Abstract
| Language | English |
|---|---|
| Pages | 136-145 |
| Number of pages | 10 |
| Journal | Analytica Chimica Acta |
| Volume | 1066 |
| Early online date | 15 Mar 2019 |
| DOIs | |
| Publication status | Published - 20 Aug 2019 |
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Keywords
- Mixed mode ultra-high performance liquid chromatography
- Hydrophilic interaction chromatography
- Weak anion exchange chromatography
- Biocatalysis
- Bio-based glucaric acid
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Mixed-mode liquid chromatography for the rapid analysis of biocatalytic glucaric acid reaction pathways. / Petroll, Kerstin; Care, Andrew; Waterstraat, Martin; Bergquist, Peter; Sunna, Anwar.
In: Analytica Chimica Acta, Vol. 1066, 20.08.2019, p. 136-145.Research output: Contribution to journal › Article › Research › peer-review
TY - JOUR
T1 - Mixed-mode liquid chromatography for the rapid analysis of biocatalytic glucaric acid reaction pathways
AU - Petroll, Kerstin
AU - Care, Andrew
AU - Waterstraat, Martin
AU - Bergquist, Peter
AU - Sunna, Anwar
PY - 2019/8/20
Y1 - 2019/8/20
N2 - 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 method developed was used for monitoring the enzymatic synthesis of the final step in the GlucA pathway, and showed that GlucA was produced from GA with near 100% conversion and a titre of 9.2 g L−1.
AB - 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 method developed was used for monitoring the enzymatic synthesis of the final step in the GlucA pathway, and showed that GlucA was produced from GA with near 100% conversion and a titre of 9.2 g L−1.
KW - Mixed mode ultra-high performance liquid chromatography
KW - Hydrophilic interaction chromatography
KW - Weak anion exchange chromatography
KW - Biocatalysis
KW - Bio-based glucaric acid
UR - http://www.scopus.com/inward/record.url?scp=85063255074&partnerID=8YFLogxK
U2 - 10.1016/j.aca.2019.03.023
DO - 10.1016/j.aca.2019.03.023
M3 - Article
VL - 1066
SP - 136
EP - 145
JO - Analytica Chimica Acta
T2 - Analytica Chimica Acta
JF - Analytica Chimica Acta
SN - 0003-2670
ER -