Borate ion-assisted stabilization of beta-galactosidase from Aspergillus oryzae by polyhydroxy compounds in water-miscible organic solvents.
The stability of beta-galactosidase from Aspergillus oryzae in water-miscible organic solvents in different buffers at various pH values ranging from 4.6 to 8.0 was studied. The stability of the enzyme in all six organic solvents studied was dependent on pH and on the type of buffer ions present. At a given pH, destabilization by organic solvents was highest in sodium borate buffer. The destabilization of beta-galactosidase by these solvents could be reversed by addition of sugars or polyhydroxy compounds exclusively in sodium borate, suggesting a role of borate ions in stabilization. A similar effect of addition of mannitol was observed on deactivation of beta-galactosidase by N, N-dimethylformamide (DMF). Exclusively in sodium borate, at pH 8.0, the addition of mannitol (0.02 M) not only prevented the deactivation by DMF (8%, v/v) but increased the enzyme activity to the level at its optimum pH. Since beta-galactosidase from Aspergillus oryzae is a glycoprotein, complexation of the borate ions to the carbohydrate part may result in change in protein conformation, which, without leading to denaturation or inactivation of the enzyme, may facilitate interaction of the organic solvents with the enzyme leading to its denaturation. Such a denaturation is probably prevented by addition of polyhydroxy compounds, which appear to compete favorably with the carbohydrate moiety of the protein in complexing with borate ions. This should result in the enzyme regaining its native conformation.
Shubhada
S
S
Centre for Protein Engineering and Biomedical Research, Voluntary Health Services, Madras, India.
Sundaram
P V
PV
D001236:Aspergillus oryzae / Q000201:enzymology*; D001709:Biotechnology; D001881:Borates / Q000494:pharmacology; D002021:Buffers; D004795:Enzyme Stability / Q000187:drug effects; D006863:Hydrogen-Ion Concentration; D008353:Mannitol / Q000494:pharmacology; D011108:Polymers / Q000494:pharmacology; D012997:Solvents; D014867:Water; D001616:beta-Galactosidase / Q000378:metabolism*
D001881:Borates; D002021:Buffers; D011108:Polymers; D012997:Solvents; C024617:polyol; D014867:Water; D008353:Mannitol; C010634:sodium borate; D001616:beta-Galactosidase