ENZYME ENGINEERING Volume 2 A Continuation Order Plan is available for this series. A continuation order will bring delivery of each new volume immediately upon publication. Volumes are billed only upon actual shipment. For further information please contact the publisher. ENZYME ENGINEERING Volume 2 Edited by E. Kendall Pye Department of Biochemistry School of Medicine University of Pennsylvania Philadelphia, Pennsylvania and Lemuel B.Wingard, Jr. Department of Pharmacology School of Medicine University of Pittsburgh Pittsburgh, Pennsylvania PLENUM PRESS • NEW YORK AND LONDON Library of Congress Cataloging in Publication Data Engineering Foundation Conference on Enzyme Engineering, 2d, Henniker, N. H., 1973. Enzyme engineering. Called volume 2 in continuation of a volume with the same title published in 1972, which contains the papers of the 1st Engineering Foundation Con ference on Enzyme Engineering. 1. Enzymes - Industrial applications - Congresses. I. Pye, E. KendalL ed. II. Wingard, L., ed. III. Title. [DNLM: 1. Biomedical engineering - Con gresses. 2. Enzymes - Congresses. W3 EN696] TP248.E5E53 1973 660'.63 74-13768 ISBN-13: 978-1-4615-8899-3 e-ISBN-13: 978-1-4615-8897-9 DOl: 10.1007/978-1-4615-8897-9 ACKNOWLEDGMENTS Permission has been received from the various publishers for reproduction of the following figures. p. 25, Fig. 2 Exp. Cell Res. (Academic Press, Inc.! p. 28, Fig. 5 Science (Amer. Assoc. Advanc. Sci.) p. 84-86, Fig. 1, 2, 3 Enzyme (S. Karger AG, Basel) p. 84-87, Fig. 1,2,4 Biotechnol. Bioeng. (John Wiley & Sons, Inc.) p. 118, Fig. 2 Nature (Macmillan Journals Ltd., London) p. 226-227, Fig. 1,2 Biotechnol. Bioeng. (John Wiley & Sons, Inc.) p. 344-348, Fig. 1-7 Die Starke (Verlag Chemie GMBH) p. 428, Fig. 1 J. Polymer Sci. (John Wiley & Sons, Inc.) This volume contains most of the papers and research reports presented at the Second Engineering Foundation conference on Enzyme Engineering held at Henniker, New Hampshire, August 5-10, 1973. © 1974 Plenum Press, New York Softcover reprint of the hardcover I st edition 1973 A Division of Plenum Publishing Corporation 227 West 17th Street, New York, N.Y. 10011 United Kingdom edition published by Plenum Press, London A Division of Plenum Publishing Company, Ltd. 4a Lower John Street, London WIR 3PD, England 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 ORGANIZATION OF THE CONFERENCE Sponsor: The Engineering Foundation 345 East 47th Street New York, N.Y. 10017 USA Financial Support: Corning Glass Works Corning, N.Y. 14830 USA and The Engineering Foundation Executive Chairman of the Conference: Lemuel B. Wingard, Jr. Program Chairman of the Conference: E. Kendall Pye Advisory Board: Theodore Cayle William H. McMullen III Thomas M.S. Chang Robert S. Melville Ichiro Chib at a Klaus Mosbach Peter Dunnill E. Kendall Pye Victor Edwards Daniel I.C. Wang Richard Falb Howard H. Weetall George G. Guilbault Lemuel B. Wingard, Jr. Arthur E. Humphrey ORGANIZATION FOR FUTURE ENZYME ENGINEERING CONFERENCES Summer 1975 -- Engineering Foundation International Conference tentatively scheduled for USA Executive Committee: E. Kendall Pye Executive Chairman Howard H. Weetall -- Program Chairman Leon Goldstein Lemuel B. Wingard, Jr. Advisory Board: Thomas M.S. Chang Yehuda Levin Ichiro Chibata Georg Manecke Leland C. Clark, Jr. David L. Marshall Clark K. Colton William H. McMullen Alkis Constantinides K. Mosbach Peter Dunnill David Novelli George G. Guilbault David F. Ollis William E. Hornby Hiritoshi Samejima C.G. Heden Bern J. Schnyder Csaba Horvath P.V. Sundaram Arthur E. Humphrey Shuichi Suzuki D. Jarowek Daniel I.C. Wang Summer 1977 -- International Conference tentatively scheduled for Europe PREFACE Considerable worldwide interest has arisen in recent years in the controlled use of enzymes as catalysts in industrial processing, analytical chemistry and medical therapy. This interest has genera ted the new interdisciplinary field of Enzyme Engineering, which includes both the scientific and technologic aspects of the produc tion, purification, immobilization, and application of enzymes in a variety of situations and reactor configurations. A series of Engineering Foundation conferences on Enzyme Engineering was initia ted to provide an international forum for the exchange of ideas and information over the entire range of this new field. The outstanding success of the first two conferences attests to the vigor and poten tial of this field to contribute significantly to a better under standing