BIOPROCESSENGINEERINGPRINCIPLES

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 2013660.6’3—dc23 2012007234

British Library Cataloguing-in-Publication Data

A catalogue record for this book is available from the British Library.

For information on all Academic Press publications visitour Web site at www.elsevierdirect.com

Printed in the United Kingdom

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

CONTENTS

Preface vii

PART 1INTRODUCTION

1. Bioprocess Development: AnInterdisciplinary Challenge 3

1.1 Steps in Bioprocess Development: A TypicalNew Product from Recombinant DNA 7

1.2 A Quantitative Approach 9

2. Introduction to EngineeringCalculations 13

2.1 Physical Variables, Dimensions, and Units 142.2 Units 192.3 Force and Weight 222.4 Measurement Conventions 232.5 Standard Conditions and Ideal Gases 292.6 Physical and Chemical Property Data 312.7 Stoichiometry 322.8 Methods for Checking and Estimating

Results 35Summary of Chapter 2 37References 44Suggestions for Further Reading 44

3. Presentation and Analysis of Data 45

3.1 Errors in Data and Calculations 453.2 Presentation of Experimental Data 543.3 Data Analysis 553.4 Graph Paper with Logarithmic

Coordinates 653.5 General Procedures for Plotting Data 693.6 Process Flow Diagrams 70Summary of Chapter 3 73References 82Suggestions for Further Reading 82

PART 2MATERIAL AND ENERGY

BALANCES

4. Material Balances 87

4.1 Thermodynamic Preliminaries 874.2 Law of Conservation of Mass 894.3 Procedure for Material Balance

Calculations 914.4 Material Balance Worked Examples 944.5 Material Balances with Recycle, Bypass,

and Purge Streams 1144.6 Stoichiometry of Cell Growth and Product

Formation 116Summary of Chapter 4 127References 136Suggestions for Further Reading 136

5. Energy Balances 139

5.1 Basic Energy Concepts 1395.2 General Energy Balance Equations 1415.3 Enthalpy Calculation Procedures 1445.4 Enthalpy Change in Nonreactive

Processes 1455.5 Steam Tables 1505.6 Procedure for Energy Balance Calculations

without Reaction 1515.7 Energy Balance Worked Examples without

Reaction 1515.8 Enthalpy Change Due to Reaction 1565.9 Heat of Reaction for Processes with Biomass

Production 1595.10 Energy Balance Equation for Cell Culture 1645.11 Cell Culture Energy Balance Worked

Examples 165Summary of Chapter 5 170References 176Suggestions for Further Reading 176

iii

6. Unsteady-State Material and EnergyBalances 177

6.1 Unsteady-State Material BalanceEquations 177

6.2 Unsteady-State Energy Balance Equations 1816.3 Solving Differential Equations 1826.4 Solving Unsteady-State Mass Balances 1836.5 Solving Unsteady-State Energy Balances 189Summary of Chapter 6 192References 197Suggestions for Further Reading 197

PART 3PHYSICAL PROCESSES

7. Fluid Flow 201

7.1 Classification of Fluids 2017.2 Fluids in Motion 2027.3 Viscosity 2087.4 Momentum Transfer 2107.5 Non-Newtonian Fluids 2117.6 Viscosity Measurement 2137.7 Rheological Properties of Fermentation

Broths 2177.8 Factors Affecting Broth Viscosity 2187.9 Turbulence 223Summary of Chapter 7 248References 252Suggestions for Further Reading 253

8. Mixing 255

8.1 Functions of Mixing 2558.2 Mixing Equipment 2568.3 Flow Patterns in Stirred Tanks 2618.4 Impellers 2658.5 Stirrer Power Requirements 2828.6 Power Input by Gassing 2928.7 Impeller Pumping Capacity 2938.8 Suspension of Solids 2958.9 Mechanisms of Mixing 2988.10 Assessing Mixing Effectiveness 3008.11 Scale-Up of Mixing Systems 3048.12 Improving Mixing in Fermenters 3058.13 Multiple Impellers 306

