title: Improved activity of α-chymotrypsin on silica particles - A high-pressure stopped-flow study.
abstract: Pressure is well known to affect the catalytic rate of enzymes dissolved in solution. To better understand enzyme kinetics at aqueous-solid interfaces, we have carried out a high-pressure stopped-flow activity study of α-chymotrypsin (α-CT) that is adsorbed on silica particles and, for comparison, dissolved in solution. The enzyme reaction was modulated using pressures up to 2000bar and recorded using the high-pressure stopped-flow technique. The results indicate an 8-fold enhancement of the turnover number upon α-CT adsorption and a further increase of the catalytic rate in the pressure range up to 1000bar. From the pressure dependence of the catalytic rate, apparent activation volumes have been determined. In the adsorbed state of α-CT, a pronounced change of the activation volume is found with increasing pressure. Furthermore, owing to suppression of its autolysis, a significantly longer storage time of α-CT can be achieved when the enzyme is adsorbed on silica particles. The results obtained are discussed in terms of a surface-induced selection of conformational substates of the enzyme-substrate complex.
authors: TU Dortmund University, Department of Chemistry and Chemical Biology, Otto-Hahn-Str. 4a, D-44227 Dortmund, Germany.
authors: Vitor
authors: V
authors: Schuabb
authors: TU Dortmund University, Department of Chemistry and Chemical Biology, Otto-Hahn-Str. 4a, D-44227 Dortmund, Germany.
authors: Roland
authors: R
authors: Winter
authors: TU Dortmund University, Department of Chemistry and Chemical Biology, Otto-Hahn-Str. 4a, D-44227 Dortmund, Germany. Electronic address: claus.czeslik@uni-dortmund.de.
authors: Claus
authors: C
authors: Czeslik
mesh_terms: D000327:Adsorption; D002918:Chymotrypsin / Q000378:metabolism*; D004800:Enzymes, Immobilized / Q000378:metabolism*; D007700:Kinetics; D011312:Pressure; D012822:Silicon Dioxide; D012996:Solutions; D013379:Substrate Specificity
chemical_list: D004800:Enzymes, Immobilized; D012996:Solutions; D012822:Silicon Dioxide; D002918:Chymotrypsin; C117608:alpha-chymotrypsin
keywords: Enzyme; Interface; Pressure; Silica; Stopped-flow method