title: Overproduction of beta-glucosidase in active form by an Escherichia coli system coexpressing the chaperonin GroEL/ES.
abstract: beta-Glucosidase from Cellvibrio gilvus was successfully overproduced in soluble form in Escherichia coli, with the coexpression of GroEL/ES. Without the GroEL/ES protein, the beta-glucosidase overexpressed in E. coli constituted a huge amount (80%) of the total cellular protein, but was localized in the insoluble fraction, and little activity was detected in the soluble fraction. Coexpression of the E. coli GroEL/ES had a drastic impact on the proper folding of the beta-glucosidase; 20% of the overexpressed enzyme was recovered in the soluble fraction in active form. In addition, the synergistic effect of GroEL/ES and the low induction temperature led to 70% solubilization of the total expressed target protein and more than a 20-fold increase in activity. Similar effects of GroEL/ES were also observed on the overexpressed beta-glucosidase from Agrobacterium tumefaciens.
authors.affiliation: National Food Research Institute, Ministry of Agriculture, Forestry and Fishery, Ibaraki, Japan. lili@nfri.affrc.go.jp
authors.forename: S
authors.initials: S
authors.lastname: Machida
authors.forename: Y
authors.initials: Y
authors.lastname: Yu
authors.forename: S P
authors.initials: SP
authors.lastname: Singh
authors.forename: J D
authors.initials: JD
authors.lastname: Kim
authors.forename: K
authors.initials: K
authors.lastname: Hayashi
authors.forename: Y
authors.initials: Y
authors.lastname: Kawata
mesh_terms: D018834:Chaperonin 60 / Q000235:genetics*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D005798:Genes, Bacterial / Q000235:genetics; D016946:Gram-Negative Aerobic Rods and Cocci / Q000201:enzymology; D013696:Temperature; D001617:beta-Glucosidase / Q000235:genetics*
chemical_list: D018834:Chaperonin 60; D001617:beta-Glucosidase