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Immobilization of Enzymes and Cells

SECOND EDITION

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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

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Immobilization of

Enzymes and Cells

S

ECOND

E

DITION

Edited by

Jose M. Guisan

Institute of Catalysis, CSIC

Campus UAM–Cantoblanco

Madrid, Spain

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© 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 BiotechnologyTMis 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, Immobi-lized. QU 135 I3141 2006]

TP248.65.I45I46 2006 660.6'34—dc22

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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

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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.

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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,

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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

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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

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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

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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

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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

´

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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

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