A Polysaccharide Utilization Locus from an Uncultured Bacteroidetes Phylotype Suggests Ecological Adaptation and Substrate Versatility
ABSTRACT Recent metagenomic analyses have identified uncultured bacteria that are abundant in the rumen of herbivores and that possess putative biomass-converting enzyme systems. Here we investigate the saccharolytic capabilities of a polysaccharide utilization locus (PUL) that has been reconstructe...
Published in: | Applied and Environmental Microbiology |
---|---|
Main Authors: | , , , , , , , , |
Other Authors: | |
Format: | Article in Journal/Newspaper |
Language: | English |
Published: |
American Society for Microbiology
2015
|
Subjects: | |
Online Access: | http://dx.doi.org/10.1128/aem.02858-14 https://journals.asm.org/doi/pdf/10.1128/AEM.02858-14 |
id |
crasmicro:10.1128/aem.02858-14 |
---|---|
record_format |
openpolar |
spelling |
crasmicro:10.1128/aem.02858-14 2024-06-23T07:50:46+00:00 A Polysaccharide Utilization Locus from an Uncultured Bacteroidetes Phylotype Suggests Ecological Adaptation and Substrate Versatility Mackenzie, A. K. Naas, A. E. Kracun, S. K. Schückel, J. Fangel, J. U. Agger, J. W. Willats, W. G. T. Eijsink, V. G. H. Pope, P. B. Drake, H. L. 2015 http://dx.doi.org/10.1128/aem.02858-14 https://journals.asm.org/doi/pdf/10.1128/AEM.02858-14 en eng American Society for Microbiology https://journals.asm.org/non-commercial-tdm-license Applied and Environmental Microbiology volume 81, issue 1, page 187-195 ISSN 0099-2240 1098-5336 journal-article 2015 crasmicro https://doi.org/10.1128/aem.02858-14 2024-05-27T12:59:18Z ABSTRACT Recent metagenomic analyses have identified uncultured bacteria that are abundant in the rumen of herbivores and that possess putative biomass-converting enzyme systems. Here we investigate the saccharolytic capabilities of a polysaccharide utilization locus (PUL) that has been reconstructed from an uncultured Bacteroidetes phylotype (SRM-1) that dominates the rumen microbiome of Arctic reindeer. Characterization of the three PUL-encoded outer membrane glycoside hydrolases was performed using chromogenic substrates for initial screening, followed by detailed analyses of products generated from selected substrates, using high-pressure anion-exchange chromatography with electrochemical detection. Two glycoside hydrolase family 5 (GH5) endoglucanases (GH5_g and GH5_h) demonstrated activity against β-glucans, xylans, and xyloglucan, whereas GH5_h and the third enzyme, GH26_i, were active on several mannan substrates. Synergy experiments examining different combinations of the three enzymes demonstrated limited activity enhancement on individual substrates. Binding analysis of a SusE-positioned lipoprotein revealed an affinity toward β-glucans and, to a lesser extent, mannan, but unlike the two SusD-like lipoproteins previously characterized from the same PUL, binding to cellulose was not observed. Overall, these activities and binding specificities correlated well with the glycan content of the reindeer rumen, which was determined using comprehensive microarray polymer profiling and showed an abundance of various hemicellulose glycans. The substrate versatility of this single PUL putatively expands our perceptions regarding PUL machineries, which so far have demonstrated gene organization that suggests one cognate PUL for each substrate type. The presence of a PUL that possesses saccharolytic activity against a mixture of abundantly available polysaccharides supports the dominance of SRM-1 in the Svalbard reindeer rumen microbiome. Article in Journal/Newspaper Arctic Svalbard svalbard reindeer ASM Journals (American Society for Microbiology) Arctic Svalbard Applied and Environmental Microbiology 81 1 187 195 |
institution |
Open Polar |
collection |
ASM Journals (American Society for Microbiology) |
op_collection_id |
crasmicro |
language |
English |
description |
