title: New GH16 β-glucanase from Paenibacillus barcinonensis BP-23 releases a complex pattern of mixed-linkage oligomers from barley glucan.
abstract: The gene coding for a lichenase from Paenibacillus barcinonensis BP-23, a powerful carbohydrate-degrading strain, was obtained using a genome walking strategy and expressed in Escherichia coli for further characterization. The amino acid sequence deduced from lic16A revealed that the lichenase is a single-domain enzyme belonging to the GH16 family. Purified recombinant Lic16A showed exclusive activity on β-1,3-1,4-glucans, showing a Km of 16.88 mg/mL and a Vmax of 266.09 U/mg using lichenan as a substrate. Lic16A was stable at 55 °C for at least 3 H in moderate pH conditions. Thin-layer chromatography analysis showed that the enzyme released a complex mixture of hydrolysis products, which consisted of different length oligosaccharides of intermediate mobility among cellooligomers. The health benefits of β-glucans's consumption and the increased interest for the use of their oligomers as prebiotics add interest to the study of Lic16A for the production of β-glucan-derived oligosaccharides and the evaluation of their biotechnological potential. This is the first report on β-1,3-1,4-glucanase produced by P. barcinonensis.
authors.affiliation: Department of Microbiology, Faculty of Biology, University of Barcelona, Barcelona, Spain.
authors.forename: Liliana Alexandra
authors.initials: LA
authors.lastname: Cerda
authors.affiliation: Department of Microbiology, Faculty of Biology, University of Barcelona, Barcelona, Spain.
authors.forename: Susana Valeria
authors.initials: SV
authors.lastname: Valenzuela
authors.affiliation: Department of Microbiology, Faculty of Biology, University of Barcelona, Barcelona, Spain.
authors.forename: Pilar
authors.initials: P
authors.lastname: Diaz
authors.affiliation: Department of Microbiology, Faculty of Biology, University of Barcelona, Barcelona, Spain.
authors.forename: Francisco I Javier
authors.initials: FI
authors.lastname: Pastor
mesh_terms: D003001:Cloning, Molecular; D005936:Glucans / Q000737:chemistry / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001467:Hordeum / Q000737:chemistry / Q000378:metabolism*; D006868:Hydrolysis; D056507:Paenibacillus / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D005936:Glucans; D011994:Recombinant Proteins; D006026:Glycoside Hydrolases; C028020:licheninase
keywords: Paenibacillus; lichenase; oligomers; prebiotics; purification; β-glucan