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dc.contributor.authorSekiguchi, Satoshija
dc.contributor.authorHashida, Yasuhikoja
dc.contributor.authorYasukawa, Kiyoshija
dc.contributor.authorInouye, Kuniyoja
dc.contributor.alternative井上, 國世ja
dc.description.abstractBovine intestine alkaline phosphatase (BIALP) is widely used as a signaling enzyme in sensitive assays such as enzyme immunoassay (EIA). In this study, we evaluated the effects of various aminoalcohols and amines on the activity of BIALP in the hydrolysis of p-nitrophenyl phosphate (pNPP) at pH 9.8, at 20 °C. The Kcat values at 0.05 M diethanolamine, 0.1 M triethanolamine, and 0.2 M N-methylethanolamine were 190 ± 10, 840 ± 30, and 500 ± 10 s[−1], respectively. The Kcat values increased with increasing concentrations of diethanolamine, triethanolamine, and N-methylethanolamine and reached 1240 ± 60, 1450 ± 30, and 2250 ± 80 s[−1], respectively, at 1.0 M. On the other hand, the Kcat values at 0.05-1.0 M ethanolamine, ethylamine, methylamine, and dimethylamine were in the range of 100–600 s[−1]. These results indicate that diethanolamine, triethanolamine and N-methylethanolamine highly activate BIALP and might be suitable as a dilution buffer of BIALP in EIA. Interestingly, the Km values increased with increasing concentrations of diethanolamine and N-methylethanolamine, but not triethanolamine: the Km value at 1.0 M diethanolamine (0.83 ± 0.15 mM) was 12-fold higher than that at 0.05 M (0.07 ± 0.01 mM), and that at 1.0 M N-methylethanolamine (2.53 ± 0.20 mM) was 14-fold higher than that at 0.2 M (0.18 ± 0.02 mM), while that at 1.0 M triethanolamine (0.31 ± 0.01 mM) was similar as that at 0.2 M (0.25 ± 0.01 mM), suggesting that the mechanisms of BIALP activation are different between the aminoalcohols.ja
dc.publisherElsevier Inc.ja
dc.rights© 2011 Elsevier Inc.ja
dc.rightsThis is not the published version. Please cite only the published version. この論文は出版社版でありません。引用の際には出版社版をご確認ご利用ください。ja
dc.subjectAlkaline phosphataseja
dc.subjectBovine intestine alkaline phosphataseja
dc.subjectEnzyme immunoassayja
dc.titleEffects of amines and aminoalcohols on bovine intestine alkaline phosphatase activityja
dc.type.niitypeJournal Articleja
dc.identifier.jtitleEnzyme and Microbial Technologyja
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