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PHYSICS AND CHEMISTRY BASIS OF BIOTECHNOLOGY VOLUME 7 FOCUS ON BIOTECHNOLOGY Volume7 SeriesEditors MARCELHOFMAN Centrefor Veterinary andAgrochemical Research, Tervuren, Belgium JOZEF ANNÉ Rega Institute, University of Leuven, Belgium Volume Editors MARCEL DE CUYPER Katholieke Universiteit, Leuven Interdisciplinaire Research Center, Kortrijk, Belgium JEFF W.M. BULTE National Institutes of Health, Bethesda, MD, U.S.A. COLOPHON Focus on Biotechnology is an open-ended series of reference volumes produced by Kluwer Academic Publishers BV in co-operation with the Branche Belge de la Société de Chimie Industrielle a.s.b.1. The initiative has been taken in conjunction with the Ninth European Congress on Biotechnology. ECB9 has been supported by the Commission of the European Communities, the General Directorate for Technology, Research and Energy of the Wallonia Region, Belgium and J. Chabert, Minister for Economy of the Brussels Capital Region. Physics and Chemistry Basis of Biotechnology Volume 7 Edited by MARCEL DE CUYPER Katholieke Universiteit Leuven, Interdisciplinaire Research Center, Kortrijk, Belgium and JEFF W.M. BULTE National Institutes of Health, Bethesda, MD, U.S.A. KLUWER ACADEMIC PUBLISHERS NEW YORK / BOSTON / DORDRECHT / LONDON / MOSCOW eBook ISBN: 0-306-46891-3 Print ISBN: 0-792-37091-0 ©2002 Kluwer Academic Publishers New York, Boston, Dordrecht, London, Moscow Print ©2001 Kluwer Academic Publishers Dordrecht All rights reserved No part of this eBook may be reproduced or transmitted in any form or by any means, electronic, mechanical, recording, or otherwise, without written consent from the Publisher Created in the United States of America Visit Kluwer Online at: http://kluweronline.com and Kluwer's eBookstore at: http://ebooks.kluweronline.com EDITORS PREFACE At the end of the 20th century, a tremendous progress was made in biotechnology in its widest sense. This progress was largely possible as a result of joint efforts of top academic researchers in both pure fundamental sciences and applied research. The surplus value of such interdisciplinary approaches was clearly highlighted during the 9th European Congress on Biotechnology that was held in Brussels, Belgium (11-15 July, 1999). The present volume in the ‘Focus on Biotechnology’ series, entiteld ‘Physics and Chemistry Basis for Biotechnology’ contains selected presentations from this meeting, A collection of experts has made serious efforts to present some of the latest developments in various scientific fields and to unveil prospective evolutions on the threshold of the new millenium. In all contributions the emphasis is on emerging new areas of research in which physicochemical principles form the foundation. In reading the different chapters, it appears that more than ever significant advances in biotechnology very often depend on breakthroughs in the biotechnology itself (e.g. new instruments, production devices, detection methods), which - in turn - can be realized by implementing the appropriate physical and chemical principles into the new application. This ‘common’ pattern is illustrated in the different chapters. Some highly relevant, next generation scientific topics that are treated deal with de novo synthesis of materials for gene transfection, imaging contrast agents, radiotherapy, aroma measurements, psychrophilic environments, biomimetic materials, bioradicals, biosensors, and more. Given the diversity of the selected topics, we are confident that scientists with an open mind, who are looking for new frontiers, will find several chapters of particular interest. Some of the topics will give useful, up-to-date information on scientific aspects that may be either right in, or at the interface of their own field of research. We would like to thank the many authors who did such an excellent job in writing and submitting their papers to us. It was enjoyable to interact with them, and there is no question that the pressure we put on many of them was worthwhile. Marcel De Cuyper Jeff W.M. Bulte This page intentionally left blank. TABLE OF CONTENTS EDITORS PREFACE ................................................................................................ V Biomimetic materials synthesis ................................................................................... 9 Aleksey Nedoluzhko and Trevor Douglas ........................................................ 9 Abstract ................................................................................................................. 9 1 . Introduction ....................................................................................................... 9 2 . Principles ........................................................................................................... 1 0 2.1. Nucleation .................................................................................................... 1 1 2.2. Growth .......................................................................................................... 1 1 2.3. Biomolecules and supramolecular assemblies as templates for crystal growth .............................................................................................................. 12 3 . Examples .............................................................................................................. 12 3.1. Proteins ...................................................................................................................12 3.1 . 1. Ferritin ................................................................................................ 13 3.1.2. Bacterial S-Layers ........................................................................... ......... 1 4 3.1.3. Anisotropic Structures -Tobacco Mosaic Virus ................................... 