Identification of a histidyl residue in the active center of endoglucanase D from Clostridium thermocellum.

@article{Tomme1991IdentificationOA,
  title={Identification of a histidyl residue in the active center of endoglucanase D from Clostridium thermocellum.},
  author={Peter Tomme and Sylvie Chauvaux and Pierre B{\'e}guin and J L Millet and J. P. Aubert and Marc Claeyssens},
  journal={The Journal of biological chemistry},
  year={1991},
  volume={266 16},
  pages={10313-8}
}
Diethylpyrocarbonate modification of endoglucanase D from Clostridium thermocellum, cloned in Escherichia coli, resulted in a rapid but partial (maximally 70-80%) loss of activity. The second-order rate constant of inactivation proved to be exceptionally high (3210 M-1.min-1). A 3-fold reduction of the kcat and a 2-fold increase of the Km for 2'-chloro-4'-nitrophenyl beta-cellobioside were observed. Spectrophotometric analysis indicate the presence of one rapidly (k = 0.45 min-1) and two slower… CONTINUE READING