ENZYME ENGINEERING

Volume 6

A Continu.tlon Ord.r PI.n i. .v.iI.bl. for thl. aerl... A continu.tion ord.r will bring d.liv.ry of •• ch n.w volum. Imm.dl.t.ly upon publlc.tlon. Volum ••• re blll.d only upon .ctu.1 shlpm.nt. For furth.r Informallon pl •••• cont.ct th. publl.h.r.

ENZYME ENGINEERING Volume 6

Edited by

Iehiro Chibata Research Laboratory of Applied Biochemistry Tanabe Seiyaku Company Osaka, Japan

Saburo Fukui Department of Industrial Chemistry Faculty of Engineering Kyoto University Kyoto, Japan

and

Lemuel B. Wingard, Jr. Department of Pharmacology School of Medicine University of Pittsburgt. Pittsburgh, Pennsylvania

PLENUM PRESS · NEW YORK AND LONDON

The Library of Congress cataloged the second volume of this title as follows:

Engineering Foundation Conference on Enzyme Engineering, 2d, Henniker, N. H., 1973. Enzyme engineering; [papers] Edited by E. Kendall Pye and Lemuel B. Wingard, Jr.

New York, Plenum Press [1974]

Called volume 2 in continuation of a volume with the same title published in 1972, which contains the papers of the 1 st Engineering Foundation Conference on Enzyme Engineering.

1. Enzymes - Industrial applications - Congresses . I. Pye, E. Kendall, ed. II. Wingard, L., ed. III. Title. [DNLM: 1. Biomedical engineering - Congresses. 2. En­zymes - Congresses. W3 EN696] TP248.E5E531973 660'.63 74-13768 ISBN 978-1-4615-9292-1 ISBN 978-1-4615-9290-7{eBook) 001 10.1007/978-1-4615-9290-7

LC 74-13768

Proceedings of the Sixth International Enzyme Engineering Conference, held September 20'- 25, 1981, in Kashikojima, Japan

© 1982 Plenum Press, New York Softcover reprint of the hardcover 1 st edition 1982

A Division of Plenum Publishing Corporation 233 Spring Street, New York, N.Y. 10013

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

COSPONSORS:

The Engineering Foundation 345 East 47th Street New York, NY 10017 U.S.A.

EXECUTIVE COMMITTEE:

Saburo Fukui • • Ichiro Chibata • David Fink • • • Jan Konecny ••• Garfield Royer Howard Weetall • Michael Weibel • Lemuel B. Wingard Jr •• Sandford Cole. • • • •

PROGRAM COMMITTEE:

I. Chibata, chm. C. Horvath H. Inoue A. 1. Laskin K. Mosbach T. Murachi

v

Japanese Society of Enzyme Engineering

c/o Tanabe Seiyaku Co. Osaka, Japan

Executive Chairman Program Chairman U.S.A. Member European Member U.S.A. Member U.S.A. Member U.S.A. Member Permanent Member Conference Director

D. Ryu H. Samejima S. Suzuki D. Thomas F. Wagner H. Yamada

vi ORGANIZATION OF THE CONFERENCE

ADVISORY BOARD:

F. Bartoli B. Mattiasson I. V. Berezin A. S. Michaels G. Broun K. Mosbach T. Cayle T. Murachi T. M. S. Chang A. H. Nishikawa F. F. Davis H. Okada H. Filippusson D. Ryu R. M. Flora H. Samejima F. Garcia-Hernandez G. Schmidt-Kastner S. Gestrelius M. Seidman H. U. Geyer B. P. Sharma C. Horvath S. Suzuki J. V. Hupkes D. Thomas H. Inoue W. R. Vieth E. Katchalski-Katzir F. Wagner J. Klein J. C. Weaver M-R. Kula M. Wilchek A. I. Laskin H. Yamada P. Linko F. Widmer G. Manecke

FINANCIAL SUPPORT:

103 Japanese companies contributed through the Japanese Society of Enzyme Engineering

Engineering Foundation New York, NY, U.S.A.

