High activity of inclusion bodies formed in Escherichia coli overproducing Clostridium thermocellum endoglucanase D

FEBS Letters

Dhurjati, Prasad¹  Béguin, Pierre¹  Millet, Jacqueline¹  Tokatlidis, Kostas¹  Aubert, Jean-Paul¹ 

[1] Unité Physiologie Cellulaire, Département des Biotechnologies and URA CNRS 1300, Institut Pasteur, rue du Dr. Roux, 75724 Paris Cedex 15, France

关键词: Inclusion body;    Clostridium thermocellum;    Endoglucanase D;    Protein folding;    EG;    endoglucanase (E.C. 3.2.1.4);    PAGE;    polyacrylamide gel electrophoresis;    p-NPC;    p-nitrophenyl-β-D-cellobioside;    p-NPCase;    p-nitrophenyl-β-D-cellobiosidase;    SDS;    sodium dodecyl sulfate;   

DOI  :  10.1016/0014-5793(91)80478-L

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The formation of cytoplasmic inclusion bodies by Escherichia coli overproducing Clostridium thermocellum endoglucanase D (EGD) was investigated. EGD was found in inclusion bodies as a 68 kDa form, whereas the size of the cytoplasmic form was 65 kDa. Upon solubilization with urea followed by dialysis, the 68 kDa form was converted to the 65 kDa species. Proteolysis occurred within the COOH-terminal, reiterated region of the 68 kDa form, which is conserved among most C. thermocellum endoglucanase, but is not required for catalytic activity. The specific activity of the enzyme embedded in inclusion bodies was close to that of the purified protein. Thus, inclusion body formation does not involve denaturation of the catalytic domain of EGD, but more likely, the participation of the reiterated, conserved region in intermolecular interactions.

【 授权许可】

Unknown