Overproduction, purification, and characterization of beta-1,3-glucanase type II in Escherichia coli

An Escherichia coli recombinant system produced soluble and full-length bet a -1,3-glucanase type II (BglII) cloned from the yeast-lytic actinomycete O erskovia xanthineolytica. The expression system was designed to produce rec ombinant BglII with a six-histidine peptide fused to the carboxy end of the protein. The expression level was optimized to produce 30% of total protei n of E. coli as the recombinant protein, releasing 75% to the extracellular space. The 43-kDa recombinant protein was purified by IMAC to homogeneity and its molecular and biochemical characteristics were studied, showing tha t there are no important functional differences with those properties descr ibed for the BglII purified from O. xanthineolytica. (C) 2001 Academic Pres s.