Immobilization of Enzymes and Cells SECOND EDITION

Immobilization of Enzymes and Cells SECOND EDITION

M E T H O D S I N B I O T E C H N O L O G Y John M. Walker, SERIES EDITOR 22. Immobilization of Enzymes and Cells, Second Edition, edited by Jose M. Guisan, 2006 21. Food-Borne Pathogens: Methods and Protocols, edited by Catherine Adley, 2006 20. Natural Products Isolation, Second Edition, edited by Satyajit D. Sarker, Zahid Latif, and Alexander I. Gray, 2006 19. Pesticide Protocols, edited by José L. Martínez Vidal and Antonia Garrido Frenich, 2006 18. Microbial Processes and Products, edited by Jose Luis Barredo, 2005 17. Microbial Enzymes and Biotransformations, edited by Jose Luis Barredo, 2005 16. Environmental Microbiology: Methods and Protocols, edited by John F. T. Spencer and Alicia L. Ragout de Spencer, 2004 15. Enzymes in Nonaqueous Solvents: Methods and Protocols, edited by Evgeny N. Vulfson, Peter J. Halling, and Herbert L. Holland, 2001 14. Food Microbiology Protocols, edited by J. F. T. Spencer and Alicia Leonor Ragout de Spencer, 2000 13. Supercritical Fluid Methods and Protocols, edited by John R. Williams and Anthony A. Clifford, 2000 12. Environmental Monitoring of Bacteria, edited by Clive Edwards, 1999 11. Aqueous Two-Phase Systems, edited by Rajni Hatti-Kaul, 2000 10. Carbohydrate Biotechnology Protocols, edited by Christopher Bucke, 1999 9. Downstream Processing Methods, edited by Mohamed A. Desai, 2000 8. Animal Cell Biotechnology, edited by Nigel Jenkins, 1999 7. Affinity Biosensors: Techniques and Protocols, edited by Kim R. Rogers and Ashok Mulchandani, 1998 6. Enzyme and Microbial Biosensors: Techniques and Protocols, edited by Ashok Mulchandani and Kim R. Rogers, 1998 5. Biopesticides: Use and Delivery, edited by Franklin R. Hall and Julius J. Menn, 1999 4. Natural Products Isolation, edited by Richard J. P. Cannell, 1998 3. Recombinant Proteins from Plants: Production and Isolation of Clinically Useful Compounds, edited by Charles Cunningham and Andrew J. R. Porter, 1998 2. Bioremediation Protocols, edited by David Sheehan, 1997 1. Immobilization of Enzymes and Cells, edited by Gordon F. Bickerstaff, 1997

M E T H O D S I N B I O T E C H N O L O G Y Immobilization of Enzymes and Cells SECOND EDITION Edited by Jose M. Guisan Institute of Catalysis, CSIC Campus UAM Cantoblanco Madrid,

2006 Humana Press Inc. 999 Riverview Drive, Suite 208 Totowa, New Jersey 07512 www.humanapress.com All rights reserved. No part of this book may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, microfilming, recording, or otherwise without written permission from the Publisher. Methods in Biotechnology TM is a trademark of The Humana Press Inc. All papers, comments, opinions, conclusions, or recommendations are those of the author(s), and do not necessarily reflect the views of the publisher. This publication is printed on acid-free paper. ANSI Z39.48-1984 (American Standards Institute) Permanence of Paper for Printed Library Materials. Cover design by Patricia F. Cleary Cover illustration: Porous solid supports used in double immobilization (Fig. 3, Chapter 6; see full caption on p. 68 and discussion on p. 67). For additional copies, pricing for bulk purchases, and/or information about other Humana titles, contact Humana at the above address or at any of the following numbers: Tel.: 973-256-1699; Fax: 973-256-8341; E-mail: orders@humanapr.com; or visit our Website: www.humanapress.com Photocopy Authorization Policy: Authorization to photocopy items for internal or personal use, or the internal or personal use of specific clients, is granted by Humana Press Inc., provided that the base fee of US $30.00 per copy is paid directly to the Copyright Clearance Center at 222 Rosewood Drive, Danvers, MA 01923. For those organizations that have been granted a photocopy license from the CCC, a separate system of payment has been arranged and is acceptable to Humana Press Inc. The fee code for users of the Transactional Reporting Service is: [1-58829-290-8/06 $30.00]. Printed in the United States of America. 10 9 8 7 6 5 4 3 2 1 eisbn 1-59745-053-7 Library of Congress Cataloging-in-Publication Data Immobilization of enzymes and cells / edited by Jose M. Guisan. 2nd ed. p. ; cm. (Methods in biotechnology ; 22) Includes bibliographical references and index. ISBN 1-58829-290-8 (alk. paper) 1. Immobilized enzymes Biotechnology. 2. Immobilized cells Biotechnology. I. Guisan, Jose M. II. Series. [DNLM: 1. Enzymes, Immobilized. 2. Biotechnology methods. 3. Cells, Immobilized. QU 135 I3141 2006] TP248.65.I45I46 2006 660.6'34 dc22 2006001106

