Carboxymethyl celluase (CMCase) was purified from cell-free extract of the recombinant Escherichia coli carrying a Bacillus sp. 79-23 CMCase gene (celS) by DEAE-Sepharose and phenyl-Sepharose column chromatography with specific activity of 400 U/mg pr...
Carboxymethyl celluase (CMCase) was purified from cell-free extract of the recombinant Escherichia coli carrying a Bacillus sp. 79-23 CMCase gene (celS) by DEAE-Sepharose and phenyl-Sepharose column chromatography with specific activity of 400 U/mg protein. The molecular mass of the purified enzyme was estimated to be approximately 37.5 daltons by sodium dodecyl sulfatepolycarylamide gel electrophoresis. The viscosity of carboxymethyl celluose was dramatically decreased by the purified enzyme identifying that the purified enzyme is a CMCase, while the enzyme was known to be active on para-nitrophenyl-β-cellobioside. The CMCase activity was markedly inhibited by cellobiose, but was not inhibited by glucose. The enzyme activity was activated by divalent metal ions such as Mn²+ and Cu²+, but was completely inhibited by Hg²+. The enzyme was very stable below 50℃ and retained 80% of its maximum activity after incubation for 3 hours at 55℃, suggesting that few problems are in handing the Bacillus sp. CMCase as industrial enzyme at ambient temperature.