Role of polyols in thermal inactivation of shark ornithine transcarbamoylase.
The ability of activity modulators of ornithine transcarbamoylase (OCT) from the liver of the thresher shark Alopias vulpinus to stabilize the enzyme against thermal denaturation was investigated in the tri-buffer at pH 7.8, at temperatures ranging from 60 to 70 (o)C, in the presence of polyhydroxylic molecules such as glycerol and sugars. The study indicated that in the presence of 0.5 M sugars and 1.6 M glycerol in the preincubation medium the OCT activity increases. When trehalose is introduced directly in the reaction mixture in a range of concentration of 0.25-0.5 M, the activity is lower than that with maltose, glycerol and buffer alone. Kinetic data for carbamoyl phosphate and ornithine with and without maltose and glycerol are similar, whereas trehalose increases the kinetic values. Arrhenius plots show an increase of activation energy due to trehalose, whereas values obtained with maltose and glycerol are similar to the control.
Belloco
E
E
Department of Organic and Biological Chemistry, University of Messina, Salita Sperone 31, Villaggio S. Agata, 98166 Messina, Italy. bellocco@isengard.unime.it
Laganà
G
G
Barreca
D
D
Ficarra
S
S
Tellone
E
E
Magazù
S
S
Branca
C
C
Kotyk
A
A
Galtieri
A
A
Leuzzi
U
U
D000818:Animals; D002241:Carbohydrates / Q000494:pharmacology*; D004734:Energy Metabolism / Q000502:physiology; D004789:Enzyme Activation / Q000502:physiology; D005990:Glycerol / Q000494:pharmacology*; D006207:Half-Life; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008320:Maltose / Q000494:pharmacology; D009954:Ornithine Carbamoyltransferase / Q000737:chemistry*; D011489:Protein Denaturation / Q000502:physiology; D012754:Sharks / Q000378:metabolism*; D013395:Sucrose / Q000494:pharmacology; D014199:Trehalose / Q000494:pharmacology
D002241:Carbohydrates; D013395:Sucrose; D008320:Maltose; D014199:Trehalose; D009954:Ornithine Carbamoyltransferase; D005990:Glycerol