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Use of the contour approach for visualizing the dynamic behavior of intermediates during O-nitrophenyl-β-d-galactoside hydrolysis by β-galactosidase

Paper ID Volume ID Publish Year Pages File Format Full-Text
35304 45085 2009 7 PDF Available
Title
Use of the contour approach for visualizing the dynamic behavior of intermediates during O-nitrophenyl-β-d-galactoside hydrolysis by β-galactosidase
Abstract

ONPG disappearance and ONP appearance were synchronously measured during ONPG hydrolysis by β-galactosidase using in situ on-line UV–vis spectroscopy. Intermediate formation was determined by the formula d[ONPG]/dt − d[ONP]/dt. The combined effects of temperature and time on νinst and νinc during the conversion of ONPG to ONP were expressed by the isogram method in which contour plots were used. Based on this approach, new insights were obtained into the irreversible-continuous conversion of ONPG to ONP during hydrolysis. The intermediate was a moving mass that flowed in three-dimensional space from the substrate to the product. The results of this study support the use of the isogram method for understanding the mechanisms of enzyme-catalyzed reactions via the dynamic resolution approach.

Keywords
ONPG, O-nitrophenyl-β-d-galactoside; ONP, O-nitrophenol; νinst, instantaneous rate; νinc, instantaneous increment; AUC, area under the curveβ-Galactosidase; Intermediate; Isograms method; Enzyme-catalyzed mechanisms
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Use of the contour approach for visualizing the dynamic behavior of intermediates during O-nitrophenyl-β-d-galactoside hydrolysis by β-galactosidase
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 44, Issue 12, December 2009, Pages 1374–1380
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