Biochemical Engineering: A Laboratory Manual

248 Pages·2021·18.569 MB·English

by  Debabrata Das, Debayan Das

Biochemical Engineering

Biochemical Engineering

A Laboratory Manual
Debabrata Das | Debayan Das
September7,2020 16:19 JSPBook-9inx6in 00-Experiments-Prelims
Published by
JennyStanfordPublishingPte.Ltd.
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Singapore039190
Email:[email protected]
Web:www.jennystanford.com
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AcataloguerecordforthisbookisavailablefromtheBritishLibrary.
Biochemical Engineering: A Laboratory Manual
Copyright �c 2021JennyStanfordPublishingPte.Ltd.
All rights reserved. This book, or parts thereof, may not be reproduced in any
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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
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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
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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
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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