Biocatalysis
An Industrial Perspective

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Catalysis Series
Editor-in-chief:

Chris Hardacre, University of Manchester, UK

Series editors:

Bert Klein Gebbink, Utrecht University, The Netherlands
Jose Rodriguez, Brookhaven National Laboratory, USA

Titles in the series:

1: Carbons and Carbon Supported Catalysts in Hydroprocessing
2: Chiral Sulfur Ligands: Asymmetric Catalysis
3: Recent Developments in Asymmetric Organocatalysis
4: Catalysis in the Refining of Fischer–Tropsch Syncrude
5: Organocatalytic Enantioselective Conjugate Addition Reactions: A
Powerful Tool for the Stereocontrolled Synthesis of Complex Molecules
6: N-Heterocyclic Carbenes: From Laboratory Curiosities to Efficient
Synthetic Tools
7: P-Stereogenic Ligands in Enantioselective Catalysis
8: Chemistry of the Morita–Baylis–Hillman Reaction
9: Proton-Coupled Electron Transfer: A Carrefour of Chemical Reactivity
Traditions

10: Asymmetric Domino Reactions
11: C–H and C–X Bond Functionalization: Transition Metal Mediation
12: Metal Organic Frameworks as Heterogeneous Catalysts
13: Environmental Catalysis Over Gold-Based Materials
14: Computational Catalysis
15: Catalysis in Ionic Liquids: From Catalyst Synthesis to Application
16: Economic Synthesis of Heterocycles: Zinc, Iron, Copper, Cobalt,
Manganese and Nickel Catalysts
17: Metal Nanoparticles for Catalysis: Advances and Applications
18: Heterogeneous Gold Catalysts and Catalysis
19: Conjugated Linoleic Acids and Conjugated Vegetable Oils
20: Enantioselective Multicatalysed Tandem Reactions
21: New Trends in Cross-Coupling: Theory and Applications
22: Atomically-Precise Methods for Synthesis of Solid Catalysts
23: Nanostructured Carbon Materials for Catalysis
24: Heterocycles from Double-Functionalized Arenes: Transition Metal
Catalyzed Coupling Reactions

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25: Asymmetric Functionalization of C–H Bonds
26: Enantioselective Nickel-catalysed Transformations
27: N-Heterocyclic Carbenes: From Laboratory Curiosities to Efficient
Synthetic Tools, 2nd edition
28: Zeolites in Catalysis: Properties and Applications
29: Biocatalysis: An Industrial Perspective

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Biocatalysis

An Industrial Perspective
Edited by

Gonzalo de Gonzalo

University of Seville, Spain
Email:
and


Pablo Domínguez de María

Sustainable Momentum, SL, Las Palmas de Gran Canaria, Spain
Email:

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Catalysis Series No. 29
Print ISBN: 978-1-78262-619-0
PDF ISBN: 978-1-78262-999-3
EPUB ISBN: 978-1-78801-246-1
ISSN: 1757-6725
A catalogue record for this book is available from the British Library
© The Royal Society of Chemistry 2018
All rights reserved
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Preface
Biocatalysis has acquired a relevant role in organic synthesis over recent
decades, stimulated by the outstanding developments in microbiology,
molecular biology, and bioinformatics, together with a mature know-how
in medium engineering and enzyme immobilization. Remarkable breakthroughs in those areas have enabled a wide accessibility to geneticallydesigned (tailored) biocatalysts as required at industrial scale, and in
many cases at acceptable prices. The accumulated knowledge has triggered
the implementation of enzyme catalysis in commercial processes – to
manufacture a broad product portfolio –, leading to highly selective reactor
set-ups, often conducted under mild conditions, and in many cases in an
environmentally-friendly manner.
The penetration of biocatalysis in industry is not, however, a straightforward task. Interdisciplinary teams need to act in a coordinated way, and
often under time-pressured deadlines. Remarkably, whilst there is ample
information available on how a biocatalytic process can (academically) be set
up, available knowledge emphasizing the link between laboratory research
and industrial processes is somewhat scarce. To enable a better transition
between these two worlds, the participation of industrialists in the academic

