High-throughput in vitro glycoside hydrolase (HIGH) screening for enzyme discovery.

A high-throughput protein-expression and screening method (HIGH method, see picture) provides a rapid approach to the discovery of active glycoside hydrolases in environmental samples. Finally, HIGH screening combines cloning, protein expression, and enzyme hydrolysis in one pot; thus, the entire process from gene expression to activity detection requires only three hours.

[1]  Charlotte K. Williams,et al.  The Path Forward for Biofuels and Biomaterials , 2006, Science.

[2]  Diane F. Davis,et al.  Improved Catalytic Efficiency and Active Site Modification of 1,4-β-d-Glucan Glucohydrolase A from Thermotoga neapolitana by Directed Evolution* , 2004, Journal of Biological Chemistry.

[3]  Himmel,et al.  Cellulase for commodity products from cellulosic biomass , 1999, Current opinion in biotechnology.

[4]  Safiyh Taghavi,et al.  Bioprospecting metagenomes: glycosyl hydrolases for converting biomass , 2009, Biotechnology for biofuels.

[5]  Michael E Himmel,et al.  Implications of cellobiohydrolase glycosylation for use in biomass conversion , 2008, Biotechnology for biofuels.

[6]  Dana C Nadler,et al.  Binding modules alter the activity of chimeric cellulases: Effects of biomass pretreatment and enzyme source , 2010, Biotechnology and bioengineering.

[7]  Kevin Burgess,et al.  Fluorescent indicators for intracellular pH. , 2010, Chemical reviews.

[8]  J. Swartz,et al.  Energizing cell-free protein synthesis with glucose metabolism. , 2005, Biotechnology and bioengineering.

[9]  Brandi L. Cantarel,et al.  The Carbohydrate-Active EnZymes database (CAZy): an expert resource for Glycogenomics , 2008, Nucleic Acids Res..

[10]  J. Swartz,et al.  Total amino acid stabilization during cell-free protein synthesis reactions. , 2006, Journal of biotechnology.

[11]  Michael C Jewett,et al.  An integrated cell-free metabolic platform for protein production and synthetic biology , 2008, Molecular systems biology.

[12]  S. Tringe,et al.  Metagenomic Discovery of Biomass-Degrading Genes and Genomes from Cow Rumen , 2011, Science.

[13]  Charles M Schroeder,et al.  Thermostable enzymes as biocatalysts in the biofuel industry. , 2010, Advances in applied microbiology.

[14]  J. Cherry,et al.  Directed evolution of industrial enzymes: an update. , 2003, Current opinion in biotechnology.