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A Versatile High Throughput Screen for Dioxygenase Activity Using Solid-Phase Digital Imaging

Joern, John M. and Sakamoto, Takeshi and Arisawa, Akira and Arnold, Frances H. (2001) A Versatile High Throughput Screen for Dioxygenase Activity Using Solid-Phase Digital Imaging. Journal of Biomolecular Screening, 6 (4). pp. 219-223. ISSN 1087-0571. https://resolver.caltech.edu/CaltechAUTHORS:20180824-101334951

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Abstract

We have developed a solid-phase, high throughput (10,000 clones/day) screen for dioxygenase activity. The cis-di- hydrodiol product of dioxygenase bioconversion is converted to a phenol by acidification or to a catechol by reaction with cis-dihydrodiol dehydrogenase. Gibbs reagent reacts quickly with these oxygenated aromatics to yield colored products that are quantifiable using a microplate reader or by digital imaging and image analysis. The method is reproducible and quantitative at product concentrations of only 30,uM, with essentially no background from media components. This method is an effective general screen for aromatic oxidation and should be a useful tool for the discovery and directed evolution of oxygenases.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1177/108705710100600403DOIArticle
ORCID:
AuthorORCID
Arnold, Frances H.0000-0002-4027-364X
Additional Information:© 2001 The Society for Biomolecular Screening. This research is supported by the National Science Foundation (Grant# BES-9981770) and by Maxygen, Inc.
Funders:
Funding AgencyGrant Number
NSFBES-9981770
Maxygen, Inc.UNSPECIFIED
Issue or Number:4
Record Number:CaltechAUTHORS:20180824-101334951
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20180824-101334951
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:89147
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:24 Aug 2018 17:26
Last Modified:03 Oct 2019 20:13

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