Kinetic modeling of cellobiose by a β-glucosidase from Aspergillus fumigatus
The final step in lignocellulose enzymatic saccharification is the cellobiose conversion toglucose by β-glucosidases (BG). In this work, a valid kinetic model to describe cellobiosedegradation for an industrial mixture of BG enzymes present in Aspergillus fumigatus isselected. Firstly, the enzyme mixture was characterised in terms of protein content andenzymatic activity on p-NPG (1326 U mLpreparation−1), determining the molecular weight ofthe only BG activity band observed in zymograms by SDS-PAGE and MALDI-TOF: 94 kDa.Subsequently, to select the correct kinetic model for the enzymatic hydrolysis of cellobiose,a combined strategy was performed: Firstly, non-linear regressions were applied to initialhydrolysis rate data for different enzyme concentrations and initial substrate and productconcentrations, observing inhibition by cellobiose and glucose. Secondly, the optimal kineticmodel was discriminated by a coupled non-linear regression-DOE numerical integrationapproach, by fitting several possible kinetic models involving different product inhibitionmechanisms to progress curve data from runs at various initial substrate concentrationsand temperatures. The best kinetic model involves non-competitive substrate inhibitionand product competitive inhibition with two binding sites for glucose.
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Format: | artículo biblioteca |
Language: | English |
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Elsevier
2018
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Subjects: | Kinetic model discrimination, Biorefinery, Cellobiose, ?-glucosidase, Enzymatic hydrolysis, Glucose, |
Online Access: | http://hdl.handle.net/20.500.12792/830 http://hdl.handle.net/10261/289256 |
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dig-inia-es-10261-2892562023-02-15T07:52:58Z Kinetic modeling of cellobiose by a β-glucosidase from Aspergillus fumigatus Wojtusik, M. Yepes, C. M. Villar, J. C. Cordes, A. Arroyo, M. Garcia-Ochoa, F. Ladero, M. Kinetic model discrimination Biorefinery Cellobiose ?-glucosidase Enzymatic hydrolysis Glucose The final step in lignocellulose enzymatic saccharification is the cellobiose conversion toglucose by β-glucosidases (BG). In this work, a valid kinetic model to describe cellobiosedegradation for an industrial mixture of BG enzymes present in Aspergillus fumigatus isselected. Firstly, the enzyme mixture was characterised in terms of protein content andenzymatic activity on p-NPG (1326 U mLpreparation−1), determining the molecular weight ofthe only BG activity band observed in zymograms by SDS-PAGE and MALDI-TOF: 94 kDa.Subsequently, to select the correct kinetic model for the enzymatic hydrolysis of cellobiose,a combined strategy was performed: Firstly, non-linear regressions were applied to initialhydrolysis rate data for different enzyme concentrations and initial substrate and productconcentrations, observing inhibition by cellobiose and glucose. Secondly, the optimal kineticmodel was discriminated by a coupled non-linear regression-DOE numerical integrationapproach, by fitting several possible kinetic models involving different product inhibitionmechanisms to progress curve data from runs at various initial substrate concentrationsand temperatures. The best kinetic model involves non-competitive substrate inhibitionand product competitive inhibition with two binding sites for glucose. 2023-02-15T07:52:58Z 2023-02-15T07:52:58Z 2018 artículo Chemical Engineering Research and Design 136: 502-512 (2018) http://hdl.handle.net/20.500.12792/830 http://hdl.handle.net/10261/289256 10.1016/j.cherd.2018.06.020 0263-8762 en none Elsevier |
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Kinetic model discrimination Biorefinery Cellobiose ?-glucosidase Enzymatic hydrolysis Glucose Kinetic model discrimination Biorefinery Cellobiose ?-glucosidase Enzymatic hydrolysis Glucose |
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Kinetic model discrimination Biorefinery Cellobiose ?-glucosidase Enzymatic hydrolysis Glucose Kinetic model discrimination Biorefinery Cellobiose ?-glucosidase Enzymatic hydrolysis Glucose Wojtusik, M. Yepes, C. M. Villar, J. C. Cordes, A. Arroyo, M. Garcia-Ochoa, F. Ladero, M. Kinetic modeling of cellobiose by a β-glucosidase from Aspergillus fumigatus |
description |
The final step in lignocellulose enzymatic saccharification is the cellobiose conversion toglucose by β-glucosidases (BG). In this work, a valid kinetic model to describe cellobiosedegradation for an industrial mixture of BG enzymes present in Aspergillus fumigatus isselected. Firstly, the enzyme mixture was characterised in terms of protein content andenzymatic activity on p-NPG (1326 U mLpreparation−1), determining the molecular weight ofthe only BG activity band observed in zymograms by SDS-PAGE and MALDI-TOF: 94 kDa.Subsequently, to select the correct kinetic model for the enzymatic hydrolysis of cellobiose,a combined strategy was performed: Firstly, non-linear regressions were applied to initialhydrolysis rate data for different enzyme concentrations and initial substrate and productconcentrations, observing inhibition by cellobiose and glucose. Secondly, the optimal kineticmodel was discriminated by a coupled non-linear regression-DOE numerical integrationapproach, by fitting several possible kinetic models involving different product inhibitionmechanisms to progress curve data from runs at various initial substrate concentrationsand temperatures. The best kinetic model involves non-competitive substrate inhibitionand product competitive inhibition with two binding sites for glucose. |
format |
artículo |
topic_facet |
Kinetic model discrimination Biorefinery Cellobiose ?-glucosidase Enzymatic hydrolysis Glucose |
author |
Wojtusik, M. Yepes, C. M. Villar, J. C. Cordes, A. Arroyo, M. Garcia-Ochoa, F. Ladero, M. |
author_facet |
Wojtusik, M. Yepes, C. M. Villar, J. C. Cordes, A. Arroyo, M. Garcia-Ochoa, F. Ladero, M. |
author_sort |
Wojtusik, M. |
title |
Kinetic modeling of cellobiose by a β-glucosidase from Aspergillus fumigatus |
title_short |
Kinetic modeling of cellobiose by a β-glucosidase from Aspergillus fumigatus |
title_full |
Kinetic modeling of cellobiose by a β-glucosidase from Aspergillus fumigatus |
title_fullStr |
Kinetic modeling of cellobiose by a β-glucosidase from Aspergillus fumigatus |
title_full_unstemmed |
Kinetic modeling of cellobiose by a β-glucosidase from Aspergillus fumigatus |
title_sort |
kinetic modeling of cellobiose by a β-glucosidase from aspergillus fumigatus |
publisher |
Elsevier |
publishDate |
2018 |
url |
http://hdl.handle.net/20.500.12792/830 http://hdl.handle.net/10261/289256 |
work_keys_str_mv |
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