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Binding Sites for Oligosaccharide Repeats from Lactic Acid Bacteria Exopolysaccharides on Bovine β-Lactoglobulin Identified by NMR Spectroscopy

[Image: see text] Lactic acid bacterial exopolysaccharides (EPS) are used in the food industry to improve the stability and rheological properties of fermented dairy products. β-Lactoglobulin (BLG), the dominant whey protein in bovine milk, is well known to bind small molecules such as fatty acids,...

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Autores principales: Birch, Johnny, Khan, Sanaullah, Madsen, Mikkel, Kjeldsen, Christian, Møller, Marie Sofie, Stender, Emil G. P., Peters, Günther H. J., Duus, Jens Ø., Kragelund, Birthe B., Svensson, Birte
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2021
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8028130/
https://www.ncbi.nlm.nih.gov/pubmed/33842774
http://dx.doi.org/10.1021/acsomega.1c00060
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author Birch, Johnny
Khan, Sanaullah
Madsen, Mikkel
Kjeldsen, Christian
Møller, Marie Sofie
Stender, Emil G. P.
Peters, Günther H. J.
Duus, Jens Ø.
Kragelund, Birthe B.
Svensson, Birte
author_facet Birch, Johnny
Khan, Sanaullah
Madsen, Mikkel
Kjeldsen, Christian
Møller, Marie Sofie
Stender, Emil G. P.
Peters, Günther H. J.
Duus, Jens Ø.
Kragelund, Birthe B.
Svensson, Birte
author_sort Birch, Johnny
collection PubMed
description [Image: see text] Lactic acid bacterial exopolysaccharides (EPS) are used in the food industry to improve the stability and rheological properties of fermented dairy products. β-Lactoglobulin (BLG), the dominant whey protein in bovine milk, is well known to bind small molecules such as fatty acids, vitamins, and flavors, and to interact with neutral and anionic polysaccharides used in food and pharmaceuticals. While sparse data are available on the affinity of EPS–milk protein interactions, structural information on BLG–EPS complexes, including the EPS binding sites, is completely lacking. Here, binding sites on BLG variant A (BLGA), for oligosaccharides prepared by mild acid hydrolysis of two EPS produced by Streptococcus thermophilus LY03 and Lactobacillus delbrueckii ssp. bulgaricus CNRZ 1187, respectively, are identified by NMR spectroscopy and supplemented by isothermal titration calorimetry (ITC) and molecular docking of complexes. Evidence of two binding sites (site 1 and site 2) on the surface of BLGA is achieved for both oligosaccharides (LY03-OS and 1187-OS) through NMR chemical shift perturbations, revealing multivalency of BLGA for EPS. The affinities of LY03-OS and 1187-OS for BLGA gave K(D) values in the mM range obtained by both NMR (pH 2.65) and ITC (pH 4.0). Molecular docking suggested that the BLGA and EPS complexes depend on hydrogen bonds and hydrophobic interactions. The findings provide insights into how BLGA engages structurally different EPS-derived oligosaccharides, which may facilitate the design of BLG–EPS complexation, of relevance for formulation of dairy products and improve understanding of BLGA coacervation.
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spelling pubmed-80281302021-04-09 Binding Sites for Oligosaccharide Repeats from Lactic Acid Bacteria Exopolysaccharides on Bovine β-Lactoglobulin Identified by NMR Spectroscopy Birch, Johnny Khan, Sanaullah Madsen, Mikkel Kjeldsen, Christian Møller, Marie Sofie Stender, Emil G. P. Peters, Günther H. J. Duus, Jens Ø. Kragelund, Birthe B. Svensson, Birte ACS Omega [Image: see text] Lactic acid bacterial exopolysaccharides (EPS) are used in the food industry to improve the stability and rheological properties of fermented dairy products. β-Lactoglobulin (BLG), the dominant whey protein in bovine milk, is well known to bind small molecules such as fatty acids, vitamins, and flavors, and to interact with neutral and anionic polysaccharides used in food and pharmaceuticals. While sparse data are available on the affinity of EPS–milk protein interactions, structural information on BLG–EPS complexes, including the EPS binding sites, is completely lacking. Here, binding sites on BLG variant A (BLGA), for oligosaccharides prepared by mild acid hydrolysis of two EPS produced by Streptococcus thermophilus LY03 and Lactobacillus delbrueckii ssp. bulgaricus CNRZ 1187, respectively, are identified by NMR spectroscopy and supplemented by isothermal titration calorimetry (ITC) and molecular docking of complexes. Evidence of two binding sites (site 1 and site 2) on the surface of BLGA is achieved for both oligosaccharides (LY03-OS and 1187-OS) through NMR chemical shift perturbations, revealing multivalency of BLGA for EPS. The affinities of LY03-OS and 1187-OS for BLGA gave K(D) values in the mM range obtained by both NMR (pH 2.65) and ITC (pH 4.0). Molecular docking suggested that the BLGA and EPS complexes depend on hydrogen bonds and hydrophobic interactions. The findings provide insights into how BLGA engages structurally different EPS-derived oligosaccharides, which may facilitate the design of BLG–EPS complexation, of relevance for formulation of dairy products and improve understanding of BLGA coacervation. American Chemical Society 2021-03-23 /pmc/articles/PMC8028130/ /pubmed/33842774 http://dx.doi.org/10.1021/acsomega.1c00060 Text en © 2021 The Authors. Published by American Chemical Society Permits non-commercial access and re-use, provided that author attribution and integrity are maintained; but does not permit creation of adaptations or other derivative works (https://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Birch, Johnny
Khan, Sanaullah
Madsen, Mikkel
Kjeldsen, Christian
Møller, Marie Sofie
Stender, Emil G. P.
Peters, Günther H. J.
Duus, Jens Ø.
Kragelund, Birthe B.
Svensson, Birte
Binding Sites for Oligosaccharide Repeats from Lactic Acid Bacteria Exopolysaccharides on Bovine β-Lactoglobulin Identified by NMR Spectroscopy
title Binding Sites for Oligosaccharide Repeats from Lactic Acid Bacteria Exopolysaccharides on Bovine β-Lactoglobulin Identified by NMR Spectroscopy
title_full Binding Sites for Oligosaccharide Repeats from Lactic Acid Bacteria Exopolysaccharides on Bovine β-Lactoglobulin Identified by NMR Spectroscopy
title_fullStr Binding Sites for Oligosaccharide Repeats from Lactic Acid Bacteria Exopolysaccharides on Bovine β-Lactoglobulin Identified by NMR Spectroscopy
title_full_unstemmed Binding Sites for Oligosaccharide Repeats from Lactic Acid Bacteria Exopolysaccharides on Bovine β-Lactoglobulin Identified by NMR Spectroscopy
title_short Binding Sites for Oligosaccharide Repeats from Lactic Acid Bacteria Exopolysaccharides on Bovine β-Lactoglobulin Identified by NMR Spectroscopy
title_sort binding sites for oligosaccharide repeats from lactic acid bacteria exopolysaccharides on bovine β-lactoglobulin identified by nmr spectroscopy
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8028130/
https://www.ncbi.nlm.nih.gov/pubmed/33842774
http://dx.doi.org/10.1021/acsomega.1c00060
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