Catalytic properties of rabbit serum esterases hydrolyzing esterified monosaccharides.
Rabbit serum and one enzyme fraction isolated from rabbit serum by column chromatography (Fraction II) were used as catalysts in regioselective hydrolysis of radiolabelled pivaloylated monosaccharides (Piv = Me3CCO). The hydrolysis of 14C-labelled methyl 2-O-pivaloyl-(2-MP)-, 6-O-pivaloyl (6-MP)-, 2,6-di-O-pivaloyl-(2,6-DP) alpha-D- glucopyranosides and methyl 2-acetamido-2-deoxy-3,6- di-O-pivaloyl-(3,6-DPNAc) alpha-D-glucopyranosides, was studied, as well as that of the non-sugar substrates butyrylthiocholine, thiophenylbutyrate, phenylacetate and paraoxon. The specific activities of 2,6-DP, 3,6-DPNAc, butyrylthiocholine and thiophenylbutyrate were higher in Fraction II than in native sera, while those of phenylacetate and paraoxon were lower. Inhibition studies were done using the substrates mentioned and five different inhibitors, namely bis(p-nitrophenyl phosphate) (BNPP), eserine, paraoxon, HgCl2 and EDTA. The hydrolysis of 2,6-DP and 3,6-DPNAc was not inhibited by HgCl2 and only slightly by EDTA. Paraoxon, eserine and BNPP were progressive inhibitors of the hydrolysis of the two sugar substrates, and the pattern of inhibition resembled closely the inhibition of butyrylthiocholine and thiophenylbutyrate hydrolysis. This result applied to both, native serum and Fraction II. It was concluded that esterases in rabbit serum which hydrolyze pivaloylated sugar substrates belong to the category of serine esterases. Kinetic parameters (KM and Vmax), effects of temperature and pH on activity of esterases from Fraction II were also determined for the hydrolysis of sugar substrates.
Tomić
S
S
Department of Chemistry, Faculty of Science, University of Zagreb, Croatia.
Trescec
A
A
Tomasić
J
J
Petrović
B
B
Rudolf
V S
VS
Skrinjarić-Spoljar
M
M
Reiner
E
E
D000818:Animals; D002384:Catalysis; D004950:Esterases / Q000037:antagonists & inhibitors / Q000097:blood / Q000302:isolation & purification* / Q000378:metabolism*; D004952:Esters / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009005:Monosaccharides / Q000378:metabolism; D011817:Rabbits; D013379:Substrate Specificity
D004952:Esters; D009005:Monosaccharides; D004950:Esterases