Article
Biochemistry & Molecular Biology
Jose R. Guimaraes, Diego Carballares, Javier Rocha-Martin, Paulo W. Tardioli, Roberto Fernandez-Lafuente
Summary: In this study, lipases from different sources were immobilized on octylagarose beads and treated with phosphate and/or metallic salts. The effects of phosphate/metal modification on enzyme features varied depending on the loading of the support. Some enzymes showed improved stability, while others showed increased activity.
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
(2022)
Article
Biochemistry & Molecular Biology
Sabrina Ait Braham, El Hocine Siar, Sara Arana-Pena, Hossein Bavandi, Diego Carballares, Roberto Morellon-Sterling, Diandra de Andrades, Jakub F. Kornecki, Roberto Fernandez-Lafuente
Summary: This study investigates the impact of glycerol on the stability of 13 immobilized enzymes, finding that its stabilizing effect is enzyme-dependent and influenced by concentration and pH. The results emphasize the importance of considering enzyme type, glycerol concentration, and pH level when using glycerol as a stabilizing agent.
PROCESS BIOCHEMISTRY
(2021)
Article
Biochemistry & Molecular Biology
Jose R. Guimaraes, Diego Carballares, Paulo W. Tardioli, Javier Rocha-Martin, Roberto Fernandez-Lafuente
Summary: Modifications with different metal phosphates greatly altered the specificity and enantiospecificity of commercial immobilized lipases. However, enzyme stability was not significantly improved.
Article
Nanoscience & Nanotechnology
Hector Sanchez-Moran, James S. Weltz, Daniel K. Schwartz, Joel L. Kaar
Summary: "The long-term goal of biotechnology is to develop design rules for stabilizing enzymes when immobilized to materials. By investigating the stabilization of lipases immobilized on PEGMA and SBMA copolymer brush surfaces, it was found that the stability of the lipases at elevated temperatures depended on the fraction of PEGMA in the brush layer. This understanding can lead to the rational tuning of enzyme-material interfaces for reliable ruggedization of enzymes under extreme conditions."
ACS APPLIED MATERIALS & INTERFACES
(2021)
Article
Biophysics
Hongbo Suo, Moju Li, Renmin Liu, Lili Xu
Summary: In this study, imidazolium-based ionic liquid was successfully grafted to magnetic polydopamine nanoparticles for lipase immobilization. The immobilized lipase showed excellent activity and stability, with increased affinity to substrate. Additionally, the immobilized lipase can be easily separated from the reaction system with a magnet, providing new ideas for further studies in this field.
COLLOIDS AND SURFACES B-BIOINTERFACES
(2021)
Article
Chemistry, Multidisciplinary
Ana Benitez-Mateos, Francesca Paradisi
Summary: A novel halotolerant purine nucleoside phosphorylase from Halomonas elongata (HePNP) was used to achieve high conversion rates in the synthesis of valuable nucleoside drugs, along with an immobilization strategy on agarose microbeads and a catch-and-release approach for recycling excess sugar donors. This study demonstrated a high reproducibility and increased sustainability of the process, while also enhancing cost-efficiency.
Article
Biochemistry & Molecular Biology
Luis Gerardo Ramirez-Ramirez, David Enrique Zazueta-Alvarez, Hector Alonso Fileto-Perez, Damian Reyes-Jaquez, Cynthia Manuela Nunez-Nunez, Juan De Dios Galindo-De la Rosa, Javier Lopez-Miranda, Perla Guadalupe Vazquez-Ortega
Summary: This research aimed to immobilize a recombinant beta-glucosidase from Trichoderma reesei using a commercial zeolite. The immobilized enzyme showed improved catalytic characteristics, stability, and a lower KM value compared to the free enzyme. The immobilization parameters obtained through simple adsorption show promising applications in various fields.
Article
Chemistry, Analytical
Monica Jimenez-Carretero, Ylenia Jabalera, Alberto Sola-Leyva, Maria P. Carrasco-Jimenez, Concepcion Jimenez-Lopez
Summary: The immobilization of enzymes on magnetic nanoparticles has attracted interest in detecting contaminants in water samples due to their magnetic control, concentration, and reuse capabilities. In this study, nanoassemblies composed of inorganic or biomimetic magnetic nanoparticles were developed for the detection of trace amounts of organophosphate pesticides and antibiotics in water. Enzyme immobilization was achieved through electrostatic interaction or covalent bonds, with covalent bonding yielding better results. The optimized nanoassemblies demonstrated the ability to detect pollutants at low concentrations and provided enhanced stability and reuse of the enzymes.