arena appears crucial to inspire an “industrial conceptual approach” to the
biocatalytic studies.
In this connection, this book has been conceived to contribute to bridging
the gap between academic studies on biocatalysis and industrial processes
related to free enzymes, cell free extracts or whole-cells systems. To provide
a comprehensive industrial vision, several companies of different sizes and
purposes have been invited to contribute with chapters on industrial biocatalysis. The fundamental question to be answered is “How do industries
think of, approach, and implement biocatalytic procedures?” Some contributions directly describe (their) industrial processes, giving hints on metrics,

 
Catalysis Series No. 29
Biocatalysis: An Industrial Perspective
Edited by Gonzalo de Gonzalo and Pablo Domínguez de María
© The Royal Society of Chemistry 2018
Published by the Royal Society of Chemistry, www.rsc.org

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Preface

viii

productivities, and focus on the required parameters to deliver a practical
implementation. In contrast, other chapters focus on the challenges and
hurdles that are often encountered in the transit from laboratory-to-market.
All in all, we believe that this book gathers useful material that it is not
easily found in other literature sources, and that it is certainly very relevant

for the establishment of novel enzymatic processes. We think that the herein
reported “industrial thoughts” may trigger research groups (whether from
academia or companies) to further undertake action in biocatalysis.
We would like to warmly thank all the authors for their outstanding
contributions – realizing the difficulty that writing from industry may bring –
as well as to the entire Royal Society of Chemistry editorial team, for having
accompanied us in this challenging but highly rewarding adventure. We also
want to thank our families for their patience, commitment and generosity,
and for their constant support throughout the development of this book.
We sincerely hope that this book will be useful for your work in biocatalysis
and sustainable chemistry.
Gonzalo de Gonzalo, Pablo Domínguez de María
Seville and Las Palmas de Gran Canaria, Spain

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Contents
Part I: Context and Challenges for Industrial Biocatalysis
Chapter 1 An Appreciation of Biocatalysis in the Swiss Manufacturing
Environment 
3
Rebecca Buller, Katrin Hecht, Marco Antonio Mirata and
Hans-Peter Meyer




















1.1 Introduction 
1.1.1 Biocatalysis in the Swiss Manufacturing
Environment 
1.1.2 Current Status 
1.1.3 Patent Analysis 
1.2 Selected Enzyme Classes Used in the Swiss
Manufacturing Environment 
1.2.1 Oxidoreductases EC1 
1.2.2 Transferases EC2 
1.3 Challenges 
1.3.1 Regulation 
1.3.2 Development Time 
1.3.3 Technological Lock-in 
1.3.4 Public Perception 
1.3.5 Education 
1.4 Opportunities 
1.4.1 Starting Materials 

1.4.2 Sustainability and Greenness 
1.4.3 Swiss Industrial Biocatalysis Consortium 
1.4.4 New Business Ideas 

 
Catalysis Series No. 29
Biocatalysis: An Industrial Perspective
Edited by Gonzalo de Gonzalo and Pablo Domínguez de María
© The Royal Society of Chemistry 2018
Published by the Royal Society of Chemistry, www.rsc.org

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1.5 Future Directions 
 cknowledgements 
A
References 

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Chapter 2 Biocatalysis – A Greener Alternative in Synthetic Chemistry 44
Madhuresh Kumar Sethi, Purbita Chakraborty
and Rohit Shukla
















2.1 Introduction 
2.2 Motivation for Industry to Use/Research on
Biocatalysis 
2.3 Challenges Faced by Biocatalysis in Industry
2.4 Prospects 
2.5 Overview of Current Enzyme-based Processes
Implemented/In-progress at Industrial, Commercial
Scale 
2.6 Our Experience in Some Chemoenzymatic Projects 
2.6.1 Protease-mediated Synthesis of Valganciclovir
Intermediate 
2.6.2 Chemoenzymatic Synthesis of Optically Pure
Rivastigmine Intermediate Using ADH from
Baker’s Yeast and KRED 
2.6.3 Preparation of Deoxynojirimycin, Key
Intermediate of an Anti-diabetic Drug 
2.7 Potential Safety Aspects 
2.8 Conclusions 
Abbreviations 
Glossary 

