Molecular cloning and nucleotide sequence of the groEL gene from the alkaliphilic Bacillus sp. strain C-125 and reactivation of thermally inactivated alpha-glucosidase by recombinant GroEL.
The groEL gene of the alkaliphilic Bacillus sp. strain C-125 was cloned in Escherichia coli and sequenced. The groEL gene encoded a polypeptide of 544 amino acids and was preceded by the incomplete groES gene, lacking its 5'-end. The sequence of the derived amino acids was 87.5% identical to that of B. subtilis, 85.4% identical to that of B. stearothemophilus, and 60.9% identical to that of E. coli. The GroEL protein was expressed in E. coli. Purified GroEL protected yeast alpha-glucosidase from irreversible aggregation at a high temperature and the addition of Mg-ATP was essential for reactivation of the alpha-glucosidase. The addition of E. coli GroES increased recovery of the enzyme activity, indicating that C-125 GroEL could function in coordination with E. coli GroES.
Xu
Y
Y
Microbiology Laboratory, Institute of Physical and Chemical Research (RIKEN), Saitama, Japan.
Kobayashi
T
T
Kudo
T
T
D000468:Alkalies; D000595:Amino Acid Sequence; D001407:Bacillus / Q000235:genetics*; D001483:Base Sequence; D018834:Chaperonin 60 / Q000096:biosynthesis / Q000235:genetics*; D003001:Cloning, Molecular; D004926:Escherichia coli; D005798:Genes, Bacterial; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis; D013696:Temperature
D000468:Alkalies; D018834:Chaperonin 60; D011994:Recombinant Proteins