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Singh, Ravindra Pal, Bhaiyya, Raja, Thakur, Raksha, Niharika, Jayashree, Singh, Chandrajeet, Latousakis, Dimitrios, Saalbach, Gerhard, Nepogodiev, Sergey A., Singh, Praveen, Sharma, Sukesh Chander, Sengupta, Shantanu, Juge, Nathalie and Field, Robert A. 
ORCID: https://orcid.org/0000-0001-8574-0275
(2022)
Biochemical basis of xylooligosaccharide utilisation by gut bacteria.
International Journal of Molecular Sciences, 23 (6).
ISSN 1661-6596
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Abstract
Xylan is one of the major structural components of the plant cell wall. Xylan present in the human diet reaches the large intestine undigested and becomes a substrate to species of the gut microbiota. Here, we characterised the capacity of Limosilactobacillus reuteri and Blautia producta strains to utilise xylan derivatives. We showed that L. reuteri ATCC 53608 and B. producta ATCC 27340 produced β-D-xylosidases, enabling growth on xylooligosaccharide (XOS). The recombinant enzymes were highly active on artificial (p-nitrophenyl β-D-xylopyranoside) and natural (xylobiose, xylotriose, and xylotetraose) substrates, and showed transxylosylation activity and tolerance to xylose inhibition. The enzymes belong to glycoside hydrolase family 120 with Asp as nucleophile and Glu as proton donor, as shown by homology modelling and confirmed by site-directed mutagenesis. In silico analysis revealed that these enzymes were part of a gene cluster in L. reuteri but not in Blautia strains, and quantitative proteomics identified other enzymes and transporters involved in B. producta XOS utilisation. Based on these findings, we proposed a model for an XOS metabolism pathway in L. reuteri and B. producta strains. Together with phylogenetic analyses, the data also revealed the extended xylanolytic potential of the gut microbiota.
| Item Type: | Article |
|---|---|
| Additional Information: | Funding Information: These studies were supported by the UK BBSRC Institute Strategic Program on Molecules from Nature—Products and Pathways (BBS/E/J/000PR9790) and the John Innes Foundation; the BBSRC and EPSRC-Newton Fund (BB/N005082/1); the Department for International Development; the BBSRC Institute Strategic Programme Grant Gut Microbes and Health (BB/R012490/1) and its constituent project BBS/E/F/000PR10353 (Theme 1, Determinants of microbe–host responses in the gut across life); the Indian Department of Biotechnology under the Newton Fund Global Research Partnership in Aquaculture programme; and the InnovateUK IBCatalyst (BB/M02903411 and EP/N033167/10). |
| Uncontrolled Keywords: | blautia producta,carbohydrate-active enzymes,glycoside hydrolase,gut microbiota,limosilactobacillus reuteri,xylan,xylooligosaccharide,β-xylosidases,catalysis,molecular biology,spectroscopy,computer science applications,physical and theoretical chemistry,organic chemistry,inorganic chemistry ,/dk/atira/pure/subjectarea/asjc/1500/1503 |
| Faculty \ School: | Faculty of Science > School of Biological Sciences Faculty of Science > School of Chemistry, Pharmacy and Pharmacology |
| Related URLs: | |
| Depositing User: | LivePure Connector |
| Date Deposited: | 03 Sep 2024 09:36 |
| Last Modified: | 12 Sep 2024 15:30 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/96461 |
| DOI: | 10.3390/ijms23062992 |
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