Downstream Processing of Nucleoside-Diphospho-Sugars from Sucrose Synthase Reaction Mixtures at Decreased Solvent Consumption

Martin Lemmerer, Katharina Schmoelzer, Alexander Gutmann, Bernd Nidetzky

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Nucleoside-diphospho-sugars (NDP-sugars) are highly demanded as specialty chemicals and as substrates for enzymatic glycosylations. Biocatalysis is efficient for their synthesis; however, downstream processing (DSP) is a bottleneck of the overall production. We describe two scalable DSP routes for recovery of NDP-glucose (NDP-glc) from sucrose synthase (SuSy) reaction mixtures. In both routes product precipitation with ethanol replaced size exclusion chromatography (SEC) which, due to high solvent consumption, constitutes the main limiting step of current DSP protocols. We also show that selective phosphomonoester hydrolysis by alkaline phosphatase, characterized in that it did not degrade the phosphodiester linkage in NDP-glc, was a useful alternative to anion-exchange chromatography (AEC) for capture and initial purification of the product. Based on comprehensive route optimization, we show uridine 5′-diphosphate glucose (UDP-glc) recovery at the ≥0.5 g scale in a yield of about 80% and in a purity of 86% (phosphatase route) and 95% (AEC route). Solvent savings of up to 38-fold compared to the conventional AEC-SEC route were achieved. There is a clear demand for both new routes, as requirements on DSP regarding purity and costs vary with the field of application. Both DSP routes seem to be applicable to other NDP-sugars and different methods of their synthesis.
LanguageEnglish
Pages3113-3122
JournalAdvanced Synthesis & Catalysis
Volume358
StatusPublished - 2016

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Sugar (sucrose)
Nucleosides
Sugars
Chromatography
Anions
Glucose
Processing
Ion exchange
Negative ions
Size exclusion chromatography
Phosphatases
Glycosylation
Recovery
Diphosphates
Uridine
Phosphoric Monoester Hydrolases
Purification
Alkaline Phosphatase
Hydrolysis
Ethanol

Fields of Expertise

  • Human- & Biotechnology

Cite this

Downstream Processing of Nucleoside-Diphospho-Sugars from Sucrose Synthase Reaction Mixtures at Decreased Solvent Consumption. / Lemmerer, Martin; Schmoelzer, Katharina; Gutmann, Alexander; Nidetzky, Bernd.

In: Advanced Synthesis & Catalysis, Vol. 358, 2016, p. 3113-3122.

Research output: Contribution to journalArticleResearchpeer-review

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AB - Nucleoside-diphospho-sugars (NDP-sugars) are highly demanded as specialty chemicals and as substrates for enzymatic glycosylations. Biocatalysis is efficient for their synthesis; however, downstream processing (DSP) is a bottleneck of the overall production. We describe two scalable DSP routes for recovery of NDP-glucose (NDP-glc) from sucrose synthase (SuSy) reaction mixtures. In both routes product precipitation with ethanol replaced size exclusion chromatography (SEC) which, due to high solvent consumption, constitutes the main limiting step of current DSP protocols. We also show that selective phosphomonoester hydrolysis by alkaline phosphatase, characterized in that it did not degrade the phosphodiester linkage in NDP-glc, was a useful alternative to anion-exchange chromatography (AEC) for capture and initial purification of the product. Based on comprehensive route optimization, we show uridine 5′-diphosphate glucose (UDP-glc) recovery at the ≥0.5 g scale in a yield of about 80% and in a purity of 86% (phosphatase route) and 95% (AEC route). Solvent savings of up to 38-fold compared to the conventional AEC-SEC route were achieved. There is a clear demand for both new routes, as requirements on DSP regarding purity and costs vary with the field of application. Both DSP routes seem to be applicable to other NDP-sugars and different methods of their synthesis.

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