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Separation and quantification of 2-Keto-3-deoxy-gluconate (KDG) a major metabolite in pectin and alginate degradation pathways

Shiny Martis B, View ORCID ProfileMichel Droux, Florelle Deboudard, View ORCID ProfileWilliam Nasser, View ORCID ProfileSam Meyer, View ORCID ProfileSylvie Reverchon
doi: https://doi.org/10.1101/2020.07.24.220400
Shiny Martis B
aUniv Lyon, Université Claude Bernard Lyon 1, INSA-Lyon, CNRS, UMR5240 MAP, F-69621, France
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Michel Droux
aUniv Lyon, Université Claude Bernard Lyon 1, INSA-Lyon, CNRS, UMR5240 MAP, F-69621, France
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  • For correspondence: michel.droux{at}univ-lyon1.fr
Florelle Deboudard
aUniv Lyon, Université Claude Bernard Lyon 1, INSA-Lyon, CNRS, UMR5240 MAP, F-69621, France
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William Nasser
aUniv Lyon, Université Claude Bernard Lyon 1, INSA-Lyon, CNRS, UMR5240 MAP, F-69621, France
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Sam Meyer
aUniv Lyon, Université Claude Bernard Lyon 1, INSA-Lyon, CNRS, UMR5240 MAP, F-69621, France
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Sylvie Reverchon
aUniv Lyon, Université Claude Bernard Lyon 1, INSA-Lyon, CNRS, UMR5240 MAP, F-69621, France
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Abstract

A rapid and sensitive High Performance Liquid Chromatography (HPLC) method with photometric and fluorescence detection is developed for routine analysis of 2-Keto-3-deoxy-gluconate (KDG), a catabolite product of pectin and alginate. These polysaccharides are primary-based compounds for biofuel production and for generation of high-value-added products. HPLC is performed, after derivatization of the 2-oxo-acid groups of the metabolite with o-phenylenediamine (oPD), using a linear gradient of trifluoroacetic acid and acetonitrile. Quantification is accomplished with an internal standard method. The gradient is optimized to distinguish KDG from its close structural analogues such as 5-keto-4-deoxyuronate (DKI) and 2,5-diketo-3-deoxygluconate (DKII). The proposed method is simple, highly sensitive and accurate for time course analysis of pectin or alginate degradation.

Highlights A fluorescent based-HPLC method report the quantification of KDG, a metabolite originating from alginate and from pectin degradation pathways, using derivatization with o-phenylenediamine (oPD)

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC 4.0 International license.
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Posted December 03, 2020.
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Separation and quantification of 2-Keto-3-deoxy-gluconate (KDG) a major metabolite in pectin and alginate degradation pathways
Shiny Martis B, Michel Droux, Florelle Deboudard, William Nasser, Sam Meyer, Sylvie Reverchon
bioRxiv 2020.07.24.220400; doi: https://doi.org/10.1101/2020.07.24.220400
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Separation and quantification of 2-Keto-3-deoxy-gluconate (KDG) a major metabolite in pectin and alginate degradation pathways
Shiny Martis B, Michel Droux, Florelle Deboudard, William Nasser, Sam Meyer, Sylvie Reverchon
bioRxiv 2020.07.24.220400; doi: https://doi.org/10.1101/2020.07.24.220400

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