A pyranose dehydrogenase-based biosensor for kinetic analysis of enzymatic hydrolysis of cellulose by cellulases

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

A pyranose dehydrogenase-based biosensor for kinetic analysis of enzymatic hydrolysis of cellulose by cellulases. / Cruys-Bagger, Nicolaj; Badino, Silke Flindt; Tokin, Radina; Gontsarik, Mark; Fathalinejad, Samin; Jensen, Kenneth; Toscano, Miguel Duarte; Sørensen, Trine Holst; Borch, Kim; Tatsumi, Hirosuke; Väljamäe, Priit; Westh, Peter.

In: Enzyme and Microbial Technology, Vol. 58-59, 2014, p. 68-74.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Cruys-Bagger, N, Badino, SF, Tokin, R, Gontsarik, M, Fathalinejad, S, Jensen, K, Toscano, MD, Sørensen, TH, Borch, K, Tatsumi, H, Väljamäe, P & Westh, P 2014, 'A pyranose dehydrogenase-based biosensor for kinetic analysis of enzymatic hydrolysis of cellulose by cellulases', Enzyme and Microbial Technology, vol. 58-59, pp. 68-74. https://doi.org/10.1016/j.enzmictec.2014.03.002

APA

Cruys-Bagger, N., Badino, S. F., Tokin, R., Gontsarik, M., Fathalinejad, S., Jensen, K., Toscano, M. D., Sørensen, T. H., Borch, K., Tatsumi, H., Väljamäe, P., & Westh, P. (2014). A pyranose dehydrogenase-based biosensor for kinetic analysis of enzymatic hydrolysis of cellulose by cellulases. Enzyme and Microbial Technology, 58-59, 68-74. https://doi.org/10.1016/j.enzmictec.2014.03.002

Vancouver

Cruys-Bagger N, Badino SF, Tokin R, Gontsarik M, Fathalinejad S, Jensen K et al. A pyranose dehydrogenase-based biosensor for kinetic analysis of enzymatic hydrolysis of cellulose by cellulases. Enzyme and Microbial Technology. 2014;58-59:68-74. https://doi.org/10.1016/j.enzmictec.2014.03.002

Author

Cruys-Bagger, Nicolaj ; Badino, Silke Flindt ; Tokin, Radina ; Gontsarik, Mark ; Fathalinejad, Samin ; Jensen, Kenneth ; Toscano, Miguel Duarte ; Sørensen, Trine Holst ; Borch, Kim ; Tatsumi, Hirosuke ; Väljamäe, Priit ; Westh, Peter. / A pyranose dehydrogenase-based biosensor for kinetic analysis of enzymatic hydrolysis of cellulose by cellulases. In: Enzyme and Microbial Technology. 2014 ; Vol. 58-59. pp. 68-74.

Bibtex

@article{1062c461f0fb4db8bbbb36adc811e8e8,

title = "A pyranose dehydrogenase-based biosensor for kinetic analysis of enzymatic hydrolysis of cellulose by cellulases",

abstract = "A novel electrochemical enzyme biosensor was developed for real-time detection of cellulase activity when acting on their natural insoluble substrate, cellulose. The enzyme biosensor was constructed with pyranose dehydrongease (PDH) from Agaricus meleagris that was immobilized on the surface of a carbon paste electrode, which contained the mediator 2,6-dichlorophenolindophenol (DCIP). An oxidation current of the reduced form of DCIP, DCIPH2, produced by the PDH-catalyzed reaction with either glucose or cellobiose, was recorded under constant-potential amperometry at +0.25V (vs. Ag/AgCl). The PDH-biosensor was shown to be anomer unspecific and it can therefore be used in kinetic studies over broad time-scales of both retaining- and inverting cellulases (in addition to enzyme cocktails). The biosensor was used for real-time measurements of the activity of the inverting cellobiohydrolase Cel6A from Hypocrea jecorina (HjCel6A) on cellulosic substrates with different morphology (bacterial microcrystalline cellulose (BMCC) and Avicel). The steady-state rate of hydrolysis increased towards a saturation plateau with increasing loads of substrate. The experimental results were rationalized using a steady-state rate equation for processive cellulases, and it was found that the turnover for HjCel6A at saturating substrate concentration (i.e. maximal apparent specific activity) was similar (0.39-0.40s(-1)) for the two substrates. Conversely, the substrate load at half-saturation was much lower for BMCC compared to Avicel. Biosensors covered with a polycarbonate membrane showed high operational stability of several weeks with daily use.",

keywords = "2,6-Dichloroindophenol, Agaricus, Alcohol Oxidoreductases, Biosensing Techniques, Calibration, Carbon, Cellulase, Cellulose, Cellulose 1,4-beta-Cellobiosidase, Computer Systems, Electrochemical Techniques, Electrodes, Equipment Design, Fungal Proteins, Hydrolysis, Hypocrea, Kinetics, Membranes, Artificial, Optical Rotation, Reproducibility of Results, Stereoisomerism, Substrate Specificity, Comparative Study, Journal Article, Research Support, Non-U.S. Gov't",

author = "Nicolaj Cruys-Bagger and Badino, {Silke Flindt} and Radina Tokin and Mark Gontsarik and Samin Fathalinejad and Kenneth Jensen and Toscano, {Miguel Duarte} and S{\o}rensen, {Trine Holst} and Kim Borch and Hirosuke Tatsumi and Priit V{\"a}ljam{\"a}e and Peter Westh",

year = "2014",

doi = "10.1016/j.enzmictec.2014.03.002",

language = "English",

volume = "58-59",

pages = "68--74",

journal = "Enzyme and Microbial Technology",

issn = "0141-0229",

publisher = "Elsevier",

}

RIS

TY - JOUR

T1 - A pyranose dehydrogenase-based biosensor for kinetic analysis of enzymatic hydrolysis of cellulose by cellulases