Review
Environmental Sciences
P. R. Yaashikaa, M. Keerthana Devi, P. Senthil Kumar
Summary: This review summarizes the basic concepts of enzyme immobilization, different immobilization techniques, and carriers used for immobilization. It also provides an in-depth overview of the potential of immobilized enzymes as bioremediation agents for the degradation of pollutants and the impact of different operating parameters. Additionally, future trends and challenges for enhancing the potential of immobilized systems for large-scale industrial wastewater treatment are discussed.
Article
Chemistry, Multidisciplinary
Xiao-Gang Yang, Ji-Rui Zhang, Xu-Ke Tian, Jian-Hua Qin, Xin-Ya Zhang, Lu-Fang Ma
Summary: Efficient enzyme immobilization is highly desirable for their recyclability and activity, and Zeolitic imidazolate framework-8 (ZIF-8) has been proved to be an effective platform for enzyme immobilization with its easy preparation and biocompatibility. However, the hydrophobic characteristic of ZIF-8 hinders its further development. This study developed a facile synthesis approach to immobilize pepsin (PEP) on ZIF-8 carrier using Ni2+ ions as an anchor, leading to remarkable catalytic activity for electrochemical oxygen evolution reaction (OER) with an ultralow overpotential.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2023)
Article
Chemistry, Physical
Diego Carballares, Roberto Morellon-Sterling, Xiaomin Xu, Frank Hollmann, Roberto Fernandez-Lafuente
Summary: This study investigated the immobilization of rAaeUPO enzyme from Agrocybe aegerita. The findings showed that the immobilization pH significantly influences the stability and activity of the immobilized enzyme.
Article
Biochemistry & Molecular Biology
Andres H. Morales, Johan S. Hero, Ana E. Ledesma, M. Alejandra Martinez, Maria C. Navarro, Maria I. Gomez, Cintia M. Romero
Summary: This study investigates the effects of pH and ionic strength fluctuations on protein-support interactions during lipase immobilization. The optimal immobilization conditions do not necessarily result in the highest hydrolytic activity. Adsorption isotherm analysis confirms this observation. Furthermore, the study demonstrates the interfacial hyperactivation of the lipase towards acidic pH and highly charged environments, and examines the mechanism of surface hydrophobic protein-carrier interactions.
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
(2023)
Article
Biochemistry & Molecular Biology
Abdel-Mohsen S. A. Ismail, Gamal El-Deen A. Helal, Doaa M. A. El-Khouly, Mona A. Esawy
Summary: This research aimed to invent a new method for cheese making using Rennin-like enzyme from fungus with high efficiency and reusability. Rhizomucor miehei (CBS: 370.65) showed promising milk clotting activity and negative mycotoxin test. The immobilized enzyme displayed better stability and could be reused more than sixteen times in cheese production. The immobilization process reduced the activation energy and calcium ions had a stimulating effect on enzyme activity. Buffalo milk and full cream milk showed the highest immobilized enzyme activity among the substrates tested.
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
(2022)
Review
Biochemistry & Molecular Biology
Veymar G. Tacias-Pascacio, Roberto Morellon-Sterling, Daniel Castaneda-Valbuena, Angel Berenguer-Murcia, Majid Rasool Kamli, Olga Tavano, Roberto Fernandez-Lafuente
Summary: This paper reviews the immobilization of papain on different supports for the first time, with a special focus on the preparation of magnetic biocatalysts and specific problems related to the immobilization of proteases. The benefits of proper immobilization and potential artifacts are discussed.
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
(2021)
Article
Pharmacology & Pharmacy
Oliwia Degorska, Daria Szada, Agata Zdarta, Wojciech Smulek, Teofil Jesionowski, Jakub Zdarta
Summary: In this study, lipase immobilized from Aspergillus niger by physical immobilization and mixed physical immobilization and ion exchange was used for kinetic resolution of the ketoprofen racemic mixture. The immobilized lipase showed improved activity and stability, and phosphate buffer at pH 7 was found to be the best solvent.
