A semi-quantitative high-throughput screening method for microbial L-tyrosine production in microtiter plates.
In this study, we developed a fluorescence-based assay for quantifying the aromatic amino acid L-tyrosine in small sample volumes employing 96-well microtiter plates. The method was based on the specific derivatisation of L-tyrosine with 1-nitroso-2-naphthol and the formation of a stable complex for the fluorescent determination of L-tyrosine concentrations. The original procedure for L-tyrosine measurements in blood or tissue samples (Waalkes and Udenfriend in J Lab Clin Med 50:733-736, 1957) was modified to a simple assay suitable for high-throughput screening of L-tyrosine producing microorganisms such as engineered Escherichia coli.
Lütke-Eversloh
Tina
T
Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Stephanopoulos
Gregory
G
D001681:Biological Assay / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005453:Fluorescence; D005818:Genetic Engineering; D017186:Titrimetry / Q000379:methods; D014443:Tyrosine / Q000032:analysis* / Q000378:metabolism
D014443:Tyrosine