3 ELSEVIER and Biotechnology Biochemical Engineering 2nd Edition Ghasem D. NajafpourBIOCHEMICAL ENGINEERING AND BIOTECHNOLOGY SECOND EDITION Grasem BL NajarmourElsevier Radarweg 29, PO Box 211, 1000 AE Amsterdam, Netherlands, The Bowlevand, Langford Lane, Kidlington, Oxford OXS 1GB, UK 225 Wyman Strect, Waltham, MA 02451, USA Copyright © 2015, 2007 Elsevier BLV. All rights reserved. No part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical, inckading photocopying. recording, or any information storage and retrieval system, without permission in writing from the publisher. Details on how to seck permission, further information about the Publisher's permissions policies and our arrangements with organizations such as the Copyright Clearance Center and the Copyright Licensing Agency, can be found at our website: wer elsevier.com / permissions. This book and the individual contributions contained in i are protected under copyright by the Publisher (other than as may be noted herein), Notices Knowledge and best practice in this field are constantly changing. As new research and experience broaden our understanding, changes in research methods, professional practices, er medical treatment may become nere-nay Practitioners and researchers must always rely on their own experience and knowledge in evaluating and using any information, methods, compounds, or experiments described herein. In ting such information or methads they should be mindful of their own satety and the safety of others, including parties for whom they have a professional responsibility. To the fullest extent of the law, neither the Publisher nor the authors, contributors, ar editors, assume any liability for any injury and/or damage to persons or property as a matter of products lability, negligence or otherwise, or from any use or operation of any methods, products, instructions, or ideas contained in the material herein, ISBN: 978-0-444-63357-6 British Library Cataloguing in Publication Data A catalogue record for this book is available from the British Library Library of Congress Cataloging-in-Publication Data A catalog record for this book is available from the Library of Congress to grow libraries in Beka developing countriesContents 1. Industrial Microbiology 1A Introduction 1 1.2 Role of Biotechnology 2 13 Role of Biosciences 5 14 Microbe Functions 5 15 Process Fermentation 7 1.6 Application of Fermentation Processes 9 17 Bropmces Products 10 1.8 Production of Lactic Acid 11 1.9 Production of Vinegar 13 1.10 Production of Amino Ackls (Lysine and Glutamic Acid) and Insulin 14 1.11 Antibiotics, Production of Penicillin. 15 12 Production of Enzymes 16 1.13 Production of Baker's Yeast 16 References 18 2. Enzyme Technology 2 Introxtuction 20 12 Enzyme Elementary Reaction Rate 2 24 Enzyme Clasifications 28 14 Enzymes Specific Function 30 15 Enzymes Actas Catalysts 30 2.6 Inhibitors of Enayme-Catalyoed Reactions 31 LT Tachutrial Application of Enzymes 34 2.3 Coenzymes 35 19 Bilect of pH om Enzyme Activities 37 AD Enzyme Unit Activities 37 2. Enzyme Deactivation 37 Nomenclature 38 112 Solve Problems 39 References 40 1.13 Case Study: Solid State Fermentation of Sugarcane Bagssse in a Tray Bisteactor foe Production of Lipase Using Ropu Oryzse 41 211 Imvoduction 41 213.2 Material and Methods 42 2.133 Results and Discussion 44 References 49 3. Gas and Liquid System (Acration and Agitation) 3.1 Introduction 53 3.2 Aeration and Agitation 53 33 Air Spanger 56 34 Agitation and Mixing Phenomena 57 435 Types ef Agitator 57 3.6 OTRinaFermenter 38 3.7 Mae Transfer ina Gas Liquid Spstem 59 3.8 GasHold.Up 60 3.9 Mass Transfer Coefficients for Stirred Tanks 61 310 Agitated Stem and Mixing Phenomena 63 3.11 Mass Transfer Limited Process, 63 Nomenclature 74 Greek Symbols 75 References 75 3.13 Cane Study: Oxygen Tranafer Rate Model in an ‘Aerated Tank for Pharmaceutical Wastewater 76 3.1341 Introduction 76 3.13.2 Material and Method 78vi coNTENTS: 3.13.3 Results and Discusion 70 3.15.4 Conchsion 81 Nomenclature 82 References 82 3.14 Case Study: Fuel_and Chemical Production from the Water Cas Shift Reaction by Fermentation Processes 83 3.14. Introduction 83 3.14.2 Kinetics of Growth in. Batch Bioreactor 84 3.143 fice of Substrate Concentration con Microbial Growth 88 3.144 Mass Transfer Phenomena 3.13.5 Kinetic of Water Gas Shift Reaction 93 3.14.6 Growth Kinetics of C0 Substrate on C. Bunglahlir 5 Nomenclature 98 Acknowledgments 9% References 