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Basic Concept of Biotechnology

Editors

Dr. K.N.Chandrashekara

Dr. Ashok Yakkaldevi

LAXMI BOOK PUBLICATION

258/34, RaviwarPeth,

Solapur-

Cell: +91 9595359435

FIRST EDITION

Rs: /-

“Basic Concept of Biotechnology”

Dr. K.N. Chandrashekara Dr. Ashok Yakkaldevi

© 2015 by Laxmi Book Publication, Solapur

All rights reserved. No part of this book may be reproduced in any

form, by mimeograph or any other means, without permission in

writing from the publisher.

ISBN-

Published by,

Laxmi Book Publication,

258/34, RaviwarPeth,

Solapur, Maharashtra, India.

Contact No. : +91 9595 359 435

Website: http://www.isrj.org

Email ID: [email protected]

individual components of living systems, we can now study the systems themselves in comprehensive scope and in exquisite molecular detail. We therefore face the tasks of effectively employing new technologies, of dealing with mountains of data, and, most important, of adjusting our thinking to understand complex systems as opposed to their individual components. Basic Concepts of Biotechnology had its origins from Dr. Ashok Yakkaldevi, Laxmi Book Publication, India. This is a book for beginners. My goal here is to familiarize the inexperienced reader with the important of tools and techniques of biochemistry and biotechnology. Thus the description of certain instrumentation and applications is not highly rigorous. This book is not intended to be a laboratory manual or a compilation of the latest techniques. There are several excellent volumes available that provide more detailed descriptions of protein analytical techniques, mass spectrometry instrumentation and techniques, and applications of these technologies. The evolution of methods and applications in this area is now so rapid that no book really could be truly up-to-date. What is exciting about my experience in introducing Basic Concepts in Biotechnology to colleagues has been the creativity with which they then apply these tools. Ultimately, the exciting potential of biotechnology rests with those who can put new technologies to work to address important questions. I would like to thank all the contribution authors, reviewers and publisher, who provided valuable suggestions, read and commented on several drafts of book chapters. I thank Dr. Ashok Yakkaldevi for excellent secretarial assistance. Finally, I thank my wife Nalina and my daughter Hamsini for their patience with me.

Dr. K. N. Chandrashekara

ACKNOWLEDGMENTS

The Dr. K. N. Chandrashekara thank Dr. Ashok Yakkaldevi for his thoroughly professional approach. His constant interest and input have had a significant impact on the final structure of this book. We are very grateful to the Scientists who have read and commented on various parts of this book: Dr. K. Chandrashekar, Dr. J. Mohan, Dr. S. Ganeshmurthy, Dr. M. K. Prasannakumar, Dr. L. N. Reddy, Dr. G. Sujan and Mr. K. N. Jagadish. All were responsible for considerable improvement to the text. We are indebted to Dr. B. Radhakrishnan, Director, UPASI TRF TRI and many colleagues who generously provided reprints of publications or information and clarified areas of confusion. Whose support and encouragement during the preparation of this book is greatly appreciated. We sincerely express our appreciation to all the authors who contributed to this book. We are thankful to all the reviewers who helped, read through and verified the existing information in this book. Without those tireless efforts and patient support this book would not have reached the readers. I am very grateful to Dr. Ashok Yakkaldevi for their thoughtful and meticulous work in efficient typesetting and proofreading the text. Suggestions from students and colleagues have been most helpful in the compiling of this book. We look forward to receiving similar input in the future. Last but not the least; we are thankful to Laxmi Book Publication for accepting our proposal to publish this book, without their efforts and support this book would not have reached the readers.

S. Vinoth Senior Research Fellow Department of Plant Science Bharathidasan University Tiruchirappalli, Tamil Nadu, India [email protected]

M.S. Suma Senior Research Fellow Department of Mechanical Engineering Indian Institute of Science Bangalore, Karnataka, India [email protected]

Shilpa R Raju Senior Research Fellow and Ph.D Scholar Department of Mechanical Engineering Indian Institute of Science Bangalore, Karnataka, India [email protected]

Madhuri Biradar Teaching Assistant Department of Applied Genetics Karnatak University Dharwad, Karnataka, India [email protected]

Harendra Modak Ph.D Scholar Department of Applied Genetics Karnatak University Dharwad, Karnataka, India [email protected]

A. R. Chidanand Research Associate and Ph.D Scholar Department of Biotechnology University of Agricultural Sciences Dharwad Dharwad, Karnataka, India [email protected]

