Enzyme engineering by tamizh

enzyme engineering
introduction
types
methods

1. ENZYME ENGINEERING Presented by: k.tamilmuhilan, I m.sc biotechnology, alagappa university .

2. CONTENT Enzymes Enzyme Engineering Introduction Objective Steps Involved Types and methods

3. ENZYMES Enzymes create chemical reaction in body. They actually speed up the rate of a chemical reaction to support life. The enzymes in your body perform very important tasks. Building muscles Destroying toxins Breaking down food particles during digestion An enzymes shape is tied to its function. Heat, disease or harsh chemical conditions can damage enzymes and change their shapes. When this happens, enzyme cannot work anymore.

4. ENZYME ENGINEERING Enzyme engineering is the process of designing enzyme by changing the sequence of amino acids through recombinant DNA technology. These products will be useful as chemical , pharmaceutical , fuel, food or agricultural additives . Since enzymes are proteins, enzyme engineering is a part of the larger activity of protein engineering. Enzyme engineering utilizes r-DNA technology to introduce the desired changes in amino acid sequences of enzymes

5. OBJECTIVES OF ENZYME ENGINEERING The chief objective of enzyme engineering is to produce an enzyme that is more useful for industrial and other applications. The various properties of an enzyme that may be modified to achieve this objective are as follows :- Improved kinetic properties. Elimination of allosteric regulation. Enhanced substrate and reaction specificity. Increased thermo stability. Alteration in optimal pH. Suitability for use in organic solvents. Increased/decreased optimal temperature.

6. Steps involved in Enzyme Engineering

7. STEPS INVOLVED:

8. METHODS: There are two general strategies for enzyme engineering : Rational design Directed evolution

9. RATIONAL DESIGN It is the earlier approach to enzyme engineering and still is widely used. In rational design, scientist uses detailed knowledge of structure and function of a protein to make desired changes. In general, it has a advantage of being inexpensive and technically easy. However, the major drawback is that a detailed structural knowledge of enzyme is often unavailable. Even when available, it can be very difficult to predict the effects of various mutations since structural information most often provide a static picture of enzyme structure. Two methods in rational design Overlap extension Whole plasmid single round PCR

10. DIRECTED EVOLUTION: Also termed In vitro evolution Directed molecular evolution Acceleration evolution Directed evolution(DE) encompasses several molecular techniques that mimic the process of Darwinian evolution in vitro, by combining random mutagenesis or recombinant Technology. It does not uses structural data of enzyme, other than its functional properties and its corresponding DNA sequence. DE is sometimes referred to irrational or semi-rational design.

11. Directed evolution Mimick recombination : DNA shuffling With restriction enzymes: 1 . Digest family of related genes 2 . DNA ligase to join fragments 3 . Chimeric genes

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