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Design of a biocatalytic cascade for the enzymatic sulfation of unsulfated chondroitin with in situ generation of PAPS
Sulfation of molecules in living organisms is a process that plays a key role in their functionality. In mammals, the sulfation of polysaccharides (glycosaminoglycans) that form the proteoglycans present in the extracellular matrix is particularly important. These polysaccharides, through their degr...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2023
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9885120/ https://www.ncbi.nlm.nih.gov/pubmed/36726741 http://dx.doi.org/10.3389/fbioe.2023.1099924 |
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author | Monterrey, Dianelis T. Benito-Arenas, Raúl Revuelta, Julia García-Junceda, Eduardo |
author_facet | Monterrey, Dianelis T. Benito-Arenas, Raúl Revuelta, Julia García-Junceda, Eduardo |
author_sort | Monterrey, Dianelis T. |
collection | PubMed |
description | Sulfation of molecules in living organisms is a process that plays a key role in their functionality. In mammals, the sulfation of polysaccharides (glycosaminoglycans) that form the proteoglycans present in the extracellular matrix is particularly important. These polysaccharides, through their degree and sulfation pattern, are involved in a variety of biological events as signal modulators in communication processes between the cell and its environment. Because of this great biological importance, there is a growing interest in the development of efficient and sustainable sulfation processes, such as those based on the use of sulfotransferase enzymes. These enzymes have the disadvantage of being 3′-phosphoadenosine 5′-phosphosulfate (PAPS) dependent, which is expensive and difficult to obtain. In the present study, a modular multienzyme system was developed to allow the in situ synthesis of PAPS and its coupling to a chondroitin sulfation system. For this purpose, the bifunctional enzyme PAPS synthase 1 (PAPSS1) from Homo sapiens, which contains the ATP sulfurylase and APS kinase activities in a single protein, and the enzyme chondroitin 4-O-sulfotransferase (C4ST-1) from Rattus norvegicus were overexpressed in E. coli. The product formed after coupling of the PAPS generation system and the chondroitin sulfation module was analyzed by NMR. |
format | Online Article Text |
id | pubmed-9885120 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-98851202023-01-31 Design of a biocatalytic cascade for the enzymatic sulfation of unsulfated chondroitin with in situ generation of PAPS Monterrey, Dianelis T. Benito-Arenas, Raúl Revuelta, Julia García-Junceda, Eduardo Front Bioeng Biotechnol Bioengineering and Biotechnology Sulfation of molecules in living organisms is a process that plays a key role in their functionality. In mammals, the sulfation of polysaccharides (glycosaminoglycans) that form the proteoglycans present in the extracellular matrix is particularly important. These polysaccharides, through their degree and sulfation pattern, are involved in a variety of biological events as signal modulators in communication processes between the cell and its environment. Because of this great biological importance, there is a growing interest in the development of efficient and sustainable sulfation processes, such as those based on the use of sulfotransferase enzymes. These enzymes have the disadvantage of being 3′-phosphoadenosine 5′-phosphosulfate (PAPS) dependent, which is expensive and difficult to obtain. In the present study, a modular multienzyme system was developed to allow the in situ synthesis of PAPS and its coupling to a chondroitin sulfation system. For this purpose, the bifunctional enzyme PAPS synthase 1 (PAPSS1) from Homo sapiens, which contains the ATP sulfurylase and APS kinase activities in a single protein, and the enzyme chondroitin 4-O-sulfotransferase (C4ST-1) from Rattus norvegicus were overexpressed in E. coli. The product formed after coupling of the PAPS generation system and the chondroitin sulfation module was analyzed by NMR. Frontiers Media S.A. 2023-01-16 /pmc/articles/PMC9885120/ /pubmed/36726741 http://dx.doi.org/10.3389/fbioe.2023.1099924 Text en Copyright © 2023 Monterrey, Benito-Arenas, Revuelta and García-Junceda. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Bioengineering and Biotechnology Monterrey, Dianelis T. Benito-Arenas, Raúl Revuelta, Julia García-Junceda, Eduardo Design of a biocatalytic cascade for the enzymatic sulfation of unsulfated chondroitin with in situ generation of PAPS |
title | Design of a biocatalytic cascade for the enzymatic sulfation of unsulfated chondroitin with in situ generation of PAPS |
title_full | Design of a biocatalytic cascade for the enzymatic sulfation of unsulfated chondroitin with in situ generation of PAPS |
title_fullStr | Design of a biocatalytic cascade for the enzymatic sulfation of unsulfated chondroitin with in situ generation of PAPS |
title_full_unstemmed | Design of a biocatalytic cascade for the enzymatic sulfation of unsulfated chondroitin with in situ generation of PAPS |
title_short | Design of a biocatalytic cascade for the enzymatic sulfation of unsulfated chondroitin with in situ generation of PAPS |
title_sort | design of a biocatalytic cascade for the enzymatic sulfation of unsulfated chondroitin with in situ generation of paps |
topic | Bioengineering and Biotechnology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9885120/ https://www.ncbi.nlm.nih.gov/pubmed/36726741 http://dx.doi.org/10.3389/fbioe.2023.1099924 |
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