title: Liquid Phase Multiplex High-Throughput Screening of Metagenomic Libraries Using p-Nitrophenyl-Linked Substrates for Accessory Lignocellulosic Enzymes.
abstract: To access the genetic potential contained in large metagenomic libraries, suitable high-throughput functional screening methods are required. Here we describe a high-throughput screening approach which enables the rapid identification of metagenomic library clones expressing functional accessory lignocellulosic enzymes. The high-throughput nature of this method hinges on the multiplexing of both the E. coli metagenomic library clones and the colorimetric p-nitrophenyl linked substrates which allows for the simultaneous screening for β-glucosidases, β-xylosidases, and α-L-arabinofuranosidases. This method is readily automated and compatible with high-throughput robotic screening systems.
authors.affiliation: Institute for Microbial Biotechnology and Metagenomics, University of the Western Cape, Cape Town, South Africa.|Centre for Bioprocess Engineering Research, University of Cape Town, Cape Town, South Africa.
authors.forename: Mariette
authors.initials: M
authors.lastname: Smart
authors.affiliation: Institute for Microbial Biotechnology and Metagenomics, University of the Western Cape, Cape Town, South Africa.|Centre for Bioprocess Engineering Research, University of Cape Town, Cape Town, South Africa.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Huddy
authors.affiliation: Institute for Microbial Biotechnology and Metagenomics, University of the Western Cape, Cape Town, South Africa.|Centre for Bioprocess Engineering Research, University of Cape Town, Cape Town, South Africa.
authors.forename: Don A
authors.initials: DA
authors.lastname: Cowan
authors.affiliation: Institute for Microbial Biotechnology and Metagenomics, University of the Western Cape, Cape Town, South Africa. ituffin@uwc.ac.za.|Centre for Bioprocess Engineering Research, University of Cape Town, Cape Town, South Africa. ituffin@uwc.ac.za.
authors.forename: Marla
authors.initials: M
authors.lastname: Trindade
mesh_terms: D044602:Cellulases / Q000235:genetics* / Q000378:metabolism*; D004789:Enzyme Activation; D015870:Gene Expression; D015723:Gene Library; D057166:High-Throughput Screening Assays; D059625:Liquid-Liquid Extraction / Q000379:methods*; D054892:Metagenome; D056186:Metagenomics / Q000379:methods; D013379:Substrate Specificity
chemical_list: D044602:Cellulases
keywords: Colorimetric substrates; Function-driven screening; High-throughput screening; Lignocellulosic enzymes; Liquid-phase screening; Metagenomics; Multiplex screening; p-Nitrophenyl substrates; α-L-Arabinofuranosidases; β-Glucosidases; β-Xylosidases