References 
Chapter 3 Biocatalytic Synthesis of Small Molecules – Past, Present
and Future 
Roland Wohlgemuth













3.1 Introduction 
3.2 Biocatalytic Conversions of Racemates 
3.2.1 Biocatalytic Resolution of Racemates 
3.2.2 Biocatalytic Deracemizations 
3.3 Biocatalytic Desymmetrizations 
3.4 Biocatalytic Asymmetric Oxidations and Reductions 
3.4.1 Biocatalytic Asymmetric Oxidations 
3.4.2 Biocatalytic Asymmetric Reductions 
3.5 Biocatalytic Asymmetric Hydrolysis and Acylation
Reactions 
3.5.1 Biocatalytic Asymmetric Hydrolysis Reactions 
3.5.2 Biocatalytic Asymmetric Acylation Reactions 


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3.6 Biocatalytic Asymmetric Transfer Reactions 
3.7 Biocatalytic Asymmetric Addition and Elimination
Reactions 
3.8 Summary and Outlook 
References 
Chapter 4 EntreChem: Building a Sustainable Business Case
in Biotechnology: From Biocatalysis to Synthetic
Biology 
Javier González Sabín and Francisco Morís















4.1 Introduction 
4.2 Biocatalysis 
4.2.1 Enantiopure Chiral Building Blocks 
4.2.2 Cascade Processes Taking Advantage of
Biocatalysis 
4.3 Drug Development 
4.3.1 Natural Products in Drug Discovery 
4.3.2 EntreChem’s Approach to Natural Products
Drug Discovery 
4.3.3 Aureolic Acids: The Quest for Clinically Viable
“Mithralogs”
4.3.4 Collismycin Analogs as Immunosuppressive
and Neuroprotective Drugs 
4.3.5 Glycosylated Indolocarbazoles as Potent and
Selective Kinase Inhibitors 
4.4 Business Models in Biocatalysis and Natural
Products Drug Discovery 
References 

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88
89
90

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112
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Part II: Biocatalysis: from Pharmaceuticals to Bulk
Chemistry
Chapter 5 Bristol-Myers Squibb: Preparation of Chiral
Intermediates for the Development of Drugs and APIs 
Ramesh N. Patel






5.1 Introduction 
5.2 Anti-Alzheimer’s Drug. Enzymatic Preparation of
(R)-5,5,5-Trifluoronorvaline 
5.3 Cholesterol Lowering Agents 
5.3.1 Enantioselective Enzymatic Acylation of
Racemic Alcohol 
5.3.2 Enzymatic Synthesis of (3S,5R)-Dihydroxy-6(benzyloxy)hexanoic Acid, Ethyl Ester 

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5.4 Calcitonin Gene-related Peptide Receptors
Antagonists (Migraine Treatment): Enzymatic
Preparation of (R)-2-Amino-3-(7-methyl-1H-indazol-
5-yl)propanoic Acid 
5.5 Antidiabetic Drugs 
5.5.1 Saxagliptin: Enzymatic Synthesis of (S)-N-
Boc-3-hydroxyadamantylglycine 
5.5.2 Saxagliptin: Enzymatic Synthesis of N-Cbz-
4,5-dehydro-L-prolineamide and N-Boc-4,5-
dehydro-L-prolineamide 
5.5.3 Saxagliptin: Enzymatic Ammonolysis of (5S)-
4,5-Dihydro-1H-pyrrole-1,5-dicarboxylic Acid,
1-(1,1-Dimethylethyl)-5-ethyl Ester 
5.5.4 GLP-1 Receptor Agonists: Enzymatic
Preparation of (S)-Amino-3-[3-{6-
(2-methylphenyl)}pyridyl]-propionic Acid 
5.6 Antihypertensive Drugs 
5.6.1 Enzymatic Synthesis of (S)-6-
Hydroxynorleucine 
5.6.2 Vanlev: Enzymatic Synthesis of Allysine
Ethylene Acetal 
5.6.3 Vanlev: Enzymatic Synthesis of Thiazepine 