Assoreni Milan, Italy

BASF Aktiengesellschaft Ludwigshafen, F. R. Germany

Bayer AG Leverkusen, F. R. Germany

Behringwerke AG Marburg (Lahn), F.R. Germany

Boehringer Mannheim GmbH. Mannheim, F. R. Germany

British Petroleum Co. Sunbury-on-Thames, England

Cetus Corp. Berkeley, CA, U.S.A.

Ciba-Geigy AG Basel, Switzerland

Corning Glass Works Corning, NY, U.S.A.

ORGANIZATION OF THE CONFERENCE

Cutter Laboratories, for Bayer/Cutter/Miles Elkhart, IN, U.S.A.

Dow Chemical Co. Midland, MI, U.S.A.

DSM, N. V. Heerlen, Netherlands

Exxon Research and Engineering Co. Florham Park, NJ, U.S.A.

FMC Corp. Princeton, NJ, U.S.A.

Genex Corp. Rockville, MD, U.S.A.

Gist-Brocades, N.V. Delft, Netherlands

Chr. Hansen's Laboratorium Copenhagen, Denmark

Henkel & Cie, GmbH. Dusseldorf, F. R. Germany

Hoechst, AG Frankfurt (Main), F. R. Germany

International Minerals & Chemicals Inc. Terre Haute, IN, U.S.A.

Kraft, Inc. Glenview, IL, U.S.A.

Miles Kali-Chemie, GmbH. Hannover, F. R. Germany

Monsanto Co. St. Louis, MO, U.S.A.

Novo Industries Bagsvaerd, Denmark

Pfizer, Inc. Groton, CT, U.S.A.

Pharmacia Inc. Piscataway, NJ, U.S.A.

G. D. Searle & Co. Skokie, IL, U.S.A.

A. E. Staley Manufacturing Co. Decatur, IL, U.S.A.

National Science Foundation Washington, DC, U.S.A.

vii

NEXT ENZYME ENGINEERING CONFERENCE

TIME: September 25-30, 1983

LOCATION: Pocono Hershey Resort Hotel White Haven, PA, U.S.A.

EXECUTIVE COMMITTEE:

Allen I. Laskin •

George T. Tsao. •

Gunter Schmidt-Kastner.

Shuichi Suzuki ••

Daniel Thomas • •

Howard H. Weetall •

Michael H. Weibel •

Lemuel B. Wingard Jr ••

(on highway Interstate 80 near Scranton, Pennsylvania)

Executive Chairman

Program Chairman

European Member

Japan Member

European Member

U.S.A. Member

U.S.A. Member

Permanent Member

Sandford S. Cole • • • • • • • • • Conference Director

viii

PREFACE

Presently, intensive and global attention is being devoted to "biotechnology"--the technology utilizing marvelous capacities of living things for human welfare. Each country is strongly promo­ting its development. In particular, enzyme engineering, whose pur­pose is to utilize efficiently enzymes, microorganisms, and cultured plant as well as animal cells as organic catalysts, is one of the main themes in the field of biotechnology.

Under these circumstances, the Sixth Enzyme Engineering Confe­rence was held at Kashikojima, Mie Prefecture, Japan from September 20 to 25, 1981, under the joint auspices of the Engineering Foun­dation of New York and the Japanese Society of Enzyme Engineering. This series of international conferences has been held biannually since 1971. The first three and the fifth conferences were held in the United States and the fourth one was in the Federal Republic of Germany. This sixth conference was the first to be held in Asia; and it was significant that a number of participant£ could visit Japany, which has produced successful achievements in the field of biotechnology.

