This fourth edition volume expands on the previous editions with new insights on important aspects to take into accounting when immobilizing enzymes and cells, illustrating outstanding examples that support those aspects, and exploring ways to fabricate and characterize heterogeneous biocatalysts including both immobilized enzymes and cells. The transformation of soluble and usually instable enzymes into heterogeneous and highly stable biocatalysts is strongly emphasized. The chapters in this book cover topics such as the importance of enzyme orientation on the support surface; application and characterization of immobilized enzymes; different functionalization chemistries for the modulation of the immobilized enzyme properties; co-immobilization of multi-enzyme systems; new analytical techniques for the characterization of heterogeneous biocatalysts; protocols for cell entrapment in alginate; preparation and characterization of biofilms; and cell encapsulation technologies. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Cutting-edge and authoritative, Immobilization of Enzymes and Cells: Methods and Protocols, Fourth Edition is a valuable resource for researchers interested in expanding their knowledge of this developing field.
1. Overview of Immobilized Enzymes Applications in Pharmaceutical, Chemical, and Food Industry
Alessandra Basso and Simona Serban
2. Parameters for the Evaluation of Immobilized Enzymes under Process Conditions
Andrés Illanes and Lorena Wilson
3. One-Point Covalent Immobilization of Enzymes on Glyoxyl-Agarose with Minimal Physico-Chemical Modification: Immobilized “Native Enzymes”
Jose M. Guisan, Fernando Lopez-Gallego, Juan M. Bolivar, Javier Rocha-Martin, and Gloria Fernandez-Lorente
4. Multi-Point Covalent Immobilization of Enzymes on Glyoxyl-Agarose with Minimal Physico-Chemical Modification: Stabilization of Industrial Enzymes
Fernando López-Gallego, Gloria Fernandez-Lorente, Javier Rocha-Martin, Juan M. Bolivar, Cesar Mateo, and Jose M. Guisan
5. Multi-Potent Covalent Immobilization of Enzymes on Supports Activated with Epoxy Groups: Stabilization of Industrial Enzymes
Cesar Mateo, Olga Abian, Gloria Fernández-Lorente, Benevides C. C. Pessela, Valeria Grazu, Jose M. Guisan, and Roberto Fernandez-Lafuente
6. Immobilization of Enzymes on Supports Activated with Glutaraldehyde: A Very Simple Immobilization Protocol
Fernando López-Gallego, Jose M. Guisán, and Lorena Betancor
7. Very Strong but Reversible Immobilization of Enzymes on Supports Coated with Ionic Polymers
Cesar Mateo, Benevides C. C. Pessela, Manuel Fuentes, Rodrigo Torres, Claudia Ortiz, Fernando López-Gallego, Lorena Betancor, Noelia Alonso-Morales, Jose M. Guisan, and Roberto Fernandez-Lafuente
8. Immobilization of Lipases by Adsorption on Hydrophobic Supports: Modulation of Enzyme Properties in Biotransformations in Anhydrous Media
Gloria Fernandez-Lorente, Javier Rocha-Martin, and Jose M. Guisan
9. Immobilization of Enzymes on Hetero-Functional Supports: Physical Adsorption Plus Additional Covalent Immobilization
Lara Trobo-Maseda, Alejandro H. Orrego, María Romero-Fernández, Jose M. Guisan, and Javier Rocha-Martin
10. Stabilization of Multimetic Enzymes via Immobilization and Further Cross-Linking with Aldehyde-Dextran
Cesar Mateo, Benevides C. C. Pessela, Manuel Fuentes, Rodrigo Torres, Lorena Betancor, Aurelio Hidalgo, Gloria Fernández-Lorente, Roberto Fernandez-Lafuente, and Jose M. Guisan
11. Immobilization of Enzymes on Magnetic Beads through Affinity Interactions
Audrey Sassolas, Akhtar Hayat, and Jean-Louis Marty
12. Immobilization of Proteins on Gold Surfaces
José M. Abad, Marcos Pita, and Víctor M. Fernández