Preface Enzymes and whole cells are able to catalyze the most complex chemical processes under the most benign experimental and environmental conditions. In this way, enzymes and cells could be excellent catalysts for a much more sustainable chemical industry. However, enzymes and cells also have some limitations for nonbiological applications: fine chemistry, food chemistry, analysis, therapeutics, and so on. Enzymes and cells may be unstable, difficult to handle under nonconventional conditions, poorly selective toward synthetic substrates, and so forth. From this point of view, the transformation from the laboratory to industry of chemical processes catalyzed by enzymes and cells may be one of the most complex and exciting goals in biotechnology. For many industrial applications, enzymes and cells have to be immobilized, via very simple and cost-effective protocols, in order to be re-used over very long periods of time. From this point of view, immobilization, simplicity, and stabilization have to be strongly related concepts. Over the last 30 years, a number of protocols for the immobilization of cells and enzymes have been reported in scientific literature. However, only very few protocols are simple and useful enough to greatly improve the functional properties of enzymes and cells, activity, stability, selectivity, and related properties. The second edition of Immobilization of Enzymes and Cells intends to complement as well as update the first edition. This volume now includes the following aspects of established and new protocols for immobilization: 1. Simple protocols for the immobilization of enzymes and cells that could be very useful for application at industrial scale. 2. Novel protocols for immobilization that can be deployed now or in the near future. 3. Immobilization protocols that can greatly improve the functional properties of enzymes and cells. 4. Different techniques for the characterization of immobilized enzymes and cells as suitable tools for the development of better immobilization techniques. 5. Protocols for the preparation of immobilized derivatives for use in very different nonconventional reaction media. 6. Different protocols for the preparation of immobilized derivatives possibly useful in varied areas of application: therapy, chemical industry, and so on. 7. New chemical reactors that overcome the limitations of a number of immobilized derivatives. There is still a long and exciting path for developing very simple and efficient protocols for the preparation, characterization, and utilization of immobilized enzymes and cells. Immobilization of Enzymes and Cells, Second Edition treats many of the very interesting results already obtained, at the same time giving readers the tools to v

vi Preface develop even more important immobilization protocols. It seems clear that successful development of excellent protocols for immobilization will promote massive implementation of enzyme and cell systems as industrial biocatalysts. Such a development could prove decisive to the emergence of much more skilled and sustainable chemical industry cost-effective production of very complex and useful molecules under the mildest conditions. The possible advances in fine chemistry, food chemistry, analysis, decontamination processes, and therapeutic applications, for example, only hint at the potential benefits. Jose M. Guisan

