Sunita Grover V. K. Batish V. Padmanabha Reddy Dairy Biotechnology Sunita Grover & V. K. Batish Dairy Microbiology Division NDRI, Karnal V. Padmanabha Reddy Dairy Microbiology Division SVVU, Tirupati Index Modules Page No Module 1. Introduction to Biotechnology Lesson 1. Definition and Scope of Biotechnology 5-6 Lesson 2. Historical development (Time line) of Biotechnology 7-15 Lesson 3. Future applications of Biotechnology 16-21 Module 2. Fundamental Biological Principles Lesson 4. Nucleic acids – Structure and function of DNA and 22-29 RNA Lesson 5. DNA replication and Genetic Code 30-34 Lesson 6. Gene expression – Transcription and Translation 35-41 Lesson 7. Gene regulation - The 'Lac operon' 42-47 Lesson 8. Mutations 48-59 Module 3. Genetic Engineering Technology/Recombinant DNA Technology Lesson 9. Molecular cloning - Tools 60-65 Lesson 10. Restriction enzymes 66-74 Lesson 11. Vectors 75-83 Lesson 12. Introduction of recombinant DNA into host cell 84-90 Lesson 13. DNA sequencing 91-95 Lesson 14. PCR and Real Time PCR 96-107 Lesson 15. DNA Microarray 108-112 Module 4. Cell Culture and Fusion Technology Lesson 16. Animal Cell Culture 113-119 Lesson 17. Protoplast/ Somatic fusion and Hybridoma 120-124 technology Lesson 18. Stem Cells and Nuclear cloning 125-134 Module 5. Application of Biotechnology in Dairying Lesson 19. Application of Genetic Engineering in dairy starter 135-140 cultures Lesson 20. Accelerated Cheese Ripening 141-146 Lesson 21. Dairy enzymes and whole cell immobilization 147-155 Lesson 22. Cloning and expression of genes in bacteria and 156-164 yeast Lesson 23. Large scale production and downstream processing 165-176 of recombinant proteins Lesson 24. Designer Milk 177-183 Lesson 25. Application and impact of Biotechnology on food 184-201 industry Lesson 26. Application of biotechnology for disposal of whey 202-205 and dairy effluents Module 6. Bioinformatics and ‘Omics’ Revolution Lesson 27. Bioinformatics and Genomics, Transcriptomics and 206-220 Proteomics Lesson 28. Metabolomics, Structural Biology, Nutrigenomics 221-229 and Pharmacogenomics Lesson 29. Ethical issues related to use of Genetically 230-235 Modified foods Lesson 30. Biosafety levels and biosafety boards operating in 236-242 India Reference 243 Dairy Biotechnology Module 1. Introduction to biotechnology Lesson 1 DEFINITION AND SCOPE OF BIOTECHNOLOGY 1.1 Introduction Biotechnology has emerged as a wonder technology with immense potential with innumerable applications in almost all walks of our life by providing a tangible solution to the problems confronting human life. This technology holds a great promise to improve the quality of life on the earth by poverty alleviation and raising the standard of living at the global level particularly in third world countries including India. Biotechnology can help us in meeting our basic needs such as food, clothing, shelter, health and safety. 1.2 What is Biotechnology? The literal meaning of Biotechnology as implied from this word is the study of tools from living things based on the split words contained therein i.e. Bios - life Teuchos - tool Logos - Study of or essence 1.3 Definition of Biotechnology There is no universal and complete definition of Biotechnology due to its wide range of usage. Although, Biotechnology has been defined differently in different countries, the most widely used and comprehensive definition of ‘Biotechnology is any technique that uses living organisms or substances from these organisms to make or modify a product, to improve plants or animals or to develop micro- organisms for specific uses’ (Office of technology Assessment of the United States Congress). Biotechnology is a multi-disciplinary concept involving many disciplines or branches of learning including all areas of life sciences such as Microbiology, Genetics, Molecular Biology, Biochemistry, Fermentation technology or Bio-process engineering and Bioinformatics etc. 5 www.AgriMoon.Com Dairy Biotechnology Biotechnology is not a new technology as it had been traditionally used by man since time immemorial without knowing the scientific principles involved in the process. The basics and advanced principles behind this powerful technology were unraveled and recognized only during the last few decades due to the tremendous progress and advancements made in the area of genetics, molecular biology that included the discovery of restriction endonucleases, polymerase chain reaction and completion of human genome project etc. 1.4 Scope of Biotechnology Biotechnology as explained above has the newest roots in the science of molecular biology, genetic engineering and microbiology. Advances in these areas have been exploited in a variety of ways both for production of industrially important bio-chemicals