Brief Introduction to Protein Engineering-Biotechnology-Unit 1- B. Pharm 6 Sem-Lect.6-Part-2
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methods of protein engineering
1) methods of protein engineering site directed mutagenesis
2) methods of protein engineering by use of gene families
3) methods of protein engineering through chemical modification
1) Methods of protein engineering site directed mutagenesis -in this there will be change in DNA sequence by substitution insertion or deletion takes place. in site directed mutagenesis M13 virus or bacteria phage are used.
2) methods of protein engineering by use of gene families - in this method isolation of genes from each species and then creation of hybrid will be generated. by this different combination are prepared, while applying this approach to subtilisin which are received from 26 species were mixed to create a library among these four enzymes were found to have improved properties.
3) methods of protein engineering through chemical modification - in this method stability of protein to high temperature and organic solvents are increased . the amino acid lysine residue can be cross linked by use of chemical linkers, for this glutaraldehyde as a protein linker are extensively used. by using glutaraldehyde different proteins like hemoglobin, insulin , phosphofructokinase, lactate dehydrogenase has been stabilized.
different approaches of protein engineering-
1) increasing the stability and biological activity of protein- bi addition of disulfide bonds- through this(a) T4 lysozyme stability are increased (b) xylanase- by introduction of disulfide bond the functional efficiency of it are improved
I) by reducing the free sulphur hydryl group- through this approach human beta interferon properties are increased
ii) by applying single amino acid changes- through this alpha antitrypsin , insulin, tissue plasminogen activator, hirudin , dihydrofolate reductase and T4 lysozyme properties are improved
2) improving kinetic properties of enzyme -by protein engineering km property of different enzymes are improved like subtilisin, asparaginase, tyrosyl synthetase , restriction endonucleases.
application of protein engineering-
1) application of protein engineering in medical 2) application of protein engineering in food industry
3) application of protein engineering in different industrial processes
4) application of protein engineering in environment.
#Pharmaceutical_Biotechnology
methods of protein engineering
1) methods of protein engineering site directed mutagenesis
2) methods of protein engineering by use of gene families
3) methods of protein engineering through chemical modification
1) Methods of protein engineering site directed mutagenesis -in this there will be change in DNA sequence by substitution insertion or deletion takes place. in site directed mutagenesis M13 virus or bacteria phage are used.
2) methods of protein engineering by use of gene families - in this method isolation of genes from each species and then creation of hybrid will be generated. by this different combination are prepared, while applying this approach to subtilisin which are received from 26 species were mixed to create a library among these four enzymes were found to have improved properties.
3) methods of protein engineering through chemical modification - in this method stability of protein to high temperature and organic solvents are increased . the amino acid lysine residue can be cross linked by use of chemical linkers, for this glutaraldehyde as a protein linker are extensively used. by using glutaraldehyde different proteins like hemoglobin, insulin , phosphofructokinase, lactate dehydrogenase has been stabilized.
different approaches of protein engineering-
1) increasing the stability and biological activity of protein- bi addition of disulfide bonds- through this(a) T4 lysozyme stability are increased (b) xylanase- by introduction of disulfide bond the functional efficiency of it are improved
I) by reducing the free sulphur hydryl group- through this approach human beta interferon properties are increased
ii) by applying single amino acid changes- through this alpha antitrypsin , insulin, tissue plasminogen activator, hirudin , dihydrofolate reductase and T4 lysozyme properties are improved
2) improving kinetic properties of enzyme -by protein engineering km property of different enzymes are improved like subtilisin, asparaginase, tyrosyl synthetase , restriction endonucleases.
application of protein engineering-
1) application of protein engineering in medical 2) application of protein engineering in food industry
3) application of protein engineering in different industrial processes
4) application of protein engineering in environment.
#Pharmaceutical_Biotechnology
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