BIOPROCESS

ENGINEERING
PRINCIPLES
SECOND EDITION

PAULINE M. DORAN

AMSTERDAM • BOSTON • HEIDELBERG • LONDON

NEW YORK • OXFORD • PARIS • SAN DIEGO
SAN FRANCISCO • SINGAPORE • SYDNEY • TOKYO
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Library of Congress Cataloging-in-Publication Data
Doran, Pauline M.
Bioprocess engineering principles / Pauline M. Doran. — 2nd ed.
p. cm.
Includes bibliographical references and index.
ISBN 978-0-12-220851-5 (pbk.)
1. Biochemical engineering. I. Title.
TP248.3.D67 2013
660.6’3—dc23 2012007234
British Library Cataloguing-in-Publication Data
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Printed in the United Kingdom

12 13 14 15 16 10 9 8 7 6 5 4 3 2 1
 CONTENTS

Preface vii PART 2

MATERIAL AND ENERGY
PART 1 BALANCES
INTRODUCTION 4. Material Balances 87
1. Bioprocess Development: An 4.1 Thermodynamic Preliminaries 87
4.2 Law of Conservation of Mass 89
Interdisciplinary Challenge 3
4.3 Procedure for Material Balance
1.1 Steps in Bioprocess Development: A Typical Calculations 91
New Product from Recombinant DNA 7 4.4 Material Balance Worked Examples 94
1.2 A Quantitative Approach 9 4.5 Material Balances with Recycle, Bypass,
and Purge Streams 114
2. Introduction to Engineering 4.6 Stoichiometry of Cell Growth and Product
Calculations 13 Formation 116
2.1 Physical Variables, Dimensions, and Units 14 Summary of Chapter 4 127
2.2 Units 19 References 136
2.3 Force and Weight 22 Suggestions for Further Reading 136
2.4 Measurement Conventions 23
2.5 Standard Conditions and Ideal Gases 29 5. Energy Balances 139
2.6 Physical and Chemical Property Data 31 5.1 Basic Energy Concepts 139
2.7 Stoichiometry 32 5.2 General Energy Balance Equations 141
2.8 Methods for Checking and Estimating 5.3 Enthalpy Calculation Procedures 144
Results 35 5.4 Enthalpy Change in Nonreactive
Summary of Chapter 2 37 Processes 145
References 44 5.5 Steam Tables 150
Suggestions for Further Reading 44 5.6 Procedure for Energy Balance Calculations
without Reaction 151
3. Presentation and Analysis of Data 45
5.7 Energy Balance Worked Examples without
3.1 Errors in Data and Calculations 45 Reaction 151
3.2 Presentation of Experimental Data 54 5.8 Enthalpy Change Due to Reaction 156
3.3 Data Analysis 55 5.9 Heat of Reaction for Processes with Biomass
3.4 Graph Paper with Logarithmic Production 159
Coordinates 65 5.10 Energy Balance Equation for Cell Culture 164
3.5 General Procedures for Plotting Data 69 5.11 Cell Culture Energy Balance Worked
3.6 Process Flow Diagrams 70 Examples 165
Summary of Chapter 3 73 Summary of Chapter 5 170
References 82 References 176
Suggestions for Further Reading 82 Suggestions for Further Reading 176

