title: Biomolecular characterization of the levansucrase of Erwinia amylovora, a promising biocatalyst for the synthesis of fructooligosaccharides.
abstract: Erwinia amylovora is a plant pathogen that affects Rosaceae, such as apple and pear. In E. amylovora the fructans, produced by the action of a levansucrase (EaLsc), play a role in virulence and biofilm formation. Fructans are bioactive compounds, displaying health-promoting properties in their own right. Their use as food and feed supplements is increasing. In this study, we investigated the biomolecular properties of EaLsc using HPAEC-PAD, MALDI-TOF MS, and spectrophotometric assays. The enzyme, which was heterologously expressed in Escherichia coli in high yield, was shown to produce mainly fructooligosaccharides (FOSs) with a degree of polymerization between 3 and 6. The kinetic properties of EaLsc were similar to those of other phylogenetically related Gram-negative bacteria, but the good yield of FOSs, the product spectrum, and the straightforward production of the enzyme suggest that EaLsc is an interesting biocatalyst for future studies aimed at producing tailor-made fructans.
authors.affiliation: Laboratory of Bioorganic Chemistry and Crystallography, Faculty of Science and Technology, Free University of Bolzano , Piazza Università 5, 39100 Bolzano, Italy.
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Caputi
authors.forename: Sergey A
authors.initials: SA
authors.lastname: Nepogodiev
authors.forename: Mickael
authors.initials: M
authors.lastname: Malnoy
authors.forename: Martin
authors.initials: M
authors.lastname: Rejzek
authors.forename: Robert A
authors.initials: RA
authors.lastname: Field
authors.forename: Stefano
authors.initials: S
authors.lastname: Benini
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D055162:Biocatalysis; D044084:Erwinia amylovora / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D006602:Hexosyltransferases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D009844:Oligosaccharides / Q000096:biosynthesis / Q000737:chemistry*
chemical_list: D001426:Bacterial Proteins; D009844:Oligosaccharides; C116580:fructooligosaccharide; D006602:Hexosyltransferases; C021513:levansucrase