5.6.4 Captopril: Enzymatic Preparation of (S)-3-
Benzoylthio-2-methylpropanoic Acid 
5.7 Antiviral Drugs. Case Study: Atazanavir 
5.7.1 Atazanavir: Enzymatic Synthesis of
(S)-Tertiary-leucine 
5.7.2 Atazanavir: Enzymatic Preparation of (1S,2R)-
[3-Chloro-2-hydroxy-1-(phenylmethyl)propyl]
carbamic Acid, 1,1-Dimethyl-ethyl Ester 
5.8 Antianxiety Drug. Buspirones: Enzymatic
Preparation of 6-Hydroxybuspirone 
5.9 Antiviral Drugs. Hepatitis B Viral (HBV) Inhibitor:
Enzymatic Asymmetric Hydrolysis and Acetylation 
5.10 Chemokine Receptor Modulators: Enzymatic
Desymmetrization of Dimethyl Ester 
5.11 Anticancer Drugs 
5.11.1 Paclitaxel Semisynthetic Process 
5.11.2 Water-soluble Taxane Derivatives 
5.11.3 Epothilones: Epothilone B and
Epothilone F 
5.11.4 IGF-1 Receptor Inhibitor: Enzymatic
Preparation of (S)-2-Chloro-1-
(3-chlorophenyl)ethanol 
5.11.5 Retinoic Acid Receptor Agonist:
Enzymatic Preparation of 2-(R)-Hydroxy-
2-(1′,2′,3′,4′-tetrahydro-1′,1′,4′,4′-
tetramethyl-6′-naphthalenyl)acetate 
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5.12 Microbial Hydroxylation of Mutilin and
Pleuromutilin 
5.13 Conclusions 
Acknowledgements 
References 
Chapter 6 Johnson Matthey: A Technology Provider Perspective to
Biocatalysis in the Fine Chemicals Industry 
Elina Siirola, Ahir Pushpanath, Desmond M. Schofield
and Paolo Braiuca















6.1 Introduction 
6.2 Commercial Considerations 
6.2.1 Technology Value 

6.2.2 Manufacturing 
6.2.3 Market Analysis 
6.2.4 Catalyst Portfolio 
6.3 Technical Considerations 
6.3.1 Enzyme Recruitment 
6.3.2 Enzyme Engineering 
6.3.3 Process Improvement 
6.4 Conclusions 
Acknowledgements 
References 
Chapter 7 EnzymeWorks: Recent Advances in Enzyme
Engineering for Chemical Synthesis 
Kui K. Chan, Ju Xin, Xiaoliang Liang, Lizeng Peng,
Bin Sun and Junhua Tao















7.1 Introduction to EnzymeWorks 

7.1.1 Current Status of Biocatalyst Development 
7.2 Biocatalysis in the Food and Beverage Industry 
7.2.1 Introduction of Stevia Development 
7.2.2 Plant Family 1 UDP-glycosyltransferase
Applications 
7.2.3 Chemoenzymatic Synthesis of
Rebaudioside M 
7.2.4 Enzyme Immobilization and Whole Cell
Biosynthesis Development 
7.2.5 Future Perspectives on Biocatalysis in the
Food and Beverage Industry 
7.3 Ketoreductase (KRED) Applications 
7.3.1 Ibrutinib Development 
7.3.2 Future Perspectives on Ketoreductase
(KRED) Biocatalysis 
7.4 Biocatalysis in the Antibiotic Industry 
7.4.1 Introduction to Cephalosporin C Acylase (CCA) 
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7.4.2 Gene Expression, Structure and Catalytic
Mechanism of Acylases 
7.4.3 Recent Advances in Cephalosporin C Acylase
(CCA) Development 
7.4.4 Future Perspectives on Acylase Biocatalysis 
7.4.5 Introduction to Deacetoxycephalosporin C
Synthase 
7.4.6 Deacetoxycephalosporin C Synthase
Structure and Mechanism 
7.4.7 Recent Advances in Deacetoxycephalosporin
C Synthase Development 
7.4.8 Future Perspectives of Antibiotic
Biocatalysis 
7.5 Future Perspectives of Biocatalyst Development 
References 
Chapter 8 Almac: An Industrial Perspective of Ene Reductase
(ERED) Biocatalysis 
Gareth Brown, Thomas S. Moody, Megan Smyth and
Stephen J. C. Taylor