This conference had 203 participants from 23 countries. There were participants from China for the first time, thus enlarging the international scope of the conference series. Besides the opening and closing lectures, 36 plenary lectures and 96 poster papers were presented; and most of them are included in this vol­ume. It is worthy of notice that some topics of genetic engineer­ing now share common interests with enzyme engineering, so that scientists and engineers in both fields should work together in the future. If these two areas can be combined successfully, it will surely lead to greater contributions and further attainments of biotechnology.

This successful conference was aided markedly by the contin­ued efforts of Dr. Sandford S. Cole of the Engineering Foundation, members of the Executive Committee, and the General Secretaries of the Japanese Society of Enzyme Engineering. Also, we are deeply indebted to the many industrial firms in Europe, the United

ix

x PREFACE

States, and Japan that contributed financial support. We heartily express our gratitude to all of them. We also express our appre­ciation to MS. Hall of the University of Pittsburgh for her fine efforts in retyping all of the edited manuscripts and to the pro­duction staff at Plenum Press for their assistance in the publi­cation of this volume.

rchiro Chibata Saburo Fukui Lemuel B. Wingard, Jr.

April, 1982

CONFERENCE HISTORY

The First Ten Years L.B. Wingard Jr.

KEYNOTE PAPER •••

CONTENTS

Recent Studies on Antibiotics and Low Molecular Weight Enzyme Inhibitors ••••••••••••

H. Umezawa

SESSION I. LARGE SCALE PRODUCTION AND PURIFICATION OF BIOMOLECULES

Chairmen: H. Weetall and H. Yamada

Integration of Large Scale Production and Purifi-

1

3

9

11

35

cation of Biomolecules • • • • • • • • •• •••• 37 K. Venkatasubramanian

Role of Active Transport of Inducer in Enzyme Biosynthesis and Process Scale-up • • • • • • 45

W.R. Vieth, K. Kaushik and K. Venkatasubraman-ian

Multi Enzyme Systems in Membrane Reactors • • • • • 61 c. Wandrey, R. Wichmann and A.S. Jandel

Scale-Up of Protein Purification by Liquid-Liquid Extraction . . . • . . . . . . . . . . . . . . . 69

M.-B. Kula, K.H. Kroner, H. Hustedt and H. Schutte

Rapid and Large-Scale Purification of Angiotensin-I Forming Enzymes and Milk Clotting Enzymes by Affinity Chromatography •• • • • • • • • • • • • • 75

K. Murakami, s. Hirose and H. Kobayashi

xi

xii CONTENTS

separation and Purification of Enzymes Via contin-uous parametric pumping • • • • • • • • • • • 77

S.Y. Huang, C.K. Lin and L.Y. Juang

SESSION II. APPLICATIONS OF BIOCATALYSTS FOR NEW REACTIONS AND ORGANIC SYNTHESIS

~hairmen: A.L. Laskin and I. Chibata

Enzymatic Synthesis of Penicillins and Cephalo­sporins by Penicillin Acylase • • • • • • • • •

R. Okachi, Y. Hashimoto, M. Kawamori, R. Katsumata, K. Takayama and T. Nara

Enzymatic Acyl Transfer in Penicillin and Cephalo-sporin Chemistry •••••••••••••••

J. Konecny, A. Schneider and M. Sieber

Enzymatic Processes for the Synthesis of Optically

79

81

91

Active Amino Acids • • • • • • • • • • • • • • • • • 97 H. Yamada

Horse Liver Alcohol Dehydrogenase: An Illustrative Example of the Potential of Enzymes in Organic Synthesis. • • • • • • • • • • • • • • • • • • • • . 107

J.B. Jones

Immobilized Enzymes and Synzymes: Applications in Organic Synthesis •••••••••••••• 117

G.P. Royer

Immobilization of Living Microbial Cells and Trans-formation of Steroids ••••••••• • • • • 123

K.A. Koshcheyenko and G.K. Skryabin

Steroid Bioconversion by Immobilized Cells 125 C. Glomon, P. Germain, A. Miclo and J.M. Engasser

Steroid 61-DehYdrogenase Isoenzymes in terium Species Chol 73 T 191 • • • • •

W.-R. Mueller, W. Preuss and R.D.