and resolution of some of the major problems faced by man kind. The first conference, which was held August 9-13, 1971, at Henniker, New Hampshire, U.S.A., aided significantly in molding the several traditional disciplines that interact to form the field of Enzyme Engineering. The conference was highly successful mainly because many of the key scientists and engineers from the several facets of Enzyme Engineering were brought together for the first time at a single residential meeting. The result was an exchange of ideas and "education" of one another in the pertinent principles of the diverse disciplines which contribute to this field. The second conference, held August 5-10, 1973, at Henniker, New Hampshire, U.S.A., was equally successful, with 190 participants ~rom over 350 applicants) attending. Eighteen countries and a wide variety of academic disciplines and occupational specialties were represented. The two major emphases of the 1973 conference were the application of immobilized enzymes and the problems in utilizing enzyme systems that require cofactors. This volume contains most of the papers and research reports presented at the 1973 conference. In addition the results of a sur vey taken at the conference and the report of an ad hoc committee that met during the conference are included. The names of the session cochairmen are included in the "Table of Contents" in appreciation of their efforts in making the 1973 conference a success. vii viii PREFACE Many thanks are due to Dr. Sandford S. Cole and his staff at the Engineering Foundation Conferences office in New York for making these conferences possible and to the Corning Glass Works for their continued financial support. E. Kendall Pye Lemuel B. Wingard, Jr. January, 1974 CONTENTS . . . . . . . . . . . . . . . . . . . . . . . . . . Preface vii DEVELOPMENTS AND CHALLENGE OF ENZYME ENGINEERING 3 LemueZ B. Wingarod~ Jr>. 1973 HENNIKER DELPHI STUDY 9 CaroZ-GOr>an Heden SESSION 1. ENZYMES: REGULATION AND NEW SOURCES. • • • • •• 15 Chairmen: K. Aunst1'Up and A. L. Demain Stability of Enzymes from Thermophilic Microorganisms 17 AZfr>ed R. Doig~ Jr>. Polypeptide Hormones from Tissue Culture • • • • , 23 Marotin Posner> Design Consideration for Animal Cell Cultures as Sources of Enzymes • • • • • • • • • • • • • • •• 31 L. K. Nyi!'i SESSION II. NEW PURIFICATION TECHNIQUES ••••••• 41 Chairmen: P. DunniZt and G.D. NoveUi Recent Developments in Enzyme Isolation Processes 43 P. DunniZZ and M.D. LiZZy Protein Purification by Immunoadsorption • • • • 47 N. G. Ander>son~ D. W. HoUaday~ J.E. Caton and J. W. HoUeman Purification of Urokinase by Affinity Chromatography 55 T. Maciag~ M.K. WeibeZ and E.K. Pye Short Report: Derivatives of Controlled Pore Glass for Affinity Chromatography • • • • • • • • 63 Geor>ge Baum ix x CONTENTS SESSION III. NEW IMMOBILIZATION TECHNIQUES AND SUPPORTS 65 Chairmen: R. FaZb and L. GoZdstein Review of Recent Enzyme Immobilization Techniques 67 Richard D. FaZb Encapsulation of Enzymes in Liquid Membrane Emulsions 77 S.W. May and N.N. Li An Organic Millieu in Immobilized Enzyme Synthesis and Catalysis ••• • • • • • • • • • • • • • • • 83 H.D. Brown~ G.J. BartZing and S.K. Chattopadhyay Immobilization of Enzymes on Phenol-Formaldehyde Resins. 91 A.C. OZson and W.L. StanZey Derivatized Nylon: A New Support for the Immobilization of Enzymes • • • • • • • • • • • • • • • • • • • • • •• 97 L. A. Freeman and M. SokoZovasky GoZdstein~ New Immobilization Techniques and Supports • • • 105 D. J01J)orek Stabilization and Immobilization of Enzymes with lmidoesters • • • • • • • • • • 115 Oskar Zaborsky Complexation of Enzymes or Whole Cells with Collagen 123 S.S. W.R. Vieth and A. Constantinides Wang~ Short Reports: Immobilized Enzymes Using Resinous Carriers 131 H. Sa:mejima and K. Kimura Covalent Coupling of Small Molecules and Proteins to Poly (2-Rydroxyethyl Methacrylate Methacrylic Acid) Hydrogels ..................... 137 T.P. T.A. A.S. Hoffman and Mate~ Horbett~ B.D. Ratner SESSION IV. IMMOBILIZED MULTI-STEP ENZYME SYSTEMS: PHYSICAL METHODS FOR EXAMINING IMMOBILIZED ENZYMES 139 Chairmen: K. Mosbach and E.K. Pye Theoretical and Practical Aspects of Immobilized Multi-Step Enzyme Systems • • • • • • • • 143 K. B. S. GestreZius and Mosbach~ Mattiasson~ P.A. Srere Enzyme Reactions for Preparative Scale Synthesis . 153 B.K. Ha:miZton~ J.P. Montgomery and D.I.C. Wang Application of Circular Dichroism to the Study of Enzyme Polymer Conjugates •• • • • . • • • • • . 161 Oskar R. Zaborsky