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Glycomic profiling identifies key-structural differences in three arabinoxylan fractions from sugarcane culms.

Carolina Victal Garbelotti1, Adriana Grandis2, Eduardo Crevelin1

  • 1Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP CEP 14040-901, Brazil.

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Summary

Understanding sugarcane biomass structure is key for biofuels. Different pretreatment methods affect how easily enzymes break down arabinoxylans, impacting sugar release for biorefineries.

Keywords:
Chemometric analysisHemicellulolytic enzymesHemicelluloseMass spectroscopyOligosaccharides extractionSugarcane biomass

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Area of Science:

  • Biomass processing
  • Enzymatic hydrolysis
  • Plant cell wall structure

Background:

  • Sugarcane biomass is a valuable resource for bioenergy and biorefineries.
  • Complex plant cell walls, particularly arabinoxylans, require pretreatment for efficient enzymatic hydrolysis.
  • Arabinoxylans need multiple enzymes to break down their backbone and side branches for xylose release.

Purpose of the Study:

  • To investigate how different sugarcane arabinoxylan structures influence xylooligosaccharide (XOS) release using various xylanolytic enzymes.
  • To identify the impact of side-branch decorations and pretreatment methods on enzyme accessibility and hydrolysis efficiency.
  • To develop strategies for optimizing enzyme combinations to overcome biomass recalcitrance.

Main Methods:

  • Extraction of three distinct arabinoxylan fractions from sugarcane culms using sodium chlorite, DMSO, and alkaline treatments.
  • Enzymatic hydrolysis using combinations of up to five xylanolytic enzymes.
  • Analysis of reducing sugar release and Liquid Chromatography-Mass Spectrometry (LC-MS) detection of xylooligosaccharides.
  • Application of chemometric analysis to identify XOS profiles and enzyme-substrate relationships.

Main Results:

  • Distinct xylooligosaccharide profiles were observed for each arabinoxylan extract after enzymatic treatment, as identified by LC-MS and chemometric analysis.
  • The degree and position of arabinoxylan side-branch decorations significantly affected enzyme activity and XOS diversity.
  • Alkaline-extracted arabinoxylans were most accessible, while post-sodium chlorite extracts were most recalcitrant, with acetyl substituents identified as a major recalcitrance factor.
  • GH10 xylanase hydrolysis products were found to be substrates for other enzymes, such as α-glucuronidase, highlighting synergistic enzyme actions.

Conclusions:

  • Arabinoxylan structure, influenced by pretreatment, dictates enzyme accessibility and the resulting xylooligosaccharide profile.
  • Acetyl substituents are a key factor contributing to the recalcitrance of sugarcane biomass.
  • The study provides a framework for selecting optimal enzyme combinations to enhance biomass processing, pretreatment selection, and sugar release for biorefinery applications.