Fundamentals of Food Biotechnology

Fundamentals of Food

Biotechnology

Fundamentals of Food

Biotechnology

Second Edition

Byong H. Lee

Distinguished Professor, School of Biotechnology

Jiangnan University, Wuxi, China

Invited Distinguished Professor, Department of Food Science &

Biotechnology, Kangwon

National University, Chuncheon, Korea

Adjunct Professor, Department of Food Science & Agric Chemistry McGill

University, Montreal, Quebec, Canada

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Library of Congress Cataloging-in-Publication Data

Lee, B. H. (Byong H.)

Fundamentals of food biotechnology / Byong H. Lee. – Second edition.

pages cm

Includes bibliographical references and index.

ISBN 978-1-118-38495-4 (cloth)

1. Food–Biotechnology. I. Title.

TP248.65.F66L44 2015

664 – dc23

2014032719

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

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may not be available in electronic books.

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Typeset in 9/11pt TimesTenLTStd by Laserwords Private Limited, Chennai, India

1 2015

Contents

Preface xi

What Is Biotechnology? xiii

What Is Food Biotechnology? xvii

Part I New Trends and Tools of Food Biotechnology 1

1 Fundamentals and New Aspects 3

1.1 Biotechnological applications of animals, plants, and microbes 3

1.2 Cellular organization and membrane structure 6

1.3 Bacterial growth and fermentation tools 11

1.3.1 Classification and reproduction of biotechnologically important bacterial system 11

1.3.2 Bacterial growth 12

1.3.3 Environmental factors affecting bacterial growth 16

1.4 Fungal growth and fermentation tools 19

1.5 Classical strain improvement and tools 22

1.5.1 Natural selection and mutation 22

1.5.2 Recombination 27

Summary 30

1.6 Systems/synthetic biology and metabolic engineering 31

Summary 36

1.7 Bioengineering and scale-up process 36

1.7.1 Microbial and process engineering factors affecting performance and economics 38

1.7.2 Fermentor and bioreactor systems 39

1.7.3 Mass transfer concept 50

1.7.4 Heat transfer concept 53

1.7.5 Mass and heat transfer practice 57

1.7.6 Scale-up and scale-down of fermentations 71

1.7.7 Scale-up challenges 81

Summary 84

1.8 Molecular thermodynamics for biotechnology 85

1.8.1 Protein folding and stability 85

Summary 92

1.8.2 Downstream processes on crystallization and chromatography 93

Summary 96

1.9 Protein and enzyme engineering 96

Summary 100

1.10 Genomics, proteomics, and bioinformatics 100

Summary 108

vi CONTENTS

1.11 Biosensors and nanobiotechnology 109

1.11.1 Biosensor 109

1.11.2 Nanobiotechnology and nanobiosensor 113

Summary 116

1.12 Quorum sensing and quenching 116

Summary 120

1.13 Micro- and nano-encapsulations 120

1.13.1 Microencapsulation 122

1.13.2 Nanoencapsulation 129

Summary 138

Bibliography 140

2 Concepts and Tools for Recombinant DNA Technology 147

2.1 Concepts of macromolecules: function and synthesis 147

2.1.1 DNA replication 147

2.1.2 Roles of RNA 150

2.1.3 Detailed aspects of protein synthesis 153

2.2 Concepts of recombinant DNA technology 161

2.2.1 Restriction endonucleases 162

2.2.2 Plasmid vectors 164

2.2.3 Purpose of gene cloning 168

2.3 DNA sequencing 180

2.4 Polymerase chain reaction (PCR) 180

2.5 Manipulation techniques of DNA 183

2.5.1 Isolation and purification of nucleic acids 183

2.5.2 Agarose gel electrophoresis 184

2.5.3 Blotting and hybridization 185

2.6 Gene cloning and production of recombinant proteins 186

2.6.1 Cloning and expression of bacterial β-galactosidase in E. coli 186

2.6.2 Cloning, expression, and production of bovine chymosin (rennet) in yeast K. lactis 188

Summary 190

Bibliography 191

Part I Questions and Answers 193

Part II Applications of Biotechnology to Food Products 205

3 Yeast-Based Processes and Products 207

3.1 Food yeasts and derivatives 207

3.1.1 Introduction 207

3.1.2 Industrial processes 207

Summary 212

3.2 Alcoholic beverages 212

3.2.1 Introduction 212

3.2.2 Production and sales of major alcoholic beverages 212

