SOLID PHASE ENZYME KINETICS SCREENING IN MICROCOLONIES

A MicroColonylmager instrument and solid phase methods to screen cells expressing mutagenized enzymes for enhanced activity. The MicroColonylmager instument and methods permit high throughput screening of enzyme libraries by time course analyses of single-pixels, using either absorption, fluorescenc...

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Hauptverfasser: COLEMAN, WILLIAM J, DILWORTH, MICHAEL R, SILVA, CHRISTOPHER M, YANG, MARY, YOUVAN, DOUGLAS C, BYLINA, EDWARD J
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creator COLEMAN, WILLIAM J
DILWORTH, MICHAEL R
SILVA, CHRISTOPHER M
YANG, MARY
YOUVAN, DOUGLAS C
BYLINA, EDWARD J
description A MicroColonylmager instrument and solid phase methods to screen cells expressing mutagenized enzymes for enhanced activity. The MicroColonylmager instument and methods permit high throughput screening of enzyme libraries by time course analyses of single-pixels, using either absorption, fluorescence or FRET. This high throughput assay can detect small differences in enzyme rates within microcolonies grown at a nearly confluent density on an assay disk. Each microcolony is analyzed simultaneously at single-pixel resolution, requiring less than 100 nl substrate/measurement. By simultaneously assaying different substrates tagged with spectrally distinct chromogenic or fluorogenic reporters, the substrate specificity of an enzyme can be changed.
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subjects APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY
BEER
BIOCHEMISTRY
CHEMISTRY
COMPOSITIONS OR TEST PAPERS THEREFOR
COMPOSITIONS THEREOF
CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES
CULTURE MEDIA
ENZYMOLOGY
INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES
MEASURING
MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS
METALLURGY
MICROBIOLOGY
MICROORGANISMS OR ENZYMES
MUTATION OR GENETIC ENGINEERING
PHYSICS
PROCESSES OF PREPARING SUCH COMPOSITIONS
PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS
SPIRITS
TESTING
VINEGAR
WINE
title SOLID PHASE ENZYME KINETICS SCREENING IN MICROCOLONIES
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