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A New, Quick, and Simple Protocol to Evaluate Microalgae Polysaccharide Composition
In this work, a new methodological approach, relying on the high specificity of enzymes in a complex mixture, was developed to estimate the composition of bioactive polysaccharides produced by microalgae, directly in algal cultures. The objective was to set up a protocol to target oligomers commonly...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7916578/ https://www.ncbi.nlm.nih.gov/pubmed/33578865 http://dx.doi.org/10.3390/md19020101 |
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author | Decamp, Antoine Michelo, Orane Rabbat, Christelle Laroche, Céline Grizeau, Dominique Pruvost, Jérémy Gonçalves, Olivier |
author_facet | Decamp, Antoine Michelo, Orane Rabbat, Christelle Laroche, Céline Grizeau, Dominique Pruvost, Jérémy Gonçalves, Olivier |
author_sort | Decamp, Antoine |
collection | PubMed |
description | In this work, a new methodological approach, relying on the high specificity of enzymes in a complex mixture, was developed to estimate the composition of bioactive polysaccharides produced by microalgae, directly in algal cultures. The objective was to set up a protocol to target oligomers commonly known to be associated with exopolysaccharides’ (EPS) nutraceutical and pharmaceutical activities (i.e., rhamnose, fucose, acidic sugars, etc.) without the constraints classically associated with chromatographic methods, while maintaining a resolution sufficiently high to enable their monitoring in the culture system. Determination of the monosaccharide content required the application of acid hydrolysis (2 M trifluoroacetic acid) followed by NaOH (2 M) neutralization. Quantification was then carried out directly on the fresh hydrolysate using enzyme kits corresponding to the main monosaccharides in a pre-determined composition of the polysaccharides under analysis. Initial results showed that the enzymes were not sensitive to the presence of TFA and NaOH, so the methodology could be carried out on fresh hydrolysate. The limits of quantification of the method were estimated as being in the order of the log of nanograms of monosaccharides per well, thus positioning it among the chromatographic methods in terms of analytical performance. A comparative analysis of the results obtained by the enzymatic method with a reference method (high-performance anion-exchange chromatography) confirmed good recovery rates, thus validating the closeness of the protocol. Finally, analyses of raw culture media were carried out and compared to the results obtained in miliQ water; no differences were observed. The new approach is a quick, functional analysis method allowing routine monitoring of the quality of bioactive polysaccharides in algal cultures grown in photobioreactors. |
format | Online Article Text |
id | pubmed-7916578 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-79165782021-03-01 A New, Quick, and Simple Protocol to Evaluate Microalgae Polysaccharide Composition Decamp, Antoine Michelo, Orane Rabbat, Christelle Laroche, Céline Grizeau, Dominique Pruvost, Jérémy Gonçalves, Olivier Mar Drugs Article In this work, a new methodological approach, relying on the high specificity of enzymes in a complex mixture, was developed to estimate the composition of bioactive polysaccharides produced by microalgae, directly in algal cultures. The objective was to set up a protocol to target oligomers commonly known to be associated with exopolysaccharides’ (EPS) nutraceutical and pharmaceutical activities (i.e., rhamnose, fucose, acidic sugars, etc.) without the constraints classically associated with chromatographic methods, while maintaining a resolution sufficiently high to enable their monitoring in the culture system. Determination of the monosaccharide content required the application of acid hydrolysis (2 M trifluoroacetic acid) followed by NaOH (2 M) neutralization. Quantification was then carried out directly on the fresh hydrolysate using enzyme kits corresponding to the main monosaccharides in a pre-determined composition of the polysaccharides under analysis. Initial results showed that the enzymes were not sensitive to the presence of TFA and NaOH, so the methodology could be carried out on fresh hydrolysate. The limits of quantification of the method were estimated as being in the order of the log of nanograms of monosaccharides per well, thus positioning it among the chromatographic methods in terms of analytical performance. A comparative analysis of the results obtained by the enzymatic method with a reference method (high-performance anion-exchange chromatography) confirmed good recovery rates, thus validating the closeness of the protocol. Finally, analyses of raw culture media were carried out and compared to the results obtained in miliQ water; no differences were observed. The new approach is a quick, functional analysis method allowing routine monitoring of the quality of bioactive polysaccharides in algal cultures grown in photobioreactors. MDPI 2021-02-10 /pmc/articles/PMC7916578/ /pubmed/33578865 http://dx.doi.org/10.3390/md19020101 Text en © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Decamp, Antoine Michelo, Orane Rabbat, Christelle Laroche, Céline Grizeau, Dominique Pruvost, Jérémy Gonçalves, Olivier A New, Quick, and Simple Protocol to Evaluate Microalgae Polysaccharide Composition |
title | A New, Quick, and Simple Protocol to Evaluate Microalgae Polysaccharide Composition |
title_full | A New, Quick, and Simple Protocol to Evaluate Microalgae Polysaccharide Composition |
title_fullStr | A New, Quick, and Simple Protocol to Evaluate Microalgae Polysaccharide Composition |
title_full_unstemmed | A New, Quick, and Simple Protocol to Evaluate Microalgae Polysaccharide Composition |
title_short | A New, Quick, and Simple Protocol to Evaluate Microalgae Polysaccharide Composition |
title_sort | new, quick, and simple protocol to evaluate microalgae polysaccharide composition |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7916578/ https://www.ncbi.nlm.nih.gov/pubmed/33578865 http://dx.doi.org/10.3390/md19020101 |
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