| Estimation of Cellobiohydrolase I Activity by Numerical Differentiation of Dynamic Ultraviolet Spectroscopy |
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| 作者姓名： | Bin WU Yue ZHAO Pei-Ji GAO The State Key Laboratory of Microbial Technology Shandong University Jinan China |
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| 作者单位： | Bin WU Yue ZHAO Pei-Ji GAO The State Key Laboratory of Microbial Technology，Shandong University，Jinan 250100，China |
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| 基金项目： | This study was supported the grants from the National Natural Science Foundation of China(No.30370013)，Natlonal Basic Research Program of China(No.2003CB716006 and No.2004CB719702) |
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| 摘 要： | 1,4-β-D-glucan cellobiohydrolase I (CBH I),p-nitrophenyl β-D-cellobioside,p-nitrophenol andcellobiose show distinct ultraviolet spectra,allowing the design of an assay to track the dynamic process ofp-nitrophenyl β-D-cellobioside hydrolysis by CBH I.Based on the linear relationship between p-nitrophenolformation in the hydrolysate and its first derivative absorption Curve of AUC340_400_(nm)(area under the curve),a new sensitive assay for the determination of CBH I activity was developed.The dynamic parameters ofcatalysis reaction,such as Vm and k_(cat),can all be derived from this result.The influence of β-glucosidase andendoglucanase in crude enzyme sample on the assay was discussed in detail.This approach is useful foraccurate determination of the activity of CBHs.
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Estimation of Cellobiohydrolase I Activity by Numerical Differentiation of Dynamic Ultraviolet Spectroscopy |
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| Bin WU Yue ZHAO Pei-Ji GAO The State Key Laboratory of Microbial Technology,Shandong University,Jinan ,China.Estimation of Cellobiohydrolase I Activity by Numerical Differentiation of Dynamic Ultraviolet Spectroscopy[J].Acta Biochimica et Biophysica Sinica,2006(6). |
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| Authors: | Bin WU Yue ZHAO Pei-Ji GAO The State Key Laboratory of Microbial Technology Shandong University Jinan China |
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| Institution: | Bin WU Yue ZHAO Pei-Ji GAO The State Key Laboratory of Microbial Technology,Shandong University,Jinan 250100,China |
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| Abstract: | |
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| Keywords: | cellobiohydrolase dynamic spectrum area under the curve(AUC) |
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