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- Title
Estimation of Cellobiohydrolase I Activity by Numerical Differentiation of Dynamic Ultraviolet Spectroscopy.
- Authors
Bin Wu; Yue Zhao; Pei-Ji Gao
- Abstract
1,4-β- D-glucan cellobiohydrolase I (CBH I), p-nitrophenyl β- D-cellobioside, p-nitrophenol and cellobiose show distinct ultraviolet spectra, allowing the design of an assay to track the dynamic process of p-nitrophenyl β- D-cellobioside hydrolysis by CBH I. Based on the linear relationship between p-nitrophenol formation in the hydrolysate and its first derivative absorption curve of AUC340_400nm (area under the curve), a new sensitive assay for the determination of CBH I activity was developed. The dynamic parameters of catalysis reaction, such as Vm and kcat, can all be derived from this result. The influence of β-glucosidase and endoglucanase in crude enzyme sample on the assay was discussed in detail. This approach is useful for accurate determination of the activity of CBHs. Edited by Yi LIANG
- Subjects
CELLULOSE 1,4-beta-cellobiosidase; CELLULASE; ULTRAVIOLET spectra; HYDROLYSIS; CATALYSIS; NUMERICAL differentiation
- Publication
Acta Biochimica et Biophysica Sinica, 2006, Vol 38, Issue 6, p372
- ISSN
1672-9145
- Publication type
Article
- DOI
10.1111/j.1745-7270.2006.00179.x