Inactivation of a subtilisin in colloidal systems.
The aim of the present study is to establish the relation between the inactivation of the proteolytic enzyme Savinase and its adsorption at different types of solid-liquid interfaces. The loss of activity has been determined both in solution and in the presence of colloidal particles, which provide a surface area for adsorption of 25% of the enzyme population. Analysis of the remaining solution at different periods of incubation of the various systems shows that the intact protein is converted into autolytic degradation products at the expense of biological activity. The different particles, however, deactivate the enzymes to a different extent. Under the experimental conditions the half-life of the enzymatic activity in solution is 3.5 hours. In the presence of particles that have hydrophobic surface properties (teflon- or polystyrene latex) the half-life is reduced to 0.7 hours. On the contrary, hydrophilic silica particles stabilize the enzyme against autolysis as compared to the inactivation in solution. Polystyrene latex particles which are chemically grafted with short poly(ethylene oxide) chains ([EO]8) are, for steric reasons, also mild with respect to the reduction of enzymatic stability. It is thus concluded that the type of surface determines the mode in which the enzyme is adsorbed on a particle which, in turn, affects the autocatalytic rate.
Maste
M C
MC
Department of Physical and Colloid Chemistry, Wageningen Agricultural University, The Netherlands.
Rinia
H A
HA
Brands
C M
CM
Egmond
M R
MR
Norde
W
W
D000327:Adsorption; D003102:Colloids; D004795:Enzyme Stability / Q000187:drug effects; D011092:Polyethylene Glycols / Q000494:pharmacology; D011137:Polystyrenes / Q000494:pharmacology; D011138:Polytetrafluoroethylene / Q000494:pharmacology; D012697:Serine Endopeptidases / Q000737:chemistry*; D012822:Silicon Dioxide / Q000494:pharmacology; D013381:Subtilisins / Q000737:chemistry*
D003102:Colloids; D011137:Polystyrenes; D011092:Polyethylene Glycols; D012822:Silicon Dioxide; D011138:Polytetrafluoroethylene; D012697:Serine Endopeptidases; D013381:Subtilisins; C032923:microbial serine proteinases