4. Fermentation Process Control 4.1 Introduction 104 4.2 Bioreactor Controlling Probes 105 4.3 Characteristics of Bioreactor Sensors 106 44 Temperature Measurement and Control 108 4.5 Dissolved Oxygen Measurement and Control 109 4.6 pHRedox Measurement and Control 110 4.7 Detection and Prevention of Foam 112 48 Bien 113 Nomenclature 124 References 14 5. Growth Kinetics 5. Inroduction 129 5.2 Indirect Measurements of Cell Growth 12 5.4 Cell Growth in Batch Culture 131 34 Gromth Phases 131 5.5 Kinetics of Batch Culture 132 546 Growth Kinetics for Continuous Culture 133 5.7 Material Balance for Comins Stised Tank Reactor (CSTR) 137 3.8 Enoyime Reactiom Kinetics 143 5.9 Unstructured Kinetic Model 173. Nomenclature 179) References 180) 5.11 Case Stady- Enoyme Kinetic Models for Resolution of Racemic Ibuprofen Esters ima Membrane Reactor 181 S11 Introduction 181 5.11.2 Enzyme Kinetics 182 5.11.3 Enzyme Kinetics for Rapid Equilibrium. System ((Qhuasi-Equaliberum) | 187 5.114 Derivation of Enzymatic Rate Equation from Rapid Equilibrium, Assumpeicn 187 5.11.5 Verification of Kinetic Mechanism 189 References 191 6. Bioreactor Design, 6.1 Inimduction 193 6.2 Bioreactors: Background 194 63 Type of Bioreactor 195 6.4 Stirred Tank Bioreactor 198 6.5 Bubble Column Fermenter 201 66 Airlift Booreactors 202 6.7 Heat Transfer 203 6.8 Design Equations for CSTR Fermenter 103 609 Temperature Effect on Rate Constant 209 6.10 Scale-Up of Stinved-Tank Bioreactor 210 G11 Beclogscal Transport of Oxygen through Cells 217 Nomenclature 223 References 223 7. Downstream Processing 7A Intmduction 228 7.2 Downstream Processing 228 7.3 Filkraticn 231 74 Centrifugation 235 75 Sedimentation 235 7.6 Flotation 237 7.7 Emerging Technology foe Cell Recovery 238 78 Call Disruption 238 7.9 Solvent Extraction 239 TO Adsorption 142 TAL Chromatography 244 7.12 Crystallization Process 253 7.13 Freese.Drying 255 Nomenclature 255 References 256CONTENTS. vii 8. Immobilization of Microbial Cells for the Production of Organic Acid and Ethanol BLL Introduction 250 8.2 Immobilized Microbial Cells 260 83 ICR Experiments 263 84 FCR Rate Model 263 Nomenclature 266 References 266 5.6 Case Stay: Ethanol Fermentation in an Immobilized Cell Reactor Using Saccharomyces cerevisiae 267 86.1 Introduction 267 9.6.2 Materials and Methods 268 9163 Results and Decussion 272 864 Conclusion 281 Nomenclature 282 Acknowledgment 282 References 282 8.7 Furdhmentals of Immabilisation Techaolewy, and Mathematical Model for ICR Performance 254 7.1 Immobilization of Microorganisms by Covalent Bonds 234 87.2 Onygen Transfer to Immobilized Micmcrganisms 284 8.7.3 Substrate Transfer to Emmnobilized Microorganisms 285 8.74 Growth and Colony Formation of Immobilized Microorginoms 286 87.5 Immobilised Systems foe Ethanol Production 188 Reference 25% 9. Material and Elemental Balance 911 Inteduction 21 9.2 Media Preparation for Fermentation 292 913 Growth of Stoichiometry anal Elemental Balances 403 ‘94 Energy Balance for Comtinucus Ethanol Fermentation 305 9.5 Mass Balance for Biological Processes 306 916 Conservation of Mass Principle 309 97 Embden Meyerhof Pamas Pathway 318 References 326 10. Application of Fermentation Processes 10.1 Introduction 330 10.2 Prexduction of Ethanol by Fermentation 330 10.3 Benefits from Bacethanal Fuel 331 104 Stoichiometry of Biochemical Reaction 331 10.5 Optical Cell Density 332 10.6 Kinetics of Growth and Product Formation 333 10.7 Preparation of Stock Culture 34 10.8 Inoculum Preparation 335 10.9 Inoculation of Seed Culture 336 1010 Analytical Method for Sugar Analysis 337 1011 Analytical Method for Ethanol Amilysis 338 10.12 Refractive Index Determination 338 10.13 Cell Dry Weight Measurements 339 10.14 Yield Caleulation 339 10.15 Batch Fermentation Experiment ¥40 10.16 Continuous Fermentation Experiment ML 10.17 Media Sterilization 342 1018 Batch Experiment 342 10.19 Expected Results 343 References 344 LL, Production of Antibiotics Iniraduction 346 1L1 Herball Medicines and Chemical Agents H7 AL3 The History of Penicillin 348 Production of Penicillin 349 Microorganisms and Media 4 16 Inoculum Preparation 49 17 Filtration and Extraction of Penicillin 352 LS Experimental Procedure 152 1.9 Fermenter Description 352 LAO Analytical Method for Basassay and Detection of Antibiotic 352 LLU Antibiogram and Biological Asay 353 LLA2 Submerged Culruse 354 1113 Bioreactor Design and Control 355 LL14 Estimation of Dimension of Fermenter 336 1115 Determination of the