8.14 Retrofitting 3118.15 Effect of Rheological Properties on

Mixing 3128.16 Role of Shear in Stirred Fermenters 315Summary of Chapter 8 322References 329Suggestions for Further Reading 332

9. Heat Transfer 333

9.1 Heat Transfer Equipment 3339.2 Mechanisms of Heat Transfer 3409.3 Conduction 3409.4 Heat Transfer Between Fluids 3469.5 Design Equations for Heat Transfer

Systems 3519.6 Application of the Design Equations 3649.7 Hydrodynamic Considerations with

Cooling Coils 369Summary of Chapter 9 371References 377Suggestions for Further Reading 377

10. Mass Transfer 379

10.1 Molecular Diffusion 38010.2 Role of Diffusion in Bioprocessing 38210.3 Film Theory 38310.4 Convective Mass Transfer 38410.5 Oxygen Uptake in Cell Cultures 39310.6 Factors Affecting Oxygen Transfer

in Fermenters 40010.7 Measuring Dissolved Oxygen

Concentration 40710.8 Estimating Oxygen Solubility 40910.9 Mass Transfer Correlations for Oxygen

Transfer 41110.10 Measurement of kLa 41310.11 Measurement of the Specific Oxygen Uptake

Rate, qO 42510.12 Practical Aspects of Oxygen Transfer

in Large Fermenters 42710.13 Alternative Methods for Oxygenation

without Sparging 42910.14 Oxygen Transfer in Shake Flasks 430Summary of Chapter 10 433References 442Suggestions for Further Reading 443

iv CONTENTS

11. Unit Operations 445

11.1 Overview of Downstream Processing 44511.2 Overview of Cell Removal

Operations 45011.3 Filtration 45211.4 Centrifugation 46011.5 Cell Disruption 46711.6 The Ideal Stage Concept 46911.7 Aqueous Two-Phase Liquid

Extraction 47011.8 Precipitation 47311.9 Adsorption 48411.10 Membrane Filtration 49311.11 Chromatography 52611.12 Crystallisation 53811.13 Drying 563Summary of Chapter 11 578References 592Suggestions for Further Reading 593

PART 4REACTIONS AND REACTORS

12. Homogeneous Reactions 599

12.1 Basic Reaction Theory 59912.2 Calculation of Reaction Rates from

Experimental Data 60712.3 General Reaction Kinetics for Biological

Systems 61212.4 Determining Enzyme Kinetic Constants

from Batch Data 62112.5 Regulation of Enzyme Activity 62312.6 Kinetics of Enzyme Deactivation 62912.7 Yields in Cell Culture 63212.8 Cell Growth Kinetics 63512.9 Growth Kinetics with Plasmid

Instability 64012.10 Production Kinetics in Cell Culture 64312.11 Kinetics of Substrate Uptake in Cell

Culture 64512.12 Effect of Culture Conditions on Cell

Kinetics 64812.13 Determining Cell Kinetic Parameters

from Batch Data 64812.14 Effect of Maintenance on Yields 65112.15 Kinetics of Cell Death 65312.16 Metabolic Engineering 657Summary of Chapter 12 688

References 701Suggestions for Further Reading 702

13. Heterogeneous Reactions 705

13.1 Heterogeneous Reactions inBioprocessing 706

13.2 Concentration Gradients and ReactionRates in Solid Catalysts 707

13.3 Internal Mass Transfer and Reaction 71013.4 The Thiele Modulus and Effectiveness

Factor 72213.5 External Mass Transfer 73613.6 Liquid�Solid Mass Transfer

Correlations 73913.7 Experimental Aspects 74113.8 Minimising Mass Transfer Effects 74213.9 Evaluating True Kinetic

Parameters 74713.10 General Comments on Heterogeneous

Reactions in Bioprocessing 748Summary of Chapter 13 750References 757Suggestions for Further Reading 759

