Possible Candidates for Future Protein Engineering,Molecular Memory,Synthesis of Phenoxy Propionic,An Azole Antifungal Agent,Bromo Propionic

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Protein and Enzymes Engineering Protein and Enzymes Engineering - Introduction Engineering of Macromolecules Basic Outline Protein Engineering vs Enzyme Engineering for a Biocatalyst Protein Engineering Rationale of Protein Enzyme Engineering Basic Assumptions for Protein Engineering Key Biocatalyst Properties that are Altered Through Protein Engineering Some Examples of Improved Kinetic Parameters of Enzymes Turnover Determined by k_cat Selectivity and Specificity Some Technologies Available for Enchancing Biocatalyst Characteristics Molecular Stability Catalytic Diversity Acquiring Knowledge for Protein Engineering Study of Protein Structure Three Dimensional Protein Modeling Perturbation Theory

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Possible Candidates for Future Protein Engineering
Fol1owing are some of the proteins, whose structures are known and for which cloned genes are available, so that they can be used for protein engineering: (i) immunoglobulins, (ii) glucose isomerases, (iii) amylases, (iv) p-hydroxybenzoate hydroxylase, (v) Ribulose – 1, 5, bisphosphate carboxylase (RUBP), (vi) repressor proteins, and (g) restriction edonucleases.

Enzymes in organic solvents: molecular memory, or imprinting
Organic solvent also exhibit memory effect (also called imprinting) which is absent in aqueous medium. For instance, lyophilized α-chymotrypsin, first dissolved in water and then diluted 100-fold with t-amyl alcohol has a much higher specific activity than if the lyophilized enzyme is suspended in solvent containing same 1 % of water. In another case, if subtilisin is lyophilized from aq. solution containing various inhibitors (followed by their removal by anhydrous extraction), it is upto 100 times more active in anhydrous solvents, than if the enzyme is lyophilized in the absence of ligands. The enzyme memory disappears, if enzyme is dissolved in water.

Molecular imprinting due to an anhydrous solvent

Molecular Imprinting Due to an Anhydrous Solvent, Following Anhydrous Extraction of a Ligand, Which Brings About a Conformational Change in enzyme in Water

1. Ligand

2. In water (a conformational change)

3. Lyophilization

4. Anhydrous extractionof the ligand

5. Molecular imprint

6. Placementin

7. Anhydrious Solvent

8. Imprint retained

9. Aqueous solution

10. Imprint Disappears

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Practical applications of enzymes in organic solvents. (i) Synthesis of phenoxy-propionic herbicides and drugs.
Enantiopure 2-chloro- and 2 bromo-propionic acids as intermediates are catalysed by yeast lipase through enantioselective butanolysis in anhydrous solvents (this process is scaled up to multi-kilogram level in Austria) (ii) An azole antifungal agent has been synthesized in hundred kilogram quantity due to acetylation of a symmetrical diol that involved a stereoselective step catalysed by yeast lipase in acetonitrile. The compound was in phase III trials in 2001.

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Methods for Protein Engineering Site Directed Mutagenesis in Vivo Gene Modification Through Oligonucleotide Synthesis Breeding a Better Catalyst Using Random Events Artificial Random Mutagenesis Random DNA Shuffing Using Recombination Combination of Structure Based and Envolutionary Approaches Multienzyme Systems bi and Polyfunctional Enzymes by Gene Fusion Chemical Modification of Enzymes Production of Site Specific Nucleases Production of Artifical Semi Synthetic Oxido Reductases Flavo Enzyme Hybrid Enzymes Combinatorial Chemistry Organic Synthesis Libraries De Novo Design of Catalysts Some Early Achievements of Protein Engineering Possible Candidates for Future Protein Engineering Improving Enzymes by Using Organic Solvents