Titlebook: Directed Enzyme Evolution; Screening and Select Frances H. Arnold,George Georgiou Book 2003 Humana Press 2003

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Reverie

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Solid-Phase Screening Using Digital Image Analysise of its relative ease and high throughput compared to liquid-phase screening in multi-well plates. As with any high throughput screening approach, a suitable colorimetric or fluorimetric assay must exist (or be developed) for the enzyme function in question. There are two major requirements that mu

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Screening for Thermostabilityxquisite specificity and selectivity, they are often limited by insufficient stability. Improvements in enzyme activity through protein engineering often come at the cost of reduced stability. This is likely a result of both natural drift and a tradeoff that often exists between activity and stabili

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Spongy-Bone

High-Throughput Carbon Monoxide Binding Assay for Cytochromes P450(.). They contain a noncovalently bound protoporphyrin IX, giving these enzymes characteristic spectral properties. This heme has an available sixth coordination ligand that is able to bind carbon monoxide. Difference spectroscopy yields a spectral peak at approx 450 nm when comparing bound and unbo

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Bureaucracy

High-Throughput Screens Based on NAD(P)H Depletionic) substrates are used and no simple and reliable high-throughput method for quantitative analysis of the respective reaction product is available. The screening procedure should also be generally applicable for a certain class of enzymes. Generation of “surrogate substrates” by derivatization with