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Characterisation of the Effect of the Spatial Organisation of Hemicellulases on the Hydrolysis of Plant Biomass Polymer
- Source :
- International Journal of Molecular Sciences, Vol 21, Iss 4360, p 4360 (2020), International Journal of Molecular Sciences, International Journal of Molecular Sciences, MDPI, 2020, 21 (12), pp.4360. ⟨10.3390/ijms21124360⟩, Volume 21, Issue 12, International Journal of Molecular Sciences, 2020, 21 (12), pp.4360. ⟨10.3390/ijms21124360⟩
- Publication Year :
- 2020
- Publisher :
- MDPI AG, 2020.
-
Abstract
- International audience; Synergism between enzymes is of crucial importance in cell metabolism. This synergism occurs often through a spatial organisation favouring proximity and substrate channelling. In this context, we developed a strategy for evaluating the impact of the geometry between two enzymes involved in nature in the recycling of the carbon derived from plant cell wall polymers. By using an innovative covalent association process using two protein fragments, Jo and In, we produced two bi-modular chimeric complexes connecting a xylanase and a xylosidase, involved in the deconstruction of xylose-based plant cell wall polymer. We first show that the intrinsic activity of the individual enzymes was preserved. Small Angle X-rays Scattering (SAXS) analysis of the complexes highlighted two different spatial organisations in solution, affecting both the distance between the enzymes (53 Å and 28 Å) and the distance between the catalytic pockets (94 Å and 75 Å). Reducing sugar and HPAEC-PAD analysis revealed different behaviour regarding the hydrolysis of Beechwood xylan. After 24 h of hydrolysis, one complex was able to release a higher amount of reducing sugar compare to the free enzymes (i.e., 15,640 and 14,549 µM of equivalent xylose, respectively). However, more interestingly, the two complexes were able to release variable percentages of xylooligosaccharides compared to the free enzymes. The structure of the complexes revealed some putative steric hindrance, which impacted both enzymatic efficiency and the product profile. This report shows that controlling the spatial geometry between two enzymes would help to better investigate synergism effect within complex multi-enzymatic machinery and control the final product.
- Subjects :
- 0106 biological sciences
0301 basic medicine
Oligosaccharides
Xylose
Protein Engineering
01 natural sciences
Enzyme engineering
lcsh:Chemistry
chemistry.chemical_compound
X-Ray Diffraction
Biomass
lcsh:QH301-705.5
Spectroscopy
Plant Proteins
2. Zero hunger
chemistry.chemical_classification
Hydrolysis
General Medicine
Plants
Computer Science Applications
Reducing sugar
Xylosidases
xylosidase
Xylanase
Glycoside Hydrolases
Recombinant Fusion Proteins
Context (language use)
spatial proximity
Xylosidase
Article
Catalysis
Carbon Cycle
Inorganic Chemistry
03 medical and health sciences
Protein Domains
010608 biotechnology
synergism
Scattering, Small Angle
Génie chimique
[SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology
Physical and Theoretical Chemistry
Génie des procédés
Molecular Biology
xylanase
Organic Chemistry
Synergism
Substrate (chemistry)
Protein engineering
Xylan
Spatial proximity
enzyme engineering
030104 developmental biology
chemistry
lcsh:Biology (General)
lcsh:QD1-999
Bio Molecular Welding
Biophysics
Subjects
Details
- Language :
- English
- ISSN :
- 16616596 and 14220067
- Volume :
- 21
- Issue :
- 4360
- Database :
- OpenAIRE
- Journal :
- International Journal of Molecular Sciences
- Accession number :
- edsair.doi.dedup.....0a720e220fe4cfca3a85672d4a427695