as Sephadex beads and Sephacryl beads), and eluted efficiently with water-soluble dextran. The purity of the eluted proteins after this one-step affinity purification was ∼90% or better. The results suggest that DBD can be used as a powerful carrier for purification of various recombinant proteins.

【 授权许可】

Unknown