3.2.3 Production processes 213

Summary 225

3.3 Industrial alcohols 225

3.3.1 Introduction 225

3.3.2 Raw materials and microorganisms 226

3.3.3 Production processes 230

3.3.4 Economics 231

Summary 232

3.4 Bread and related products 232

3.4.1 Introduction 232

CONTENTS vii

3.4.2 Ingredients and formulations 233

3.4.3 Production processes 234

3.4.4 New developments 236

Summary 237

Bibliography 237

4 Bacteria-Based Processes and Products 241

4.1 Dairy products 241

4.1.1 Introduction 241

4.1.2 Basic knowledge of manufacture of dairy products 244

4.1.3 Metabolic systems in lactic acid bacteria 249

4.1.4 Genetic modification of lactic acid bacteria 252

4.1.5 Applications of genetic engineering 254

Summary 262

4.2 Meat and fish products 262

4.2.1 Introduction 262

4.2.2 Fermented meat products 263

4.2.3 New developments 267

4.2.4 Fermented fish products 267

Summary 270

4.3 Vegetable products 270

4.3.1 Introduction 270

4.3.2 Fermented vegetable products 271

4.3.3 Fermented soy products 275

4.3.4 New developments 280

Summary 280

4.4 Vinegar and other organic acids 281

4.4.1 Introduction 281

4.4.2 Acetic acid 281

4.4.3 Citric acid 283

4.4.4 Lactic acid 284

4.4.5 Malic acid 285

4.4.6 Fumaric acid 286

Summary 286

4.5 Bacterial biomass 287

4.5.1 Introduction 287

4.5.2 Microorganisms for the production of biomass 288

4.5.3 Raw materials for the production of biomass 289

4.5.4 Production process 292

4.5.5 Nutritional aspects 293

4.5.6 Economics and new developments 294

Summary 295

4.6 Polysaccharides 296

4.6.1 Introduction 296

4.6.2 Microbial polysaccharides 297

4.6.3 Fermentation process 298

4.6.4 Bacterial polysaccharides 299

4.6.5 Other polysaccharides 304

Summary 304

Bibliography 306

5 Other Organism-Based Processes and Products 313

5.1 Enzymes 313

5.1.1 Introduction 313

5.1.2 Production of enzymes 315

5.1.3 Applications 317

viii CONTENTS

5.1.4 New developments and protein engineering 326

5.1.5 Economics 328

Summary 328

5.2 Sweeteners 329

5.2.1 Introduction 329

5.2.2 Nutritive sweeteners 329

5.2.3 High-intensity sweeteners 333

5.2.4 Low calorie sweeteners 337

5.2.5 New developments 338

Summary 339

5.3 Flavors and amino acids 339

5.3.1 Introduction 339

5.3.2 Microbial flavors 340

5.3.3 Enzymatic flavor generation 347

5.3.4 Amino acids 348

5.3.5 Economics 350

Summary 351

5.4 Vitamins and pigments 352

5.4.1 Introduction 352

5.4.2 Production of vitamins 352

5.4.3 Production of pigments 356

5.4.4 Economics 359

Summary 359

5.5 Mushrooms 360

5.5.1 Introduction 360

5.5.2 Cultivation 361

5.5.3 Culture preservation 363

Summary 363

5.6 Cocoa, tea, and coffee fermentation 364

5.6.1 Introduction 364

5.6.2 Cocoa fermentation 364

5.6.3 Coffee fermentation 367

5.6.4 Tea fermentation 369

Summary 372

5.7 Bacteriocins 372

5.7.1 Introduction 372

5.7.2 Classification 373

5.7.3 Mode of action 375

5.7.4 Bioengineering of bacteriocins 376

5.7.5 Applications of bacteriocins 379

5.7.6 Commercial production of bacteriocins 382

Summary 383

5.8 Functional foods and nutraceuticals 383

5.8.1 Probiotics and prebiotics 384

5.8.2 Health claim regulation 396

Summary 397

Bibliography 397

Part II Questions and Answers 411

Part III Other Potential Applications of the New Technology 431

6 Plant Biotechnology, Animal Biotechnology, and Safety Assessment 433

6.1 Plant biotechnology 433

6.1.1 Introduction 433

6.1.2 Plant cell and tissue cultivation 435

CONTENTS ix

6.1.3 Plant breeding 437

6.1.4 Application of plant cell and tissue culture 441

Summary 448

6.2 Animal biotechnology 449

6.2.1 Introduction 449

6.2.2 Transgenic animals 449

6.2.3 Animal cell culture 453

Summary 463

6.3 Food safety issues of new biotechnologies 464

6.3.1 Introduction 464

6.3.2 Safety evaluation of novel food products 465

6.3.3 Genetically modified microorganisms and their products 467

6.3.4 Genetically modified plants and their products 469

6.3.5 Genetically modified animals and their products 473

6.3.6 Detection methods of GM crops 475

