Functional Metagenomics: Construction and High-Throughput Screening of Fosmid Libraries for Discovery of Novel Carbohydrate-Active Enzymes

Methods Mol Biol. 2016:1399:257-71. doi: 10.1007/978-1-4939-3369-3_15.

Abstract

Activity-based metagenomics is one of the most efficient approaches to boost the discovery of novel biocatalysts from the huge reservoir of uncultivated bacteria. In this chapter, we describe a highly generic procedure of metagenomic library construction and high-throughput screening for carbohydrate-active enzymes. Applicable to any bacterial ecosystem, it enables the swift identification of functional enzymes that are highly efficient, alone or acting in synergy, to break down polysaccharides and oligosaccharides.

Keywords: Carbohydrate-active enzymes; Complex glycans; Fosmidic libraries; High-throughput screening; Metagenomic DNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteria / enzymology*
  • Bacteria / genetics
  • Carbohydrates / genetics
  • Enzyme Activators / metabolism
  • Enzymes / genetics
  • Enzymes / isolation & purification*
  • Enzymes / metabolism
  • High-Throughput Screening Assays / methods*
  • Metagenomics / methods*
  • Plasmids

Substances

  • Carbohydrates
  • Enzyme Activators
  • Enzymes