From 13C-lignin to 13C-mycelium: Agaricus bisporus uses polymeric lignin as a carbon source

Katharina Duran; Michael Kohlstedt; Gijs van Erven; Cynthia E. Klostermann; Antoine H.P. America; Edwin Bakx; Johan J.P. Baars; Antonie Gorissen; Ries de Visser; Ronald P. de Vries; Christoph Wittmann; Rob N.J. Comans; Thomas W. Kuyper; Mirjam A. Kabel

doi:10.1126/sciadv.adl3419

From ¹³C-lignin to ¹³C-mycelium: Agaricus bisporus uses polymeric lignin as a carbon source

Katharina Duran
, Michael Kohlstedt
, Gijs van Erven
, Cynthia E. Klostermann
, Antoine H.P. America
, Edwin Bakx
, Johan J.P. Baars
, Antonie Gorissen
, Ries de Visser
, Ronald P. de Vries
, Christoph Wittmann
, Rob N.J. Comans
, Thomas W. Kuyper
, Mirjam A. Kabel^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Plant biomass conversion by saprotrophic fungi plays a pivotal role in terrestrial carbon (C) cycling. The general consensus is that fungi metabolize carbohydrates, while lignin is only degraded and mineralized to CO2. Recent research, however, demonstrated fungal conversion of 13C-monoaromatic compounds into proteinogenic amino acids. To unambiguously prove that polymeric lignin is not merely degraded, but also metabolized, carefully isolated 13C-labeled lignin served as substrate for Agaricus bisporus, the world's most consumed mushroom. The fungus formed a dense mycelial network, secreted lignin-active enzymes, depolymerized, and removed lignin. With a lignin carbon use efficiency of 0.14 (g/g) and fungal biomass enrichment in 13C, we demonstrate that A. bisporus assimilated and further metabolized lignin when offered as C-source. Amino acids were high in 13C-enrichment, while fungal-derived carbohydrates, fatty acids, and ergosterol showed traces of 13C. These results hint at lignin conversion via aromatic ring-cleaved intermediates to central metabolites, underlining lignin's metabolic value for fungi.

Original language	English
Article number	eadl3419
Number of pages	13
Journal	Science advances
Volume	10
Issue number	16
DOIs	https://doi.org/10.1126/sciadv.adl3419
Publication status	Published - 19 Apr 2024

Bibliographical note

Publisher Copyright:
Copyright © 2024 The Authors, some rights reserved.

Funding

Acknowledgments: We thank A. van Peer for providing the A. bisporus strain, t. van Gaal for carrying out preliminary fungal growth experiments, v. Boerkamp for expertise in nMR, n. hutnik for carbohydrate analysis, and W. van Berkel for discussions. Funding: this research received funding by the German Ministry of education and Research through grant MiSSiOn (031B0611A) and the netherlands Organization for Scientific Research (nWO) by an nWO Graduate School Green top Sector grant (GSGt.GSGt.2018.018), which is conducted in collaboration with cnc Grondstoffen and isolife bv. M.K. acknowledges support through a young investigator award, sponsored by the hansandRuthGiessen Foundation, St. ingbert, Germany. Author contributions: conceptualization: K.D., R.n.J.c., t.W.K., M.A.K., and G.v.e. Methodology: K.D., M.K., G.v.e., A.h.P.A., R.d.v., R.P.d.v., A.G., c.W., J.J.P.B., R.n.J.c., M.A.K., and e.B. investigation: K.D., M.K., G.v.e., c.e.K., A.h.P.A., c.W., and e.B. visualization: K.D. Supervision: t.W.K. and M.A.K. Writing\u2013original draft: K.D. and M.A.K. Writing\u2013review and editing: M.K., R.P.d.v., t.W.K., c.W., M.A.K., K.D., and G.v.e. Funding acquisition: c.W., J.J.P.B., M.A.K., and K.D. Data curation: K.D. and M.K. Resources: M.A.K. validation: M.A.K. and K.D. Formal analysis: K.D. Project administration: K.D. and M.A.K. Competing interests: R.d.v. and A.G. are founders of isolife Bv, who produced 13cwheat straw. the other authors declare that they have no competing interests. Data and materials availability: the MS proteomics data have been deposited to the ProteomeXchange consortium via the PRiDe (48) partner repository with the dataset identifier PXD045077 and 10.6019/PXD045077. the A. bisporus strain A15 used can be provided by Wageningen UR (Plant breeding; A. van Peer) pending scientific review and a completed material transfer agreement. Requests for the A. bisporus strain A15 should be submitted to [email protected]. All other data needed to evaluate the conclusions in this paper are present in the paper and/or the Supplementary Materials. We thank A. van Peer for providing the A. bisporus strain, T. van Gaal for carrying out preliminary fungal growth experiments, V. Boerkamp for expertise in NMR, N. Hutnik for carbohydrate analysis, and W. van Berkel for discussions. This research received funding by the German Ministry of Education and Research through grant MISSION (031B0611A) and the Netherlands Organization for Scientific Research (NWO) by an NWO Graduate School Green Top Sector grant (GSGT.GSGT.2018.018), which is conducted in collaboration with CNC Grondstoffen and IsoLife bv. M.K. acknowledges support through a young investigator award, sponsored by the Hans-and-Ruth-Giessen Foundation, St. Ingbert, Germany.

