A FLUOROMETRIC ASSAY FOR DETERMINING VARIATION OF LYSINE IN WHEAT GLIADIN PROTEINS

An effective method for estimating lysine in wheat gliadin proteins could contribute to increasing lysine in wheat. Wheat gliadin proteins were separated and collected by reverse phase high performance liquid chromatography (RP-HPLC). A fluorimetric assay with o-phthalaldehyde (OPA) was used to determine the lysine content of wheat gliadin proteins. The OPA reagent reacts specifically with the amino group of lysine in protein. Twenty fractions of wheat gliadins were collected and analyzed by the fluorimetric assay. Nine of these fractions were also analyzed for lysine content by an amino acid analyzer. The results obtained from the fluorimetric assay were significantly related to the results obtained from the amino acid analyzer (R = 0.93 for quadratic regression of the nine selected gliadin fractions). Lysine content of the wheat gliadins varied from 0.6 to 1.4 percent of the protein. This study determined that the fluorimetric assay could accurately estimate lysine in wheat gliadin proteins. Identification of high-lysine gliadin subunits could be implemented into a program of breeding for increased lysine in wheat.