Immunohistological visualization of the effect of soybean processing on Kunitz trypsin inhibitor
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Immunohistological visualization of the effect of soybean processing on Kunitz trypsin inhibitor. / Mravec, Jozef; Jørgensen, Bodil; Pedersen, Ninfa.
I: Animal Feed Science and Technology, Bind 292, 115410, 2022.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - Immunohistological visualization of the effect of soybean processing on Kunitz trypsin inhibitor
AU - Mravec, Jozef
AU - Jørgensen, Bodil
AU - Pedersen, Ninfa
N1 - Publisher Copyright: © 2022 The Authors
PY - 2022
Y1 - 2022
N2 - Anti-nutritional factors such as trypsin inhibitors represent a major hindrance in the usage of soybean-based products as feed for non-ruminant feedstock. Here we studied the effect of four different methods of soybean processing to generate soybean meal (oil-extraction with a solvent, toasting, mechanical extraction, and fermentation) on the Kunitz trypsin inhibitor and soy protein. We visualized both proteins in situ using indirect immunofluorescence on resin sections with specific rabbit-raised polyclonal antibodies and laser-scanning confocal microscopy We characterized their cellular localization in raw soybeans and show that Kunitz trypsin inhibitor and soy protein occupy different domains related to protein globoids. Semi-quantification of the signal intensities showed an only moderate reduction in detectable Kunitz trypsin inhibitor epitopes after any type of processing. On the other hand, a considerable loss of trypsin inhibitor activity was shown with a standardized assay. The effect of toasting can be seen as significantly increased green fluorescence possibly due to the production of advanced glycosylation end products. This work introduces antibody-based in situ visualization as a new promising way to study anti-nutritional factors in soybean-derived products.
AB - Anti-nutritional factors such as trypsin inhibitors represent a major hindrance in the usage of soybean-based products as feed for non-ruminant feedstock. Here we studied the effect of four different methods of soybean processing to generate soybean meal (oil-extraction with a solvent, toasting, mechanical extraction, and fermentation) on the Kunitz trypsin inhibitor and soy protein. We visualized both proteins in situ using indirect immunofluorescence on resin sections with specific rabbit-raised polyclonal antibodies and laser-scanning confocal microscopy We characterized their cellular localization in raw soybeans and show that Kunitz trypsin inhibitor and soy protein occupy different domains related to protein globoids. Semi-quantification of the signal intensities showed an only moderate reduction in detectable Kunitz trypsin inhibitor epitopes after any type of processing. On the other hand, a considerable loss of trypsin inhibitor activity was shown with a standardized assay. The effect of toasting can be seen as significantly increased green fluorescence possibly due to the production of advanced glycosylation end products. This work introduces antibody-based in situ visualization as a new promising way to study anti-nutritional factors in soybean-derived products.
KW - Anti-nutritional factors
KW - Immunofluorescence
KW - Laser-scanning confocal microscopy
KW - Maillard reaction
KW - Soybean meal
KW - Trypsin inhibitor
U2 - 10.1016/j.anifeedsci.2022.115410
DO - 10.1016/j.anifeedsci.2022.115410
M3 - Journal article
AN - SCOPUS:85136502353
VL - 292
JO - Animal Feed Science and Technology
JF - Animal Feed Science and Technology
SN - 0377-8401
M1 - 115410
ER -
ID: 319476048