Kinetics and mechanism of a reaction catalyzed by PST-01 protease from Pseudomonas aeruginosa PST-01.
The initial rates of carboxybenzoyl-alanyl-l-leucyl-amide (Z-L-Ala-L-Leu-NH(2)) synthesis from carboxybenzoyl-L-alanine (Z-L-Ala) and L-leucineamide (L-Leu-NH(2)) and Z-L-Ala-L-Leu-NH(2) hydrolysis in a homogeneous dimethyl sulfoxide-aqueous buffer solution [1:1 (v/v)] system catalyzed by PST-01 protease from Pseudomonas aeruginosa were measured under a wide range of Z-L-Ala, L-Leu-NH(2) and Z-L-Ala-L-Leu-NH(2) concentrations. The initial rates of the synthetic reaction, in which Z-L-Ala-L-Leu-NH(2) was produced from Z-L-Ala and L-Leu-NH(2), were inhibited by the substrates. Furthermore, the initial rates of the synthetic reaction were not inhibited by the product Z-L-Ala-L-Leu-NH(2), and those of the hydrolytic reaction were inhibited by Z-L-Ala and L-Leu-NH(2). All the initial rate data of the synthetic and hydrolytic reactions were well correlated with the rate equation derived based on the proposed reaction scheme.
Bobe
Iulian M
IM
Department of Chemical Engineering, Osaka Prefecture University; 1-1 Gakuen-cho, Sakai, Osaka 599-8531, Japan.
Abdelmoez
Wael
W
Ogino
Hiroyasu
H
Yasuda
Masahiro
M
Ishimi
Kosaku
K
Ishikawa
Haruo
H
D002384:Catalysis; D002850:Chromatography, Gel; D004151:Dipeptides / Q000138:chemical synthesis / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000737:chemistry* / Q000378:metabolism; D006868:Hydrolysis; D007700:Kinetics; D008970:Molecular Weight; D011550:Pseudomonas aeruginosa / Q000737:chemistry / Q000201:enzymology*; D012997:Solvents / Q000032:analysis; D013379:Substrate Specificity
D004151:Dipeptides; D012997:Solvents; D010450:Endopeptidases