M. Ranjith Kumar Post Doctoral Researcher Biotechnology Lab Department of Horticulture

Prakash M Navale Research Associate and Ph.D Scholar Department of Biotechnology

Sunchon National University Suncheon, South Korea [email protected]

Indian Institute of Horticultural Research Hessarghatta Lake Post Bangalore, Karnataka, India [email protected]

H. D. Sowmya Senior Research Fellow and Ph.D Scholar Department of Biotechnology Indian Institute of Horticultural Research Hessarghatta Lake Post Bangalore, Karnataka, India [email protected]

Sandeep Telkar Senior Research Fellow and Ph.D Scholar Department of Biotechnology Kuvempu University Shimoga, Karnataka, India [email protected]

Mr. S. Ashokraj Ph.D Scholar Biotechnology Lab Department of Horticulture Sunchon National University Suncheon, South Korea [email protected]

V. Brindha Priyadarisini Assistant Professor Department of Microbial Biotechnology Bharathiar University Coimbatore, Tamil Nadu, India [email protected]

Nalina. M and Chandrashekara. K.N.

Tools and Techniques in

Biotechnology

Harendra Modak, Chidanand Rabinal, Madhuri Biradar, Nalina. M and Chandrashekara. K.N.

Antibiotics: Microbial

sources, Production and

Optimization

Ranjith Kumar. M,

Ashokraj. S and

Brindha

Priyadarisini. V

Biotechnology in

Forensic Sciences

Harendra Modak and Madhuri Biradar

Basic Concept of Biotechnology Computer Applications and Biostatistics

Chapter 1

Biomolecules

Nalina. M, Raghavendra. A and Chandrashekara K.N

A living system grows, sustains and reproduces itself. The most amazing thing about a living system is that it is composed of non-living atoms and molecules. The pursuit of knowledge of what goes on chemically within a living system falls in the domain of Biochemistry. Even though there are thousands of different types of molecules in a cell, there are only a few basic classes of bimolecular like carbohydrates, proteins, nucleic acids, lipids, etc. Proteins and carbohydrates are essential constituents of our food. In addition, some simple molecules like vitamins and mineral salts also play an important role in the functions of organisms. The complexity of even the simplest of life forms, the single cell, cannot be overstated. Nevertheless, from a chemical perspective, cellular components can be segregated into macromolecules (DNA, RNA, proteins, etc.), relatively simple molecules (amino acids, monosaccharide’s, and lipids), and their precursors: CO2, H2O, and NH3. In general, the macromolecules tend to be polymers of small bimolecular; however, each of these molecules, whether simple or complex, is involved in a myriad of intricate metabolic reactions. A case in point is the monosaccharide glucose which is synthesized from H2O and CO2. When degraded to its precursors, it provides the cell with its energy requirements for such diverse processes as macroscopic movement as well as the synthesis of complex macromolecules. In addition, glucose is the fundamental building block of macromolecules

Basic Concept of Biotechnology Computer Applications and Biostatistics

cellulose, hemicelluloses and lignin. Carbohydrates comprise a comprehensive group of naturally occurring substances, which include innumerable sugars and sugar derivatives, as well as high-molecular weight carbohydrates (polysaccharides) like starch and cellulose in plants and glycogen in animals. A polysaccharide molecule is composed of a large number of sugar or sugar-like units. Carbohydrates are of great importance in biology. The unique reaction, which makes life possible on Earth, namely the assimilation of the green plants, produces sugar, from which originate, not only all carbohydrates but, indirectly, also all other components of living organisms. Carbohydrates form a very large group of naturally occurring organic compounds which play a vital role in daily life. They are produced in plants by the process of photosynthesis. The most common carbohydrates are glucose, fructose, sucrose, starch, cellulose etc. Chemically, the carbohydrates may be defined as polyhydroxy aldehydes or ketones or substances which give such molecules on hydrolysis. Many carbohydrates are sweet in taste and all sweet carbohydrates are called as sugars. The chemical name of the most commonly used sugar in our home is sucrose.

