Overproduction, purification, and characterization of beta-1,3-glucanase type II in Escherichia coli.
An Escherichia coli recombinant system produced soluble and full-length beta-1,3-glucanase type II (BglII) cloned from the yeast-lytic actinomycete Oerskovia xanthineolytica. The expression system was designed to produce recombinant BglII with a six-histidine peptide fused to the carboxy end of the protein. The expression level was optimized to produce 30% of total protein 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 that there are no important functional differences with those properties described for the BglII purified from O. xanthineolytica.
Salazar
O
O
Centre for Biochemical Engineering and Biotechnology, Department of Chemical Engineering, Millennium Institute for Advanced Studies in Cell Biology and Biotechnology, University of Chile, Beauchef 861, Santiago, Chile. osalazar@cec.uchile.cl
Molitor
J
J
Lienqueo
M E
ME
Asenjo
J A
JA
D000192:Actinomycetales / Q000201:enzymology*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D043326:Glucan 1,3-beta-Glucosidase; D011993:Recombinant Fusion Proteins; D001617:beta-Glucosidase / Q000235:genetics*
D011993:Recombinant Fusion Proteins; D001617:beta-Glucosidase; D043326:Glucan 1,3-beta-Glucosidase