A polysaccharide utilization locus from an uncultured Bacteroidetes phylotype suggests ecological adaptation and substrate versatility

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A polysaccharide utilization locus from an uncultured Bacteroidetes phylotype suggests ecological adaptation and substrate versatility. / Mackenzie, A.K.; Naas, A.E.; Kracun, Stjepan Kresimir; Schückel, Julia; Fangel, Jonatan Ulrik; Agger, J.W.; Willats, William George Tycho; Eijsink, V.G.H.; Pope, P.B.

In: Applied and Environmental Microbiology, Vol. 81, No. 1, 2015, p. 187-195.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Mackenzie, AK, Naas, AE, Kracun, SK, Schückel, J, Fangel, JU, Agger, JW, Willats, WGT, Eijsink, VGH & Pope, PB 2015, 'A polysaccharide utilization locus from an uncultured Bacteroidetes phylotype suggests ecological adaptation and substrate versatility', Applied and Environmental Microbiology, vol. 81, no. 1, pp. 187-195. https://doi.org/10.1128/AEM.02858-14

APA

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. (2015). A polysaccharide utilization locus from an uncultured Bacteroidetes phylotype suggests ecological adaptation and substrate versatility. Applied and Environmental Microbiology, 81(1), 187-195. https://doi.org/10.1128/AEM.02858-14

Vancouver

Mackenzie AK, Naas AE, Kracun SK, Schückel J, Fangel JU, Agger JW et al. A polysaccharide utilization locus from an uncultured Bacteroidetes phylotype suggests ecological adaptation and substrate versatility. Applied and Environmental Microbiology. 2015;81(1):187-195. https://doi.org/10.1128/AEM.02858-14

Author

Mackenzie, A.K. ; Naas, A.E. ; Kracun, Stjepan Kresimir ; Schückel, Julia ; Fangel, Jonatan Ulrik ; Agger, J.W. ; Willats, William George Tycho ; Eijsink, V.G.H. ; Pope, P.B. / A polysaccharide utilization locus from an uncultured Bacteroidetes phylotype suggests ecological adaptation and substrate versatility. In: Applied and Environmental Microbiology. 2015 ; Vol. 81, No. 1. pp. 187-195.

Bibtex

@article{7741c0ea142b4cbfa3ff1d88acaba9f7,
title = "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 reconstructed from an uncultured Bacteroidetes phylotype (SRM-1) that dominates the rumen microbiome of Arctic reindeer. CSaveharacterization 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.",
author = "A.K. Mackenzie and A.E. Naas and Kracun, {Stjepan Kresimir} and Julia Sch{\"u}ckel and Fangel, {Jonatan Ulrik} and J.W. Agger and Willats, {William George Tycho} and V.G.H. Eijsink and P.B. Pope",
note = "Copyright {\textcopyright} 2015, American Society for Microbiology. All Rights Reserved.",
year = "2015",
doi = "10.1128/AEM.02858-14",
language = "English",
volume = "81",
pages = "187--195",
journal = "Applied and Environmental Microbiology",
issn = "0099-2240",
publisher = "American Society for Microbiology",
number = "1",

}

RIS

TY - JOUR

T1 - A polysaccharide utilization locus from an uncultured Bacteroidetes phylotype suggests ecological adaptation and substrate versatility

AU - Mackenzie, A.K.

AU - Naas, A.E.

AU - Kracun, Stjepan Kresimir

AU - Schückel, Julia

AU - Fangel, Jonatan Ulrik

AU - Agger, J.W.

AU - Willats, William George Tycho

AU - Eijsink, V.G.H.

AU - Pope, P.B.

N1 - Copyright © 2015, American Society for Microbiology. All Rights Reserved.

PY - 2015

Y1 - 2015

N2 - 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. CSaveharacterization 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.

AB - 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. CSaveharacterization 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.

U2 - 10.1128/AEM.02858-14

DO - 10.1128/AEM.02858-14

M3 - Journal article

C2 - 25326301

VL - 81

SP - 187

EP - 195

JO - Applied and Environmental Microbiology

JF - Applied and Environmental Microbiology

SN - 0099-2240

IS - 1

ER -

ID: 131740856