Screening of Recombinant Lignocellulolytic Enzymes Through Rapid Plate Assays

• 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.

Ämnesord

TEKNIK OCH TEKNOLOGIER  -- Industriell bioteknik -- Bioprocessteknik (hsv//swe)

ENGINEERING AND TECHNOLOGY  -- Industrial Biotechnology -- Bioprocess Technology (hsv//eng)

Nyckelord

Agar plate assay

Screening

Biomass-degrading enzymes

Glycoside hydrolases

Carbohydrate esterases

Oxidative enzymes

Immobilized-metal affinity chromatography

Biokemisk processteknik

Biochemical Process Engineering

Publikations- och innehållstyp

vet (ämneskategori)

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