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The combined effects of temperature and assay time on the catalytic ability and stability of 1,4-β-d-glucan cellobiohydrolase I

Paper ID Volume ID Publish Year Pages File Format Full-Text
17946 42707 2008 8 PDF Available
Title
The combined effects of temperature and assay time on the catalytic ability and stability of 1,4-β-d-glucan cellobiohydrolase I
Abstract

A new method is proposed to describe the combined effects of temperature and assay time on the reaction rates of 1,4-β-d-glucan cellobiohydrolase I (CBHI). The method involves determination of the product (PNP) released from substrate (PNPC) that was monitored spectroscopically and calculated by area under curve (AUC). The combined effects of temperature and assay time on the catalytic reaction rates were expressed as the composite function of the instantaneous reaction rate (vinst)(vinst) termed c−vinstc−vinst, denoted as dy/dT = dy/dt × dt/dT  . And the total instantaneous increment of product expressed as the complete differential of c−vinstc−vinst equals to the sum of the partial derivative of temperature and time denoted as: c−vinst=(∂p/∂T)tdT+(∂p/∂t)Tdtc−vinst=(∂p/∂T)tdT+(∂p/∂t)Tdt. The kinetic curves of c−vinstc−vinst and c−vinstc−vinst can be directly derived from the UV spectra by numerical differentiation and do not require any assumption of complex rate equations. The optimum temperature (Topt) and the heat-inactivated temperature (Tinac-50) were estimated based on the relationship between the first derivative of composite function (c−vinst)(c−vinst) and the second derivative (c−vsec)(c−vsec). This approach is particularly suited for biotechnological applications, given its precision and simplicity. The unsuitability of the Michaelis–Menten equation as used for determining the effect of temperature on reaction rates was discussed.

Keywords
PNPC, p-nitrophenyl β-d-cellobioside; PNP, p-nitrophenol; AUC, area under the curve; vappvapp, apparent velocity; vinstvinst, instantaneous rate of velocity; vsecvsec, second order derivative of velocity; c−vinstc−vinst, composite function of vinstvinst;
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The combined effects of temperature and assay time on the catalytic ability and stability of 1,4-β-d-glucan cellobiohydrolase I
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 43, Issue 3, 5 September 2008, Pages 237–244
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