15 3.1.4. Spherical Virus Protein Cages ........................................................... 16 3.2. Synthetic polyamides -Dendrimers ........................................................... 17 3.3. Gels ........................................................................................................... 17 3.4. Composite materials ................................................................................. 18 3.5. Organized surfactant assemblies ............................................................... 19 3.5.1. Confined surfactant assemblies .......................................................... 19 3.5.1.1. Reverse micelles (water-in-oil microemulsions) ....................... 19 3.5.1.2. Oil-in-water micelles .................................................................. 24 3.5.1.3. Vesicles ..................................................................................... 24 3.5.2. Layered surfactant assemblies .......................................................... 25 3.5.2.1. Surfactantmonolayers andLangmuir-Blodgettfilms.............25 3.5.2.2. Self-assembled films ................................................................ 27 3.6. Synthesis of Mesoporous Materials ........................................................ 28 3.6.1. Liquid crystal templating mechanism ............................................. 28 3.6.2. Synthesisofbiomimeticmaterialswithcomplexarchitecture.......31 3.7. Synthesis of inorganic materials using polynucleotides ....................... 32 3.7.1. Synthesis not involving specific nucleotide-nucleotide interactions ................................................................................................................... 32 3.7.2. Synthesis involving nucleotide-nucleotide interactions ................. 34 3.8. Biological synthesis of novel materials .................................................. 37 3.9. Organization of Nanoparticles into Ordered Structures ......................... 37 References ......................................................................................................... 39 Dendrimers: .......................................................................................................... 47 Chemical principles and biotechnology applications ............................................ 47 L . Henry Bryant, Jr . and Jeff W.M. Bulte ..................................................... 47 1 Summary ........................................................................................................... 47 1 . Synthesis ....................................................................................................... 47 1 . 1. Divergent ................................................................................................ 48 1.2. Convergent ............................................................................................. 49 1.3. Heteroatom ............................................................................................. 49 1.4. Solid phase .............................................................................................. 5 1 1.5. Other ........................................................................................................ 5 1 2 . Characterisation ............................................................................................. 52 3 . Biotechnology applications ........................................................................... 53 3.1. Biomolecules .......................................................................................... 53 3.2. Glycobiology .......................................................................................... 55 3.3. Peptide dendrimers ................................................................................. 56 3.4. Boron neutron capture therapy ............................................................... 57 3.5. MR imaging agents ................................................................................ 58 3.6. Metal encapsulation ............................................................................... 60 3.7. Transfection agents ................................................................................ 60 3.8. Dendritic box .......................................................................................... 62 4 . Concluding remarks ...................................................................................... 63 References ......................................................................................................... 63 Rational design of P450 enzymes for biotechnology ............................................. 7 1 Sheila J . Sadeghi. Georgia E . Tsotsou, Michael Fairhead. Yergalem T . Meharenna and Gianfranco Gilardi .................................................................... 7 1 Abstract .............................................................................................................. 7 1 1 . Introduction ................................................................................................... 72 1 . 1. Interprotein Electron Transfer ................................................................ 72 1.2. Structure-function of cytochrome P450 enzymes .................................. 75 1.2.1. P450 redox chains ........................................................................... 75 1.2.2. P450 catalysis .................................................................................. 76 1.2.3. Bacterial P450s in biotechnology .................................................... 76 1.2.4. P450s in drug metabolism ............................................................... 78 1.3. Chimeras of P450 enzymes .................................................................... 