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High activity of inclusion bodies formed in Escherichia coli overproducing Clostridium thermocellum endoglucanase D
Author(s) -
Tokatlidis Kostas,
Dhurjati Prasad,
Millet Jacqueline,
Béguin Pierre,
Aubert Jean-Paul
Publication year - 1991
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(91)80478-l
Subject(s) - clostridium thermocellum , inclusion bodies , escherichia coli , biochemistry , cellulase , proteolysis , chemistry , enzyme , biology , microbiology and biotechnology , gene
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.