Open AccessOverproduction of β‐glucosidase in active form by an Escherichia coli system coexpressing the chaperonin GroEL/ES
Author(s) -
Machida Sachiko,
Yu Yong,
Singh Satya P,
Kim JongDeog,
Hayashi Kiyoshi,
Kawata Yasushi
Publication year - 1998
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1998.tb12839.x
Subject(s) - groel , escherichia coli , chaperonin , overproduction , agrobacterium tumefaciens , biochemistry , biology , enzyme , protein folding , microbiology and biotechnology , gene , transgene
β‐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 β‐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 β‐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 β‐glucosidase from Agrobacterium tumefaciens .