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...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-02-01
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| Series: | Marine Drugs |
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| Online Access: | https://www.mdpi.com/1660-3397/19/2/101 |
| _version_ | 1797411668513259520 |
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| author | Antoine Decamp Orane Michelo Christelle Rabbat Céline Laroche Dominique Grizeau Jérémy Pruvost Olivier Gonçalves |
| author_facet | Antoine Decamp Orane Michelo Christelle Rabbat Céline Laroche Dominique Grizeau Jérémy Pruvost Olivier Gonçalves |
| author_sort | Antoine Decamp |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-03-09T04:48:29Z |
| format | Article |
| id | doaj.art-08d6bb53c01a4fc7b9f65c962f1061aa |
| institution | Directory Open Access Journal |
| issn | 1660-3397 |
| language | English |
| last_indexed | 2024-03-09T04:48:29Z |
| publishDate | 2021-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Marine Drugs |
| spelling | doaj.art-08d6bb53c01a4fc7b9f65c962f1061aa2023-12-03T13:11:59ZengMDPI AGMarine Drugs1660-33972021-02-0119210110.3390/md19020101A New, Quick, and Simple Protocol to Evaluate Microalgae Polysaccharide CompositionAntoine Decamp0Orane Michelo1Christelle Rabbat2Céline Laroche3Dominique Grizeau4Jérémy Pruvost5Olivier Gonçalves6Université de Nantes, GEPEA, UMR CNRS 6144, 37 boulevard de l’Université, 44600 Saint-Nazaire, FranceUniversité de Nantes, GEPEA, UMR CNRS 6144, 37 boulevard de l’Université, 44600 Saint-Nazaire, FranceUniversité de Nantes, GEPEA, UMR CNRS 6144, 37 boulevard de l’Université, 44600 Saint-Nazaire, FranceInstitut Pascal UMR 6602, Université Clermont Auvergne, CNRS, SIGMA Clermont, 63000 Clermont-Ferrand, FranceUniversité de Nantes, GEPEA, UMR CNRS 6144, 37 boulevard de l’Université, 44600 Saint-Nazaire, FranceUniversité de Nantes, GEPEA, UMR CNRS 6144, 37 boulevard de l’Université, 44600 Saint-Nazaire, FranceUniversité de Nantes, GEPEA, UMR CNRS 6144, 37 boulevard de l’Université, 44600 Saint-Nazaire, FranceIn 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.https://www.mdpi.com/1660-3397/19/2/101enzymatic quantificationrapid and cost-effective methodeasy-to-use bioactive (exo)polysaccharide profilingmicroalgae |
| spellingShingle | Antoine Decamp Orane Michelo Christelle Rabbat Céline Laroche Dominique Grizeau Jérémy Pruvost Olivier Gonçalves A New, Quick, and Simple Protocol to Evaluate Microalgae Polysaccharide Composition Marine Drugs enzymatic quantification rapid and cost-effective method easy-to-use bioactive (exo)polysaccharide profiling microalgae |
| 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 | enzymatic quantification rapid and cost-effective method easy-to-use bioactive (exo)polysaccharide profiling microalgae |
| url | https://www.mdpi.com/1660-3397/19/2/101 |
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