iii
iv CONTENTS

6. Unsteady-State Material and Energy 8.14 Retrofitting 311

Balances 177 8.15 Effect of Rheological Properties on
Mixing 312
6.1 Unsteady-State Material Balance 8.16 Role of Shear in Stirred Fermenters 315
Equations 177 Summary of Chapter 8 322
6.2 Unsteady-State Energy Balance Equations 181 References 329
6.3 Solving Differential Equations 182 Suggestions for Further Reading 332
6.4 Solving Unsteady-State Mass Balances 183
6.5 Solving Unsteady-State Energy Balances 189 9. Heat Transfer 333
Summary of Chapter 6 192
References 197 9.1 Heat Transfer Equipment 333
Suggestions for Further Reading 197 9.2 Mechanisms of Heat Transfer 340
9.3 Conduction 340
9.4 Heat Transfer Between Fluids 346
9.5 Design Equations for Heat Transfer
PART 3 Systems 351
9.6 Application of the Design Equations 364
PHYSICAL PROCESSES 9.7 Hydrodynamic Considerations with
7. Fluid Flow 201 Cooling Coils 369
Summary of Chapter 9 371
7.1 Classification of Fluids 201
References 377
7.2 Fluids in Motion 202
7.3 Viscosity 208 Suggestions for Further Reading 377
7.4 Momentum Transfer 210
7.5 Non-Newtonian Fluids 211 10. Mass Transfer 379
7.6 Viscosity Measurement 213 10.1 Molecular Diffusion 380
7.7 Rheological Properties of Fermentation 10.2 Role of Diffusion in Bioprocessing 382
Broths 217 10.3 Film Theory 383
7.8 Factors Affecting Broth Viscosity 218 10.4 Convective Mass Transfer 384
7.9 Turbulence 223 10.5 Oxygen Uptake in Cell Cultures 393
Summary of Chapter 7 248 10.6 Factors Affecting Oxygen Transfer
References 252 in Fermenters 400
Suggestions for Further Reading 253 10.7 Measuring Dissolved Oxygen
Concentration 407
8. Mixing 255 10.8 Estimating Oxygen Solubility 409
8.1 Functions of Mixing 255 10.9 Mass Transfer Correlations for Oxygen
8.2 Mixing Equipment 256 Transfer 411
8.3 Flow Patterns in Stirred Tanks 261 10.10 Measurement of kLa 413
8.4 Impellers 265 10.11 Measurement of the Specific Oxygen Uptake
8.5 Stirrer Power Requirements 282 Rate, qO 425
8.6 Power Input by Gassing 292 10.12 Practical Aspects of Oxygen Transfer
8.7 Impeller Pumping Capacity 293 in Large Fermenters 427
8.8 Suspension of Solids 295 10.13 Alternative Methods for Oxygenation
8.9 Mechanisms of Mixing 298 without Sparging 429
8.10 Assessing Mixing Effectiveness 300 10.14 Oxygen Transfer in Shake Flasks 430
8.11 Scale-Up of Mixing Systems 304 Summary of Chapter 10 433
8.12 Improving Mixing in Fermenters 305 References 442
8.13 Multiple Impellers 306 Suggestions for Further Reading 443
 CONTENTS v
11. Unit Operations 445 References 701
Suggestions for Further Reading 702
11.1 Overview of Downstream Processing 445
11.2 Overview of Cell Removal 13. Heterogeneous Reactions 705
Operations 450
13.1 Heterogeneous Reactions in
11.3 Filtration 452
Bioprocessing 706
11.4 Centrifugation 460
13.2 Concentration Gradients and Reaction
11.5 Cell Disruption 467
Rates in Solid Catalysts 707
11.6 The Ideal Stage Concept 469
13.3 Internal Mass Transfer and Reaction 710
11.7 Aqueous Two-Phase Liquid
13.4 The Thiele Modulus and Effectiveness
Extraction 470
Factor 722
11.8 Precipitation 473
13.5 External Mass Transfer 736
11.9 Adsorption 484
13.6 Liquid Solid Mass Transfer
11.10 Membrane Filtration 493
Correlations 739
11.11 Chromatography 526
13.7 Experimental Aspects 741
11.12 Crystallisation 538
13.8 Minimising Mass Transfer Effects 742
11.13 Drying 563
13.9 Evaluating True Kinetic
Summary of Chapter 11 578
Parameters 747
References 592
13.10 General Comments on Heterogeneous
Suggestions for Further Reading 593
Reactions in Bioprocessing 748
Summary of Chapter 13 750
PART 4 References 757
Suggestions for Further Reading 759
REACTIONS AND REACTORS
12. Homogeneous Reactions 599 14. Reactor Engineering 761
14.1 Bioreactor Engineering in Perspective 762
12.1 Basic Reaction Theory 599
14.2 Bioreactor Configurations 765
12.2 Calculation of Reaction Rates from
14.3 Practical Considerations for Bioreactor
Experimental Data 607
Construction 773
12.3 General Reaction Kinetics for Biological
14.4 Monitoring and Control of Bioreactors 778
Systems 612
14.5 Ideal Reactor Operation 789
12.4 Determining Enzyme Kinetic Constants
14.6 Sterilisation 823
from Batch Data 621
14.7 Sustainable Bioprocessing 834
12.5 Regulation of Enzyme Activity 623
Summary of Chapter 14 844
12.6 Kinetics of Enzyme Deactivation 629
References 850
12.7 Yields in Cell Culture 632
Suggestions for Further Reading 852
12.8 Cell Growth Kinetics 635
12.9 Growth Kinetics with Plasmid
Instability 640
Appendices
12.10 Production Kinetics in Cell Culture 643 A. Conversion Factors 855
12.11 Kinetics of Substrate Uptake in Cell B. Ideal Gas Content 859
