Using advanced vibrational molecular spectroscopy (ATR-Ft/IRS) to study heating process induced changes on protein molecular structure of biodegradation residues in cool-climate adapted faba bean seeds: Relationship with rumen and intestinal protein digestion in ruminant systems.

The objective of this study was to evaluate the effects of heating process on protein molecular structure from ruminal degradation residues in cool-climate adapted faba bean seeds in relation to crude protein (CP), in situ degradation kinetics, rumen protein degradation and intestinal protein digestion parameters in dairy cows. Seeds of six faba bean varieties with low (Snowbird, Snowdrop, 219_16) and normal tannin (Fatima, 346_10, SSNS_1) were collected from three different locations, and were heated 3 min by microwave irradiation (MI, dry heating) or heated 1 h by steam pressure toasting (SP, moist heating) or kept raw as a control. Heat treated samples were used for rumen incubating 24, 12, 8, 4, 2, 0 h(s) in two replicate runs and then residues from 12 h of rumen degradation were used for three steps in vitro technique for determining intestinal protein digestion. Attenuated total reflectance Fourier transforms infrared spectroscopy (ATR-Ft/IRS) was used for analyzing protien molecular structure of residual faba bean seeds. The results showed that SP increased the intensities of amide I, amide II, α-helix and β-sheet but decreased amide I to amide II height and area ratio, α-helix to β-sheet height ratio from 12 and 24 h of ruminal degradation, and MI decreased all the intensities of amide I, amide II, α-helix and β-sheet and ratios except amide I to amide II area ratio of residues from 24 h of ruminal degradation. Additionally, the intensities of amide I, amide II, α-helix and β-sheet had a unique pattern of increasing first and then decreasing with the increasing ruminal digestion time for SP treatment, while amide I to amide II height and area ratio, α-helix to β-sheet height ratio were declining. For the MI groups, this pattern was not observed and the intensities were rather consistent across the digestion process. Rumen protein degradation parameters including rumen bypass crude protein (BCP) or rumen undegradable protein (RUP) and rumen degradable protein (RDP) closely correlated with protein molecular structure of to peak heights, areas and ratios. Regression equations based on residual protein molecular structure presented a good estimation power for soluble fraction (S, R = 0.79), degradable fraction (D, R = 0.805), BCP (R = 0.941), RUP (R = 0.941) and RDP (R = 0.811). Overall, heat-induced changes in rumen residual protein molecular structures were related to CP, in situ degradation kinetics, rumen protein degradation and rumen protein digestion parameters.