8.1 Introduction 
8.1.1 Almac Group 
8.1.2 Biocatalysis at Almac 
8.1.3 The Rise of Biocatalysis 
8.2 Introduction to Alkene Reduction 
8.3 An Introduction to Ene Reductases and How
They Work 
8.4 Examples of Ene Reductase Reactions Reported
in the Literature 
8.4.1 Ene Reductases as Part of a Reaction

Sequence 
8.4.2 Ene Reductases and Solvents 
8.4.3 Challenges of Co-factor Recycle 
8.4.4 Avoiding the Use of Nicotinamide
Co-factors 
8.4.5 Impact of Synthetic Biology 
8.4.6 Ene Reductases in Reverse: Oxidation 
8.4.7 Thermophilic Ene-reductases 
8.4.8 Alternative Screening Methods 
8.5 Example of Utilisation of an ERED at Industrial
Scale 
8.6 Transition of Ene Reductases to Mainstream
Biocatalytic Use 
8.7 Conclusions 
Acknowledgements 
References 
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Chapter 9 GSK: Biocatalyst Discovery and Optimisation 
Marcelo Kern, Gheorghe-Doru Roiban, Andrew Fosberry
and Radka Snajdrova











9.1 Introduction 
9.2 Biocatalyst Discovery 
9.2.1 Design of Enzyme Panels 
9.2.2 Imine Reductase Panel – Importance and
Applicability 
9.3 Biocatalyst Optimisation 
9.3.1 Nelarabine Case Study 
9.4 Conclusions 
Acknowledgements 
References 

Chapter 10 PETROBRAS: Efforts on Biocatalysis for Fuels and
Chemicals Production 
Aline Machado de Castro and José André Cavalcanti
da Silva














10.1 PETROBRAS Overview 
10.2 Hydrolysis of Lignocellulosic and Starchy
Biomass 
10.3 Synthesis of Solvents 
10.3.1 Glycerol Carbonate 
10.3.2 Butyl Acetate 
10.4 Synthesis and Degradation of Polymers 
10.4.1 Synthesis of Polyesters 
10.4.2 Depolymerization of Poly(ethylene
terephthalate) 
10.5 Synthesis of Biolubricants 
10.6 Synthesis of Biodiesel 
10.7 Concluding Remarks 
References 

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Chapter 11 MetGen: Value from Wood – Enzymatic Solutions 
Klara R. Birikh, Matti W. Heikkila, Alex Michine,
Antoine Mialon, Toni Grönroos, Petri Ihalainen, Antti
Varho, Veera Hämäläinen, Anu Suonpää and Sami-Pekka
Rantanen

298




298





11.1 Introduction 
11.1.1 METGEN – Masters of Enzyme Technology
and Genetic Engineering 
11.1.2 Biocatalysis of Wood – Motivation and
Challenges 
11.2 Enzymes in Pulp and Paper 
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11.2.1 Enzymes in Pulp & Paper Industry Sector –
Business Aspect 
11.2.2 Major Enzyme Components for Wood
Applications 
11.2.3 Enzyme Development – from Laboratory
to Industry 
11.2.4 MetZyme® LIGNO™ 
11.2.5 MetZyme® BRILA™ 
11.2.6 Concluding Remarks on Enzymes in Pulp
and Paper 
11.3 Biorefinery Enzymes 
11.3.1 Renewable Chemical Industry Segment –
Business Aspect 
11.3.2 Wood Biorefinery Concept 
11.3.3 Biomass Hydrolysis – Chemicals and Enzymes 
11.3.4 Biomass Is Not Oil; It Is More Like Soup
of the Day 
11.3.5 Beyond Sugars 
11.3.6 Biorefinery Enzymes – Concluding Remarks 
11.3.7 Wood in Pulp and Paper and Biorefinery –
Common Problems or Window for
Opportunity? 
Abbreviations 