Corynebac-

Schmid

Use of Immobilized Enzyme Requiring Cofactor Re­generation and of Immobilized Mycelium for Steroid

127

Modification • • • • • • • • • • • • • • • • • • • • 129 M.D. Legoy, F.Ergan, P. Dhulster, M.N. Kim and G. Gellf

CONTENTS

Hydroxylation of Steroids by Immobilized Microbial Cells ................... .

A. Tanaka, K. Sonomoto, M. Hoq, N. Usui, K. Nomura and S. Fukui

Enzymatic Synthesis of Sulfur and Selenium Amino Acids ..... . . . . •

K. Soda, N. Esaki and H. Tanaka

Production of Alkaloids with Immobilized Cells of . Catharanthus Roseus • • • • • •

C.A. Lambe, A. Reading, S. Roe, A. Rosevear, and A.R. Thomson

Propyleneoxide Production by Immobilized Nocardia Corallina B-276 • • • • •

K. Furuhashi, S. Uchida, I. Karube and S. Suzuki

Air-Oxidation of Linoleic Acid by Lipoxygenase­Containing Particles Suspended in Water-Insoluble Organic Solvent • • • • • • • • • • • • • • • • •

T. Yamane

xiii

131

135

137

139

141

Progress Toward Artificial Photosynthesis • • • • • • 143 D.J. Graves

Light-Sensitization of a Microbial Protease Y.Y. Lee, K.N. Kuan, and P. Melius

Enzymatic Photosensitive Materials N.F. Kazanskaya

Ester Exchange of Triglyceride by Entrapped Lipase

145

149

in Organic Solvent • • • • • • • • • • • • • • • • • 151 K. Yokozeki, T. Tanaka, S. Yamanaka, T. Takinami, Y. Hirose, K. Sonomoto, A. Tanaka and S. Fukui

Solvent Production by Clostridium Acetobutylicum in Aqueous Two-Phase Systems • • • • • • • • • • • • • • 153

B. Mattiasson, M. Suominen, E. Andersson, L. Haggstrom, P.-A. Albertsson and B. Hahn-Hagerdal

Enzyme Kinetics and Mass-Transfer on Two Liquid Phase Heterogeneous Systems •••••••••••• 157

J.M.C. Duarte

xiv CONTENTS

Physico-Chemical Means of Increasing the Yield of End Products in Biocatalysis •••• • • • • • • • • 159

r. v. Berezin

Heat-Stable L-Lactate Dehydrogenase and its Applica-tion to an Enzyme Reactor • • • • • • • • • • • • • • 161

T. Ohta, H. Taguchi and H. Matsuzaswa

Multienzymes and Cofactors Immobilized within Lipid­Polyamide Membrane Microcapsules for Sequential Substrate Conversion • • • • • • • • • • • • • • • • 163

Y.T. Yu and T.M.S. Chang

Tyrosine and 3,4-Dihydroxyphenyl-Alanine Synthesis by Citrobacter Freundii • • • • • • • • • • • • • • • 165

N.S. Egorov, M.B. Koupletskaja, and E.N. Kondratieva

Enzyme Catalysts for the Stereo and Regio Selective Oxyfunctionalization of Organic Substrates ••••• 167

S.W. May

SESSION III. NEW IMMOBILIZATION TECHNIQUES OF BIo­MATERIALS AND THEIR APPLICATIONS

Chairmen: D. Thomas and N. Ise 171

High-Rate, Continuous Waste Processor for the Produc-tion of High BTU Gas Using Immobilized Microbes • • • 173

R.A. Messing

New Developments in the preparation and Characteriza-tion of Polymerbound Biocatalysts 181

J. Klein and G. Manecke

Bioconversion of Lipophilic or Water Insoluble Com­pounds by Immobilized Biocatalysts in Organic Solvent Systems • • • • • • • • • • • • • • • • • • • • 191

