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

Petroll, K; Care, A; Waterstraat, M; Bergquist, PL; Sunna, A

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

Petroll, K Care, A

Waterstraat, M Bergquist, PL Sunna, A

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Publisher:: ELSEVIER SCIENCE BV
Publication Type:: Journal Article
Citation:: Anal Chim Acta, 2019, 1066, pp. 136-145
Issue Date:: 2019-08-20

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Field	Value	Language
dc.contributor.author	Petroll, K
dc.contributor.author	Care, A https://orcid.org/0000-0002-0035-7961
dc.contributor.author	Waterstraat, M
dc.contributor.author	Bergquist, PL
dc.contributor.author	Sunna, A
dc.date.accessioned	2022-03-25T04:23:34Z
dc.date.available	2019-03-12
dc.date.available	2022-03-25T04:23:34Z
dc.date.issued	2019-08-20
dc.identifier.citation	Anal Chim Acta, 2019, 1066, pp. 136-145
dc.identifier.issn	0003-2670
dc.identifier.issn	1873-4324
dc.identifier.uri	http://hdl.handle.net/10453/155555
dc.description.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 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.
dc.format	Print-Electronic
dc.language	eng
dc.publisher	ELSEVIER SCIENCE BV
dc.relation.ispartof	Anal Chim Acta
dc.relation.isbasedon	10.1016/j.aca.2019.03.023
dc.rights	info:eu-repo/semantics/closedAccess
dc.subject	0301 Analytical Chemistry, 0399 Other Chemical Sciences
dc.subject.classification	Analytical Chemistry
dc.subject.mesh	Aldehyde Oxidoreductases
dc.subject.mesh	Biocatalysis
dc.subject.mesh	Carbohydrate Conformation
dc.subject.mesh	Chromatography, Liquid
dc.subject.mesh	Escherichia coli
dc.subject.mesh	Glucaric Acid
dc.subject.mesh	Rhizobiaceae
dc.subject.mesh	Rhizobiaceae
dc.subject.mesh	Escherichia coli
dc.subject.mesh	Glucaric Acid
dc.subject.mesh	Aldehyde Oxidoreductases
dc.subject.mesh	Chromatography, Liquid
dc.subject.mesh	Carbohydrate Conformation
dc.subject.mesh	Biocatalysis
dc.title	Mixed-mode liquid chromatography for the rapid analysis of biocatalytic glucaric acid reaction pathways.
dc.type	Journal Article
utslib.citation.volume	1066
utslib.location.activity	Netherlands
utslib.for	0301 Analytical Chemistry
utslib.for	0399 Other Chemical Sciences
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Life Sciences
utslib.copyright.status	closed_access	*
dc.date.updated	2022-03-25T04:23:33Z
pubs.publication-status	Published
pubs.volume	1066

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 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.

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http://hdl.handle.net/10453/155555