ABSTRACT Recent metagenomic analyses have identified uncultured bacteria that are abundant in the rumen of herbivores and that possess putative biomass-converting enzyme systems. Here we investigate the saccharolytic capabilities of a polysaccharide utilization locus (PUL) that has been reconstructed from an uncultured Bacteroidetes phylotype (SRM-1) that dominates the rumen microbiome of Arctic reindeer. Characterization of the three PUL-encoded outer membrane glycoside hydrolases was performed using chromogenic substrates for initial screening, followed by detailed analyses of products generated from selected substrates, using high-pressure anion-exchange chromatography with electrochemical detection. Two glycoside hydrolase family 5 (GH5) endoglucanases (GH5_g and GH5_h) demonstrated activity against β-glucans, xylans, and xyloglucan, whereas GH5_h and the third enzyme, GH26_i, were active on several mannan substrates. Synergy experiments examining different combinations of the three enzymes demonstrated limited activity enhancement on individual substrates. Binding analysis of a SusE-positioned lipoprotein revealed an affinity toward β-glucans and, to a lesser extent, mannan, but unlike the two SusD-like lipoproteins previously characterized from the same PUL, binding to cellulose was not observed. Overall, these activities and binding specificities correlated well with the glycan content of the reindeer rumen, which was determined using comprehensive microarray polymer profiling and showed an abundance of various hemicellulose glycans. The substrate versatility of this single PUL putatively expands our perceptions regarding PUL machineries, which so far have demonstrated gene organization that suggests one cognate PUL for each substrate type. The presence of a PUL that possesses saccharolytic activity against a mixture of abundantly available polysaccharides supports the dominance of SRM-1 in the Svalbard reindeer rumen microbiome. |
author2 |
Drake, H. L. |
format |
Article in Journal/Newspaper |
author |
Mackenzie, A. K. Naas, A. E. Kracun, S. K. Schückel, J. Fangel, J. U. Agger, J. W. Willats, W. G. T. Eijsink, V. G. H. Pope, P. B. |
spellingShingle |
Mackenzie, A. K. Naas, A. E. Kracun, S. K. Schückel, J. Fangel, J. U. Agger, J. W. Willats, W. G. T. Eijsink, V. G. H. Pope, P. B. A Polysaccharide Utilization Locus from an Uncultured Bacteroidetes Phylotype Suggests Ecological Adaptation and Substrate Versatility |
author_facet |
Mackenzie, A. K. Naas, A. E. Kracun, S. K. Schückel, J. Fangel, J. U. Agger, J. W. Willats, W. G. T. Eijsink, V. G. H. Pope, P. B. |
author_sort |
Mackenzie, A. K. |
title |
A Polysaccharide Utilization Locus from an Uncultured Bacteroidetes Phylotype Suggests Ecological Adaptation and Substrate Versatility |
title_short |
A Polysaccharide Utilization Locus from an Uncultured Bacteroidetes Phylotype Suggests Ecological Adaptation and Substrate Versatility |
title_full |
A Polysaccharide Utilization Locus from an Uncultured Bacteroidetes Phylotype Suggests Ecological Adaptation and Substrate Versatility |
title_fullStr |
A Polysaccharide Utilization Locus from an Uncultured Bacteroidetes Phylotype Suggests Ecological Adaptation and Substrate Versatility |
title_full_unstemmed |
A Polysaccharide Utilization Locus from an Uncultured Bacteroidetes Phylotype Suggests Ecological Adaptation and Substrate Versatility |
title_sort |
polysaccharide utilization locus from an uncultured bacteroidetes phylotype suggests ecological adaptation and substrate versatility |
publisher |
American Society for Microbiology |
publishDate |
2015 |
url |
http://dx.doi.org/10.1128/aem.02858-14 https://journals.asm.org/doi/pdf/10.1128/AEM.02858-14 |
geographic |
Arctic Svalbard |
geographic_facet |
Arctic Svalbard |
genre |
Arctic Svalbard svalbard reindeer |
genre_facet |
Arctic Svalbard svalbard reindeer |
op_source |
Applied and Environmental Microbiology volume 81, issue 1, page 187-195 ISSN 0099-2240 1098-5336 |
op_rights |
https://journals.asm.org/non-commercial-tdm-license |
op_doi |
https://doi.org/10.1128/aem.02858-14 |
container_title |
Applied and Environmental Microbiology |
container_volume |
81 |
container_issue |
1 |
container_start_page |
187 |
op_container_end_page |
195 |
_version_ |
1802641676850692096 |