Molecular chemical structure of barley proteins revealed by ultra‐spatially resolved synchrotron light sourced FTIR microspectroscopy: Comparison of barley varieties

Barley protein structure affects the barley quality, fermentation, and degradation behavior in both humans and animals among other factors such as protein matrix. Publications show various biological differences among barley varieties such as Valier and Harrington, which have significantly different degradation behaviors. The objectives of this study were to reveal the molecular structure of barley protein, comparing various varieties (Dolly, Valier, Harrington, LP955, AC Metcalfe, and Sisler), and quantify protein structure profiles using Gaussian and Lorentzian methods of multi‐component peak modeling by using the ultra‐spatially resolved synchrotron light sourced Fourier transform infrared microspectroscopy (SFTIRM). The items of the protein molecular structure revealed included protein structure α‐helices, β‐sheets, and others such as β‐turns and random coils. The experiment was performed at the National Synchrotron Light Source in Brookhaven National Laboratory (BNL, US Department of Energy, NY). The results showed that with the SFTIRM, the molecular structure of barley protein could be revealed. Barley protein structures exhibited significant differences among the varieties in terms of proportion and ratio of model‐fitted α‐helices, β‐sheets, and others. By using multi‐component peaks modeling at protein amide I region of 1710–1576 cm −1 , the results show that barley protein consisted of approximately 18–34% of α‐helices, 14–25% of β‐sheets, and 44–69% others. AC Metcalfe, Sisler, and LP955 consisted of higher ( P < 0.05) proportions of α‐helices (30–34%) than Dolly and Valier (α‐helices 18–23%). Harrington was in between which was 25%. For protein β‐sheets, AC Metcalfe, and LP955 consisted of higher proportions (22–25%) than Dolly and Valier (13–17%). Different barley varieties contained different α‐helix to β‐sheet ratios, ranging from 1.4 to 2.0, although the difference were insignificant ( P > 0.05). The ratio of α‐helices to others (0.3 to 1.0, P < 0.05) and that of β‐sheets to others (0.2 to 0.8, P < 0.05) were different among the barley varieties. It needs to be pointed out that using a multi‐peak modeling for protein structure analysis is only for making relative estimates and not exact determinations and only for the comparison purpose between varieties. The principal component analysis showed that protein amide I Fourier self‐deconvolution spectra were different among the barley varieties, indicating that protein internal molecular structure differed. The above results demonstrate the potential of the SFTIRM to localize relatively pure protein areas in barley tissues and reveal protein molecular structure. The results indicated relative differences in protein structures among the barley varieties, which may partly explain the biological differences among the barley varieties. Further study is needed to understand the relationship between barley molecular chemical structure and biological features in terms of nutrient availability and digestive behavior. © 2006 Wiley Periodicals, Inc. Biopolymers 85:308–317, 2007. This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com