1. |
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Introduction
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Biocatalysis, green chemistry, why proteins need to be engineered |
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Introduction ProSAR approach
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Biocatalysis, green chemistry, why proteins need to be engineered |
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2. |
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Enzyme Stability
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Measuring protein stability |
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Engineer Stable Proteins
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Strategies to rationally engineer more stable proteins |
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Irreversible unfolding via MG
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Strategies to rationally engineer more stable proteins |
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3. |
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Making enzymes faster
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Measuring reaction rates (MM kinetics) / Strategies to increase reaction rates |
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4. |
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Measure Selectivity
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Measuring enzyme selectivity |
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Engineer Selectivity
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Strategies to increase selectivity |
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5. |
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Computer Modeling Overview
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Viewing protein structures/
Molecular mechanics approach to modeling |
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Critical evaluation of computer model
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Extending molecular mechanics to model protein stability & pH dependence / Modeling selectivity (conformations & molecular dynamics) / Modeling reactivity (QM and other approaches) |
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6. |
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Mutagenesis Single Substitution
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Overview of directed evolution: screening, random mutagenesis / Mutagenesis using error prone PCR - methods, polymerase fidelity |
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Better NNK
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Saturation mutagenesis approaches |
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Circular permuation
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Saturation mutagenesis approaches |
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7. |
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Multiple Substitutions
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Bioinformatics approaches / Stepwise combinations of substitutions /Adding multiple mutations simultaneously |
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8. |
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Recombination Methods
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Recombination methods - limitations, homology |
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9. |
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Genomic Approaches
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Other approaches to mutagenesis (e.g. circular permutations, neutral drift libraries) |
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10. |
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Screening
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Screening strategies |
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Screening
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Screening strategies |
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Selection
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Selection strategies |
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11. |
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open topic
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Expression Vectors |
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12. |
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open topic
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Metabolic Engineer |
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