Funders	Funder number
hansandRuthGiessen Foundation
Hans-and-Ruth-Giessen Foundation
Bundesministerium für Bildung und Forschung	031B0611A
Nederlandse Organisatie voor Wetenschappelijk Onderzoek	GSGT.GSGT.2018.018
nWO Graduate School Green top Sector	GSGt.GSGt.2018.018

Access to Document

10.1126/sciadv.adl3419Licence: CC BY

sciadv.adl3419Final published version, 2.86 MBLicence: CC BY

Cite this

Duran, K., Kohlstedt, M., van Erven, G., Klostermann, C. E., America, A. H. P., Bakx, E., Baars, J. J. P., Gorissen, A., de Visser, R., de Vries, R. P., Wittmann, C., Comans, R. N. J., Kuyper, T. W., & Kabel, M. A. (2024). From ¹³C-lignin to ¹³C-mycelium: Agaricus bisporus uses polymeric lignin as a carbon source. Science advances, 10(16), Article eadl3419. https://doi.org/10.1126/sciadv.adl3419

@article{3979748cafa7465291f23621750f78ac,

title = "From 13C-lignin to 13C-mycelium: Agaricus bisporus uses polymeric lignin as a carbon source",

abstract = "Plant biomass conversion by saprotrophic fungi plays a pivotal role in terrestrial carbon (C) cycling. The general consensus is that fungi metabolize carbohydrates, while lignin is only degraded and mineralized to CO2. Recent research, however, demonstrated fungal conversion of 13C-monoaromatic compounds into proteinogenic amino acids. To unambiguously prove that polymeric lignin is not merely degraded, but also metabolized, carefully isolated 13C-labeled lignin served as substrate for Agaricus bisporus, the world's most consumed mushroom. The fungus formed a dense mycelial network, secreted lignin-active enzymes, depolymerized, and removed lignin. With a lignin carbon use efficiency of 0.14 (g/g) and fungal biomass enrichment in 13C, we demonstrate that A. bisporus assimilated and further metabolized lignin when offered as C-source. Amino acids were high in 13C-enrichment, while fungal-derived carbohydrates, fatty acids, and ergosterol showed traces of 13C. These results hint at lignin conversion via aromatic ring-cleaved intermediates to central metabolites, underlining lignin's metabolic value for fungi.",