Contents Preface... v Contributors...xi 1 Immobilization of Enzymes as the 21st Century Begins: An Already Solved Problem or Still an Exciting Challenge? Jose M. Guisan... 1 2 Immobilization of Enzymes: A Literature Survey Beatriz M. Brena and Francisco Batista-Viera... 15 3 Cross-Linked Enzyme Aggregates Roger A. Sheldon, Rob Schoevaart, and Luuk M. van Langen... 31 4 Immobilization Stabilization of Enzymes by Multipoint Covalent Attachment on Supports Activated With Epoxy Groups Cesar Mateo, Olga Abian, Gloria Fernández-Lorente, Benevides C. C. Pessela, Valeria Grazu, Jose M. Guisan, and Roberto Fernandez-Lafuente... 47 5 Glutaraldehyde in Protein Immobilization: A Versatile Reagent Lorena Betancor, Fernando López-Gallego, Noelia Alonso-Morales, Gisella Dellamora, Cesar Mateo, Roberto Fernandez-Lafuente, and Jose M. Guisan... 57 6 Practical Protocols for Lipase Immobilization Via Sol-Gel Techniques Manfred T. Reetz... 65 7 Encapsulation of Enzymes Using Polymers and Sol-Gel Techniques Mònica Campàs and Jean-Louis Marty... 77 8 Design of Smart Biocatalysts: Immobilization of Enzymes on Smart Polymers Ipsita Roy and Munishwar N. Gupta... 87 9 Affinity Immobilization of Tagged Enzymes Silvana Andreescu, Bogdan Bucur, and Jean-Louis Marty... 97 10 Bioaffinity Immobilization Ipsita Roy and Munishwar N. Gupta... 107 11 One-Step Purification, Immobilization, and Stabilization of Poly-Histidine-Tagged Enzymes Using Metal Chelate Epoxy Supports Cesar Mateo, Benevides C. C. Pessela, Valeria Grazu, Rodrigo Torres, Fernando López-Gallego, Jose M. Guisan, and Roberto Fernandez-Lafuente... 117 vii

viii Contents 12 Stabilization of Multimeric 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... 129 13 Purification, Immobilization, Hyperactivation, and Stabilization of Lipases by Selective Adsorption on Hydrophobic Supports Jose M. Palomo, Gloria Fernández-Lorente, Cesar Mateo, Rosa L. Segura, Claudia Ortiz, Roberto Fernandez-Lafuente, and Jose M. Guisan... 143 14 Immobilization and Stabilization of Proteins by Multipoint Covalent Attachment on Novel Amino-Epoxy-Sepabeads Cesar Mateo, Benevides C. C. Pessela, Valeria Grazu, Fernando López-Gallego, Rodrigo Torres, Manuel Fuentes, Aurelio Hidalgo, Jose M. Palomo, Lorena Betancor, Gloria Fernández-Lorente, Claudia Ortiz, Olga Abian, Jose M. Guisan, and Roberto Fernandez-Lafuente... 153 15 Improved Stabilization of Chemically Aminated Enzymes Via Multipoint Covalent Attachment on Glyoxyl Supports Tamara Montes, Fernando López-Gallego, Manuel Fuentes, Cesar Mateo, Valeria Grazu, Lorena Betancor, Jose M. Guisan, and Roberto Fernandez-Lafuente... 163 16 Stabilization of New Imprint Property of Glucose Oxidase in Pure Aqueous Medium by Cross-Linked-Imprinting Approach Alankar Vaidya and Lutz Fischer... 175 17 Reversible Covalent Immobilization of Enzymes Via Their Thiol Groups Francisco Batista-Viera, Karen Ovsejevi, and Carmen Manta... 185 18 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... 205 19 Immobilization of Enzymes on Magnetic Particles v Martina Koneracká, Peter Kopcanský, Milan Timko, Chenyl Nynitapal Ramchand, Zainul M. Saiyed, Michael Trevan, and Anil de Sequeira... 217 20 Immobilization of Proteins on Gold Surfaces José M. Abad, Marcos Pita, and Víctor M. Fernández... 229 21 Immobilization of Enzymes on Electrodes Gilvanda Silva Nunes and Jean-Louis Marty... 239

