Immobilization of Enzymes and Cells (4^th Ed., 4th ed. 2020) Methods and Protocols Methods in Molecular Biology Series, Vol. 2100

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.

Preface…

Table of Contents…

Contributing Authors…

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

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

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

17. Environmentally-Friendly Enzyme Immobilization onto MOF Materials

Victoria Gascón and Manuel Sánchez-Sánchez

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

31. Therapeutic Applications of Encapsulated Cells

Argia Acarregui, Gorka Orive, José Luis Pedraz, and Rosa María Hernández