Chapter 15 Preparative HPLC of carbohydrates

Hicks, Kevin B.; Hotchkiss, Arland T.

doi:10.1016/s0301-4770(02)80040-9

Cited by 1 publication

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“…Depending on the application, a trade-off should be considered between purity and product yield, decreasing reaction time if the former is favored and increasing the reaction time if the latter is preferred. Alternatively, either preparative high-performance liquid chromatography (HPLC) [ 36 , 37 ] or simulated moving bed chromatography might be used to separate monosaccharides, as established by the production of psicose from sucrose with a three-enzyme cascade reaction [ 9 ]. A more elegant solution might also be provided by adding an additional enzyme to the reaction mixture, creating a coupled process.…”

Section: Discussionmentioning

confidence: 99%

Converting Galactose into the Rare Sugar Talose with Cellobiose 2-Epimerase as Biocatalyst

et al. 2018

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Cellobiose 2-epimerase from Rhodothermus marinus (RmCE) reversibly converts a glucose residue to a mannose residue at the reducing end of β-1,4-linked oligosaccharides. In this study, the monosaccharide specificity of RmCE has been mapped and the synthesis of d-talose from d-galactose was discovered, a reaction not yet known to occur in nature. Moreover, the conversion is industrially relevant, as talose and its derivatives have been reported to possess important antimicrobial and anti-inflammatory properties. As the enzyme also catalyzes the keto-aldo isomerization of galactose to tagatose as a minor side reaction, the purity of talose was found to decrease over time. After process optimization, 23 g/L of talose could be obtained with a product purity of 86% and a yield of 8.5% (starting from 4 g (24 mmol) of galactose). However, higher purities and concentrations can be reached by decreasing and increasing the reaction time, respectively. In addition, two engineering attempts have also been performed. First, a mutant library of RmCE was created to try and increase the activity on monosaccharide substrates. Next, two residues from RmCE were introduced in the cellobiose 2-epimerase from Caldicellulosiruptor saccharolyticus (CsCE) (S99M/Q371F), increasing the kcat twofold.

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Section: Discussionmentioning

confidence: 99%