Understanding the impact of steam pretreatment severity on cellulose ultrastructure, recalcitrance, and hydrolyzability of Norway spruce

Fabio Caputo, Basel Al-Rudainy, Polina Naidjonoka, Ola Wallberg, Lisbeth Olsson, Vera Novy

Research output: Contribution to journalArticlepeer-review


The efficient use of softwood in biorefineries requires harsh pretreatment conditions to overcome biomass recalcitrance. Following harsh pretreatments, the hemicellulose is solubilized. Here, we studied the material characteristics of Norway spruce following steam pretreatment at six different severities, relating chemical and structural information to the enzymatic hydrolyzability. Steam pretreatment conditions were defined by two different temperatures (180 °C and 210 °C), with and without the addition of various acids (CH3COOH, H3PO4, H2SO4, SO2). Structural knowledge of the streams is a cornerstone for developing an efficient saccharification process.

This study combines advanced structural characterizations to gain fundamental understanding of the influence of severity of pretreatment on spruce. Structural knowledge is a cornerstone in developing an effective saccharification process by modulating pretreatment conditions and enzymes employed.

Overall structural properties were assessed by scanning electron microscopy. The effect of stream pretreatment severity on lignin and lignin-carbohydrate bonds was investigated by two-dimensional heteronuclear single quantum correlation nuclear magnetic resonance. Finally, cellulose ultrastructure was studied by applying small/wide-angle X-ray scattering. The structural characteristics of the six pretreated softwood substrates were related to the enzymatic hydrolyzability.

With increasing pretreatment severity, surface defibrillation, and lignin depolymeryzation were observed. Further, lignin-carbohydrate complexes signals were detected. Cellulose analysis revealed the rearrangement of microfibrils leading to the formation of larger microfibril aggregates. This microfibril rearrangement likely contributed to the observed increase in enzymatic hydrolysis yields as better enzyme accessibility resulted.

Original languageEnglish
JournalBiomass Conversion and Biorefinery
Publication statusE-pub ahead of print - 2022 Nov 23

Subject classification (UKÄ)

  • Paper, Pulp and Fiber Technology

Free keywords

  • cellulose accessibility
  • SEM
  • Lignin carbohydrate bonds
  • Softwood


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