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
Immobilization of
Enzymes and Cells
S
ECOND
E
DITION
Edited by
Jose M. Guisan
Institute of Catalysis, CSIC
Campus UAM–Cantoblanco
Madrid, Spain
© 2006 Humana Press Inc. 999 Riverview Drive, Suite 208 Totowa, New Jersey 07512
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Cover illustration: Porous solid supports used in “double immobilization” (Fig. 3, Chapter 6; see full caption on p. 68 and discussion on p. 67).
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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, Immobi-lized. QU 135 I3141 2006]
TP248.65.I45I46 2006 660.6'34—dc22
v
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
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.
Contents
vii
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,
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
Martina Koneracká, Peter Kopcansk´y , 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
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
xi
Contributors
JOSÉ M. ABAD • Institute of Catalysis, CSIC, Campus UAM-Cantoblanco, Madrid,
Spain
OLGA ABIAN • Institute of Catalysis, CSIC, Campus UAM-Cantoblanco, Madrid,
Spain
PATRICK ADLERCREUTZ • Department of Biotechnology, Lund University, Lund,
Sweden
NOELIA ALONSO-MORALES • Institute of Catalysis, CSIC, Campus UAM-Cantoblanco,
Madrid, Spain
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, Spain
ANA BLANDINO • Department of Chemical Engineering, Food Technology, and
Environmental Technologies, Facultad de Ciencias, University of Cadiz, Cadiz, Spain
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, Spain
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, Spain
TERESA DE DIEGO • Departamento de Bioquímica y Biología Molecular B e
Inmunología, Facultad de Química, Universidad de Murcia, Campus del Espinardo, Spain
AMAIA ESQUISABEL • Laboratorio de Farmacia y Tecnologia Farmacéutica, Facultad
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, Spain
ROBERTO FERNANDEZ-LAFUENTE • Institute of Catalysis, CSIC, Campus
UAM-Cantoblanco, Madrid, Spain
GLORIA FERNÁNDEZ-LORENTE • Institute of Catalysis, CSIC, Campus
UAM-Cantoblanco, Madrid, Spain
LUTZ FISCHER • Institute of Food Technology, Department of Biotechnology,
University of Hohenheim, Stuttgart, Germany
MANUEL FUENTES • Institute of Catalysis, CSIC, Campus UAM-Cantoblanco,
Madrid, Spain
ALICIA RODRÍGUEZ GASCÓN • Laboratorio de Farmacia y Tecnologia Farmacéutica,
Facultad de Farmacia, Universidad del País Vasco (EHU-UPV), Vitoria-Gasteiz, Spain
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,
Spain
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,
Spain
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, Spain
AURELIO HIDALGO • Institute of Catalysis, CSIC, Campus UAM-Cantoblanco,
Madrid, Spain
JOSÉ L. IBORRA • Departamento de Bioquímica y Biología Molecular B e
Inmunología, Facultad de Química, Universidad de Murcia, Campus del Espinardo, Spain
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
PETER KOPCANSKY • Institute of Experimental Physics, Slovak Academy of Sciences,
Kosice, Slovakia
FERNANDO LÓPEZ-GALLEGO • Institute of Catalysis, CSIC, Campus
UAM-Cantoblanco, Madrid, Spain
PEDRO LOZANO • Departamento de Bioquímica y Biología Molecular B e
Inmunología, Facultad de Química, Universidad de Murcia, Campus del Espinardo, Spain
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,
Spain
TAMARA MONTES • Institute of Catalysis, CSIC, Campus UAM-Cantoblanco, Madrid,
Spain
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, Spain
CLAUDIA ORTIZ • Institute of Catalysis, CSIC, Campus UAM-Cantoblanco, Madrid,
Spain
IGNACIO DE ORY • Department of Chemical Engineering, Food Technology, and
Environmental Technologies, Facultad de Ciencias, University of Cadiz, Cadiz, Spain
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,
Spain
JOSÉ LUIS PEDRAZ • Laboratorio de Farmacia y Tecnologia Farmacéutica, Facultad
de Farmacia, Universidad del País Vasco (EHU-UPV), Vitoria-Gasteiz, Spain
BENEVIDES C. C. PESSELA • Institute of Catalysis, CSIC, Campus UAM-Cantoblanco,
Madrid, Spain
MARCOS PITA • Institute of Catalysis, CSIC, Campus UAM-Cantoblanco, Madrid, Spain
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, Spain
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,
Spain
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
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,
Spain
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