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Screening of Recombinant Lignocellulolytic Enzymes Through Rapid Plate Assays
Industrial Biotechnology and Biocatalysis Group, Biotechnology Laboratory, Department of Synthesis and Development of Industrial Processes, School of Chemical Engineering, National Technical University of Athens, Athens, Greece.
Industrial Biotechnology and Biocatalysis Group, Biotechnology Laboratory, Department of Synthesis and Development of Industrial Processes, School of Chemical Engineering, National Technical University of Athens, Athens, Greece.
Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.ORCID-id: 0000-0003-0079-5950
Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik. Industrial Biotechnology and Biocatalysis Group, Biotechnology Laboratory, Department of Synthesis and Development of Industrial Processes, School of Chemical Engineering, National Technical University of Athens, Athens, Greece.ORCID-id: 0000-0003-0078-5904
2021 (Engelska)Ingår i: Protein Downstream Processing: Design, Development, and Application of High and Low-Resolution Methods / [ed] Nikolaos E. Labrou, Springer Nature, 2021, 2, s. 479-503Kapitel i bok, del av antologi (Övrigt vetenskapligt)
Abstract [en]

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 a 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, hemicellulolytic, and ligninolytic activities and their isolation using immobilized-metal affinity chromatography.

Ort, förlag, år, upplaga, sidor
Springer Nature, 2021, 2. s. 479-503
Serie
Methods in Molecular Biology, ISSN 1064-3745, E-ISSN 1940-6029
Nyckelord [en]
Agar plate assay, Screening, Biomass-degrading enzymes, Glycoside hydrolases, Carbohydrate esterases, Oxidative enzymes, Immobilized-metal affinity chromatography
Nationell ämneskategori
Bioprocessteknik
Forskningsämne
Biokemisk processteknik
Identifikatorer
URN: urn:nbn:se:ltu:diva-81306DOI: 10.1007/978-1-0716-0775-6_30PubMedID: 33128767Scopus ID: 2-s2.0-85094982532OAI: oai:DiVA.org:ltu-81306DiVA, id: diva2:1487621
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ISBN för värdpublikation: 978-1-0716-0774-9,  978-1-0716-0775-6

Tillgänglig från: 2020-11-03 Skapad: 2020-11-03 Senast uppdaterad: 2023-09-05Bibliografiskt granskad

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Christakopoulos, PaulTopakas, Evangelos

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