Enzymatic Hydrolysis of Wheat Arabinoxylan by a Recombinant “Minimal” Enzyme Cocktail Containing β‐Xylosidase and Novel endo ‐1,4‐β‐Xylanase and α‐ l ‐Arabinofuranosidase Activities
Abstract
This study describes the identification of the key enzyme activities required in a “minimal” enzyme cocktail able to catalyze hydrolysis of water‐soluble and water‐insoluble wheat arabinoxylan and whole vinasse, a fermentation effluent resulting from industrial ethanol manufacture from wheat. The optimal arabinose‐releasing and xylan‐depolymerizing enzyme activities were identified from data obtained when selected, recombinant enzymes were systematically supplemented to the different arabinoxylan substrates in mixtures; this examination revealed three novel α‐ l ‐arabinofuranosidase activities: (i) one GH51 enzyme from Meripilus giganteus and (ii) one GH51 enzyme from Humicola insolens , both able to catalyze arabinose release from singly substituted xylose; and (iii) one GH43 enzyme from H. insolens able to catalyze the release of arabinose from doubly substituted xylose. Treatment of water‐soluble and water‐insoluble wheat arabinoxylan with an enzyme cocktail containing a 20%:20%:20%:40% mixture and a 25%:25%:25%:25% mixture, respectively, of the GH43 α‐ l ‐arabinofuranosidase from H. insolens (Abf II), the GH51 α‐ l ‐arabinofuranosidase from M. giganteus (Abf III), a GH10 endo ‐1,4‐β‐xylanase from H. insolens (Xyl III), and a GH3 β‐xylosidase from Trichoderma reesei (β‐xyl) released 322 mg of arabinose and 512 mg of xylose per gram of water‐soluble wheat arabinoxylan dry matter and 150 mg of arabinose and 266 mg of xylose per gram of water‐insoluble wheat arabinoxylan dry matter after 24 h at pH 5, 50 °C. A 10%:40%:50% mixture of Abf II, Abf III, and β‐xyl released 56 mg of arabinose and 91 mg of xylose per gram of vinasse dry matter after 24 h at pH 5, 50 °C. The optimal dosages of the “minimal” enzyme cocktails were determined to be 0.4, 0.3, and 0.2 g enzyme protein per kilogram of substrate dry matter for the water‐soluble wheat arabinoxylan, the water‐insoluble wheat arabinoxylan, and the vinasse, respectively. These enzyme protein dosage levels were ∼14, ∼18, and ∼27 times lower than the dosages used previously, when the same wheat arabinoxylan substrates were hydrolyzed with a combination of Ultraflo L and Celluclast 1.5 L, two commercially available enzyme preparations produced by H. insolens and T. reesei .
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Only various crude, commercial enzyme mixtures designed for plant material processing have been used as hydrolysis aids for extraction of brown seaweed biofunctional polysaccharide fractions, compared with the advanced stage of designed enzyme mixtures for complex plant materials or heterogeneous plant cell wall polysaccharides.
“However, as compared to the quite advanced stage of development of designed enzyme mixtures for hydrolysis of complex plant materials or heterogenous plant cell wall polysaccharides, 39,40 only various crude, commercial enzyme mixtures, designed for plant material processing, have been used as hydrolysis aids for extraction of brown seaweed biofunctional polysaccharide fractions.5,41”
From Fucoidans from brown seaweeds: an update on structures, extraction techniques and use of enzymes as tools for structural elucidation (2013) · cites this paper as [39]
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- Authors
- Sørensen, H. R., Pedersen, S., Jørgensen, C. T., & Meyer, A. S.
- Affiliations
- Novozymes (Denmark) (DK)
- Technical University of Denmark (DK)
- Corresponding author
- Meyer, Anne S.
- ORCID
- Meyer, Anne S.
- Venue
- Biotechnology Progress
- Publisher
- Wiley
- Published
- 2007-01
- Volume
- 23
- Issue
- 1
- Pages
- 100-107
- ISSN
- 1520-6033 (ISSN-L)
- 8756-7938
- OpenAlex
- W1986802076
- Cited by
- 113
- References
- 29
- Citations by year
- 2026: 2 · 2025: 3 · 2024: 4 · 2023: 5 · 2022: 4 · 2021: 4 · 2020: 7 · 2019: 6 · 2018: 3 · 2017: 2 · 2016: 6 · 2015: 7 · 2014: 7 · 2013: 6 · 2012: 10
- FWCI
- 6.93
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Biotechnology Progress (publishedVersion)
- PubMed (publishedVersion)
- Technical University of Denmark, DTU Orbit (Technical University of Denmark, DTU) (submittedVersion)
- Topics
- Physical Sciences › Engineering › Biomedical Engineering › Biofuel production and bioconversion
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Enzyme Production and Characterization
- Life Sciences › Agricultural and Biological Sciences › Plant Science › Polysaccharides and Plant Cell Walls
- Keywords
- wheat arabinoxylan, enzymatic hydrolysis, β-xylosidase, xylanase, α-L-arabinofuranosidase, Meripilus giganteus, Humicola insolens, Trichoderma reesei, vinasse, xylose release, recombinant enzyme, xylan depolymerization, lignocellulosic biomass
- MeSH terms
- Enzyme Activation, Glycoside Hydrolases, Hydrolysis, Plant Proteins, Recombinant Proteins, Triticum, Xylans, Xylosidases, Protein Engineering, Endo-1,4-beta Xylanases
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