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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Bibliographische Detailangaben
Hauptverfasser:	COLEMAN, WILLIAM J, DILWORTH, MICHAEL R, SILVA, CHRISTOPHER M, YANG, MARY, YOUVAN, DOUGLAS C, BYLINA, EDWARD J
Format:	Patent
Sprache:	eng ; fre
Schlagworte:	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
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Beschreibung
Zusammenfassung:	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.