title: Acceleration of enzymatic reaction of trypsin through the formation of water-soluble complexes with poly(ethylene glycol)-block-poly(alpha,beta-aspartic acid).
abstract: The amidase activity of bovine pancreas trypsin in water-soluble complexes with poly(ethylene glycol)-block-poly(alpha,beta-aspartic acid) (PEG-PAA) was evaluated by a colorimetric assay using L-lysine p-nitroanilide as a substrate. The enzymatic reaction of trypsin was accelerated through the complexation with PEG-PAA. By determining the kinetic parameters of the enzymatic reaction of trypsin, it was confirmed that the catalytic rate constant of the complexed trypsin was 15 times higher than that of the native trypsin. From the evaluation of pH dependence of initial reaction rate, it was indicated that this acceleration was induced by a stabilization of the imidazolium ion of the His residue in the catalytic site, the Asp-His-Ser triad, of trypsin due to the Asp units of PEG-PAA. The hydrogen bonded Asp-His pairs are critical constituents in several key enzymatic reactions including serine protease and apurinic endonucleases, and it was expected that the acceleration of the catalytic reaction might occur for other enzymes by the formation of water-soluble complexes with PEG-PAA.
authors.affiliation: Department of Applied Materials Science, Graduate School of Engineering, Osaka Prefecture University, 1-1 Gakuen-cho, Sakai, Osaka 599-8531, Japan.
authors.forename: Akifumi
authors.initials: A
authors.lastname: Kawamura
authors.forename: Yuriko
authors.initials: Y
authors.lastname: Yoshioka
authors.forename: Atsushi
authors.initials: A
authors.lastname: Harada
authors.forename: Kenji
authors.initials: K
authors.lastname: Kono
mesh_terms: D000596:Amino Acids; D000818:Animals; D020134:Catalytic Domain; D002417:Cattle; D003124:Colorimetry; D007700:Kinetics; D011092:Polyethylene Glycols / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*; D012995:Solubility; D014357:Trypsin / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000596:Amino Acids; D011506:Proteins; C493635:polyethylene glycol-block-polyaspartic acid; D011092:Polyethylene Glycols; D014357:Trypsin