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Home » Biology Homework Help » Biotechnology » Enzyme Engineering
Enzyme Engineering
Improvement is the activity and usefulness of an existing enzyme or creation of new enzyme activity by making suitable changes in its amino acid sequence is called enzyme engineering. When this approach is used to modify the properties of any protein, whether enzyme or non-enzyme, it is termed as protein engineering. Since enzymes are proteins, enzyme engineering is a part of the larger activity of protein engineering. Enzyme engineering utilizes recombinant DNA technology to introduce the desired changes in the amino acid sequences of enzymes.

Recombinant DNA technology is also used to transfer encoding useful enzymes from dangerous, unapproved, slow growing or low producing micro-organisms into safe, fast growing and high producing micro-organisms. In addition, of the gene into the concerned organism. For example, E. coli gene encoding penicillin-G-oxidase, was integrated in pBR 322 and the recombinant plasmid was introduced into E. coli. Upto 50 copies per cell of the plasmid were produced thereby amplifying the gene 50-fold. The recombinant strain produced the enzyme in considerably higher quantities than the original/parent strain, which reduced recombinant DNA technology are likely to be much more fruitful as well as numerous than that modifying amino acid sequences of enzymes. But it should be kept in mind that these are not examples of enzyme or protein engineering which must rest on modification of the amino acid sequence of the concerned enzyme or protein.

Objectives

The chief objective of enzyme engineering is to produce an enzyme that is more useful for industrial and/or other applications. The various properties of an enzyme that may be modified to achieve this objective are as follows: (i) improved kinetic properties, (ii) elimination of allosteric regulation, (iii) enhanced substrate and reaction specificity, (iv) increase a thermostability, (v) alteration in optimal pH, (vi) suitability for use in organic solvents, (vii) increased/decreased optimal temperature etc.

Principles

The structure and function of an enzyme molecule, for that matter of any protein molecule, are chiefly determined by its amino acid sequences, i.e. its primary structure. Therefore, any change in the properties of an enzyme is always reflected in its primary structure. Conversely, a change in the amino acid sequence should alter the properties of the enzyme. But this is not always the case because the enzymatic properties etc. are changed only when amino acid changes and introduced in certain critical regions of the protein. Therefore, it is of great importance to know the critical regions for the various functions of an enzyme, and to be able to predict the effect of specific amino acid changes in these areas on the various functions. However, the present knowledge of the relationships between amino acid sequence, and three-dimensional structure and properties of enzymes, obtained from a large database, is only partially operative. It allows an explanation of changes in structure and function on the basis of changes in amino acid sequence, but it does not allow a dependable prediction of the influences of specific amino acid changes on the structure and function of enzymes.

It may, however, be reasonable to anticipate that as elaborate databases and improved softwares become available, it should become possible to predict with for greater confidence the structural and functional changes in enzymes produced by specified changes in their amino acid sequences. The effectiveness of enzyme engineering will be greatly enhanced then, and this activity may have a tremendous influence on enzyme technology.

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