Membrane-enclosed multienzyme (MEME) synthesis of 2,7-anhydro-sialic acid derivatives

• Published in: Carbohydrate research 2017-11, Vol.451, p.110-117
• Main Authors: Monestier, Marie, Latousakis, Dimitrios, Bell, Andrew, Tribolo, Sandra, Tailford, Louise E., Colquhoun, Ian J., Le Gall, Gwenaelle, Yu, Hai, Chen, Xi, Rejzek, Martin, Dedola, Simone, Field, Robert A., Juge, Nathalie
• Format: Article
• Language: English
• Subjects: 2,7-anhydro-Neu5Ac; 2,7-anhydro-Neu5Gc; Glycoproteins - metabolism; Intramolecular trans-sialidase; N-Acetylneuraminic Acid - analogs & derivatives; N-Acetylneuraminic Acid - chemistry; N-Acetylneuraminic Acid - metabolism; Neuraminidase - metabolism; Ruminococcus - enzymology; Ruminococcus - metabolism; Ruminococcus gnavus; Sialic acid enzymatic synthesis
• ISSN: 0008-6215; 1873-426X
• DOI: 10.1016/j.carres.2017.08.008

Naturally occurring 2,7-anhydro-alpha-N-acetylneuraminic acid (2,7-anhydro-Neu5Ac) is a transglycosylation product of bacterial intramolecular trans-sialidases (IT-sialidases). A facile one-pot two-enzyme approach has been established for the synthesis of 2,7-anhydro-sialic acid derivatives including those containing different sialic acid forms such as Neu5Ac and N-glycolylneuraminic acid (Neu5Gc). The approach is based on the use of Ruminoccocus gnavus IT-sialidase for the release of 2,7-anhydro-sialic acid from glycoproteins, and the conversion of free sialic acid by a sialic acid aldolase. This synthetic method, which is based on a membrane-enclosed enzymatic synthesis, can be performed on a preparative scale. Using fetuin as a substrate, high-yield and cost-effective production of 2,7-anhydro-Neu5Ac was obtained to high-purity. This method was also applied to the synthesis of 2,7-anhydro-Neu5Gc. The membrane-enclosed multienzyme (MEME) strategy reported here provides an efficient approach to produce a variety of sialic acid derivatives. Membrane-enclosed multienzyme (MEME) approach for the production of 2,7-anhydro-sialic acid derivatives. [Display omitted] •A novel membrane enclosed multiple enzyme approach was developed for the production of 2,7-anhydro-sialic acid derivatives.•2,7-anhydro-Neu5Ac was produced in high-yield and high-purity.•Neu5Gc-rich glycoprotein was produced using a one-pot two-step enzymatic synthesis.•The method allowed the synthesis of a novel compound, 2,7-anhydro-Neu5Gc.