Evolution of the Cellulose Microfibril through Gamma-Valerolactone-Assisted Co-Solvent and Enzymatic Hydrolysis

Elise B. Gilcher, Nathaniel Kuch, Joshua T. Del Mundo, Samantha F. Ausman, Leoncio Santiago-Martinez, Catherine F.M. Clewett, Esther W. Gomez, Enrique D. Gomez, Thatcher W. Root, Brian G. Fox, James A. Dumesic

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

Biomass recalcitrance during deconstruction remains a key bottleneck to affordable biomass processing technologies. A clear connection between the cell wall structure and biomass deconstruction is necessary to understand how lignocellulosic material is broken down to valuable monomeric components. Here, we monitor changes in the cellulose microfibril domains of poplar, sorghum, and switchgrass throughout gamma-valerolactone (GVL)-water co-solvent pretreatment and enzymatic hydrolysis using solid-state 13C cross-polarization magic angle spinning nuclear magnetic resonance spectroscopy (CP/MAS 13C-NMR) and wide-angle X-ray scattering (WAXS). Spectral fitting of NMR peaks corresponding to different cellulose microenvironments at the C4 carbon center suggests that a mildly acidic GVL-water co-solvent pretreatment of poplar leads to nearly full removal of xylan-cellulose linkages, which primes the cellulose for enzymatic attack. The spectral fitting also suggests that the pretreatment causes significant depletion of the inaccessible fibril surface domains with an increase in more thermally stable crystalline resonances (Iβ). WAXS confirmed a decrease in the lattice spacing between (200) crystalline planes with increasing co-solvent pretreatment severity. These results are interpreted as an opening of bound microfibril surfaces previously inaccessible to the co-solvent system, which leaves behind a more thermally stable, crystalline domain that is potentially prone to relaxation and recrystallization. Full conversion of residual GVL-pretreated biomass was achieved after the GVL co-solvent pretreatment at 140 °C using a commercial enzyme cocktail, CTec2, which contains different cellulases and other enzymes. Spectral fitting of enzymatically hydrolyzed samples by a single engineered cellulase, CelR, suggests that the residual cellulose recalcitrance is mainly due to the inability of CelR to digest the Iβ crystalline domain present in pretreated samples. This work helps to provide new information regarding the structure of the cell wall and recalcitrance throughout GVL-water mild acidolysis and CelR enzymatic biomass deconstruction by tracking the evolution of structural domains within the cellulose microfibril. This work further directs recommendations for improving the conversion and sugar yields in future studies. Our findings inform inquiry into larger questions of cellulose recalcitrance through GVL pretreatment and CelR enzymatic hydrolysis and give insight into subsequent required steps for full cellulose conversion with attention to the most recalcitrant cellulose structures.

Original languageEnglish (US)
Pages (from-to)3270-3283
Number of pages14
JournalACS Sustainable Chemistry and Engineering
Volume11
Issue number8
DOIs
StatePublished - Feb 27 2023

All Science Journal Classification (ASJC) codes

  • General Chemistry
  • Environmental Chemistry
  • General Chemical Engineering
  • Renewable Energy, Sustainability and the Environment

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