S. Fukui and A. Tanaka

Multiple Carrier Regeneration and Enzyme Immobiliza-tion In Situ •••••• • • • • • • • • • • 201

J.E. Prenosil and E. Stuker

Viability and Biosynthetic Capacity of Immobilized Plant Cells • • • • • • • • • • • • • • • • 203

P. Brodelius, F. Constabel and W.G.W. Kurz

CONTENTS

Immobilized Enzymes of Heterogenous Structure K. Nakamura, K. Hibino and Y. Yano

xv

205

Fibrous Support for Immobilization of Enzymes • • •• 207 H. Ichijo, T. Suehiro, A. Yamauchi, S. Ogawa, M. Sakurai and N. Fujii

High Temperature Cell-Trapped Ultrafiltration Mem-branes

E. Drioli, G. Iorio, M. DeRosa, A. Gambacorta, and B. Nicolaus

209

Immobilized Modifiers for Proteins (HiPs) • . • . •• 211 ~l.H. Scouten, C. Lewis, A. Barnett, R. Haller and and W. Iobst

Enzyme Immobilization in Porous Supports J.E. Bailey

Immobilization of Microbial Cells Using Gelatin and Glutaraldehyde • • • • • • • • • • • •

Q. Wang, X. Ji and Z. Yuan

Iw~obilization of S-Galactosidase from E. Coli CSH36 and Its Microbial Cells Using Cellulose Beads • • • •

Y. Hong, S. Kwon, M. Chun and M. Sernetz

Hydrophobic Immobi~ization of Enzymes and Polynuc­leotides on Trityl Agarose •••••

P. Cashion, A. Javed, V. Lentini, D. Harrison, J. Seeley and G. Sathe

Influence of the Activation Degree of the Support on the Properties of Agarose-Nuclease • • • • • • •

A. Ballesteros, J.M. Guisan, and J. Serrano

Immobilization of Biofunctional Components by Radia­tion Polymerization and Applications • • • • • • • •

I. Kaetsu, M. Kumakura, T. Fujimura, M. Yoshida, F. Yoshii, M. Asano, M. Tamada, and N. Kasai

Immobilization of Whole Cell Glucose Isomerase within Soybean Protein

C.L. Lai

Enzyme Stabilization in High Macromolecular Concen-

213

215

217

219

223

225

227

tration Environments ••••• • • • • • • • 229 G. Greco Jr., G. Marrucci, and L. Gianfreda

xvi CONTENTS

Intrinsic Stability of Thermophilic Enzymes: 6-Phos­phogluconate Dehydrogenase from Bacillus Stearo-thermophilus and Yeast •••••••••••••• • 233

F.M. Veronese, E. Boccu and A. Fontana

Stabilization of Penicillin Amidohydrolase Immobi-lized on Eupergit C • • • • • • • • • • • • • • • • • 235

K. Sauber and D.M. Kramer

Stabilization of Fumarase Activity of Brevibacterium Flavum Cells by Immobilization with K Carrageenan and Polyethyleneimine • • • • • • • • • • • • • • • 237

T. Tosa, I. Takata, and I. Chibata

Application of Polyethylene Glycol-Bound NAD Deriva­tive and Thermostable Dehydrogenase in a Model Enzyme Reactor • • • • • • • • • • • • • • • • • • • • • • • 239

I. Urabe, N. Katayama, and H. Okada

Use of the Porous Mineral Spherosil as a Carrier for Enzymes: Fixation and Purification • • • • • 241

B. Mirabel

SESSION IV. INDUSTRIAL APPLICATIONS OF IMMOBILIZED BIOMATERIALS

Chairmen: A. Michaels and H. Samajima

Potential Application of Immobilized Viable Cells in the Food Industry: Malolactic Fermentation of Wine •

S. Gestrelius

Production of L-Tryptophan with Immobilized Cells F. Wagner, S. Lang, W.-G. Bang, K.D. Vorlop, and J. Klein

Applications of Immobilized Tannin fOr Protein and Metal Adsorption • • • • • • • • • • • • • • •

I. Chibata, T. Tosa, T. Mori, T. Watanabe, K. Yamashita and N. Sakata

Industrial Applications of Immobilized Biomaterials in China . . . . . . . • . . . . . . . . . . . .

