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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,...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical Society
2021
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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. |
format | Online Article Text |
id | pubmed-8028130 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | American Chemical Society |
record_format | MEDLINE/PubMed |
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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