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Efficient methane production from agro-industrial residues using anaerobic fungal-rich consortia

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Anaerobic digestion (AD) emerges as a pivotal technique in climate change mitigation, transforming organic materials into biogas, a renewable energy form. This process significantly impacts energy production and waste management, influencing greenhouse gas emissions. Traditional research has largely focused on anaerobic bacteria and methanogens for methane production. However, the potential of anaerobic lignocellulolytic fungi for degrading lignocellulosic biomass remains less explored. In this study, buffalo rumen inocula were enriched and acclimatized to improve lignocellulolytic hydrolysis activity. Two consortia were established: the anaerobic fungi consortium (AFC), selectively enriched for fungi, and the anaerobic lignocellulolytic microbial consortium (ALMC). The consortia were utilized to create five distinct microbial cocktails–AF0, AF20, AF50, AF80, and AF100. These cocktails were formulated based on varying of AFC and ALMC by weights (w/w). Methane production from each cocktail of lignocellulosic biomasses (cassava pulp and oil palm residues) was evaluated. The highest methane yields of CP, EFB, and MFB were obtained at 337, 215, and 54 mL/g VS, respectively. Cocktails containing a mix of anaerobic fungi, hydrolytic bacteria (Sphingobacterium sp.), syntrophic bacteria (Sphaerochaeta sp.), and hydrogenotrophic methanogens produced 2.1–2.6 times higher methane in cassava pulp and 1.1–1.2 times in oil palm empty fruit bunch compared to AF0. All cocktails effectively produced methane from oil palm empty fruit bunch due to its lipid content. However, methane production ceased after 3 days when oil palm mesocarp fiber was used, due to long-chain fatty acid accumulation. Anaerobic fungi consortia showed effective lignocellulosic and starchy biomass degradation without inhibition due to organic acid accumulation. These findings underscore the potential of tailored microbial cocktails for enhancing methane production from diverse lignocellulosic substrates.

Key Points

Enhanced methane production from diverse lignocellulosic biomasses using enriched microbial cocktails.

Efficient degradation of lignocellulosic biomass and starch by consortia of anaerobic fungi constituent.

The number of anaerobic fungi affects the amount of methane production.

Importance of tailored microbial cocktails for maximizing methane production.

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Data availability

The metagenomic data pertaining to ALMC were archived in the Sequence Read Archive (SRA) of the National Centre for Biotechnology Information (NCBI) under the BioProject accession number PRJNA609587, while the corresponding data for AFC were stored under the accession number PRJNA666510.


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The authors acknowledge Prof. Dr. Anna Schnürer of the Department of Molecular Sciences, Swedish University of Agricultural Sciences, Sweden, for providing support in molecular techniques. The laboratory facility was facilitated by ECoWaste.


This work was financial supported by King Mongkut’s University of Technology Thonburi (KMUTT) through the Petchra Pra Jom Klao Doctoral Academic Scholarship for Ph.D. Program and KMUTT Post-Doctoral Fellowship. The research funding in this project is supported by Thailand Science Research and Innovation (TSRI), National Science, Research, and Innovation Fund (NSRF) Fiscal year 2024 and KMUTT.

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All authors contributed to the study conception and design. Material preparation and data collection were performed by NT. Data analysis was done by all authors. The first draft of the manuscript was written by NT. All authors read and approved the final manuscript.

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Correspondence to Pawinee Chaiprasert.

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Thongbunrod, N., Chaiprasert, P. Efficient methane production from agro-industrial residues using anaerobic fungal-rich consortia. World J Microbiol Biotechnol 40, 239 (2024).

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