References 

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Part III: Biocatalyst Optimization with Industrial
Perspectives
Chapter 12 LentiKat’s: Industrial Biotechnology, Experiences
and Visions 
Radek Stloukal, Jarmila Watzková and Kristýna Turková












12.1 Introduction 
12.2 Lentikats Biotechnology 
12.2.1 Potential of Lentikats Biotechnology 
12.2.2 Properties of Lentikats Biotechnology 
12.2.3 Production of Lentikats Biocatalyst 
12.3 Experiences in Wastewater Treatment 
12.3.1 Municipal Wastewater Treatment 
12.3.2 Industrial Wastewater Treatment 
12.3.3 Special Applications 
12.3.4 Advantages of Lentikats Biotechnology in
Wastewater Treatment 

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12.3.5 Product Lines 
12.3.6 Wastewater Treatment Applications 
12.4 Experiences in the Pharmaceutical & Food Industry 
12.4.1 Food Technology Industry 
12.4.2 Pharmaceutical Industry 
12.4.3 Bio-based Chemicals Industry 
12.4.4 Advantages of Lentikats Biotechnology in
the Pharmaceutical & Food Industry 
12.4.5 Application Examples in the Pharmaceutical
& Food Industry 
12.4.6 Pharmaceutical & Food Applications 
12.5 Vision 

References 

Chapter 13 EziG: A Universal Platform for Enzyme Immobilisation 
Karim Engelmark Cassimjee and Hans-Jürgen Federsel





















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13.1 Introduction 
345
13.2 A General Methodology for Enzyme Reuse 
348
13.2.1 The Potential of Biocatalysis by Far Exceeds
Its Current Exploitation 
348
13.2.2 Unlocking the Potential of Enzymes 
349
13.2.3 Immobilised Enzymes for the
Pharmaceutical Industry 
350
13.2.4 The Reusable Enzyme Utopia – Enzymes
Anchored in Space 
350
13.2.5 The EziG Technology 
351
13.2.6 Standardised Procedure for Immobilisation  354
13.2.7 Lower Cost Materials versus High
Performance 
354
13.3 Case Studies 

355
13.3.1 In-reactor Enzyme Immobilisation 
355
13.3.2 Two-phase System in Flow for in situ
Product Removal 
356
13.3.3 Candida antarctica Lipase B (CalB) 
357
13.3.4 Co-immobilisation for Cascade Reactions 
358
13.4 Prospects 
359
13.4.1 Stability versus Activity – Replacing Low
Cost Catalysts 
359
13.4.2 Biocatalysis in Flow – Towards Manufacturing
Processes in Continuous Mode 
360
13.5 Conclusions 
360
Acknowledgements 
361
References 
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Chapter 14 Cross-linked Enzyme Aggregates (CLEAs): From
Concept to Industrial Biocatalyst 
Roger A. Sheldon






