Culture 645 C. Physical and Chemical Property
12.12 Effect of Culture Conditions on Cell
Kinetics 648 Data 861
12.13 Determining Cell Kinetic Parameters D. Steam Tables 879
from Batch Data 648 E. Mathematical Rules 887
12.14 Effect of Maintenance on Yields 651
F. U.S. Sieve and Tyler Standard Screen
12.15 Kinetics of Cell Death 653
12.16 Metabolic Engineering 657 Series 895
Summary of Chapter 12 688 Index 899
 Preface to the Second Edition
As originally conceived, this book is in-
tended as a text for undergraduate and
postgraduate students with little or no engi-
neering background. It seeks to close the gap
of knowledge and experience for students
trained or being trained in molecular biology,
biotechnology, and related disciplines who
are interested in how biological discoveries
are translated into commercial products and
services. Applying biology for technology
development is a multidisciplinary challenge
requiring an appreciation of the engineering
aspects of process analysis, design, and scale-
up. Consistent with this overall aim, basic
biology is not covered in this book, as a
biology background is assumed. Moreover,
although most aspects of bioprocess engi-
neering are presented quantitatively, priority
has been given to minimising the use of com-
plex mathematics that may be beyond the
comfort zone of nonengineering readers.
Accordingly, the material has a descriptive
focus without a heavy reliance on mathemati-
cal detail.
Following publication of the first edition
of Bioprocess Engineering Principles, I was
delighted to find that the book was also
being adopted in chemical, biochemical,
and environmental engineering programs
that offer bioprocess engineering as a curric-
ulum component. For students with several
years of engineering training under their
belts, the introductory nature and style of
the earlier chapters may seem tedious and
vii
inappropriate. However, later in the book,
topics such as fluid flow and mixing, heat
and mass transfer, reaction engineering, and
downstream processing are presented in
detail as they apply to bioprocessing, thus
providing an overview of this specialty
stream of traditional chemical engineering.
Because of its focus on underlying scien-
tific and engineering principles rather than on
specific biotechnology applications, the mate-
rial presented in the first edition remains rele-
vant today and continues to provide a sound
basis for teaching bioprocess engineering.
However, since the first edition was pub-
lished, there have been several important
advances and developments that have signifi-
cantly broadened the scope and capabilities
of bioprocessing. New sections on topics such
as sustainable bioprocessing and metabolic
engineering are included in this second edi-
tion, as these approaches are now integral to
engineering design procedures and commer-
cial cell line development.
Expanded coverage of downstream pro-
cessing operations to include membrane fil-
tration, protein precipitation, crystallisation,
and drying is provided. Greater and more
in-depth treatment of fluid flow, turbulence,
mixing, and impeller design is also available
in this edition, reflecting recent advances in
our understanding of mixing processes and
their importance in determining the perfor-
mance of cell cultures. More than 100 new
illustrations and 150 additional problems
viii
and worked examples have been included
in this updated edition. A total of over 340
problems now demonstrate how the funda-
mental principles described in the text are
applied in areas such as biofuels, bioplas-
tics, bioremediation, tissue engineering,
site-directed mutagenesis, recombinant
protein production, and drug develop-
ment, as well as for traditional microbial
fermentation.
I acknowledge with gratitude the feed-
back and suggestions received from many
users of the first edition of Bioprocess Engi-
neering Principles over the last 15 years or so.
Your input is very welcome and has helped
shape the priorities for change and elaboration
in the second edition. I would also like to
thank Robert Bryson-Richardson and Paulina
Mikulic for their special and much appreciated
PREFACE
assistance under challenging circumstances in
2011. Bioprocess engineering has an important
place in the modern world. I hope that this
book will make it easier for students and grad-
uates from diverse backgrounds to appreciate
the role of bioprocess engineering in our lives
and to contribute to its further progress and
development.
Pauline M. Doran
Swinburne University of Technology
Melbourne, Australia
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