Microbial sensors for small molecules: development of a mevalonate biosensor.
We describe a novel biosensor strain for detection and quantification of a small molecule, mevalonate. The biosensor strain is an Escherichia coli mevalonate auxotroph that expresses the green fluorescent protein and reports on the mevalonate concentration in the growth medium through a change in growth rate. A model describing the growth rate dependence on mevalonate was developed in order to use the biosensor strain for high-throughput screening (HTS) and quantitative measurement of mevalonate in the extracellular environment. In general, this method should be applicable to the quantification of any small molecule for which an auxotroph can be developed and will be useful for HTS of evolved metabolic pathways for which there is no readily available screen or selection.
Pfleger
Brian F
BF
Department of Chemical Engineering, University of California-Berkeley, Berkeley, CA 94720-1462, USA.
Pitera
Douglas J
DJ
Newman
Jack D
JD
Martin
Vincent J J
VJ
Keasling
Jay D
JD
D019747:Aldose-Ketose Isomerases / Q000502:physiology; D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods*; D004926:Escherichia coli / Q000378:metabolism*; D008401:Gas Chromatography-Mass Spectrometry; D008798:Mevalonic Acid / Q000032:analysis*; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000502:physiology; D010088:Oxidoreductases / Q000502:physiology; D013729:Terpenes / Q000378:metabolism
D009097:Multienzyme Complexes; D013729:Terpenes; D010088:Oxidoreductases; C114231:1-deoxy-D-xylulose 5-phosphate reductoisomerase; D019747:Aldose-Ketose Isomerases; D008798:Mevalonic Acid