Contents ix 22 Immobilization of Enzymes for Use in Organic Media Patrick Adlercreutz... 251 23 Immobilization of Enzymes for Use in Ionic Liquids Pedro Lozano, Teresa de Diego, and José L. Iborra... 257 24 Immobilization of Enzymes for Use in Supercritical Fluids Pedro Lozano, Teresa de Diego, and José L. Iborra... 269 25 Immobilized Enzymes for Biomedical Applications Amaia Esquisabel, Rosa María Hernández, Alicia Rodríguez Gascón, and José Luis Pedraz... 283 26 Characterization of Immobilized Enzymes by Microcalorimetry Ezio Battistel and Giovanni Rialdi... 295 27 Use of Immobilized Biocatalysts in Fluidized Bed Format Ipsita Roy and Munishwar N. Gupta... 311 28 Taylor Couette Vortex Flow in Enzymatic Reactors Roberto Campos Giordano and Raquel de Lima Camargo Giordano... 321 29 A Novel Immobilization Method for Entrapment: LentiKats Marc Schlieker and Klaus-Dieter Vorlop... 333 30 Encapsulation of Cells in Alginate Gels Gorka Orive, Rosa María Hernández, Alicia Rodríguez Gascón, and José Luis Pedraz... 345 31 Immobilization of Cells on Polyurethane Foam Ignacio de Ory, Gema Cabrera, Martin Ramirez, and Ana Blandino... 357 32 Immobilization of Cells With Transition Metal Pedro Fernandes... 367 33 Immobilization of Microalgae Nirupama Mallick... 373 34 Bioluminescence in Immobilized Cells for Biomass Detection and Biosensor Applications v Marián Navrátil, Juraj Svitel, Peter Gemeiner... 393 35 A Proteomic Approach to Biofilm Cell Physiology Laurent Coquet, Sébastien Vilain, Pascal Cosette, Thierry Jouenne, and Guy-Alain Junter... 403 36 Encapsulation of Bacteria for Biodegradation of Gasoline Hydrocarbons Peyman Moslemy, Serge R. Guiot, and Ronald J. Neufeld... 415 37 Biomedical Applications of Immobilized Cells Gorka Orive, Rosa María Hernández, Alicia Rodríguez Gascón, and José Luis Pedraz... 427 Index... 439

Contributors JOSÉ M. ABAD Institute of Catalysis, CSIC, Campus UAM-Cantoblanco, Madrid, OLGA ABIAN Institute of Catalysis, CSIC, Campus UAM-Cantoblanco, Madrid, PATRICK ADLERCREUTZ Department of Biotechnology, Lund University, Lund, Sweden NOELIA ALONSO-MORALES Institute of Catalysis, CSIC, Campus UAM-Cantoblanco, Madrid, SILVANA ANDREESCU Department of Chemistry, Clarkson University, Potsdam, New York FRANCISCO BATISTA-VIERA Cátedra de Bioquímica, Facultad de Química, Universidad de la República, Montevideo, Uruguay EZIO BATTISTEL Istituto Guido Donegani, Polimeri Europa SpA, Novara, Italy LORENA BETANCOR Institute of Catalysis, CSIC, Campus UAM-Cantoblanco, Madrid, ANA BLANDINO Department of Chemical Engineering, Food Technology, and Environmental Technologies, Facultad de Ciencias, University of Cadiz, Cadiz, BEATRIZ M. BRENA Cátedra de Bioquímica, Facultad de Química, Universidad de la República, Montevideo, Uruguay BOGDAN BUCUR BIOMEM, Centre de Phytopharmacie, Université de Perpignan, Perpignan Cedex, France GEMA CABRERA Department of Chemical Engineering, Food Technology and Environmental Technologies, Facultad de Ciencias, University of Cadiz, Cadiz, MÒNICA CAMPÀS BIOMEM, Centre de Phytopharmacie, Université de Perpignan, Perpignan Cedex, France LAURENT COQUET Laboratory of Polymers, Biopolymers, and Membranes (PBM), UMR 6522 CNRS and European Institute for Peptide Research (IFRMP 23), University of Rouen Cedex, France PASCAL COSETTE Laboratory of Polymers, Biopolymers, and Membranes (PBM), UMR 6522 CNRS and European Institute for Peptide Research (IFRMP 23), University of Rouen Cedex, France GISELLA DELLAMORA Institute of Catalysis, CSIC, Campus UAM-Cantoblanco, Madrid, TERESA DE DIEGO Departamento de Bioquímica y Biología Molecular B e Inmunología, Facultad de Química, Universidad de Murcia, Campus del Espinardo, AMAIA ESQUISABEL Laboratorio de Farmacia y Tecnologia Farmacéutica, Facultad de Farmacia, Universidad del País Vasco (EHU-UPV), Vitoria-Gasteiz, xi

