Cargando…

Identification and characterization of novel cellulolytic and hemicellulolytic genes and enzymes derived from German grassland soil metagenomes

Soil metagenomes represent an unlimited resource for the discovery of novel biocatalysts from soil microorganisms. Three large-inserts metagenomic DNA libraries were constructed from different grassland soil samples and screened for genes conferring cellulase or xylanase activity. Function-driven sc...

Descripción completa

Detalles Bibliográficos
Autores principales: Nacke, Heiko, Engelhaupt, Martin, Brady, Silja, Fischer, Christiane, Tautzt, Janine, Daniel, Rolf
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Netherlands 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3298741/
https://www.ncbi.nlm.nih.gov/pubmed/22187078
http://dx.doi.org/10.1007/s10529-011-0830-2
_version_ 1782226034796527616
author Nacke, Heiko
Engelhaupt, Martin
Brady, Silja
Fischer, Christiane
Tautzt, Janine
Daniel, Rolf
author_facet Nacke, Heiko
Engelhaupt, Martin
Brady, Silja
Fischer, Christiane
Tautzt, Janine
Daniel, Rolf
author_sort Nacke, Heiko
collection PubMed
description Soil metagenomes represent an unlimited resource for the discovery of novel biocatalysts from soil microorganisms. Three large-inserts metagenomic DNA libraries were constructed from different grassland soil samples and screened for genes conferring cellulase or xylanase activity. Function-driven screening identified a novel cellulase-encoding gene (cel01) and two xylanase-encoding genes (xyn01 and xyn02). From sequence and protein domain analyses, Cel01 (831 amino acids) belongs to glycoside hydrolase family 9 whereas Xyn01 (170 amino acids) and Xyn02 (255 amino acids) are members of glycoside hydrolase family 11. Cel01 harbors a family 9 carbohydrate-binding module, previously found only in xylanases. Both Xyn01 and Xyn02 were most active at 60°C with high activities from 4 to 10 and optimal at pH 7 (Xyn01) and pH 6 (Xyn02). The cellulase gene, cel01, was expressed in E. coli BL21 and the recombinant enzyme (91.9 kDa) was purified. Cel01 exhibited high activity with soluble cellulose substrates containing β-1,4-linkages. Activity with microcrystalline cellulose was not detected. These data, together with the analysis of the degradation profiles of carboxymethyl cellulose and barley glucan indicated that Cel01 is an endo 1,4-β-glucanase. Cel01 showed optimal activity at 50°C and pH 7 being highly active from pH range 5 to 9 and possesses remarkable halotolerance. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s10529-011-0830-2) contains supplementary material, which is available to authorized users.
format Online
Article
Text
id pubmed-3298741
institution National Center for Biotechnology Information
language English
publishDate 2011
publisher Springer Netherlands
record_format MEDLINE/PubMed
spelling pubmed-32987412012-03-21 Identification and characterization of novel cellulolytic and hemicellulolytic genes and enzymes derived from German grassland soil metagenomes Nacke, Heiko Engelhaupt, Martin Brady, Silja Fischer, Christiane Tautzt, Janine Daniel, Rolf Biotechnol Lett Original Research Paper Soil metagenomes represent an unlimited resource for the discovery of novel biocatalysts from soil microorganisms. Three large-inserts metagenomic DNA libraries were constructed from different grassland soil samples and screened for genes conferring cellulase or xylanase activity. Function-driven screening identified a novel cellulase-encoding gene (cel01) and two xylanase-encoding genes (xyn01 and xyn02). From sequence and protein domain analyses, Cel01 (831 amino acids) belongs to glycoside hydrolase family 9 whereas Xyn01 (170 amino acids) and Xyn02 (255 amino acids) are members of glycoside hydrolase family 11. Cel01 harbors a family 9 carbohydrate-binding module, previously found only in xylanases. Both Xyn01 and Xyn02 were most active at 60°C with high activities from 4 to 10 and optimal at pH 7 (Xyn01) and pH 6 (Xyn02). The cellulase gene, cel01, was expressed in E. coli BL21 and the recombinant enzyme (91.9 kDa) was purified. Cel01 exhibited high activity with soluble cellulose substrates containing β-1,4-linkages. Activity with microcrystalline cellulose was not detected. These data, together with the analysis of the degradation profiles of carboxymethyl cellulose and barley glucan indicated that Cel01 is an endo 1,4-β-glucanase. Cel01 showed optimal activity at 50°C and pH 7 being highly active from pH range 5 to 9 and possesses remarkable halotolerance. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s10529-011-0830-2) contains supplementary material, which is available to authorized users. Springer Netherlands 2011-12-21 2012 /pmc/articles/PMC3298741/ /pubmed/22187078 http://dx.doi.org/10.1007/s10529-011-0830-2 Text en © The Author(s) 2011 https://creativecommons.org/licenses/by-nc/4.0/ This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.
spellingShingle Original Research Paper
Nacke, Heiko
Engelhaupt, Martin
Brady, Silja
Fischer, Christiane
Tautzt, Janine
Daniel, Rolf
Identification and characterization of novel cellulolytic and hemicellulolytic genes and enzymes derived from German grassland soil metagenomes
title Identification and characterization of novel cellulolytic and hemicellulolytic genes and enzymes derived from German grassland soil metagenomes
title_full Identification and characterization of novel cellulolytic and hemicellulolytic genes and enzymes derived from German grassland soil metagenomes
title_fullStr Identification and characterization of novel cellulolytic and hemicellulolytic genes and enzymes derived from German grassland soil metagenomes
title_full_unstemmed Identification and characterization of novel cellulolytic and hemicellulolytic genes and enzymes derived from German grassland soil metagenomes
title_short Identification and characterization of novel cellulolytic and hemicellulolytic genes and enzymes derived from German grassland soil metagenomes
title_sort identification and characterization of novel cellulolytic and hemicellulolytic genes and enzymes derived from german grassland soil metagenomes
topic Original Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3298741/
https://www.ncbi.nlm.nih.gov/pubmed/22187078
http://dx.doi.org/10.1007/s10529-011-0830-2
work_keys_str_mv AT nackeheiko identificationandcharacterizationofnovelcellulolyticandhemicellulolyticgenesandenzymesderivedfromgermangrasslandsoilmetagenomes
AT engelhauptmartin identificationandcharacterizationofnovelcellulolyticandhemicellulolyticgenesandenzymesderivedfromgermangrasslandsoilmetagenomes
AT bradysilja identificationandcharacterizationofnovelcellulolyticandhemicellulolyticgenesandenzymesderivedfromgermangrasslandsoilmetagenomes
AT fischerchristiane identificationandcharacterizationofnovelcellulolyticandhemicellulolyticgenesandenzymesderivedfromgermangrasslandsoilmetagenomes
AT tautztjanine identificationandcharacterizationofnovelcellulolyticandhemicellulolyticgenesandenzymesderivedfromgermangrasslandsoilmetagenomes
AT danielrolf identificationandcharacterizationofnovelcellulolyticandhemicellulolyticgenesandenzymesderivedfromgermangrasslandsoilmetagenomes