6.3.7 Detection methods of transgenic animals and fish 480

6.3.8 Containment: physical and biological 481

6.3.9 Promises and limitations 481

Summary 482

Bibliography 483

Part III Questions and Answers 491

Index 497

Preface

In the past decade, major breakthroughs have happened and enormous progress has been

made in all aspects of genetic engineering and biotechnology. This is clearly reflected in the

voluminous publications of original research, patents, peer reviewed books, and symposia.

However, an exciting account of how this new biotechnology can affect traditional meth￾ods of producing foods and beverages is the need of the hour. Many professional reference

texts on food biotechnology are now available, but none of it is appropriate as classroom

text. Most such volumes are the work of multiple contributors and the normal didactic cri￾teria required to explain terms, flowcharts and frames of reference are lacking. No attempt

has been made to explain the translation of basic scientific information into practical appli￾cations. Moreover, biotechnology has become a fashionable subject and, as one of the most

abused buzz words of the decade, it now comprises a huge body of information. The very

scope of this knowledge presents serious problems to instructors and students. Which facts

are the most important for them to learn and which are less important? How can they assess

the significance of food systems and food products? In writing this book, I have tried to

keep these problems at the forefront and have therefore aimed at making the treatment of

food biotechnology comprehensible rather than comprehensive. I see that separate pieces

of a puzzle eventually fit together to form a picture that is clearer and more readily etched

in memory than the design on the individual pieces. Experience in teaching this subject has

made clear to me the importance of explaining the basic concepts of biotechnology, which

is essentially multidisciplinary, to students who may have limited backgrounds in the scale

up of bioengineering and rapidly developing new tools.

I hope that this book will prove valuable to both students and instructors as well as

to research and industrial practitioners in specific aspects of the field who seek a broad

view on food biotechnology. This book aims to give readers, general science students,

and practicing researchers, an overview of the essential features of food biotechnology

not covered in other institutions as typical science curriculum. The treatment of subjects

is necessarily selective, but the volume seeks to balance the traditional biotechnologies

with the new, and science and engineering with their industrial applications and potential.

Because of the interdisciplinary nature of the subject and the overlapping nature of the

principles of biochemistry, microbiology, and biochemical engineering, the second edition

does not include this part. Instead, the New Trends and Tools of Food Biotechnology

section in Part I (Fundamentals and New Aspects) has included Systems/Synthetic Biology

and Metabolic Engineering, Bioengineering and Scale-Up Process, Molecular Thermody￾namics for Biotechnology, Protein and Enzyme Engineering, Genomics, Proteomics and

Bioinformatics, Biosensors and Nanobiotechnology, Quorum Sensing and Quenching,

and Micro- and Nanoencapsulations. For the Concepts and Tools for Recombinant DNA

Technology (Chapter 2), examples of Gene Cloning and Production of Recombinant

Proteins have been included. In Chapter 5 on Other Microorganisms-Based Processes

and Products, a new section on Bacteriocins and Functional Foods and Nutraceuticals was

xii PREFACE

supplemented and the Waste Management and Food Processing section was deleted; it

will be included in my forthcoming book entitled: “Advanced Fermentation Technology.”

In Part III, Chapter 6 included Plant Biotechnology, Animal Biotechnology, and Safety

Assessment and Detection Methods and other sections were detailed. Up-to-date reading

materials as well as questions and answers have been included in all parts.

