화학공학소재연구정보센터
Journal of Fermentation and Bioengineering, Vol.77, No.2, 199-201, 1994
Molecular-Cloning of Thermostable Beta-Glucosidase Gene from a Thermophilic Anaerobe Na10 and Its High Expression in Escherichia-Coli
A gene encoding beta-glucosidase was cloned from a thermophilic anaerobe strain NA10 into Escherichia coli. The 4.6 kb of HindIII fragment was shown to direct the synthesis of beta-glucosidase. A homologous DNA sequence was found to be present in strain NA10 by Southern blot analysis. The beta-glucosidase was purified from the recombinant E. coli. The molecular weight of the enzyme was estimated to be 50.1 kDal using SDS-polyacrylamide gel electrophoresis. The enzyme was heat stable; the optimum pH and temperature were 5.5 and 85-degrees-C, respectively. The enzyme could hydrolyse low molecular weight beta-1,3 and beta-1,4 glucans. The beta-glucosidase was produced in the cytosol of the recombinant E. coli (7-20% of total protein), without fusion to any E. coli promoter sequences.