Methods for discovery of novel cellulosomal cellulases using genomics and biochemical tools

Yonit Ben-David, Bareket Dassa, Lizi Bensoussan, Edward A. Bayer, Sarah Moraïs

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

Abstract

Cell wall degradation by cellulases is extensively explored owing to its potential contribution to biofuel production. The cellulosome is an extracellular multienzyme complex that can degrade the plant cell wall very efficiently, and cellulosomal enzymes are therefore of great interest. The cellulosomal cellulases are defined as enzymes that contain a dockerin module, which can interact with a cohesin module contained in multiple copies in a noncatalytic protein, termed scaffoldin. The assembly of the cellulosomal cellulases into the cellulosomal complex occurs via specific protein–protein interactions. Cellulosome systems have been described initially only in several anaerobic cellulolytic bacteria. However, owing to ongoing genome sequencing and metagenomic projects, the discovery of novel cellulosome-producing bacteria and the description of their cellulosomal genes have dramatically increased in the recent years. In this chapter, methods for discovery of novel cellulosomal cellulases from a DNA sequence by bioinformatics and biochemical tools are described. Their biochemical characterization is also described, including both the enzymatic activity of the putative cellulases and their assembly into mature designer cellulosomes.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages67-84
Number of pages18
DOIs
StatePublished - 1 Jan 2018
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume1796
ISSN (Print)1064-3745

Keywords

  • Biofuels
  • Biomass degradation
  • Carbohydrate-binding module (CBM)
  • Cellulases
  • Cellulosomes
  • Cohesin
  • Dockerin
  • Glycoside hydrolases
  • Scaffoldin

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