Table Of ContentBiochemical Engineering
Biochemical Engineering
A Laboratory Manual
Debabrata Das | Debayan Das
September7,2020 16:19 JSPBook-9inx6in 00-Experiments-Prelims
Published by
JennyStanfordPublishingPte.Ltd.
Level34,CentennialTower
3TemasekAvenue
Singapore039190
Email:[email protected]
Web:www.jennystanford.com
British Library Cataloguing-in-Publication Data
AcataloguerecordforthisbookisavailablefromtheBritishLibrary.
Biochemical Engineering: A Laboratory Manual
Copyright �c 2021JennyStanfordPublishingPte.Ltd.
All rights reserved. This book, or parts thereof, may not be reproduced in any
form or by any means, electronic or mechanical, including photocopying,
recording or any information storage and retrieval system now known or to
be invented, without written permission from the publisher.
For photocopying of material in this volume, please pay a copying
fee through the Copyright Clearance Center, Inc., 222 Rosewood Drive,
Danvers, MA 01923, USA. In this case permission to photocopy is not
requiredfromthepublisher.
ISBN978-981-4877-36-7(Hardcover)
ISBN978-1-003-11105-4(eBook)
September7,2020 16:19 JSPBook-9inx6in 00-Experiments-Prelims
Contents
Preface xiii
List of Symbols xvii
Greek Letters xxi
Laboratory Report Format xxiii
1 Layout of a Fermenter with Controller 1
1.1 Introduction 1
1.2 AccessoriesofFermenter 2
1.2.1 AerationSystem 2
1.2.2 MechanicalSeal 3
1.2.3 FermenterVessel 4
1.2.4 FeedPorts 4
1.2.5 Baffles 4
1.2.6 Impeller 4
1.2.7 Sparger 5
1.2.8 Outlet 5
1.2.9 SafetyValve 5
1.3 TemperatureLoop 5
1.3.1 Heater 5
1.3.2 ColdFinger 5
1.3.3 SolenoidValve 6
1.4 MonitoringandControlLoop 6
1.4.1 TemperatureProbe 6
1.4.2 pHProbe 6
1.4.3 O Sensor 7
2
1.4.4 Motor 8
1.5 PeristalticPump 8
1.6 FoamControl 9
September7,2020 16:19 JSPBook-9inx6in 00-Experiments-Prelims
vi Contents
2 Enzymatic Reaction Kinetics 11
2.1 EnzymeActivity 11
2.1.1 Alpha-AmylaseActivity 11
2.2 EnzymaticReactionKinetics 12
2.2.1 Michaelis–MentenKinetics 12
2.2.2 LimitationoftheMichaelis–MentenEquation 16
2.2.3 DeterminationofKineticConstants 16
2.3 CalibrationCurveoftheSubstrate 18
2.4 PreparationofEnzymeSolution 18
2.4.1 Procedure 18
2.5 Observation 19
2.6 Results 19
2.6.1 SampleCalculations 20
2.7 Discussion 20
3 Kinetics of the Immobilized Enzymatic Reaction 23
3.1 PreparationofEnzymeSolution 23
3.1.1 DefinitionofEnzymeImmobilization 23
3.2 KineticTheory 23
3.2.1 EffectivenessFactorandDamko¨hlerNumber 26
3.2.1.1 Determinationofthefactoraffects
overallrateofthereaction 28
3.3 PreparationofImmobilizedEnzyme 28
3.3.1 PreparationofEnzymaticReactionMixture 29
3.4 Observation 29
3.5 CalculatedData 29
3.6 Discussion 31
4 Cell Growth Kinetics in Controlled Fermenter 33
4.1 Introduction 33
4.2 Theory 33
4.3 DataTable 36
4.3.1 CalculatedData 36
4A Cell Growth Kinetics of Bacteria in a Batch Process 39
4A.1 Purpose 39
4A.2 ApparatusRequired 39
4A.3 MediumPreparation 40
4A.3.1 CompositionoftheMedium 40
September7,2020 16:19 JSPBook-9inx6in 00-Experiments-Prelims
Contents vii
4A.3.2 ProcedureforMediumPreparation 40