Classification of Carbohydrates Carbohydrates are classified into three groups depending upon their behavior on hydrolysis. (i) Monosaccharide’s: A polyhydroxy aldehyde or ketone which cannot be hydrolyzed further to a smaller molecule containing these functional groups is known as a monosaccharide. About 20 monosaccharides occur in nature and glucose is the most common amongst them. Monosaccharides are further classified on the basis of the number of carbon atoms and the functional group present in them. If a monosaccharide contains an aldehyde group, it is known as an aldose and if it contains a keto group,

Basic Concept of Biotechnology Computer Applications and Biostatistics

it is known as a ketose. The number of carbon atoms present is also included while classifying the compound as is evident from the examples given in Table 1. Table 1: Classification of monosaccharides No. of carbon atoms Type of monosaccharide Aldose Ketose

Three

Aldotriose (Glyceraldehyde) Ketotriose Four Aldotetrose ((Xylose) Ketotetrose Five Aldopentose(Erythrose) Ketopentose Six Aldohexose (Glucose) Ketohexose Seven Aldoheptose Ketoheptose

Glucose occurs freely in nature as well as in the combined form. It is present in sweet fruits and honey. Ripe grapes also contain glucose in large amounts. (ii) Disaccharides: Carbohydrates which give two monosaccharide molecules on hydrolysis are called disaccharides e.g. sucrose, maltose, lactose etc. (iii) Polysaccharides: Carbohydrates which yield a large number of monosaccharide units on hydrolysis e.g. starch, glycogen, cellulose etc

Structure of Monosaccharide’s Although a large number of monosaccharide’s are found in nature, we will confine our discussion here to four of them only viz. D- glucose, D-fructose, D-ribose and 2-deoxy-D-ribose. D-Glucose (an aldohexose) is the monomer for many other carbohydrates. Alone or in combination, glucose is probably the most abundant organic compound on the earth. D-Fructose (a ketohexose) is a sugar that is found with

Basic Concept of Biotechnology Computer Applications and Biostatistics

The α- and ß-forms of other sugars also exist in the cyclic form. D-Ribose forms a five member ring structure as shown below

D-before the name of above example indicates the configuration of particular stereoisomer. Stereoisomers are assigned relative configurations as D– or L –. This system of assigning the relative

Basic Concept of Biotechnology Computer Applications and Biostatistics

configuration refers to their relation with glyceraldehydes. Glyceraldehydes contain one asymmetric carbon atom so exists in two enantiomeric forms as shown below.

All those compounds which can be correlated to (+) - glyceraldehyde are said to have D-configuration and those can be correlated to (–) -glyceraldehyde are said to have L–configuration. In monosaccharides it is the lowest asymmetric carbon atom (shown in the box) by which the correlation is made. As in (+) glucose the lowest asymmetric carbon atom has – OH group on the right side which matches with (+) glyceraldehyde hence it is assigned D-configuration.

Basic Concept of Biotechnology Computer Applications and Biostatistics

Cellulose is another natural polysaccharide which is the main component of wood and other plant materials. It consists of long chain of ß-D-glucose molecules.

Importance of carbohydrates Carbohydrates are of great importance in biology. The unique reaction, which makes life possible on the Earth, namely the assimilation of the green plants, produces sugar, from which originate, not only all carbohydrates but, directly or indirectly, all other components of living

Basic Concept of Biotechnology Computer Applications and Biostatistics

organisms. The carbohydrates are a major source of metabolic energy, both for plants and for animals that depend on plants for food. Aside from the sugars and starch that meet this vital nutritional role, carbohydrates also serve as a structural material (cellulose), a component of the energy transport compound ATP, recognition sites on cell surfaces, and one of three essential components of DNA and RNA. Importance can be considered under following headings;

Metabolic/Nutritional The important role of carbohydrates, generally, in the metabolism of living organisms, is well known. The biological breakdown of carbohydrates (often spoken of as "combustion") supplies the principal part of the energy that every organism needs for various processes. Carbohydrates and their metabolism has been the subject of biochemical and medical research for a long time. Carbohydrates play a major role in promoting health fitness, form a major part of food and help a great deal in building body strength, by generating energy. They are one among the three prominent macronutrients that serve as excellent energy providers, the other two being fats and proteins. Carbohydrate intake can take place in different forms like sugar, starch, fibers etc., which are a dietary staple in most parts of the world, and the oxidation of carbohydrates is the central energy-yielding pathway in most non-photosynthetic organisms. The functions of carbohydrates are multiple and it is owing to this fact that it becomes all the more necessary to incorporate carbohydrates in our meal. For instant for energy generation, sugars and starch act as the perfect fuel that enables us to carry out our physical activities efficiently and effectively. Fiber does wonders in keeping your bowel function going smooth. Carbohydrates add on to the taste and appearance of food item, thus making the dish tempting and mouth watering. They are sometimes used