Cholesterol oxidase from Streptomyces hygroscopicus and Brevibacterium sterolicum: effect of surfactants and organic solvents on activity.
We have studied systematically the effect of the non-ionic surfactants Thesit and Triton X-100, and of propan-2-ol (used as a substrate solubilizer) on the activity of the cholesterol oxidases from Streptomyces hygroscopicus (SCO) and Brevibacterium sterolicum (BCO). Low concentrations of Thesit lead to an activity increase with both enzymes; at higher surfactant concentrations the opposite effect occurs. Triton X-100 inactivates both enzymes at all concentrations. It is deduced that these surfactants exert their effects by interaction with the enzymes and not by affecting micellar phenomena. The effect of propan-2-ol on SCO, in contrast with that on BCO, depends on the buffer concentration (potassium phosphate). Other organic solvents induce results similar to those obtained with SCO and propan-2-ol. A significant difference between the two cholesterol oxidases emerges when stability is tested at 25 degrees C and in the presence of different concentrations of propan-2-ol: BCO activity is rapidly inactivated, whereas SCO still has 70% of the initial activity after 5 h in the presence of 30% propan-2-ol. From our results, SCO seems to be the catalyst of choice in comparison with BCO for the exploitation of cholesterol oxidases in biotechnology and applied biochemistry.
Pollegioni
L
L
Department of Structural and Functional Biology, University of Insubria, via J.H. Dunant 3, 21100 Varese, Italy.
Gadda
G
G
Ambrosius
D
D
Ghisla
S
S
Pilone
M S
MS
D001951:Brevibacterium / Q000378:metabolism*; D002021:Buffers; D002789:Cholesterol Oxidase / Q000378:metabolism*; D004795:Enzyme Stability; D012997:Solvents / Q000494:pharmacology*; D013050:Spectrometry, Fluorescence; D013302:Streptomyces / Q000201:enzymology*; D013501:Surface-Active Agents / Q000494:pharmacology*
D002021:Buffers; D012997:Solvents; D013501:Surface-Active Agents; D002789:Cholesterol Oxidase