Management of the diffusion of 4-methylumbelliferone across phases in microdroplet-based systems for in vitro protein evolution
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Wu, Nan; Courtois, Fabienne; Zhu, Yonggang; Oakeshott, John Graham; Easton, Christopher; Abell, Christopher
Description
Fluorongenic reagents based on 4-methylumbelliferone (4-MU) have been widely used for the detection of phosphatase, sulfatase, esterase, lipase and glycosidase activities in conventionally formatted enzyme assay systems. However, the sensitivity of assays based on these substrates is also potentially very useful in the microdroplet formats now being developed for high throughput in vitro evolution experiments. In this article, we report the investigation of diffusion of 4-MU as a model dye from...[Show more]
dc.contributor.author | Wu, Nan | |
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dc.contributor.author | Courtois, Fabienne | |
dc.contributor.author | Zhu, Yonggang | |
dc.contributor.author | Oakeshott, John Graham | |
dc.contributor.author | Easton, Christopher | |
dc.contributor.author | Abell, Christopher | |
dc.date.accessioned | 2015-12-10T22:41:09Z | |
dc.identifier.issn | 0173-0835 | |
dc.identifier.uri | http://hdl.handle.net/1885/57761 | |
dc.description.abstract | Fluorongenic reagents based on 4-methylumbelliferone (4-MU) have been widely used for the detection of phosphatase, sulfatase, esterase, lipase and glycosidase activities in conventionally formatted enzyme assay systems. However, the sensitivity of assays based on these substrates is also potentially very useful in the microdroplet formats now being developed for high throughput in vitro evolution experiments. In this article, we report the investigation of diffusion of 4-MU as a model dye from water-in-oil droplets and the internal aqueous phase of water-in-oil-in-water droplets in microfluidics. The effect of BSA in the aqueous phase on the diffusion of 4-MU is also discussed. Based on these results, we provided here proof-of-concept of the reaction of the enzyme OpdA with the substrate coumaphos in water-in-oil-in-water droplets. In this double-emulsion system, the reaction of OpdA and coumaphos was achieved by allowing coumaphos to diffuse from the continuous aqueous phase across the oil phase into the internal aqueous droplets. | |
dc.publisher | Wiley-VCH Verlag GMBH | |
dc.source | Electrophoresis | |
dc.subject | Keywords: coumafos; hymecromone; phosphotriesterase; water oil cream; article; diffusion; enzyme substrate complex; microfluidic analysis; phase transition; Animals; Cattle; Coumaphos; Diffusion; Emulsions; Enzyme Assays; Fluorescent Dyes; Hymecromone; Micelles; Mi Directed evolution; Microfluidics; Single- and double-emulsion droplets; Substrate diffusion | |
dc.title | Management of the diffusion of 4-methylumbelliferone across phases in microdroplet-based systems for in vitro protein evolution | |
dc.type | Journal article | |
local.description.notes | Imported from ARIES | |
local.identifier.citationvolume | 31 | |
dc.date.issued | 2010 | |
local.identifier.absfor | 030302 - Nanochemistry and Supramolecular Chemistry | |
local.identifier.ariespublication | u4005981xPUB414 | |
local.type.status | Published Version | |
local.contributor.affiliation | Wu, Nan, College of Physical and Mathematical Sciences, ANU | |
local.contributor.affiliation | Courtois, Fabienne, University of Cambridge | |
local.contributor.affiliation | Zhu, Yonggang, CSIRO Materials Science and Engineering | |
local.contributor.affiliation | Oakeshott, John Graham, CSIRO Division of Entomology | |
local.contributor.affiliation | Easton, Christopher, College of Physical and Mathematical Sciences, ANU | |
local.contributor.affiliation | Abell, Christopher, University of Cambridge | |
local.description.embargo | 2037-12-31 | |
local.bibliographicCitation.issue | 18 | |
local.bibliographicCitation.startpage | 3121 | |
local.bibliographicCitation.lastpage | 3128 | |
local.identifier.doi | 10.1002/elps.201000140 | |
local.identifier.absseo | 970103 - Expanding Knowledge in the Chemical Sciences | |
dc.date.updated | 2016-02-24T10:23:40Z | |
local.identifier.scopusID | 2-s2.0-77956940537 | |
local.identifier.thomsonID | 000283389900015 | |
Collections | ANU Research Publications |
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