14.1 Introduction: Biocatalysis is Green and
Sustainable 
14.2 Immobilisation of Enzymes 

14.3 The CLEA Technology 
14.3.1 The Concept 
14.3.2 Preparation of CLEAs 
14.3.3 Physical Properties of CLEAs 
14.3.4 Advantages and Limitations of CLEAs 
14.3.5 Reactor Design 
14.4 Scope of the CLEA Technology 
14.4.1 Hydrolase CLEAs 
14.4.2 Oxidoreductase and Lyase CLEAs 
14.5 Multi- and Combi-CLEAs 
14.6 Magnetic CLEAs: The New Frontier 
14.7 Applications of CLEAs, Combi-CLEAs and
mCLEAs 
14.7.1 1G and 2G Biofuels Production 
14.7.2 Food and Beverages Processing 
14.7.3 Synthesis of Semi-synthetic Penicillin
and Cephalosporin Antibiotics 
14.7.4 Removal of Dyes, Pharma Residues and
Endocrine Disruptors from Waste
Water 
14.7.5 Other Potential Applications 
14.8 Conclusions and Future Prospects 
References 

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Chapter 15 SynBiocat: Protein Purification, Immobilization and
Continuous-flow Processes 
397
Diána Weiser, Zoltán Boros, József Nagy, Gábor Hornyánszky,
Evelin Bell, Péter Sátorhelyi and László Poppe











15.1 Introduction 
15.2 SynBiocat – From Protein Purification to
Continuous-flow Processes 
15.2.1 Enzyme Production and
Purification 
15.2.2 Enzyme Immobilization 
15.2.3 Desktop Bioreactor Applications 
15.3 Conclusion 
List of Abbreviations 
Acknowledgements 
References 
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Part IV: Emerging Industrial Biocatalysis
Chapter 16 Microvi: MicroNiche Engineering™ for Biocatalysis
in the Water and Chemical Industries 
Ameen Razavi and Fatemeh Shirazi







16.1 Introduction to Microvi 
16.2 Microvi’s MicroNiche Engineering™ Platform 
16.3 Case Study: MicroNiche Biocatalysts for Water
Purification 
16.4 Producing Case Study: MicroNiche Biocatalysts
for Biobased Chemicals 
16.5 Conclusions 
References 

Chapter 17 Nofima: Peptide Recovery and Commercialization by
Enzymatic Hydrolysis of Marine Biomass 
Birthe Vang, Themis Altintzoglou, Ingrid Måge, Sileshi G.
Wubshet, Nils K. Afseth and Ragnhild D. Whitaker















17.1 Nofima: The Company 
17.2 Hydrolysis of Marine Biomass 
17.2.1 Chemical Hydrolysis of Marine Biomass 
17.2.2 Enzymatic Hydrolysis of Marine Biomass 
17.3 Enzymes Used for Bioconversion 
17.4 Quality and Classification of Marine Biomass 
17.5 Functional Properties and Bioactivities of
Hydrolyzed Marine Biomass 
17.6 Commercialization of Products from Marine
Biomass 
17.7 Conclusions 
17.8 Case Examples 
17.8.1 Marealis – Producing a Nutraceutical from
Shrimp Peels 
17.8.2 Polybait AS – Producing Fishing Bait
from Fisheries By-products 
References 

Chapter 18 CO2 Solutions: A Biomimetic Approach to Mitigate CO2
Emissions – The Use of Carbonic Anhydrase in an
“Industrial Lung” 

Eric Madore and Sylvie Fradette



18.1 Introduction 
18.2 Conventional Post-combustion CO2 Capture
Technologies 

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18.3 CSI’ Technology: An Industrial Lung 

18.4 Selection and Development of a Robust CA 

18.4.1 Elevated Ionic Strength 

18.4.2 High pH 

18.4.3 Temperatures Above 60 °C 

18.4.4 Effect of High Surface Volume Ratio 

18.4.5 Effect of High Shear Stress 

18.4.6 Effect of Contaminants 

18.4.7 Effect of Solid–Liquid Interface 


18.4.8 Carbonic Anhydrase Development 

18.5 Technology Validation/Demonstration at Pilot Scale 

18.6 Conclusions 

Acknowledgements 

References 
Subject Index 

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Part I
Context and Challenges for Industrial
Biocatalysis

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Chapter 1

An Appreciation of Biocatalysis
in the Swiss Manufacturing
Environment
Rebecca Buller*a, Katrin Hechta, Marco Antonio
Miratab and Hans-Peter Meyerc
a