79 1.3.1, Bacterial P450-P450-reductase fusion protein systems ................. 80 1.3.2. Plant P450-P450-reductase fusion proteins ..................................... 80 1.3.3. Plant/mammalianP450-P450-reductasefusionproteins................80 1.3.4. Mammalian fusion proteins .............................................................. 8 1 1.4. Biosensing ............................................................................................... 8 1 2 . Engineering artificial redox chains ............................................................... 84 3 . Screening methods for P450 activity ............................................................ 90 3.1. Assay methods for P450-linked activity ................................................ 90 3.2. Development of a new high-through-put screening method for NAD(P)H linked activity ........................................................................ 91 3.3. Validity of the new screening method .................................................... 93 4 . Designing a human/bacterial 2E1-BM3 P450 enzyme ................................. 94 4.1. Modelling ............................................................................................... 95 4.2. Construction ........................................................................................... 96 2 4.3. Expression and functionality .................................................................. 96 5 . Conclusions ................................................................................................... 97 Acknowledgements ........................................................................................... 98 References ........................................................................................................ 98 Amperometric enzyme-based biosensors for application in food and beverage industry .............................................................................................................. .105 Elisabeth Csöregi, Szilveszter GÁspÁr, Mihaela Niculescu, Bo Mattiasson. Wolfgang Schuhmann ..................................................................................... 105 Summary ......................................................................................................... 105 1 . Biosensors -Fundamentals ......................................................................... 106 2 . Prerequisites for application of biosensors in food industry ....................... 107 3.Existingbiosensorconfigurations andrelatedelectron-transferpathways 107 3.1. Biosensors based on O2or H2O2detection ........................................... 109 3.2. Biosensors based on free-diffusing redox mediators ........................... 110 3.3. Integrated sensor designs (reagentless biosensors) .............................. 113 4 . Selected practical examples ........................................................................ 115 4.1. Redox hydrogel integrated peroxidase based hydrogen peroxide biosensors .................................................................................................... 115 4.2. Amine oxidase-based biosensors for monitoring of fish freshness ...... 117 4.3. Alcohol biosensors based on alcohol dehydrogenase .......................... 119 5 . Enzyme-based amperometric biosensors for monitoring in different biotechnological processes .............................................................................. 1 23 6. Conclusions . . . .. . . . .. . . . .. . . . .. . . . .. . . . .. . . . .. . . . .. . . . .. . . . .. . . . .. . . . .. . . . .. . . . . .. . . . .. . . . .. . . . .. . . . .. . . . .. 125 Acknowledgements ......................................................................................... 1 25 References ....................................................................................................... 1 26 Supportedlipidmembranesforreconstitutionofmembraneproteins............... 131 Britta Lindholm-Sethson ............................................................................. 131 Abstract ........................................................................................................... 131 1 .Introduction ................................................................................................ 131 2 . Objective ..................................................................................................... 1 32 2.1. The plasma membrane ......................................................................... 132 2.2. The artificial cell membrane ..................................................................... 132 2.2.1. Unsupported artificial bilayer membranes .................................... 133 2.2.2. Supported artificial bilayer membranes (s-BLMs) ........................ 1 34 2.2.2.1. Formation of s-BLMs ............................................................. 1 34 2.2.2.2. Reconstitution of membrane proteins into the membrane ..... 1 34 2.2.3. Various methods of investigation ................................................. . 1 34 3 . s-BLMs in close contact with the solid support .........................................136 3.1. Langmuir-Blodgett films on solid supports .......................................... 136 3.1.1. Pure phospholipid films ................................................................ 137 3.1.2. s-BLM as receptor surface ............................................................ 138 3.1.3. s-BLM with ion channels and/or ionophores ................................ 138 3.1.4. s-BLM with other integral membrane proteins ............................. 138 3.2. Vesicle fusion ....................................................................................... 139 3.2.1. LB/vesicle method and/or direct fusion ........................................ 139 3