Article
Biochemistry & Molecular Biology
Daniel Castaneda-Valbuena, Angel Berenguer-Murcia, Roberto Fernandez-Lafuente, Roberto Morellon-Sterling, Veymar G. Tacias-Pascacio
Summary: The fishing industry generates valuable waste that contains a high amount of proteins. Processing these proteins can lead to products with nutritional, pharmacological, and technological benefits due to the presence of peptides. This review compiles investigations on the production of peptides from fish proteins using pepsin as a catalyst, highlighting their biological properties and potential uses.
PROCESS BIOCHEMISTRY
(2022)
Article
Biochemistry & Molecular Biology
Diego Carballares, Roberto Fernandez-Lafuente, Javier Rocha-Martin
Summary: This paper presents different methodologies for the immobilization of D-amino acid oxidase (DAAO). The results suggest that immobilization on glutaraldehyde support at pH 7 provides the best stability and activity for the enzyme.
PROCESS BIOCHEMISTRY
(2022)
Article
Chemistry, Physical
Diego Carballares, Javier Rocha-Martin, Roberto Fernandez-Lafuente
Summary: The study revealed a significant impact of buffer nature and concentration on the stability of DAAO enzyme, especially at pH 7 and 9. The dissociation process of the enzyme plays a crucial role in the buffer solution, but using immobilized DAAO biocatalyst can mitigate the effects of buffer on enzyme stability.
Article
Chemistry, Multidisciplinary
Thays N. da Rocha, Diego Carballares, Jose R. Guimaraes, Javier Rocha-Martin, Paulo W. Tardioli, Luciana R. B. Goncalves, Roberto Fernandez-Lafuente
Summary: Lipases A and B from Candida antarctica, Thermomyces lanuginosus, and Candida rugosa were immobilized on different carriers and exhibited different specificity and optimal activities. The stability of the preparations was strongly influenced by the substrate and conditions for residual activity determination, not necessarily corresponding to the optimal temperatures.
SUSTAINABLE CHEMISTRY AND PHARMACY
(2022)
Article
Biochemistry & Molecular Biology
Thays N. da Rocha, Roberto Morellon-Sterlling, Javier Rocha-Martin, Juan M. Bolivar, Luciana R. B. Goncalves, Roberto Fernandez-Lafuente
Summary: Penicillin G acylase from Escherichia coli was successfully immobilized on vinyl sulfone agarose beads. The immobilization was optimized with the purpose of obtaining a stable and active biocatalyst. The study found that vinyl sulfone agarose beads have heterofunctionality, offering new possibilities for enzyme immobilization.
Article
Chemistry, Physical
Thays N. da Rocha, Roberto Morellon-Sterling, Luciana R. B. Goncalves, Juan M. Bolivar, Andres R. Alcantara, Javier Rocha-Martin, Roberto Fernandez-Lafuente
Summary: In this study, MANAE-vinyl sulfone (VS) agarose beads were found to be a better support for immobilizing PGA enzyme compared to agarose-VS beads. The modified beads allowed for rapid and covalent immobilization of PGA, resulting in improved stability compared to low ionic strength VS-agarose.
Article
Chemistry, Physical
Jose R. Guimaraes, Diego Carballares, Javier Rocha-Martin, Andres R. Alcantara, Paulo W. Tardioli, Roberto Fernandez-Lafuente
Summary: The lipase from Thermomyces lanuginosus (TLL) was immobilized on a methacrylate macroporous resin coated with octadecyl groups for enhanced stability. Covalent immobilization of the enzyme was achieved by activating the support with divinyl sulfone. The use of different blocking agents affected the functional features of the biocatalyst.
Article
Chemistry, Physical
Amalie Vang Host, Roberto Morellon-Sterling, Diego Carballares, John M. Woodley, Roberto Fernandez-Lafuente
Summary: Enzymes have excellent catalytic features, but stability remains an issue. Co-immobilization of enzymes can further improve enzyme properties, but it introduces the challenge of different enzyme stabilities. This paper investigates the effect of co-immobilized enzymes with varying stabilities on operational performance through simulations and discusses strategies to overcome these challenges.
