title: Definition and characterization of enzymes for maximal biocatalytic solubilization of prebiotic polysaccharides from potato pulp.
abstract: Potato pulp is a high-volume co-processing product resulting from industrial potato starch manufacturing. Potato pulp is particularly rich in pectin, notably galactan branched rhamnogalacturonan I polysaccharides, which are highly bifidogenic when solubilized. The objective of the present study was to characterize and compare four homogalacturonan degrading enzymes capable of catalyzing the required solubilization of these pectinaceous polysaccharides from potato pulp in a 1 min reaction. An additional purpose was to assess the influence of the pH and the potential buffer chelating effects on the release of these polysaccharides from the potato pulp. The pH and temperature optima of two selected pectin lyases from Emericella nidulans (formerly known as Aspergillus nidulans) and Aspergillus niger were determined to 8.6 and 4.0, respectively, at ≥100 °C within 1 min of reaction. The optima for the two selected polygalacturonases from E. nidulans and Aspergillus aculeatus were determined to pH 4.4 and 46 °C, and pH 3.7 and ≥80 °C, respectively. The polygalacturonase from A. aculeatus was 4-42 times more heat-resistant at 50 °C than the other enzymes. The difference in pH optima of the pectin lyases and the exceptional thermal stabilities of some of the enzymes are proposed to be related to specific amino acid substitutions, stabilizing hydrogen bonding and structural traits of the enzymes. The K(M) and V(max) values ranged from 0.3-0.6g/L and 0.5-250.5 U/mg protein, respectively. Phosphate buffer induced release of a higher amount of dry matter than Tris-acetate buffer at pH 6, indicating a chelating effect of the phosphate. Moreover, the phosphate had a higher chelating effect at pH 6 than at pH 4. The optimal conditions for a high yield of polysaccharides from potato pulp were therefore: 1% (w/w) potato pulp treated with 1% (w/w) enzyme/substrate (E/S) pectin lyase from E. nidulans and 1% (w/w) E/S polygalacturonase from A. aculeatus at pH 6.0 and 60 °C for 1 min.
authors: Center for BioProcess Engineering, Department of Chemical and Biochemical Engineering, Technical University of Denmark, 2800 Kgs. Lyngby, Denmark.
authors: Lise V
authors: LV
authors: Thomassen
authors: Dorte M
authors: DM
authors: Larsen
authors: Jørn D
authors: JD
authors: Mikkelsen
authors: Anne S
authors: AS
authors: Meyer
mesh_terms: D001230:Aspergillus / Q000201:enzymology*; D001233:Aspergillus nidulans / Q000201:enzymology; D001234:Aspergillus niger / Q000201:enzymology; D055162:Biocatalysis; D002021:Buffers; D002614:Chelating Agents; D019649:Food Industry / Q000379:methods; D005656:Fungal Proteins / Q000302:isolation & purification / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D009005:Monosaccharides / Q000032:analysis; D010710:Phosphates; D010936:Plant Extracts / Q000737:chemistry; D011096:Polygalacturonase / Q000302:isolation & purification / Q000378:metabolism*; D011133:Polysaccharide-Lyases / Q000302:isolation & purification / Q000378:metabolism*; D011134:Polysaccharides / Q000378:metabolism*; D055550:Protein Stability; D011198:Solanum tuberosum / Q000737:chemistry*; D013045:Species Specificity; D013696:Temperature
chemical_list: D002021:Buffers; D002614:Chelating Agents; D005656:Fungal Proteins; D009005:Monosaccharides; D010710:Phosphates; D010936:Plant Extracts; D011134:Polysaccharides; D011096:Polygalacturonase; D011133:Polysaccharide-Lyases; C027349:pectin lyase