Open AccessAssessment of Colorimetric Reporter Enzymes in the PURE System
Author(s) -
Caitlin E Sharpes,
J. Barry McManus,
Steven M Blum,
Glory E Mgboji,
Matthew W. Lux
Publication year - 2021
Publication title -
acs synthetic biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.156
H-Index - 66
ISSN - 2161-5063
DOI - 10.1021/acssynbio.1c00360
Subject(s) - multiplex , enzyme , biosensor , substrate (aquarium) , analyte , computational biology , pairing , computer science , reporter gene , combinatorial chemistry , biochemistry , chemistry , biological system , nanotechnology , biochemical engineering , biology , gene , gene expression , materials science , chromatography , bioinformatics , physics , engineering , ecology , superconductivity , quantum mechanics
Colorimetric reporter enzymes are useful for generating eye-readable biosensor readouts that do not require a device to interpret, an attractive property for applications in remote or developing parts of the world. The use of cell-free gene expression further facilitates such applications via amenability to lyophilization and incorporation into materials like paper. Currently, detection of multiple analytes simultaneously with these systems requires multiple reactions or a device. Here we evaluate seven enzymes and 15 corresponding substrates for functionality in a particular cell-free expression system known as PURE. We report eight enzyme/substrate pairs spanning four enzymes that are compatible with PURE. Of the four enzymes, three pairings exhibit no cross-reactivity. We finally show that at least one pairing can be used to create a third color when both are present, highlighting the potential use of these reporters for multiplex sensing.