AU - Cruys-Bagger, Nicolaj

AU - Badino, Silke Flindt

AU - Tokin, Radina

AU - Gontsarik, Mark

AU - Fathalinejad, Samin

AU - Jensen, Kenneth

AU - Toscano, Miguel Duarte

AU - Sørensen, Trine Holst

AU - Borch, Kim

AU - Tatsumi, Hirosuke

AU - Väljamäe, Priit

AU - Westh, Peter

PY - 2014

Y1 - 2014

N2 - A novel electrochemical enzyme biosensor was developed for real-time detection of cellulase activity when acting on their natural insoluble substrate, cellulose. The enzyme biosensor was constructed with pyranose dehydrongease (PDH) from Agaricus meleagris that was immobilized on the surface of a carbon paste electrode, which contained the mediator 2,6-dichlorophenolindophenol (DCIP). An oxidation current of the reduced form of DCIP, DCIPH2, produced by the PDH-catalyzed reaction with either glucose or cellobiose, was recorded under constant-potential amperometry at +0.25V (vs. Ag/AgCl). The PDH-biosensor was shown to be anomer unspecific and it can therefore be used in kinetic studies over broad time-scales of both retaining- and inverting cellulases (in addition to enzyme cocktails). The biosensor was used for real-time measurements of the activity of the inverting cellobiohydrolase Cel6A from Hypocrea jecorina (HjCel6A) on cellulosic substrates with different morphology (bacterial microcrystalline cellulose (BMCC) and Avicel). The steady-state rate of hydrolysis increased towards a saturation plateau with increasing loads of substrate. The experimental results were rationalized using a steady-state rate equation for processive cellulases, and it was found that the turnover for HjCel6A at saturating substrate concentration (i.e. maximal apparent specific activity) was similar (0.39-0.40s(-1)) for the two substrates. Conversely, the substrate load at half-saturation was much lower for BMCC compared to Avicel. Biosensors covered with a polycarbonate membrane showed high operational stability of several weeks with daily use.

AB - A novel electrochemical enzyme biosensor was developed for real-time detection of cellulase activity when acting on their natural insoluble substrate, cellulose. The enzyme biosensor was constructed with pyranose dehydrongease (PDH) from Agaricus meleagris that was immobilized on the surface of a carbon paste electrode, which contained the mediator 2,6-dichlorophenolindophenol (DCIP). An oxidation current of the reduced form of DCIP, DCIPH2, produced by the PDH-catalyzed reaction with either glucose or cellobiose, was recorded under constant-potential amperometry at +0.25V (vs. Ag/AgCl). The PDH-biosensor was shown to be anomer unspecific and it can therefore be used in kinetic studies over broad time-scales of both retaining- and inverting cellulases (in addition to enzyme cocktails). The biosensor was used for real-time measurements of the activity of the inverting cellobiohydrolase Cel6A from Hypocrea jecorina (HjCel6A) on cellulosic substrates with different morphology (bacterial microcrystalline cellulose (BMCC) and Avicel). The steady-state rate of hydrolysis increased towards a saturation plateau with increasing loads of substrate. The experimental results were rationalized using a steady-state rate equation for processive cellulases, and it was found that the turnover for HjCel6A at saturating substrate concentration (i.e. maximal apparent specific activity) was similar (0.39-0.40s(-1)) for the two substrates. Conversely, the substrate load at half-saturation was much lower for BMCC compared to Avicel. Biosensors covered with a polycarbonate membrane showed high operational stability of several weeks with daily use.

KW - 2,6-Dichloroindophenol

KW - Agaricus

KW - Alcohol Oxidoreductases

KW - Biosensing Techniques

KW - Calibration

KW - Carbon

KW - Cellulase

KW - Cellulose

KW - Cellulose 1,4-beta-Cellobiosidase

KW - Computer Systems

KW - Electrochemical Techniques

KW - Electrodes

KW - Equipment Design

KW - Fungal Proteins

KW - Hydrolysis

KW - Hypocrea

KW - Kinetics

KW - Membranes, Artificial

KW - Optical Rotation

KW - Reproducibility of Results

KW - Stereoisomerism

KW - Substrate Specificity

KW - Comparative Study

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1016/j.enzmictec.2014.03.002

DO - 10.1016/j.enzmictec.2014.03.002

M3 - Journal article

C2 - 24731827

VL - 58-59

SP - 68

EP - 74

JO - Enzyme and Microbial Technology

JF - Enzyme and Microbial Technology

SN - 0141-0229

ER -

ID: 167806961