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Effects of amines and aminoalcohols on bovine intestine alkaline phosphatase activity

Paper ID Volume ID Publish Year Pages File Format Full-Text
17402 42666 2011 6 PDF Available
Title
Effects of amines and aminoalcohols on bovine intestine alkaline phosphatase activity
Abstract

Bovine 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.

Keywords
ALP, alkaline phosphatase; BIALP, bovine intestine alkaline phosphatase; EIA, enzyme immunoassay; pNPP, p-nitrophenyl phosphateAlkaline phosphatase; Amine; Aminoalcohol; Bovine intestine alkaline phosphatase; Enzyme immunoassay
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Effects of amines and aminoalcohols on bovine intestine alkaline phosphatase activity
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 49, Issue 2, 10 July 2011, Pages 171–176
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us