Screening of Recombinant Lignocellulolytic Enzymes Through Rapid Plate Assays
2021 (English)In: Protein Downstream Processing: Design, Development, and Application of High and Low-Resolution Methods / [ed] Nikolaos E. Labrou, Springer Nature, 2021, 2, p. 479-503Chapter in book (Other academic)
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.
Place, publisher, year, edition, pages
Springer Nature, 2021, 2. p. 479-503
Series
Methods in Molecular Biology, ISSN 1064-3745, E-ISSN 1940-6029
Keywords [en]
Agar plate assay, Screening, Biomass-degrading enzymes, Glycoside hydrolases, Carbohydrate esterases, Oxidative enzymes, Immobilized-metal affinity chromatography
National Category
Bioprocess Technology
Research subject
Biochemical Process Engineering
Identifiers
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
Note
ISBN för värdpublikation: 978-1-0716-0774-9, 978-1-0716-0775-6
2020-11-032020-11-032023-09-05Bibliographically approved