Zurich University of Applied Sciences, School of Life Sciences and Facility
Management, Institute of Chemistry and Biotechnology, Einsiedlerstrasse
31, 8820 Wädenswil, Switzerland; bLonza AG, 3930 Visp, Switzerland;
c
HES-SO Valais-Wallis, Institute of Life Technologies, Route du Rawyl 64,
1950 Sion, Switzerland
*E-mail:


1.1 Introduction
Relative to its size and population, Switzerland is the global number one
in the field of large and small molecule pharmaceuticals. Despite (or perhaps due to) being a small country without a colonial history or noteworthy 
natural resources with the exception of water, a remarkable landscape, and
the brains of its inhabitants, Switzerland is one of the biotechnology hotspots
in Europe. Once the rural poorhouse, Switzerland became one of the first
countries to be industrialized. Chocolate, watches and banks are the typical
enumerations of someone asked about the economic activities of Switzerland.
The pharmaceutical and chemical industries are usually not mentioned,
although they have become a major economic driver and source of wealth.
 
Catalysis Series No. 29
Biocatalysis: An Industrial Perspective
Edited by Gonzalo de Gonzalo and Pablo Domínguez de María
© The Royal Society of Chemistry 2018
Published by the Royal Society of Chemistry, www.rsc.org

3

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Chapter 1

4

As in England, the seed for industrialization was the textile industry,
which led to two other key industries in Switzerland: the machine industry
and the chemical industry. The machine industry was effectively triggered
by the continental blockade by Napoleon I in the early 19th century, which

forced the local ironmongers to develop and construct their own looms,
as they were cut off from English machines and spare parts. Indirectly the
food industry, as represented by Nestlé, also profited, as industrialization
favored food products that responded to the shorter time the working class
had available for household chores. As elsewhere, the dyes required for the 
textile industry brought about the industrialization of organic chemistry. The
subsequent expansion of the production of dyes to organic chemicals, small
molecule pharmaceuticals and ultimately biopharmaceuticals was straightforward due to strategic foresight, visionary entrepreneurs, the openness of
Switzerland to foreign capital, and entrepreneurial immigrants. The banks
founded at that time also played a key role in capitalizing this industrialization. Unlike in other countries,1 these developments were not hampered by
absolutist rulers who clung to the political and societal status quo up to the
second half of the 19th century.
Today, the Swiss chemical industry has moved up the value chain from
the production of basic chemicals to the manufacture of fine chemicals.
For example, although Lonza still successfully operates a cracker in Switzerland, which was built in the 1960s, competition has increased substantially
as classic oil-producing countries have started to install on-site chemical
production complexes to keep the first part of the value chain in their own
country. Consequently, the chemical and pharmaceutical industries have
continuously refocused on core competencies and high-value products,
resulting in an entirely new landscape, structures and companies between
1970 and today.2 The Swiss chemical industry is heavily dependent on foreign trade as only ∼5% of the sales are in the home market while 95% of
the industry's products are exported. Furthermore, most raw materials
for chemical and pharmaceutical production in Switzerland have to be
imported, over 80% of them from the EU. The chemical and pharmaceutical
industry is now the country’s leading exporter,3 generating 4% of its gross
domestic product and selling products to other countries valued at around
CHF 79 billion annually (roughly 40% of total Swiss exports). About 90%
of roughly 3000 products manufactured by the Swiss chemical industry are
chemical specialties.4 The industry employs ∼65 000 people in Switzerland
and over 355 000 globally.

Switzerland has been ranked the most innovative country for the sixth
consecutive year by the Global Innovation Index.5 Roche and Novartis rank
among the top ten companies with the largest R&D investments, alongside
other industrial giants such as Microsoft, Samsung, Toyota and Amazon.6
The Swiss chemical industry is focused on life science and chemical specialties, and the world-wide sales of the top ten Swiss companies is split as
follows: pharmaceuticals 63%, fine and specialty chemicals 13%, crop protection 9%, diagnostics 8% and vitamins, flavors and fragrances 7%. Since

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