Crude gliadins isolated from three wheat varieties (Karl, Tam 107 and CS-
93) were subjected to in vitro hydrolysis by the extracellular enzymes pepsin,
peptidase and pancreatin. Gliadins from one variety (Karl) also were exposed
to the intracellular enzymes cathepsins Band D. A reverse phase high
performance liquid chromatography (RP-HPLC) method was developed to
separate unhydrolyzed and hydrolyzed gliadins. The unhydrolyzed crude
gliadins were resolved into 12-14 peaks, with at least five peaks that appeared
to be common to all three wheat varieties. Gliadin peptides were resolved
into between 44 and 71 peaks, suggesting that a large numbers of peptides are
derived from proteins present in more than one of the four major gliadin
fractions (i.e.; α, β, γ, and ω - gliadins). No major differences were detected
between chromatograms of extracellular digests and those of
extracellular /intracellular digests indicating that cathepsins Band D may not
contribute to more complete gliadin digestion. The molecular weights and
amino acid sequences of gliadin peptides will need to be determined by HPLC mass
spectrometry (HPLC-MS) for accurate qualitative and/or quantitative
comparisons of individual digests. Our in vitro hydrolysis/RP-HPLC
methods may be applicable, however, in the generation of celiac active
peptides for future toxicity testing.
next to an author's name indicates that all files or directories associated with their ETD are accessible from the Virginia Tech campus network only.
![[VT]](http://scholar.lib.vt.edu/images/ETD-db/restricted.gif)
