초록
<▼1><P>Naturally occurring 2,7-anhydro-alpha-N-acetylneuraminic acid (2,7-anhydro-Neu5Ac) is a transglycosylation product of bacterial intramolecular <I>trans</I>-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 <I>Ruminoccocus gnavus</I> 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.</P></▼1><▼2><P><B>Highlights</B></P><P>•<P>A novel membrane enclosed multiple enzyme approach was developed for the production of 2,7-anhydro-sialic acid derivatives.</P>•<P>2,7-anhydro-Neu5Ac was produced in high-yield and high-purity.</P>•<P>Neu5Gc-rich glycoprotein was produced using a one-pot two-step enzymatic synthesis.</P>•<P>The method allowed the synthesis of a novel compound, 2,7-anhydro-Neu5Gc.</P></P></▼2>