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Fungus-derived protein particles as cell-adhesive matrices for cell-cultivated food

Cell-adhesive factors mediate adhesion of cells to substrates via peptide motifs such as the Arg–Gly–Asp (RGD) sequence. With the onset of sustainability issues, there is a pressing need to find alternatives to animal-derived cell-adhesive factors, especially for cell-cultivated food applications. I...

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Autores principales: Teo, Yu Xing, Lee, Kah Yin, Goh, Corinna Jie Hui, Wang, Loo Chien, Sobota, Radoslaw M., Chiam, Keng-Hwee, Du, Chan, Wan, Andrew C. A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10344888/
https://www.ncbi.nlm.nih.gov/pubmed/37443321
http://dx.doi.org/10.1038/s41538-023-00209-y
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author Teo, Yu Xing
Lee, Kah Yin
Goh, Corinna Jie Hui
Wang, Loo Chien
Sobota, Radoslaw M.
Chiam, Keng-Hwee
Du, Chan
Wan, Andrew C. A.
author_facet Teo, Yu Xing
Lee, Kah Yin
Goh, Corinna Jie Hui
Wang, Loo Chien
Sobota, Radoslaw M.
Chiam, Keng-Hwee
Du, Chan
Wan, Andrew C. A.
author_sort Teo, Yu Xing
collection PubMed
description Cell-adhesive factors mediate adhesion of cells to substrates via peptide motifs such as the Arg–Gly–Asp (RGD) sequence. With the onset of sustainability issues, there is a pressing need to find alternatives to animal-derived cell-adhesive factors, especially for cell-cultivated food applications. In this paper, we show how data mining can be a powerful approach toward identifying fungal-derived cell-adhesive proteins and present a method to isolate and utilize these proteins as extracellular matrices (ECM) to support cell adhesion and culture in 3D. Screening of a protein database for fungal and plant proteins uncovered that ~5.5% of the unique reported proteins contain RGD sequences. A plot of fungi species vs RGD percentage revealed that 98% of the species exhibited an RGD percentage > = 1%. We observed the formation of protein particles in crude extracts isolated from basidiomycete fungi, which could be correlated to their stability towards particle aggregation at different temperatures. These protein particles were incorporated in 3D fiber matrices encapsulating mouse myoblast cells, showing a positive effect on cell alignment. We demonstrated a cell traction stress on the protein particles (from Flammulina velutipes) that was comparable to cells on fibronectin. A snapshot of the RGD-containing proteins in the fungal extracts was obtained by combining SDS-PAGE and mass spectrometry of the peptide fragments obtained by enzymatic cleavage. Therefore, a sustainable source of cell-adhesive proteins is widely available in the fungi kingdom. A method has been developed to identify candidate species and produce cell-adhesive matrices, applicable to the cell-cultivated food and healthcare industries.
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spelling pubmed-103448882023-07-15 Fungus-derived protein particles as cell-adhesive matrices for cell-cultivated food Teo, Yu Xing Lee, Kah Yin Goh, Corinna Jie Hui Wang, Loo Chien Sobota, Radoslaw M. Chiam, Keng-Hwee Du, Chan Wan, Andrew C. A. NPJ Sci Food Article Cell-adhesive factors mediate adhesion of cells to substrates via peptide motifs such as the Arg–Gly–Asp (RGD) sequence. With the onset of sustainability issues, there is a pressing need to find alternatives to animal-derived cell-adhesive factors, especially for cell-cultivated food applications. In this paper, we show how data mining can be a powerful approach toward identifying fungal-derived cell-adhesive proteins and present a method to isolate and utilize these proteins as extracellular matrices (ECM) to support cell adhesion and culture in 3D. Screening of a protein database for fungal and plant proteins uncovered that ~5.5% of the unique reported proteins contain RGD sequences. A plot of fungi species vs RGD percentage revealed that 98% of the species exhibited an RGD percentage > = 1%. We observed the formation of protein particles in crude extracts isolated from basidiomycete fungi, which could be correlated to their stability towards particle aggregation at different temperatures. These protein particles were incorporated in 3D fiber matrices encapsulating mouse myoblast cells, showing a positive effect on cell alignment. We demonstrated a cell traction stress on the protein particles (from Flammulina velutipes) that was comparable to cells on fibronectin. A snapshot of the RGD-containing proteins in the fungal extracts was obtained by combining SDS-PAGE and mass spectrometry of the peptide fragments obtained by enzymatic cleavage. Therefore, a sustainable source of cell-adhesive proteins is widely available in the fungi kingdom. A method has been developed to identify candidate species and produce cell-adhesive matrices, applicable to the cell-cultivated food and healthcare industries. Nature Publishing Group UK 2023-07-13 /pmc/articles/PMC10344888/ /pubmed/37443321 http://dx.doi.org/10.1038/s41538-023-00209-y Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Teo, Yu Xing
Lee, Kah Yin
Goh, Corinna Jie Hui
Wang, Loo Chien
Sobota, Radoslaw M.
Chiam, Keng-Hwee
Du, Chan
Wan, Andrew C. A.
Fungus-derived protein particles as cell-adhesive matrices for cell-cultivated food
title Fungus-derived protein particles as cell-adhesive matrices for cell-cultivated food
title_full Fungus-derived protein particles as cell-adhesive matrices for cell-cultivated food
title_fullStr Fungus-derived protein particles as cell-adhesive matrices for cell-cultivated food
title_full_unstemmed Fungus-derived protein particles as cell-adhesive matrices for cell-cultivated food
title_short Fungus-derived protein particles as cell-adhesive matrices for cell-cultivated food
title_sort fungus-derived protein particles as cell-adhesive matrices for cell-cultivated food
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10344888/
https://www.ncbi.nlm.nih.gov/pubmed/37443321
http://dx.doi.org/10.1038/s41538-023-00209-y
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