14. Reactor Engineering 761

14.1 Bioreactor Engineering in Perspective 76214.2 Bioreactor Configurations 76514.3 Practical Considerations for Bioreactor

Construction 77314.4 Monitoring and Control of Bioreactors 77814.5 Ideal Reactor Operation 78914.6 Sterilisation 82314.7 Sustainable Bioprocessing 834Summary of Chapter 14 844References 850Suggestions for Further Reading 852

AppendicesA. Conversion Factors 855

B. Ideal Gas Content 859

C. Physical and Chemical Property

Data 861

D. Steam Tables 879

E. Mathematical Rules 887

F. U.S. Sieve and Tyler Standard Screen

Series 895

Index 899

vCONTENTS

Preface to the Second Edition

As originally conceived, this book is in-tended as a text for undergraduate andpostgraduate students with little or no engi-neering background. It seeks to close the gapof knowledge and experience for studentstrained or being trained in molecular biology,biotechnology, and related disciplines whoare interested in how biological discoveriesare translated into commercial products andservices. Applying biology for technologydevelopment is a multidisciplinary challengerequiring an appreciation of the engineeringaspects of process analysis, design, and scale-up. Consistent with this overall aim, basicbiology is not covered in this book, as abiology background is assumed. Moreover,although most aspects of bioprocess engi-neering are presented quantitatively, priorityhas been given to minimising the use of com-plex mathematics that may be beyond thecomfort zone of nonengineering readers.Accordingly, the material has a descriptivefocus without a heavy reliance on mathemati-cal detail.

Following publication of the first editionof Bioprocess Engineering Principles, I wasdelighted to find that the book was alsobeing adopted in chemical, biochemical,and environmental engineering programsthat offer bioprocess engineering as a curric-ulum component. For students with severalyears of engineering training under theirbelts, the introductory nature and style ofthe earlier chapters may seem tedious and

inappropriate. However, later in the book,topics such as fluid flow and mixing, heatand mass transfer, reaction engineering, anddownstream processing are presented indetail as they apply to bioprocessing, thusproviding an overview of this specialtystream of traditional chemical engineering.

Because of its focus on underlying scien-tific and engineering principles rather than onspecific biotechnology applications, the mate-rial presented in the first edition remains rele-vant today and continues to provide a soundbasis for teaching bioprocess engineering.However, since the first edition was pub-lished, there have been several importantadvances and developments that have signifi-cantly broadened the scope and capabilitiesof bioprocessing. New sections on topics suchas sustainable bioprocessing and metabolicengineering are included in this second edi-tion, as these approaches are now integral toengineering 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 morein-depth treatment of fluid flow, turbulence,mixing, and impeller design is also availablein this edition, reflecting recent advances inour understanding of mixing processes andtheir importance in determining the perfor-mance of cell cultures. More than 100 newillustrations and 150 additional problems

vii

and worked examples have been includedin this updated edition. A total of over 340problems now demonstrate how the funda-mental principles described in the text areapplied in areas such as biofuels, bioplas-tics, bioremediation, tissue engineering,site-directed mutagenesis, recombinantprotein production, and drug develop-ment, as well as for traditional microbialfermentation.

I acknowledge with gratitude the feed-back and suggestions received from manyusers of the first edition of Bioprocess Engi-neering Principles over the last 15 years or so.Your input is very welcome and has helpedshape the priorities for change and elaborationin the second edition. I would also like tothank Robert Bryson-Richardson and PaulinaMikulic for their special and much appreciated

assistance under challenging circumstances in2011. Bioprocess engineering has an importantplace in the modern world. I hope that thisbook will make it easier for students and grad-uates from diverse backgrounds to appreciatethe role of bioprocess engineering in our livesand to contribute to its further progress anddevelopment.

Pauline M. Doran

Swinburne University of Technology

Melbourne, Australia

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text for their courses, additional teachingresources are available by registering atwww.textbooks.elsevier.com.

viii PREFACE