13. Immobilization of Enzymes in Protein Films
Daniel Sánchez-de Alcázar, Mantas Liutkus, and Aitziber L. Cortajarena
14. Encapsulation of Enzymes in Porous Capsules via particle Templating
Mikhail V. Zyuzin, Pedro Ramos-Cabrer, and Susana Carregal-Romero
15. Enzyme Immobilization in Wall-Coated Flow Microreactors
Donya Valikhani, Juan M. Bolivar, and Bernd Nidetzky
16. In Situ Immobilization of Enzymes in Biomimetic Silica
Erienne Jackson, Sonali Correa, and Lorena Betancor
18. Co-Immobilization and Co-Localization of Multi-Enzymes Systems on Porous Materials
Alejandro H. Orrego, Fernando López-Gallego, Gloria Fernandez-Lorente, José M. Guisan, and Javier Rocha-Martin
19. Single-Particle Kinetics of Immobilized Enzymes by Harnessing the Autofluorescence of Co-Immobilized Cofactors
Ana I. Benítez-Mateos
20. Intraparticle pH Sensing within Immobilized Enzymes: Immobilized Yellow Fluorescent Protein as Optical Sensor for Spatiotemporal Mapping of pH Inside Porous Particles
Tanja Consolati, Juan M. Bolivar, Zdenek Petrasek, Jose Berenguer, Aurelio Hidalgo, Jose M. Guisán, and Bernd Nidetzky
21. Manufacturing of Protein-Based Biomaterials Coupling Cell-Free Protein Synthesis with Protein Immobilization
Fernando López-Gallego and Ana I. Benítez-Mateos
22. Immobilization of Enzymes as Cross-Linked Enzyme Aggregates: General Strategy to Obtain Robust Biocatalysts
Susana Velasco-Lozano
23. NMR OF Immobilized Enzymes
Linda Cerofolini, Enrico Ravera, Marco Fragai, and Claudio Luchinat
24. Whole Cell Entrapment Techniques
Jorge A. Trelles and Cintia W. Rivero
25. Immobilization of INS1E Insulin-Producing Cells within Injectable Alginate Hydrogels
Albert Espona-Noguera, Jesús Ciriza, Alberto Cañibano-Hernández, Laura Saenz del Burgo, and Jose Luis Pedraz
26. Immobilization of Cells on Polyurethane Foam
Ignacio de Ory, Gema Cabrera, Martin Ramirez, and Ana Blandino
27. Covalent Immobilization of Microbial Cells on Microchannel Surfaces
Gorazd Stojkovič and Polona Žnidaršič-Plazl
28. Immobilization of Cells by Magnetic Nanoparticles
Alireza Ebrahiminezhad, Seyedeh-Masoumeh Taghizadeh, Younes Ghasemi, and Aydin Berenjian
29. Cultivation of Productive Biofilms in Flow-Reactors and their Characterization by CLSM
Christian David, Ingeborg Heuschkel, Katja Bühler, and Rohan Karande
30. Immobilization of Microalgae
Nirupama Mallick
31. Therapeutic Applications of Encapsulated Cells
Argia Acarregui, Gorka Orive, José Luis Pedraz, and Rosa María Hernández
This fourth edition volume expands on the previous editions with new insights on important aspects to take into accounting when immobilizing enzymes and cells, illustrating outstanding examples that support those aspects, and exploring ways to fabricate and characterize heterogeneous biocatalysts including both immobilized enzymes and cells. The transformation of soluble and usually instable enzymes into heterogeneous and highly stable biocatalysts is strongly emphasized. The chapters in this book cover topics such as the importance of enzyme orientation on the support surface; application and characterization of immobilized enzymes; different functionalization chemistries for the modulation of the immobilized enzyme properties; co-immobilization of multi-enzyme systems; new analytical techniques for the characterization of heterogeneous biocatalysts; protocols for cell entrapment in alginate; preparation and characterization of biofilms; and cell encapsulation technologies. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Cutting-edge and authoritative, Immobilization of Enzymes and Cells: Methods and Protocols, Fourth Edition is a valuable resource for researchers interested in expanding their knowledge of this developing field.