TY - JOUR
T1 - Development of a Multi-Enzymatic Biocatalytic System through Immobilization on High Quality Few-Layer bio-Graphene
AU - Alatzoglou, Christina
AU - Patila, Michaela
AU - Giannakopoulou, Archontoula
AU - Spyrou, Konstantinos
AU - Yan, Feng
AU - Li, Wenjian
AU - Chalmpes, Nikolaos
AU - Polydera, Angeliki C.
AU - Rudolf, Petra
AU - Gournis, Dimitrios
AU - Stamatis, Haralambos
N1 - Funding Information:
This project was co-financed by the European Regional Development Fund of the European Union and Greek national funds through the Operational Program Competitiveness, Entrepreneurship, and Innovation, under the call RESEARCH CREATE INNOVATE (project T2EDK-02171). This work also received support from the Advanced Materials research program of the Zernike National Research Centre under the Bonus Incentive Scheme of the Dutch Ministry for Education, Culture and Science.
Publisher Copyright:
© 2022 by the authors.
PY - 2023/1
Y1 - 2023/1
N2 - In this work, we report the green production of few-layer bio-Graphene (bG) through liquid exfoliation of graphite in the presence of bovine serum albumin. Microscopic characterization evaluated the quality of the produced nanomaterial, showing the presence of 3–4-layer graphene. Moreover, spectroscopic techniques also confirmed the quality of the resulted bG, as well as the presence of bovine serum albumin on the graphene sheets. Next, for the first time, bG was used as support for the simultaneous covalent co-immobilization of three enzymes, namely β-glucosidase, glucose oxidase, and horseradish peroxidase. The three enzymes were efficiently co-immobilized on bG, demonstrating high immobilization yields and activity recoveries (up to 98.5 and 90%, respectively). Co-immobilization on bG led to an increase of apparent KM values and a decrease of apparent Vmax values, while the stability of the nanobiocatalysts prevailed compared to the free forms of the enzymes. Co-immobilized enzymes exhibited high reusability, preserving a significant part of their activity (up to 72%) after four successive catalytic cycles at 30 °C. Finally, the tri-enzymatic nanobiocatalytic system was applied in three-step cascade reactions, involving, as the first step, the hydrolysis of p-Nitrophenyl-β-D-Glucopyranoside and cellobiose.
AB - In this work, we report the green production of few-layer bio-Graphene (bG) through liquid exfoliation of graphite in the presence of bovine serum albumin. Microscopic characterization evaluated the quality of the produced nanomaterial, showing the presence of 3–4-layer graphene. Moreover, spectroscopic techniques also confirmed the quality of the resulted bG, as well as the presence of bovine serum albumin on the graphene sheets. Next, for the first time, bG was used as support for the simultaneous covalent co-immobilization of three enzymes, namely β-glucosidase, glucose oxidase, and horseradish peroxidase. The three enzymes were efficiently co-immobilized on bG, demonstrating high immobilization yields and activity recoveries (up to 98.5 and 90%, respectively). Co-immobilization on bG led to an increase of apparent KM values and a decrease of apparent Vmax values, while the stability of the nanobiocatalysts prevailed compared to the free forms of the enzymes. Co-immobilized enzymes exhibited high reusability, preserving a significant part of their activity (up to 72%) after four successive catalytic cycles at 30 °C. Finally, the tri-enzymatic nanobiocatalytic system was applied in three-step cascade reactions, involving, as the first step, the hydrolysis of p-Nitrophenyl-β-D-Glucopyranoside and cellobiose.
KW - bio-Graphene
KW - bovine serum albumin
KW - cascade reactions
KW - green synthesis
KW - multi-enzymatic immobilization
KW - nanobiocatalytic system
UR - http://www.scopus.com/inward/record.url?scp=85145831061&partnerID=8YFLogxK
U2 - 10.3390/nano13010127
DO - 10.3390/nano13010127
M3 - Article
AN - SCOPUS:85145831061
SN - 2079-4991
VL - 13
JO - Nanomaterials
JF - Nanomaterials
IS - 1
M1 - 127
ER -