Mixed-mode liquid chromatography for the rapid analysis of biocatalytic glucaric acid reaction pathways.

Mixed-mode liquid chromatography for the rapid analysis of biocatalytic glucaric acid reaction pathways.
Anal Chim Acta. 2019 Aug 20;1066:136-145
Authors: Petroll K, Care A, Waterstraat M, Bergquist PL, Sunna A
Abstract
Glucaric acid (GlucA) has been identified as one of the top 10 potential bio-based chemicals for replacement of oil-based chemicals. Several synthetic enzyme pathways have been engineered in bacteria and yeast to produce GlucA from glucose and myo-inositol. However, the yields and titres achieved with these systems remain too low for the requirements of a bio-based GlucA industry. A major limitation for the optimisation of GlucA production via synthetic enzymatic pathways are the laborious analytical procedures required to detect the final product (GlucA) and pathway intermediates. We have developed a novel method for the simple and simultaneous analysis of GlucA and pathway intermediates to address this limitation using mixed mode (MM) HILIC and weak anion exchange chromatography (WAX), referred to as MM HILIC/WAX, coupled with RID. Isocratic mobile phase conditions and the sample solvent were optimised for the separation of GlucA, glucose-1-phosphate (G1P), glucose-6-phosphate (G6P), inositol-1-phosphate (I1P), myo-inositol and glucuronic acid (GA). The method showed good repeatability, precision and excellent accuracy with detection and quantitation limits (LOD and LOQ) of 1.5-2 and 577 mM, respectively. The meth...

Source: Analytica Chimica Acta - April 29, 2019 Category: Chemistry Authors: Petroll K, Care A, Waterstraat M, Bergquist PL, Sunna A Tags: Anal Chim Acta Source Type: research