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The endoglucanase CelA from Clostridium thermocellum was strongly expressed in Bacillus subtilis. The enzyme was purified by Ni2+-affinity chromatography. Site-directed substitution of D278 with an asparagine or an alanine residue surprisingly did not decrease the apparent kcat value. Further substitutions of two other potentially critical residues, Y215 and D152, resulted in a 2-fold decrease in...
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