Article
Biotechnology & Applied Microbiology
Pedro Abellanas-Perez, Diego Carballares, Javier Rocha-Martin, Roberto Fernandez-Lafuente
Summary: This article analyzes the interactions between enzyme crowding and its chemical modification on enzyme activity and stability. The results show that the effects of chemical modifications vary depending on the enzyme support loading. Lowly loaded enzymes increase their activity after modification, while the opposite is true for highly loaded enzymes. TNBS modification increases the stability of highly loaded enzymes but decreases their activity.
BIOTECHNOLOGY PROGRESS
(2023)
Article
Engineering, Chemical
David Gonzalez-Miranda, Diego Carballares, Tomas Pedregal, Roberto Fernandez-Lafuente, Miguel Ladero, Juan M. Bolivar
Summary: This study achieved efficient synthesis of glycerol carbonate using immobilized lipases under solventless conditions. The use of immobilized C. rugosa lipase showed the highest turnover numbers and allowed for multiple reaction cycles. The results of this research highlight the significance of reaction engineering approaches in developing efficient sustainable processes for synthesizing valuable chemicals.
INDUSTRIAL & ENGINEERING CHEMISTRY RESEARCH
(2023)
Article
Biochemistry & Molecular Biology
Pedro Abellanas-Perez, Diego Carballares, Roberto Fernandez-Lafuente, Javier Rocha-Martin
Summary: This study immobilized two lipases, CALB and TLL, on octyl agarose, and observed the stability of the enzymes under different treatment methods and buffers. It was found that the stability of the enzymes varied depending on the treatment method and buffer used. The presence of intermolecular crosslinkings enhanced stability for the highly loaded enzymes, while intramolecular crosslinkings played a role in stabilizing the lowly loaded enzymes.
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
(2023)
Article
Biochemistry & Molecular Biology
El-Hocine Siar, Roberto Morellon-Sterling, Diego Carballares, Javier Rocha-Martin, Oveimar Barbosa, Juan M. Bolivar, Roberto Fernandez-Lafuente
Summary: The immobilization of ficin extract on glyoxyl agarose beads stabilized the enzyme, but the stability was highest when immobilized for 3 hours. The biocatalyst prepared for 24 hours was less stable when thermally inactivated in the presence of air. Incubating the thermally inactivated biocatalysts with dithiothreitol allowed for the recovery of almost total activity, with better recovery observed for the 24 hour biocatalyst. This suggests that the faster enzyme inactivation of the 24 hour biocatalyst was due to Cys oxidation.
PROCESS BIOCHEMISTRY
(2023)
Article
Nanoscience & Nanotechnology
Keyvan Jodeiri, Aleksandra Foerster, Gustavo F. Trindade, Jisun Im, Diego Carballares, Roberto Fernandez-Lafuente, Marcos Pita, Antonio L. De Lacey, Christopher Parmenter, Christopher Tuck
Summary: The drive for miniaturization in enzyme-based bioelectronics has led to the development of 3D microstructured electrodes. Additive manufacturing combined with electroless metal plating allows the production of these electrodes with high surface area. The challenge lies in ensuring strong adhesion between the metal layer and polymer structure, which has been achieved in this study through the introduction of an interfacial adhesion layer.
ACS APPLIED MATERIALS & INTERFACES
(2023)
Article
Chemistry, Multidisciplinary
Leandros Paschalidis, Sara Arana-Pena, Volker Sieber, Jakob Burger
Summary: Advances in enzymatic cascade reactions have led to increased interest in enzyme co-immobilization in porous particles. The optimal conditions for different spatial immobilization distributions (SIDs) are not fully understood, and there is a lack of simulation and optimization methods for these systems. This study provides a theoretical framework for modeling enzymatic cascade reactions with enzymes immobilized in porous particles. Different SIDs were analyzed, and it was found that homogeneous co-immobilization outperforms individual immobilization when diffusion is slow compared to reaction rates. Heterogeneous co-immobilization with enzymes positioned at the entry of the pore is advantageous when reaction rates or substrate diffusion is slow.
REACTION CHEMISTRY & ENGINEERING
(2023)