S. Zhang

Production of L-Alanine from Ammonium Fumarate Using Two Types of Immobilized Microbial Cells ••••••

T. Sato, S. Takamatsu, K. Yamamoto, I. umemura, T. Tosa, and I. Chibata

243

r-, ,245) ~

:-251

259

265

271

CONTENiS

New Process for Production of High Fructose Corn Syrup Using Combined Adsorption and an Enzyme Re-

xvii

actor . . . . . . . . . . . . . . . . . 273 K. Hashimoto, S. Adachi and H. Noujima

Immobilized Multienzyme Systems for Starch Processing A. Lindroos, Y.Y. Linko, and P. Linko 275

Glucoamylase Covalently Coupled to Porous Glass 279 G. Li, J. Huang, X. Kou and S. zhang

Rotary Multidisc Reactor of Collagen Supported Immo-bilized Glucoamylase • • • • • • • • • • • • • • • • 281

S. Gondo, H. Koya and M. Morishita

Kinetics for the Hydrolysis of Soluble Starch by Glucoamylase and Application to an Immobilized Enzyme System • • • • • • • • • • • • • • • • • • • • • • • 283

K. Kusunoki and K. Kawakami

Reduced Thermostability of Modified Mucor Miehei Rennet

S. Branner-Jorgensen, P. Eigtved and P. Schneider

287

Application of Immobilized Enzymes to Milk Curdling • 289 S. Shimizu, K. Ohmiya, S.-E. Yun and T. Kobayashi

Production of 7-Aminodesacetoxycephalosporanic Acid by Immobilized E. coli Cells • • • • 291

Z. Wang, H. Yuo, M. Wang, Q. Jiao, W. Han, W. Sun and Q. Zhang

Technical Applications of Lactase and Aminoacid Acylase Immobilized to Form Plexazym • . • . 293

H. Plainer, B.G. Spbrssler and H. Uhlig

Continuous Hydrolysis of Lactose in Skim Milk and Acid Whey by Immobilized Lactase of Aspergillus Oryzae • • • • • • • • • • • • • • • • • • • • • 295

H. Hirohara, H. Yamamoto, E. Kawano and T. Nagase

~actic Acid Fermentation with Immobilized Lacto-bacillus Sp. •••••••••••••••••••• 299

S.L. Stenroos, Y.Y. Linko, P. Linko, M. Harju, and M. Heikonen

xviii CONTENTS

Production of I-Malic Acid with Immobilized Thermo-philic Bacterium, T.ber.mus Rubens Nov. sp. ••••• 303

Y. Ado, '1'. Kawamoto, I. Masunaga, K. '1'akayama, S. '1'akasawa, and K. Kimura .

Continuous Hydrolysis of Concentrated Sucrose Solu-tions by Immobilized Invertase • • • • • • • • 305

D. Combes and P. Monsan

A Series of Covalently Sonded Enzymes and Their Applications •••••••••• • • • • • • • • 307

S. Liu, Z. Yuan, O. Nang, J. Nang and Y. Zeng

The Application of Fluidized Beds for Improved Enzyme Reactor Performance • • • •

A. Renken, B. Flaschel and P.-F. Fauquex

Control of Continuous Coenzyme Regeneration R. Nichmann and C. Nandrey

A New Approach to Membrane Reactor Design and Opera-

309

311

tion ... . . . . . . . . . . . . . . . . . . . 313 B. Flaschel, B. Raetz and A. Renken

