Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Pivot Concepts:
Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Target Concepts:
Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Query: EC:3.2.1.21 (
beta-glucosidase
)
3,280
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
In continuation of our investigation on structure and function relationship of beta-glucosidases from mesophilic and thermophilic bacteria, we constructed a chimeric gene by shuffling 17% length in C terminal region of
beta-glucosidase
of Agrobacterium tumefaciens with the corresponding homologous region of Cellvibrio gilvus
beta-glucosidase
. The chimeric gene was overexpressed in E. coli BL21 (DE3) using pET vector. However, nearly all of the
beta-glucosidase
produced was trapped into inclusion bodies in catalytically
non-functional
state. Attempts were made to solubilize the overexpressed protein by co-expression with molecular chaperone, GroEL/ES, in vivo. The molecular chaperone assisted protein folding that had earlier yielded encouraging results, did not improve the solubilization in the present case with a chimeric
beta-glucosidase
. Further, we explored protein renaturation under in vitro conditions using various dialysis strategies. Dialysis, rapid dilution and a newly devised method of folding immobilized proteins yielded active enzyme. The usefulness of the in vitro folding methods to obtain functional enzymes from overproduced but
non-functional
proteins has been discussed.
...
PMID:Overexpression and protein folding of a chimeric beta-glucosidase constructed from Agrobacterium tumefaciens and Cellvibrio gilvus. 2290 13
