Biochemical Characterization of a Family 15 Carbohydrate Esterase from a Bacterial Marine Arctic Metagenome.
The glucuronoyl esterase enzymes of wood-degrading fungi (Carbohydrate Esterase family 15; CE15) form part of the hemicellulolytic and cellulolytic enzyme systems that break down plant biomass, and have possible applications in biotechnology. Homologous enzymes are predicted in the genomes of severa...
Published in: | PLOS ONE |
---|---|
Main Authors: | , , |
Format: | Article in Journal/Newspaper |
Language: | English |
Published: |
Public Library of Science (PLoS)
2016
|
Subjects: | |
Online Access: | https://doi.org/10.1371/journal.pone.0159345 https://doaj.org/article/02d9daacd4824dd2bdb45b266881119d |
id |
ftdoajarticles:oai:doaj.org/article:02d9daacd4824dd2bdb45b266881119d |
---|---|
record_format |
openpolar |
spelling |
ftdoajarticles:oai:doaj.org/article:02d9daacd4824dd2bdb45b266881119d 2023-05-15T15:00:54+02:00 Biochemical Characterization of a Family 15 Carbohydrate Esterase from a Bacterial Marine Arctic Metagenome. Concetta De Santi Nils Peder Willassen Adele Williamson 2016-01-01T00:00:00Z https://doi.org/10.1371/journal.pone.0159345 https://doaj.org/article/02d9daacd4824dd2bdb45b266881119d EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC4951047?pdf=render https://doaj.org/toc/1932-6203 1932-6203 doi:10.1371/journal.pone.0159345 https://doaj.org/article/02d9daacd4824dd2bdb45b266881119d PLoS ONE, Vol 11, Iss 7, p e0159345 (2016) Medicine R Science Q article 2016 ftdoajarticles https://doi.org/10.1371/journal.pone.0159345 2022-12-31T12:42:20Z The glucuronoyl esterase enzymes of wood-degrading fungi (Carbohydrate Esterase family 15; CE15) form part of the hemicellulolytic and cellulolytic enzyme systems that break down plant biomass, and have possible applications in biotechnology. Homologous enzymes are predicted in the genomes of several bacteria, however these have been much less studied than their fungal counterparts. Here we describe the recombinant production and biochemical characterization of a bacterial CE15 enzyme denoted MZ0003, which was identified by in silico screening of a prokaryotic metagenome library derived from marine Arctic sediment. MZ0003 has high similarity to several uncharacterized gene products of polysaccharide-degrading bacterial species, and phylogenetic analysis indicates a deep evolutionary split between these CE15s and fungal homologs.MZ0003 appears to differ from previously-studied CE15s in some aspects. Some glucuronoyl esterase activity could be measured by qualitative thin-layer chromatography which confirms its assignment as a CE15, however MZ0003 can also hydrolyze a range of other esters, including p-nitrophenyl acetate, which is not acted upon by some fungal homologs. The structure of MZ0003 also appears to differ as it is predicted to have several large loop regions that are absent in previously studied CE15s, and a combination of homology-based modelling and site-directed mutagenesis indicate its catalytic residues deviate from the conserved Ser-His-Glu triad of many fungal CE15s. Taken together, these results indicate that potentially unexplored diversity exists among bacterial CE15s, and this may be accessed by investigation of the microbial metagenome. The combination of low activity on typical glucuronoyl esterase substrates, and the lack of glucuronic acid esters in the marine environment suggest that the physiological substrate of MZ0003 and its homologs is likely to be different from that of related fungal enzymes. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLOS ONE 11 7 e0159345 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Medicine R Science Q |
spellingShingle |
Medicine R Science Q Concetta De Santi Nils Peder Willassen Adele Williamson Biochemical Characterization of a Family 15 Carbohydrate Esterase from a Bacterial Marine Arctic Metagenome. |
topic_facet |
Medicine R Science Q |
description |
The glucuronoyl esterase enzymes of wood-degrading fungi (Carbohydrate Esterase family 15; CE15) form part of the hemicellulolytic and cellulolytic enzyme systems that break down plant biomass, and have possible applications in biotechnology. Homologous enzymes are predicted in the genomes of several bacteria, however these have been much less studied than their fungal counterparts. Here we describe the recombinant production and biochemical characterization of a bacterial CE15 enzyme denoted MZ0003, which was identified by in silico screening of a prokaryotic metagenome library derived from marine Arctic sediment. MZ0003 has high similarity to several uncharacterized gene products of polysaccharide-degrading bacterial species, and phylogenetic analysis indicates a deep evolutionary split between these CE15s and fungal homologs.MZ0003 appears to differ from previously-studied CE15s in some aspects. Some glucuronoyl esterase activity could be measured by qualitative thin-layer chromatography which confirms its assignment as a CE15, however MZ0003 can also hydrolyze a range of other esters, including p-nitrophenyl acetate, which is not acted upon by some fungal homologs. The structure of MZ0003 also appears to differ as it is predicted to have several large loop regions that are absent in previously studied CE15s, and a combination of homology-based modelling and site-directed mutagenesis indicate its catalytic residues deviate from the conserved Ser-His-Glu triad of many fungal CE15s. Taken together, these results indicate that potentially unexplored diversity exists among bacterial CE15s, and this may be accessed by investigation of the microbial metagenome. The combination of low activity on typical glucuronoyl esterase substrates, and the lack of glucuronic acid esters in the marine environment suggest that the physiological substrate of MZ0003 and its homologs is likely to be different from that of related fungal enzymes. |
format |
Article in Journal/Newspaper |
author |
Concetta De Santi Nils Peder Willassen Adele Williamson |
author_facet |
Concetta De Santi Nils Peder Willassen Adele Williamson |
author_sort |
Concetta De Santi |
title |
Biochemical Characterization of a Family 15 Carbohydrate Esterase from a Bacterial Marine Arctic Metagenome. |
title_short |
Biochemical Characterization of a Family 15 Carbohydrate Esterase from a Bacterial Marine Arctic Metagenome. |
title_full |
Biochemical Characterization of a Family 15 Carbohydrate Esterase from a Bacterial Marine Arctic Metagenome. |
title_fullStr |
Biochemical Characterization of a Family 15 Carbohydrate Esterase from a Bacterial Marine Arctic Metagenome. |
title_full_unstemmed |
Biochemical Characterization of a Family 15 Carbohydrate Esterase from a Bacterial Marine Arctic Metagenome. |
title_sort |
biochemical characterization of a family 15 carbohydrate esterase from a bacterial marine arctic metagenome. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2016 |
url |
https://doi.org/10.1371/journal.pone.0159345 https://doaj.org/article/02d9daacd4824dd2bdb45b266881119d |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS ONE, Vol 11, Iss 7, p e0159345 (2016) |
op_relation |
http://europepmc.org/articles/PMC4951047?pdf=render https://doaj.org/toc/1932-6203 1932-6203 doi:10.1371/journal.pone.0159345 https://doaj.org/article/02d9daacd4824dd2bdb45b266881119d |
op_doi |
https://doi.org/10.1371/journal.pone.0159345 |
container_title |
PLOS ONE |
container_volume |
11 |
container_issue |
7 |
container_start_page |
e0159345 |
_version_ |
1766332953197019136 |