Biotransformation
Biocatalysis and Biotransformation
Redesigning existing enzymes by protein engineering
Metabolic engineering for multistep biotransformation
Cyanide Biotransformation
Redesigning existing enzymes by protein engineering
WHAT IS PROTEIN ENGINEERING???
It is a process of developing useful or valuable proteins which involves * Modification of existing enzymes * Production of new enzymes
In other words, the design and construction of new proteins or enzymes with novel or desired functions, through the modification of amino acid sequences using recombinant DNA technology.
What is the need??? * Native proteins are not well suited for industrial application * Native proteins are not optimized for medicinal purposes
Protein Engineering is used to: * Increase the efficiency of enzyme-catalyzed reactions * Eliminate the need for cofactor in enzymatic reaction * Change substrate binding site to increase specificity * Change the thermal tolerance * Change the pH stability * Increase proteins resistance to proteases (purification).
General Strategies…
1. Rational Design
The scientist uses detailed knowledge of the structure and function of the protein to make desired changes.
Advantages:
* Inexpensive * Technically easy
Disadvantages:
* Detailed structural knowledge of a protein is often unavailable. * Extremely difficult to predict the effects of various mutations.
Site Directed Mutagenesis * Directed to a specific site in a pre-determined way, hence the name. * Involves changing the base sequence of DNA and testing its effect on gene or DNA function. * Can be done in vitro on cloned DNA or in vivo by homologous recombination.
2. Directed Evolution * Random mutagenesis is applied to a protein, and a selection regime is used to pick out variants that have the desired qualities. * Produces superior results to
Redesigning existing enzymes by protein engineering
Metabolic engineering for multistep biotransformation
Cyanide Biotransformation
Redesigning existing enzymes by protein engineering
WHAT IS PROTEIN ENGINEERING???
It is a process of developing useful or valuable proteins which involves * Modification of existing enzymes * Production of new enzymes
In other words, the design and construction of new proteins or enzymes with novel or desired functions, through the modification of amino acid sequences using recombinant DNA technology.
What is the need??? * Native proteins are not well suited for industrial application * Native proteins are not optimized for medicinal purposes
Protein Engineering is used to: * Increase the efficiency of enzyme-catalyzed reactions * Eliminate the need for cofactor in enzymatic reaction * Change substrate binding site to increase specificity * Change the thermal tolerance * Change the pH stability * Increase proteins resistance to proteases (purification).
General Strategies…
1. Rational Design
The scientist uses detailed knowledge of the structure and function of the protein to make desired changes.
Advantages:
* Inexpensive * Technically easy
Disadvantages:
* Detailed structural knowledge of a protein is often unavailable. * Extremely difficult to predict the effects of various mutations.
Site Directed Mutagenesis * Directed to a specific site in a pre-determined way, hence the name. * Involves changing the base sequence of DNA and testing its effect on gene or DNA function. * Can be done in vitro on cloned DNA or in vivo by homologous recombination.
2. Directed Evolution * Random mutagenesis is applied to a protein, and a selection regime is used to pick out variants that have the desired qualities. * Produces superior results to