Metagenomic approaches to the discovery of cellulases

Methods Enzymol. 2012:510:375-94. doi: 10.1016/B978-0-12-415931-0.00020-3.

Abstract

Most of the microorganisms responsible for nutrient cycling in the environment have yet to be cultivated, and this could include those species responsible for the degradation of cellulose. Known cellulases are well defined at the protein sequence level, but gene variants are difficult to amplify from environmental DNA. The identification of novel cellulase genes independent of DNA amplification is made possible by adopting a direct metagenome sequencing approach to provide genes that can be cloned, expressed, and characterized prior to potential exploitation, all in the absence of any information on the species from which they originated. In this chapter, emerging strategies and methods that will enable the identification of novel cellulase genes and provide an unbiased perspective on gene expression in situ are presented.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteria / enzymology*
  • Bacteria / genetics*
  • Cellulases / genetics*
  • DNA / genetics
  • DNA / isolation & purification
  • Fungi / enzymology*
  • Fungi / genetics*
  • Gene Expression Profiling / methods
  • Gene Library
  • Metagenomics / methods*
  • RNA / genetics
  • RNA / isolation & purification
  • Transduction, Genetic

Substances

  • RNA
  • DNA
  • Cellulases