title: Screening and purification of recombinant lignocellulolytic enzymes.
abstract: In the search for novel biomass-degrading enzymes through mining microbial genomes, it is necessary to apply functional tests during high-throughput screenings, which are capable of detecting enzymatic activities directly by way of plate assay. Using the most efficient expression systems of Escherichia coli and Pichia pastoris, the production of high amount of His-tagged recombinant proteins could be thrived, allowing the one-step isolation by affinity chromatography. Here, we describe simple and efficient assay techniques for the detection of various biomass-degrading enzymatic activities on agar plates, such as cellulolytic and hemicellulolytic activities and their isolation using immobilized-metal affinity chromatography.
authors.affiliation: Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens, Athens, Greece.
authors.forename: Evangelos
authors.initials: E
authors.lastname: Topakas
authors.forename: Paul
authors.initials: P
authors.lastname: Christakopoulos
mesh_terms: D004798:Enzymes / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D006868:Hydrolysis; D008031:Lignin / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D004798:Enzymes; D011994:Recombinant Proteins; C036909:lignocellulose; D008031:Lignin