Reynolds Number 358 L116 Determination of Power Input 358 LAT Determination of Oxygen Transfer Rate 359viii coNTENTS: 11.18 Design Specification Sheet for the Bioreactor 361 References 361 12. Production of Citric Acid 12. Innresduction 363 12.2 Production of Citric Acid in Batch Bicreactors 364 12.3 Factors Affecting Mok! Grmth and the Fermentation Process 345 12.4 Starter oe Seeding an Inoculum 367 125 Seed Culture 367 12.6 Citric Acid Production 367 127 Amabtical Method 368 11.8 Processes for Recovery and Pursication of Citric Acid 340 12.9 Experimental Run 369 12.10 Kinetic Model in Batch Citric Acid Fermentation 371 Nomenclature 373 References 373 15, Bioprocess Scale-up 13 Introduction 375 13.2 Scale-up Prcedure from Laboratory Scale to Plant Scale 376 133 Bioreactor Design Criteria 384 134 Continuous Stirred Tank Reactor (Chemostat versus Tubular Plug Flow 388 135 Dynamic Model and Oxygen Transfer Rate in Activated Shake 399 136 Acrobic Wastewater Treatment 410 Nomenclature 415 References 416 14, Single-Cell Protein 14.1 Intreduction 418 14.2 Dissolved Oxygen in Single-Cell Protein Production 419 14.3 Batch and Continuous Fermentation, for Production of Single-Cell Protein 419 14.4 Batch Experment for Production of Baker's Yeast. 421 14.5 Separation of Microbsal Biomass 421 14.6 Background 422 14.7 Preduction Method 422 14.8 Media Preparation for Singgle-Cell Protein Production 424 14.9 Analytical Methods 424 14.10 Single-Cell Protein Processes 426 14.11 Nutsitional Value of Single-Cell Frotein 428 14.12 Organisms and Substrates for Single.Cell Protein Production 420 14.13 Advantages and Disadvantages of Single-Cell Protein. 432 14.14 Preparation for Experimental Run 433 References 433 15, Sterilization 15.1 Introduction 435 15.2 Conteol of Microbial Population, by Physical Agents 436 15.3 Death Rate of Living Organisms 437 154 Batch Sterilization 437 15.3 Continuous Sterilization. 439 156 Hot Plates 441 15.7 High-Temperature Sterilization 442 15.8 Stenlized Media for Microbiology 442 15.9 Dry Heat Sterilization 450 15.10 Seerilization with Fileariom 450 15.11 Microwave Sterilization 450 15.12 Electron Beam Sterilization 451 15.13 Chemical Sterilization 451 15.14 Low-Temperature Sterilization 452 Nomenclature 452 References 452 16. Membrane Reactor 161 Introduction 455 162 Membrane Biseeactors 457 163 Membrane and MBR Development 465 References 466 164 Case Study: Enhanced Ethanol Fermentation in a Continuous Membeane Bioreactor: Pervaporation Technique 468 1641 Introduction 468 1642 Experimental 470 16-43 Results and Discussion 472 16-44 Conclusion 481 Acknowledgments 481 References 481CONTENTS 165 Case Study: Inorganic Zirconia Y-Abumina- (Coated Membrane on Ceramic Support 483 165.1 Introduction 483 165.2 Materials and Methods 485 1654 Results and Discussion 491 165.4 Conclusion 492 Acknowleduments 492 References 493 17. Advanced Downstream Processing in Biotechnology 17 Introduction 497 2 Protein Products 497 173 Cell Disruption 498 17 Protein Purification 500 175 General Problems Associated With Conventional Techniques 501 6 Fluidiced Bed Adsorption 502 117 Design and Operation of Liquid Flubdized Beds 504 178 Experimental Procedure 510 17.9 Process Imegration in Protein Recovery S11 Nomenclature 513 References 513 17.11 Case Study: Biochemical Characterization of a Custom Expanded! Bel Column for Protein Purification 316 ITALA Introduction 516 17.112 Materials and Methods 517 ITAL3 Results and Discussion 518 17.114 Conchusion 524 References 525 18. Microbial Fuel Cells: A New Source of Power 18.1 Introduction 528 18.2 Beclogical Fuel Cell 529 183 Microbial Fuel Gell 531 Acknowledgment 551 References 531 19. Biological Treatment 19.1 Introduction 558 19.2 Organic Removal in Sustainable Microbial Growth 558 19.3 Microbial Metabolism 559 19.4 Microbial Growth Kinetics 561 19.5 Growels Rate and Treatment Kinetics 562 19.8 Removal Mechanisms in Biological Procenes 568 19.7 Aerobic Bisoxidation 569 198 Anaerctsc Digestion 572 19.9 Abiotic Lames 576 19.10 Volatilization 578 1911 Biological Nienfication and Denitrification 379 19112 Biological Trestment Process: Suspended and Attached Growth 586 References 394 20. Biofuel Production 201 Biofuel Production and Glatall Scenarios 597 2012 Feedstock for Biofuel Production 600 203 Procenes and Technologies 609 204 Intensification and Integration 622 205 Economic Perspective 624 References 625 Appendix: Constants and Conversion Factors 631 Index 633