Influence of Compaction in Gel-Immobilized Enzyme Packed Bed Reactors ••••••••••••

S. Furusaki, Y. Okamura and'!'. Miyauchi

SESSION V. BIOMASS CONVERSION WITH ENERGY PRODUCTION Chairmen:N. Vieth and S. SUzuki

Enzymatic Removal of Hazardous Pollutants from In-

315

317

dustrial Aqueous Effluents • • • • ~ • • • • • • • • 319 A.M. Klibanov

Enzymatic Hydrolysis of Cellulose: Effects of Struc-tural properties of Cellulose on Hydrolysis Kinetics 325

D.D.Y. Ryu and S.B. ~ee

Continuous Ethanol Fermentation by Immobilized Biocatalysts • • • • • • • • • • • • • • • 335

- P. Liilko and Y.Y. Linko

An Immobilized Yeast Cell Column for the Fermentation of Molasses • • • • • • • •

D.F. Day and D. Sarkar 343

CONTENTS

Pilot Operation for Continuous Alcohol Fermentation of Molasses in an Immobilized Bioreactor ••• • • •

S. Fukushima and S. Hanai

Enzymatic Hydrolysis of Cellulose • • • • • • Y. Harano, H. Ooshima, K. Ohmine and M. Sakata

ATP Regeneration by Enzymes of Alcohol Fermentation and Kinases of Yeast and Its Computer Simulation

R. Matsuno, M. Asada, K. Nakanishi and T. Kamikubo

Extracellular Cellulases Produced by a Yeast-Like

xix

347

349

351

Fungus • • • • • • • • • • • • • • • • • • 353 G. Larios, A. Gilbon, Y. Lara and C. Huitron

Relationship Between Extracellular Proteases and the Cellulase Complex of Trichoderma Reesei • • • • • • • 355

C.P. Dunne

SESSION VI. ANALYTICAL APPLICATIONS OF IMMOBILIZED BIOMATERIALS

Chairmen: LoB. wingard Jr. and H. Okada

Binding Assays Involving Separation in Aqueous Two­Phase Systems: Partition Affinity Ligand Assay

357

(PALA) ••••••••••••••••••••••• 359 B. Mattiasson, T.G.I. Ling, and M. Ramstorp

Use of Immobilized Enzyme Reactors in Automated Clinical Analyses • • • • • • • • • • • 369

T. Mu.r:achi

Immobilized Enzymes in Analysis: Applications and Economic Aspects • • • • • • • • • • • 177

M. Gloger, M. Nelboeck, D. Doring, and S. Klose

Microbial Sensors for Gas Analysis S. Suzuki and I. Karube

Analytical Uses of Immobilized Enzymes G.G. Guilbault

Cost Analysis and Viability of Immobilized Enzymes in

387

395

Routine Analysis • • • • • • • • • • • • • • • • • • 405 P. V. Sundaram

Multipurpose Enzyme-Collagen Membrane Electrodes D.C. Gautheron, P.R. Coulet and C. Bertrand

409

xx CONTENTS

Long-Term Stability of Air-Dried Enzyme Electrodes with Selective Enzymic Collagen Membranes • • • • • • 411

P.R. Coulet, D.C. Gautheron, and G. Bardeletti

Enzyme Electrodes Based on Insolubilized Enzyme Mem-branes Coupled with an Electrochemical Transducer 413

J.L. Romette, N.D. Tran, P. Durand, J.L. Boitieux, and J.L. Navarro

Potentiometric Glucose Sensor: Enzymatic Activity and Potentiometric Measurements • • • • • •

L. B. Wingard Jr. and J. F. Castner

Microbial Sensor for Preliminary Screening of Mutagens • • • • • • • • • •

I. Karube and S. Suzuki

Application of Microbial Electrode to Analysis of Waste Water • • • • • . • • • • • • . • . • • .

