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.