4A.3.3 ProcedurefortheOperationofthe
Fermenter 41
4A.4 DeterminationofSubstrateConcentration 41
4A.4.1 GlucoseEstimationbyDinitrosalicylicAcid
(DNS) 41
4A.4.2 DeterminationofCellMassConcentration 42
4A.5 Observation 42
4A.6 CalculatedData 45
4A.6.1 SampleCalculation 46
4A.7 Discussion 47
4A.7.1 PossibleEffectofContaminationofthe
CulturebyanOrganismwithValueof
DoublingTimeoftheCell,t 47
d
4A.7.2 ContaminationProblems 48
4A.7.3 DifferencebetweenK andK 49
m s
4A.8 Conclusion 49
4B Cell Growth Kinetics of Yeast in a Batch Process 51
4B.1 Purpose 51
4B.2 BackgroundStatement 51
4B.3 ApparatusRequired 52
4B.4 MediumPreparation 53
4B.5 CulturePreparation 53
4B.5.1 Procedure 54
4B.5.2 CellMassConcentrationEstimation 55
4B.5.3 GlucoseEstimation 55
4B.6 Observation 56
4B.7 CalculatedData 58
4B.7.1 SampleCalculation 59
4B.8 Discussion 61
4B.8.1 ContaminationProblems
61
4B.8.2 IssueofAlcoholicFermentation 61
4B.9 Conclusion 62
4C Cell Growth Kinetics of Microalgae in a Batch Process 65
4C.1 Purpose 65
4C.2 BackgroundStatement 65
September7,2020 16:19 JSPBook-9inx6in 00-Experiments-Prelims
viii Contents
4C.2.1 ModesofGrowth 66
4C.2.2 CultivationMethods 66
4C.2.2.1 Openponds 66
4C.2.2.2 Photobioreactor 66
4C.2.3 LightIntensity 66
4C.3 Theory 67
4C.3.1 Growth-LimitingSubstrate 67
4C.4 ProcessRequirement 68
4C.4.1 Medium 68
4C.5 Procedure 68
4C.5.1 CulturePreparation 68
4C.5.2 SampleCollectionProcedure 69
4C.5.3 EstimationofChlorophyllContent 69
4C.5.4 DryCellWeightEstimation 69
4C.5.5 AnalyzingSubstrateConsumptionbyGas
ChromatographyFlameIonization
Detection 70
4C.6 Observation 71
4C.6.1 SampleCalculation 74
4C.7 Discussion 75
4C.8 Conclusion 76
5 Determination of Volumetric Mass Transfer Coefficient 79
5.1 Purpose 79
5.2 Microorganism 79
5.3 Theory 80
5.4 Requirements 81
5.4.1 Medium 82
5.4.2 MediumPreparationProcedure 82
5.4.3 CulturePreparation 83
5.4.4 Procedure 83
5.5 Observation 84
5.5.1 SampleCalculations 95
5.6 Discussion 96
5.6.1 Importanceofk a Value 97
L
5.6.2 BenefitsofMeasuringk a 97
L
5.6.3 ParametersInfluencingk a Value 97
L
5.7 Conclusion 98
September7,2020 16:19 JSPBook-9inx6in 00-Experiments-Prelims
Contents ix
6 Mixing Time Determination 99
6.1 Purpose 99
6.2 Theory 99
6.2.1 MechanismofMixing 100
6.2.1.1 Distribution 100
6.2.1.2 Dispersion 100
6.2.1.3 Diffusion 101
6.2.1.4 Commonstirrers 101
6.2.1.5 Powerconsumption 102
6.3 ApparatusRequired 103
6.4 PreparationofCMCSolution 103
6.5 MixingTimeDetermination 104
6.6 Observation 105
6.7 SampleCalculation 106
6.8 ResultandDiscussion 107
6.9 Conclusion 109
7 Determination of Air Filter Efficiency 111
7.1 Purpose 111
7.2 Theory 111
7.2.1 InertialImpaction 112
7.2.2 Interception 113
7.2.3 Diffusion 113
7.2.4 CollectionEfficiencies 114
7.3 Materials 114
7.4 Procedure 115
7.5 Observation 115
7.5.1 SampleCalculations 115
7.6 Discussion 116
7.7 Conclusion 117
8 Kill Curve Determination 119
8.1 Purpose 119
8.2 BackgroundStatement 119
8.3 Theory 120
8.4 Apparatus/Requirements 123
8.5 MediaComposition 123
8.6 Procedure 123