M. Hikuma, H. Suzuki, T. Yasuda, I. Karube and S. Suzuki

Biosensors Based on Enzyme Amplification and Immuno-

415

417

419

chemical selecti vi ty • • • • • • • • • • • • • • • • 421 M. Aizawa and S. Suzuki

Application of Chemiluminescence of Cypridina Luci-fer in Analog to Immobilized Enzyme Sensors • • • • • 423

T. Kobayashi, K. Saga, S. Shimizu, and T. Goto

Use of Hydrogen Sensitive Pd-MOS Components in Bio-chemical Analysis • • • • • • • • • • • • • 425

B. Danielsson, F. Winquist, K. Mosbach and I. Lundstrom

Computer Controlled Mass Spectrometer Monitoring of Fermentations • • • • • • • • • • • • • • • 429

E. Pungor Jr., C.L. Cooney and J.C. Weaver

SESSION VII. MEDICAL APPLICATIONS OF ENZYME TECHNOLOGY Chairmen: K. Mosbach and T. Murachi

431

Immobilized Heparinase: Production, Purification, and Application in Extracorporeal Therapy • • • •

R. Langer, R.J. Linhardt, C.C. Cooney and D. Tapper

433

CONTENTS

Clinical Utility of Urokinase-Treated Polymer for Antithrombogenic Material ••••••••••

T. Ohshiro

Artificial Cell Immobilized Multienzyme Systems and Cofactors • • • • • • • • • • • • • • •

T.M.S. Chang, Y.T. Yu and J. Grunwald

Antithrombogenic Activity of Artificial Medical Materials Improved by Enzyme Immobilization Techniques • • • • • • • • • • • •

Y. Miura, S. Aoyagi and K. Miyamoto

Application of Immobilized Enzymes for Biomaterials

xxi

443

451

457

Used in the Field of Thoracic Surgery • • • • • • • • 459 S. Watanabe and T. Teramatsu

Use of Immobilization principles for the Construction of Drug Targeting Systems • • • • • • • • • • • • 461

V.P. Torchilin, A.L. Klibanov, V.R. Ber-dichevsky, V.G. Omelyanenko and V.N. Smirnov

Application of Bioreactors with Immobilized L-Asparaginase • • • • • • • • • • • • • • • • • • • 465

G. Mazzola, C. Giordano, R. Longhi, G. Vecchio and R. Esposi to

Acyl-COA Synthetase and Acyl-CoA Oxidase for Deter-mination of Serum Free Fatty Acids •••••••• 467

S. Shimizu, Y. Tani and H. Yamada

Enzyme Immunoassay for Free Thyroxine • • • • • 469 H.H. Weetall, W. Hertl, F.B. Ward and L.S. Hersh

Routine Determination of Hydrogen Peroxide in Clinical Chemistry with Immobilized Aldehyde Dehydro-genase • • • • • • • • • • • • • • • • • • • • • • • 473

P. V. Sundaram

Enzyme Electrodes for Simultaneous Determination of Creatinine and Creatine in Serum or Whole Blood 475

T. Tsuchida and K. Yoda

SESSION VIII. GENETIC ENGINEERING FOR ENZYME (OR IMPORTANT BIOLOGICAL SUBSTANCES) PRODUCTION 477

Chairmen: D. Fink and K. Sakaguchi

Construction of Various Host Vector Systems and the Variation of Enzyme Levels • • • • • • • • 479

K. Sakaguchi

xxii CONTENTS

Enzymes Active on Unnatural Synthetic Compounds: Nylon Oligomer Hydrolases Controlled by a Plasmid and Their Cloning • • • • • • • • • • • • • • • • • • 491

H. Okada, S. Negoro and S. Kinoshi ta

CLOSING SESSION 501 Chairman: G. Manecke

Recent Developments and Future Aspects of Enzyme Engineering • • • • • • • • • • • • • • • • • • • 503

B. Katchalski-Katzir

LIST OF PARTICIPANTS • 511

SUBJECT INDEX 525