xii Contributors PEDRO FERNANDES Centro de Engenharia Biologica e Quimica, Instituto Superior Tecnico, Lisboa, Portugal VÍCTOR M. FERNÁNDEZ Institute of Catalysis, CSIC, Campus UAM-Cantoblanco, Madrid, ROBERTO FERNANDEZ-LAFUENTE Institute of Catalysis, CSIC, Campus UAM- Cantoblanco, Madrid, GLORIA FERNÁNDEZ-LORENTE Institute of Catalysis, CSIC, Campus UAM- Cantoblanco, Madrid, LUTZ FISCHER Institute of Food Technology, Department of Biotechnology, University of Hohenheim, Stuttgart, Germany MANUEL FUENTES Institute of Catalysis, CSIC, Campus UAM-Cantoblanco, Madrid, ALICIA RODRÍGUEZ GASCÓN Laboratorio de Farmacia y Tecnologia Farmacéutica, Facultad de Farmacia, Universidad del País Vasco (EHU-UPV), Vitoria- Gasteiz, PETER GEMEINER Department of Glycobiotechnology, Institute of Chemistry, Slovak Academy of Sciences, Bratislava, Slovakia RAQUEL DE LIMA CAMARGO GIORDANO Departamento de Engenharia Química, Universidade Federal de São Carlos, São Carlos, Brasil ROBERTO CAMPOS GIORDANO Departamento de Engenharia Química, Universidade Federal de São Carlos, São Carlos, Brasil VALERIA GRAZU Institute of Catalysis, CSIC, Campus UAM-Cantoblanco, Madrid, SERGE R. GUIOT Groupe de bioingenierie de l environnement, Biotechnology Research Institute, National Research Council of Canada, Montreal, Quebec, Canada JOSE M. GUISAN Institute of Catalysis, CSIC, Campus UAM-Cantoblanco, Madrid, MUNISHWAR N. GUPTA Department of Chemistry, Indian Institute of Technology, Delhi, N. Delhi, India ROSA MARÍA HERNÁNDEZ Laboratorio de Farmacia y Tecnologia Farmacéutica, Facultad de Farmacia, Universidad del País Vasco (EHU-UPV), Vitoria- Gasteiz, AURELIO HIDALGO Institute of Catalysis, CSIC, Campus UAM-Cantoblanco, Madrid, JOSÉ L. IBORRA Departamento de Bioquímica y Biología Molecular B e Inmunología, Facultad de Química, Universidad de Murcia, Campus del Espinardo, THIERRY JOUENNE Laboratory of Polymers, Biopolymers and Membranes (PBM), UMR 6522 CNRS and European Institute for Peptide Research (IFRMP 23), University of Rouen Cedex, France GUY-ALAIN JUNTER Laboratory of Polymers, Biopolymers and Membranes (PBM), UMR 6522 CNRS and European Institute for Peptide Research (IFRMP 23), University of Rouen Cedex, France