I must, of course, thank all those students who have helped me by compiling materi￾als used in the class to produce this book. I greatly appreciate the contribution of many

scientists who have embellished this book by permitting me to reproduce their tables and

figures, which have been illustrated in the pages of this book. I must accept my ignorance

and limitations naturally imposed on a book of this scope when it is written by a single

individual.

A special note of thanks also goes to my previous research associates and students for the

first edition at the McGill University, Dr. S. Y. Park, Dr. J. L. Berger who helped me in typ￾ing and drawing the figures, and other associates, G. Arora, M. Torres, M. B. Habibi-Najafi,

and graduate students, M. Bellem, M. Daga, J. James, and T. Wang who helped me in

many ways.

Most of all, my thanks go to Prof. Jian Chen, the President, and Prof. Guchang Du,

the Dean for their support during my stay in the School of Biotechnology at Jiangnan

University in China and the other staff in the 9th floor: Dr. F. Fang, Dr. Z. Kang, Dr. L. Song,

Dr. J. Zhang, Dr. J. Zhou, Dr. L. Liu, and Prof J. Li for their friendship during my absence.

I would like to specially thank Dr. Gazi Sakir for his comments on a part of the bioengi￾neering/scale up section, as well as my students, Dr. Zixing Dong, Yousef Mahammad, and

Nestor Ishimwe, and all international students who took my courses on Food Biotechnol￾ogy and Advanced Fermentation Technology at the Jiangnan University.

Last but not the least, I thank my wife Young for her love and encouragement; I also

thank and appreciate my sons, Edward in Toronto and David in New York, for their

patience and support during the preparation of this second manuscript.

December, 2013 Byong H. Lee

Wuxi, China

What Is Biotechnology?

We are in the middle of another industrial revolution in which biotechnology, depending

mainly on microbes, plays a major role in the production of exotic drugs, industrial chemi￾cals, bioingredients, fuel, and even food. Although biotechnology involves the potential use

of all living forms, microorganisms have played a major role in the development of this dis￾cipline because of the ease of mass growth, the rapid growth that occurs in media consisting

of cheap waste materials, and the massive diversity of metabolic types. These characteristics

in turn allow for a diverse selection of potential products and facilitate genetic manipula￾tion to improve strains for new products.

The bio in “biotechnology” means life and refers to microbes and other living cells

including animal and plant cells. The technology comprises the growth of living cells in vats

(fermentors or bioreactors) containing nutrients and oxygen (if needed) at the specified

conditions, and the processing of biological materials produced by the cells through process

integration and optimization at top efficiency for achieving commercialization. Biotechnol￾ogy has arisen through the interaction between various parts of biology and engineering,

employing techniques derived from three well-recognized disciplines: biochemistry, micro￾biology, and biochemical engineering.

The term biotechnology is not a new one, although it has certainly become fashionable in

recent years. It had its origin in prebiblical times but was not widely used until the postwar

university boom in the 1950–1960s, when the volume of scientific and engineering research

output rose dramatically. New disciplines emerged out of increasing specialization. Thus in

the early 1960s, research groups and university departments as well as journals arose with

titles such as BioTechnology, Biochemical Engineering, and Bioengineering. “Biotechnol￾ogy” is the term that has commonly survived. Table I.1 shows that prior to the twentieth

century, biotechnology consisted almost solely of spontaneous processes. The introduction

of the fed batch in the production of baker’s yeast is probably the starting point of con￾trolled biological processes designated as biotechnological. Biotechnology thus includes

many traditional processes such as brewing, baking, wine making, and cheese making; and

the production of soy sauce, tempeh, many secondary metabolites (antibiotics, steroids,

polysaccharides, etc.), and numerous food ingredients (amino acids, flavors, vitamins, and

enzymes). Traditionally, the biotechnological process based on classical microbial fermen￾tation has been augmented by simple genetic manipulation using a mutagenic agent to

improve microorganisms for food fermentation and to enhance the production of bioin￾gredients. However, it is not possible to predetermine the gene that will be affected by a

given mutagen, and it is difficult to differentiate the few superior producers from the many

inferior producers found among the survivors of a mutation treatment.

The potential of fermentation techniques was dramatically increased in the late 1960s

and early 1970s through achievements in molecular genetics, cell fusion, and enzyme

technology. A new biotechnology was founded based on these methods. However,

additional completely novel, very powerful biotechnology techniques were developed