title: Centrifugal partition chromatography in a biorefinery context: Optimisation and scale-up of monosaccharide fractionation from hydrolysed sugar beet pulp.
abstract: The isolation of component sugars from biomass represents an important step in the bioprocessing of sustainable feedstocks such as sugar beet pulp. Centrifugal partition chromatography (CPC) is used here, as an alternative to multiple resin chromatography steps, to fractionate component monosaccharides from crude hydrolysed sugar beet pulp pectin. CPC separation of samples, prepared in the stationary phase, was carried out using an ethanol: ammonium sulphate (300gL-1) phase system (0.8:1.8v:v) in ascending mode. This enabled removal of crude feedstream impurities and separation of monosaccharides into three fractions (l-rhamnose, l-arabinose and d-galactose, and d-galacturonic acid) in a single step. Throughput was improved three-fold by increasing sample injection volume, from 4 to 16% of column volume, with similar separation performance maintained in all cases. Extrusion of the final galacturonic acid fraction increased the eluted solute concentration, reduced the total separation time by 24% and removed the need for further column regeneration. Reproducibility of the separation after extrusion was validated by using multiple stacked injections. Scale-up was performed linearly from a semi-preparative 250mL column to a preparative 950mL column with a scale-up ratio of 3.8 applied to mobile phase flow rate and sample injection volume. Throughputs of 9.4gL-1h-1of total dissolved solids were achieved at the preparative scale with a throughput of 1.9gL-1h-1of component monosaccharides. These results demonstrate the potential of CPC for both impurity removal and target fractionation within biorefinery separations.
authors.affiliation: The Advanced Centre for Biochemical Engineering, Department of Biochemical Engineering, University College London, Gower Street, London WC1E 6BT, UK.
authors.forename: David P
authors.initials: DP
authors.lastname: Ward
authors.affiliation: Advanced Bioprocessing Centre, Department of Mechanical, Aerospace & Civil Engineering, Brunel University London, Uxbridge UB8 3PH, UK.
authors.forename: Peter
authors.initials: P
authors.lastname: Hewitson
authors.affiliation: The Advanced Centre for Biochemical Engineering, Department of Biochemical Engineering, University College London, Gower Street, London WC1E 6BT, UK.
authors.forename: Max
authors.initials: M
authors.lastname: Cárdenas-Fernández
authors.affiliation: Department of Biology and Biochemistry, University of Bath, Bath, Somerset BA2 7AY, UK.
authors.forename: Charlotte
authors.initials: C
authors.lastname: Hamley-Bennett
authors.affiliation: GlaxoSmithKline R&D Ltd., Medicines Research Centre, Gunnels Wood Road, Stevenage, Hertfordshire SG1 2NY, UK.
authors.forename: Alba
authors.initials: A
authors.lastname: Díaz-Rodríguez
authors.affiliation: GlaxoSmithKline R&D Ltd., Medicines Research Centre, Gunnels Wood Road, Stevenage, Hertfordshire SG1 2NY, UK.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Douillet
authors.affiliation: GlaxoSmithKline R&D Ltd., Medicines Research Centre, Gunnels Wood Road, Stevenage, Hertfordshire SG1 2NY, UK.
authors.forename: Joseph P
authors.initials: JP
authors.lastname: Adams
authors.affiliation: Department of Biology and Biochemistry, University of Bath, Bath, Somerset BA2 7AY, UK.
authors.forename: David J
authors.initials: DJ
authors.lastname: Leak
authors.affiliation: Advanced Bioprocessing Centre, Department of Mechanical, Aerospace & Civil Engineering, Brunel University London, Uxbridge UB8 3PH, UK. Electronic address: svetlana.ignatova@brunel.ac.uk.
authors.forename: Svetlana
authors.initials: S
authors.lastname: Ignatova
authors.affiliation: The Advanced Centre for Biochemical Engineering, Department of Biochemical Engineering, University College London, Gower Street, London WC1E 6BT, UK. Electronic address: g.lye@ucl.ac.uk.
authors.forename: Gary J
authors.initials: GJ
authors.lastname: Lye
mesh_terms: D027461:Beta vulgaris / Q000737:chemistry*; D002498:Centrifugation; D005591:Chemical Fractionation / Q000379:methods*; D002853:Chromatography, Liquid / Q000379:methods*; D006603:Hexuronic Acids / Q000302:isolation & purification; D006868:Hydrolysis; D009005:Monosaccharides / Q000302:isolation & purification*; D010368:Pectins / Q000737:chemistry; D015203:Reproducibility of Results
chemical_list: D006603:Hexuronic Acids; D009005:Monosaccharides; C007819:galacturonic acid; D010368:Pectins
keywords: Biorefinery; Centrifugal partition chromatography; Monosaccharides; Scale-up; Sugar beet pulp