author = "Katharina Duran and Michael Kohlstedt and \{van Erven\}, Gijs and Klostermann, \{Cynthia E.\} and America, \{Antoine H.P.\} and Edwin Bakx and Baars, \{Johan J.P.\} and Antonie Gorissen and \{de Visser\}, Ries and \{de Vries\}, \{Ronald P.\} and Christoph Wittmann and Comans, \{Rob N.J.\} and Kuyper, \{Thomas W.\} and Kabel, \{Mirjam A.\}",

note = "Publisher Copyright: Copyright {\textcopyright} 2024 The Authors, some rights reserved.",

year = "2024",

month = apr,

day = "19",

doi = "10.1126/sciadv.adl3419",

language = "English",

volume = "10",

journal = "Science advances",

issn = "2375-2548",

publisher = "American Association for the Advancement of Science",

number = "16",

}

TY - JOUR

T1 - From 13C-lignin to 13C-mycelium

T2 - Agaricus bisporus uses polymeric lignin as a carbon source

AU - Duran, Katharina

AU - Kohlstedt, Michael

AU - van Erven, Gijs

AU - Klostermann, Cynthia E.

AU - America, Antoine H.P.

AU - Bakx, Edwin

AU - Baars, Johan J.P.

AU - Gorissen, Antonie

AU - de Visser, Ries

AU - de Vries, Ronald P.

AU - Wittmann, Christoph

AU - Comans, Rob N.J.

AU - Kuyper, Thomas W.

AU - Kabel, Mirjam A.

PY - 2024/4/19

Y1 - 2024/4/19

N2 - Plant biomass conversion by saprotrophic fungi plays a pivotal role in terrestrial carbon (C) cycling. The general consensus is that fungi metabolize carbohydrates, while lignin is only degraded and mineralized to CO2. Recent research, however, demonstrated fungal conversion of 13C-monoaromatic compounds into proteinogenic amino acids. To unambiguously prove that polymeric lignin is not merely degraded, but also metabolized, carefully isolated 13C-labeled lignin served as substrate for Agaricus bisporus, the world's most consumed mushroom. The fungus formed a dense mycelial network, secreted lignin-active enzymes, depolymerized, and removed lignin. With a lignin carbon use efficiency of 0.14 (g/g) and fungal biomass enrichment in 13C, we demonstrate that A. bisporus assimilated and further metabolized lignin when offered as C-source. Amino acids were high in 13C-enrichment, while fungal-derived carbohydrates, fatty acids, and ergosterol showed traces of 13C. These results hint at lignin conversion via aromatic ring-cleaved intermediates to central metabolites, underlining lignin's metabolic value for fungi.

AB - Plant biomass conversion by saprotrophic fungi plays a pivotal role in terrestrial carbon (C) cycling. The general consensus is that fungi metabolize carbohydrates, while lignin is only degraded and mineralized to CO2. Recent research, however, demonstrated fungal conversion of 13C-monoaromatic compounds into proteinogenic amino acids. To unambiguously prove that polymeric lignin is not merely degraded, but also metabolized, carefully isolated 13C-labeled lignin served as substrate for Agaricus bisporus, the world's most consumed mushroom. The fungus formed a dense mycelial network, secreted lignin-active enzymes, depolymerized, and removed lignin. With a lignin carbon use efficiency of 0.14 (g/g) and fungal biomass enrichment in 13C, we demonstrate that A. bisporus assimilated and further metabolized lignin when offered as C-source. Amino acids were high in 13C-enrichment, while fungal-derived carbohydrates, fatty acids, and ergosterol showed traces of 13C. These results hint at lignin conversion via aromatic ring-cleaved intermediates to central metabolites, underlining lignin's metabolic value for fungi.

UR - https://www.scopus.com/pages/publications/85191103428

U2 - 10.1126/sciadv.adl3419

DO - 10.1126/sciadv.adl3419

M3 - Article

C2 - 38640242

AN - SCOPUS:85191103428

SN - 2375-2548

VL - 10

JO - Science advances

JF - Science advances

IS - 16

M1 - eadl3419

ER -