Contributors xiii MARTINA KONERACKÁ Institute of Experimental Physics, Slovak Academy of Sciences, Kosice, Slovakia v PETER KOPCANSKY Institute of Experimental Physics, Slovak Academy of Sciences, Kosice, Slovakia FERNANDO LÓPEZ-GALLEGO Institute of Catalysis, CSIC, Campus UAM- Cantoblanco, Madrid, PEDRO LOZANO Departamento de Bioquímica y Biología Molecular B e Inmunología, Facultad de Química, Universidad de Murcia, Campus del Espinardo, NIRUPAMA MALLICK Agricultural and Food Engineering Department, Indian Institute of Technology, Kharagpur, West Bengal, India CARMEN MANTA Cátedra de Bioquímica, Facultad de Química, Universidad de la República, Montevideo, Uruguay JEAN-LOUIS MARTY BIOMEM, Centre de Phytopharmacie, Université de Perpignan, Perpignan Cedex, France CESAR MATEO Institute of Catalysis, CSIC, Campus UAM-Cantoblanco, Madrid, TAMARA MONTES Institute of Catalysis, CSIC, Campus UAM-Cantoblanco, Madrid, PEYMAN MOSLEMY Sr. Scientist, Spherics, Inc., Lincoln, RI MARIÁN NAVRÁTIL Department of Biochemical Technology, Faculty of Chemical and Food Technology Slovak University of Technology, Bratislava, Slovak Republic RONALD J. NEUFELD Department of Chemical Engineering, Queen s University, Kingston, Ontario, Canada GILVANDA SILVA NUNES Technological Chemistry Department, CCET, Federal University of Maranhão, São Luís, Maranhão, Brazil GORKA ORIVE Laboratorio de Farmacia y Tecnologia Farmacéutica, Facultad de Farmacia, Universidad del País Vasco (EHU-UPV), Vitoria-Gasteiz, CLAUDIA ORTIZ Institute of Catalysis, CSIC, Campus UAM-Cantoblanco, Madrid, IGNACIO DE ORY Department of Chemical Engineering, Food Technology, and Environmental Technologies, Facultad de Ciencias, University of Cadiz, Cadiz, KAREN OVSEJEVI Cátedra de Bioquímica, Facultad de Química, Universidad de la República, Montevideo, Uruguay JOSE M. PALOMO Institute of Catalysis, CSIC, Campus UAM-Cantoblanco, Madrid, JOSÉ LUIS PEDRAZ Laboratorio de Farmacia y Tecnologia Farmacéutica, Facultad de Farmacia, Universidad del País Vasco (EHU-UPV), Vitoria-Gasteiz, BENEVIDES C. C. PESSELA Institute of Catalysis, CSIC, Campus UAM-Cantoblanco, Madrid, MARCOS PITA Institute of Catalysis, CSIC, Campus UAM-Cantoblanco, Madrid, CHENYL NYNITAPAL RAMCHAND Kemin Nutricional Technologies Pvt Ltd., Tamil Nadu, India

xiv Contributors MARTIN RAMIREZ Department of Chemical Engineering, Food Technology, and Environmental Technologies, Facultad de Ciencias, University of Cadiz, Cadiz, MANFRED T. REETZ Max-Planck-Institut für Kohlenforschung, Mülheim/Ruhr, Germany GIOVANNI RIALDI ISMAC-Istituto per lo Studio delle Macromolecole, CNR, Sezione di Genova, Genova, Italy IPSITA ROY Department of Chemistry, Indian Institute of Technology, Delhi, N. Delhi, India ZAINUL M. SAIYED School of Mechanical and Systems Engineering, University of Newcastle upon Tyne, Newcastle, United Kingdom MARC SCHLIEKER genialab GmbH, Braunschweig, Germany ROB SCHOEVAART CLEA Technologies, Delft, The Netherlands ROSA L. SEGURA Institute of Catalysis, CSIC, Campus UAM-Cantoblanco, Madrid, ANIL DE SEQUEIRA School of Science and Environment, Bath Spa University College, Newton Park Campus, Bath, United Kingdom ROGER A. SHELDON Delft University of Technology, Biocatalysis and Organic Chemistry, Delft, The Netherlands v JURAJ SVITEL Pure and Applied Biochemistry, Center for Chemistry and Chemical Engineering, Lund University, Lund, Sweden MILAN TIMKO Institute of Experimental Physics, Slovak Academy of Sciences, Kosice, Slovakia RODRIGO TORRES Institute of Catalysis, CSIC, Campus UAM-Cantoblanco, Madrid, MICHAEL TREVAN University of Westminster, London, United Kingdom ALANKAR VAIDYA Institute of Food Technology, Department of Biotechnology, University of Hohenheim, Stuttgart, Germany LUUK M. VAN LANGEN CLEA Technologies, Delft, The Netherlands SÉBASTIEN VILAIN Laboratory of Polymers, Biopolymers, and Membranes (PBM), UMR 6522 CNRS and European Institute for Peptide Research (IFRMP 23), University of Rouen Cedex, France KLAUS-DIETER VORLOP genialab GmbH, Braunschweig, Germany