title: Fast purification method of functional LPMOs from Streptomyces ambofaciens by affinity adsorption.
abstract: A simple purification method by affinity adsorption was developed to obtain functional lytic polysaccharide monooxygenases (LPMOs). The system allows the successful purification to homogeneity of the most characterized bacterial LPMO, CBP21 from Serratia marcescens, and two LPMOs from Streptomyces ambofaciens, which have not been previously characterized. The first of these new LPMOs, named SamLPMO10B is a small enzyme (15 kDa) belonging to family 10 of auxiliary activities (AA10), showing activity on β-chitin. The second LPMO, SamLPMO10C (34.7 kDa), is a bimodular enzyme comprised of an AA10 catalytic module and a carbohydrate binding module of family CBM2. SamLPMO10C shows activity on cellulosic substrates, including agricultural fiber paper pulps. The methodology developed simplifies the purification process to a binding-elution protocol with low-grade polysaccharides including Avicel. The strategy can be a cheap, simple and fast solution for the purification of LPMOs for industrial applications, leaving out periplasmic fractionation from recombinant strains therefore, with reduction of time and costs compared to conventional processes. The activity of SamLPMO10C expands the potential of the high valued LPMOs in lignocellulosic biomass valorization, reaffirming their promising role in cellulose deconstruction.
authors.affiliation: Department of Genetics, Microbiology and Statistics, Faculty of Biology, University of Barcelona, Av. Diagonal 643, 08028 Barcelona, Spain; IN2UB, Institute of Nanoscience and Nanotechnology, University of Barcelona, Joan XXIII, s/n, 08028 Barcelona, Spain. Electronic address: susanavalenzuela@ub.edu.
authors.forename: Susana V
authors.initials: SV
authors.lastname: Valenzuela
authors.affiliation: Department of Genetics, Microbiology and Statistics, Faculty of Biology, University of Barcelona, Av. Diagonal 643, 08028 Barcelona, Spain.
authors.forename: Guillem
authors.initials: G
authors.lastname: Ferreres
authors.affiliation: Department of Genetics, Microbiology and Statistics, Faculty of Biology, University of Barcelona, Av. Diagonal 643, 08028 Barcelona, Spain.
authors.forename: Gerard
authors.initials: G
authors.lastname: Margalef
authors.affiliation: Department of Genetics, Microbiology and Statistics, Faculty of Biology, University of Barcelona, Av. Diagonal 643, 08028 Barcelona, Spain; IN2UB, Institute of Nanoscience and Nanotechnology, University of Barcelona, Joan XXIII, s/n, 08028 Barcelona, Spain.
authors.forename: F I Javier
authors.initials: FIJ
authors.lastname: Pastor
mesh_terms: D000327:Adsorption; D018533:Biomass; D005591:Chemical Fractionation / Q000379:methods*; D003001:Cloning, Molecular; D008031:Lignin / Q000378:metabolism; D006899:Mixed Function Oxygenases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D008970:Molecular Weight; D011134:Polysaccharides / Q000378:metabolism*; D013302:Streptomyces / Q000201:enzymology*; D013997:Time Factors
chemical_list: D011134:Polysaccharides; C036909:lignocellulose; D008031:Lignin; D006899:Mixed Function Oxygenases
keywords: AA10; Cellulose; Chitin; Lytic polysaccharide monooxygenase; Streptomyces