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Chemoenzymatic Synthesis of β‐ D ‐Glucosides using Cellobiose Phosphorylase from Clostridium thermocellum
Author(s) -
De Winter Karel,
Van Renterghem Lisa,
Wuyts Kathleen,
Pelantová Helena,
Křen Vladimír,
Soetaert Wim,
Desmet Tom
Publication year - 2015
Publication title -
advanced synthesis and catalysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.541
H-Index - 155
eISSN - 1615-4169
pISSN - 1615-4150
DOI - 10.1002/adsc.201500077
Subject(s) - chemistry , cellobiose , disaccharide , biocatalysis , carbohydrate synthesis , organic chemistry , glycogen phosphorylase , glycoside , stereochemistry , enzyme , ionic liquid , cellulase , catalysis
Over the past decade, disaccharide phosphorylases have been successfully applied for the synthesis of numerous α‐glucosides. In contrast, much less research has been done with respect to the production of β‐glucosides. Although cellobiose phosphorylase was already successfully used for the synthesis of various disaccharides and branched trisaccharides, its glycosylation potential towards small organic compounds has not been explored to date. Unfortunately, disaccharide phosphorylases typically have a very low affinity for non‐carbohydrate acceptors, which urges the addition of solvents. The ionic liquid AMMOENG TM 101 and ethyl acetate were identified as the most promising solvents, allowing the synthesis of various β‐glucosides. Next to hexyl, heptyl, octyl, nonyl, decyl and undecyl β‐ D ‐glucopyranosides, also the formation of vanillyl 4‐ O ‐β‐ D ‐glucopyranoside, 2‐phenylethyl β‐ D ‐glucopyranoside, β‐citronellyl β‐ D ‐glucopyranoside and 1‐ O ‐β‐ D ‐glucopyranosyl hydroquinone was confirmed by nuclear magnetic resonance spectroscopy and mass spectrometry. Moreover, the stability of cellobiose phosphorylase could be drastically improved by creating cross‐linked enzyme aggregates, while the efficiency of the biocatalyst for the synthesis of octyl β‐ D ‐glucopyranoside was doubled by imprinting with octanol. The usefulness of the latter system was illustrated by performing three consecutive batch conversions with octanol imprinted cross‐linked enzyme aggregates, yielding roughly 2 g of octyl β‐ D ‐glucopyranoside.