Conversion of the free Cellvibrio japonicus xyloglucan degradation system to the cellulosomal mode.
DNA assembly
Designer cellulosome
Hemicellulase
Multi-enzyme complex
VersaTile
Xyloglucan
Journal
Applied microbiology and biotechnology
ISSN: 1432-0614
Titre abrégé: Appl Microbiol Biotechnol
Pays: Germany
ID NLM: 8406612
Informations de publication
Date de publication:
Sep 2022
Sep 2022
Historique:
received:
14
04
2022
accepted:
06
07
2022
revised:
05
07
2022
pubmed:
23
7
2022
medline:
31
8
2022
entrez:
22
7
2022
Statut:
ppublish
Résumé
Cellulosomes are multi-enzyme complexes produced by specialised micro-organisms. The spatial proximity of synergistically acting enzymes incorporated in these naturally occurring complexes supports the efficient hydrolysis of lignocellulosic biomass. Several functional designer cellulosomes, incorporating naturally non-cellulosomal cellulases, have been constructed and can be used for cellulose saccharification. However, in lignocellulosic biomass, cellulose is tightly intertwined with several hemicelluloses and lignin. One of the most abundant hemicelluloses interacting with cellulose microfibrils is xyloglucan, and degradation of these polymers is crucial for complete saccharification. Yet, designer cellulosome studies focusing on the incorporation of hemicellulases have been limited. Here, we report the conversion of the free Cellvibrio japonicus xyloglucan degradation system to the cellulosomal mode. Therefore, we constructed multiple docking enzyme variants of C. japonicus endoxyloglucanase, β-1,2-galactosidase, α-1,6 xylosidase and β-1,4-glucosidase, using the combinatorial VersaTile technique dedicated to the design and optimisation of modular proteins. We individually optimised the docking enzymes to degrade the xyloglucan backbone and side chains. Finally, we show that a purified designer xyloglucanosome comprising these docking enzymes was able to release xyloglucan oligosaccharides, galactose, xylose and glucose from tamarind xyloglucan. KEY POINTS: • Construction of xyloglucan-degrading designer cellulosome. • Conversion of free Cellvibrio japonicus enzymes to cellulosomal mode. • Type of linker inserted between dockerin and enzyme module affects docking enzyme activity.
Identifiants
pubmed: 35869373
doi: 10.1007/s00253-022-12072-0
pii: 10.1007/s00253-022-12072-0
doi:
Substances chimiques
Bacterial Proteins
0
Glucans
0
Xylans
0
xyloglucan
37294-28-3
Cellulose
9004-34-6
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
5495-5509Subventions
Organisme : Bijzonder Onderzoeksfonds UGent
ID : BOF17/DOC/086
Organisme : Bijzonder Onderzoeksfonds UGent
ID : BOF16/STA/024
Organisme : Fonds Wetenschappelijk Onderzoek
ID : 1SE2621N
Informations de copyright
© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.
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