Fusion to an endoglucanase allows alkaline phosphatase to bind to cellulose

FEBS Letters

Miller, Robert C.¹  Warren, R.Antony J.¹  Gilkes, Neil R.¹  Greenwood, Jeffrey M.¹  Kilburn, Douglas G.¹ 

[1] Department of Microbiology, University of British Columbia, 300-6174 University Blvd, Vancouver BC V6T 1W5, Canada

关键词: Endoglucanase;    Cellulose-binding domain;    Alkaline phosphatase;    Fusion protein;    Purification;    Amp;    ampicillin;    CenA;    endoglucanase A;    CM-cellulose;    carboxymethylcellulose;    Kan;    kanamycin;    XP;    5-bromo-4-chloro-3-indolyl phosphate;   

DOI  :  10.1016/0014-5793(89)81177-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Endoglucanase CenA of Cellulomonas fimi comprises an N-terminal cellulose-binding domain and a C-terminal catalytic domain joined together by a sequence of 23 proline and threonine residues (the Pro-Thr box). The domains function independently when separated by proteolysis. TnphoA has been used to generate cenA′-′phoA fusions. CenA′-′PhoA fusion polypeptides which contain the entire cellulose-binding domain of CenA bind to cellulose, allowing their purification from periplasmic extracts in a single, facile step. This result has implications for purification or immobilisation of chimeric proteins on a cheap cellulose matrix.

【 授权许可】

Unknown