including enzymes and pharmaceutically important proteins, hormones etc. and for basic studies in molecular biology. As a result of its endless potentials, the scope and prospects of biotechnology have widened dramatically for commercial exploitation. By integrating biotechnology in the process and product development at commercial scale, biotech based industries have grown enormously all over the world including India and in the process, have created new job opportunities, human resource development and poverty alleviation. Biotechnology has now become a key issue to boost the economy of different nationalities including both the developed and developing countries. *****☺***** 6 www.AgriMoon.Com Dairy Biotechnology Module 1. Introduction to biotechnology Lesson 2 HISTORICAL DEVELOPMENT (TIME LINE) OF BIOTECHNOLOGY 2.1 Introduction The term ‘Biotechnology’ was coined in 1917 by Karl Ereky, an Hungarian engineer. The term meant all the lines of work by which products were produced from raw materials with the aid of living organisms such as bacteria. There is a common misconception that Biotechnology includes recombinant DNA technology and Genetic Engineering only. Biotechnology is NOT new. Man has been manipulating living things to solve problems and improve his way of life for millennia. Roots of Biotechnology can be traced back to 6000 B.C. when Sumerians fermented beer. In 4000 BC Egyptians used yeast to prepare bread and wine. Dahi / curd and other milk fermentations had been in use in India since times immemorial during the Vedic period. Early agriculture was concentrated on producing food. Plants and animals were selectively bred and microorganisms were used to make food items such as beverages, cheese and bread. ‘New Biotechnology’ has emerged as a Biological revolution and led to creation of a world of ―Engineered products. From historical perspective, biotechnology can be classified as ancient/old, classical and new/modern biotechnology. 2.2 Ancient Biotechnology It began with early civilization. Our ancestors were producing wine, beer, and bread by using fermentation, a natural process in which the biological activity of one-celled organisms play a critical role. They also found that by manipulating the conditions under which the fermentation took place, they could improve both the quality and the yield of the ingredients themselves. 2.3 Classical Biotechnology It followed ancient biotechnology which makes wide spread use of methods from ancient biotechnology especially fermentation methods adapted to industrial production. It produces large quantities of food products and other materials in short time to meet demands of increasing population. 2.4 Modern Biotechnology It deals with manipulating genetic information. Microscopy and advanced computer technologies are used for in-depth knowledge of science. It is based on advancements in genetics research from the mid 1800‘s. In 1859, Darwin published his theory of evolution on the ’Origin of Species’. Use of biotechnology to produce new life forms emerged in mid 1900‘s and it was made possible by rDNA technology. 2.5 Milestones that led to the Development and Advancement of Modern Biotechnology 7 www.AgriMoon.Com Dairy Biotechnology The major milestones in science that laid the foundation of the modern biotechnology and their time lines are listed below in chronological order (Table 2.1). Table 2.1 Milestones in Biotechnology 8 www.AgriMoon.Com Dairy Biotechnology 9 www.AgriMoon.Com Dairy Biotechnology •1977 onwards - Biotechnology era opened up (Dawn of Biotech era) • 1977 o Genentech, Inc. (Company founded by Herbert Boyer and Robert Swanson in 1976) produced first human protein somatostatin from a transgenic bacterium which was considered as the advent of the Age of Biotechnology. o Walter Gilbert and Allan Maxam devised a method for sequencing DNA • 1978 o Recombinant Human Insulin – produced at California by Herbert W. Boyer o David Botstein discovered Restriction Fragment Length Polymorphism (RFLP) technique for studying polymorphism • 1980 o U.S. Supreme Court ruled that genetically altered life forms could be patented, thereby, allowing Exxon oil company to patent oil eating micro-organism. o Kary Mullis and others at Cetus Corporation in Berkeley, California developed Polymerase Chain Reaction (PCR). Sold patent for $300 Million in1991 • 1981 o First transgenic mice produced with rabbit beta-globin gene • 1982 o USFDA approved sale of genetically engineered human insulin (first recombinant product to be marketed in US) o Michael Smith at the University of British Columbia, Vancouver, developed Site directed mutagenesis for making precise amino acid changes anywhere in a protein. • 1983 o An automated DNA sequencer was developed o A screening test for