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Article

The Effect of Enzymatic Disintegration Using Cellulase and Lysozyme on the Efficiency of Methane Fermentation of Sewage Sludge

by
Bartłomiej Macherzyński
1,*,
Małgorzata Wszelaka-Rylik
1,*,
Anna Marszałek
2 and
Elżbieta Popowska-Nowak
1
1
Faculty of Biology and Environmental Science, Cardinal Stefan Wyszynski University in Warsaw, Wóycickiego 1/3 Str, 01-938 Warsaw, Poland
2
Faculty of Environmental and Energy Engineering, Silesian University of Technology, Konarskiego 18 Str, 44-100 Gliwice, Poland
*
Authors to whom correspondence should be addressed.
Energies 2025, 18(21), 5597; https://doi.org/10.3390/en18215597 (registering DOI)
Submission received: 14 August 2025 / Revised: 20 October 2025 / Accepted: 23 October 2025 / Published: 24 October 2025
(This article belongs to the Special Issue Nutrient and Energy Recovery from Municipal and Industrial Wastewater)

Abstract

This study presents a novel approach to intensifying the anaerobic digestion of sewage sludge through enzymatic pretreatment using hydrolytic enzymes—cellulase and lysozyme. It aims to determine how enzymatic activation affects the efficiency of methane fermentation, defined as the degree of organic matter decomposition and yield and composition of biogas. An experiment was carried out under mesophilic conditions over 20 days, analyzing the physicochemical properties of sludge, biogas production, methane content, and sanitary parameters. The addition of cellulase and lysozyme significantly enhanced process efficiency, increasing both the rate of organic matter degradation and biogas yield. The highest biogas production values (0.73 L·g−1 d.m. for cellulase and 0.72 L·g−1 d.m. for lysozyme) were obtained at a 4% (w/w) enzyme concentration, with a corresponding increase in the degree of organic matter decomposition to 78.7% and 80.0%, respectively. The produced biogas contained 58–61% methane, exceeding the values observed in the control sample, which indicates a positive effect of enzymatic activation on methane selectivity. Enhanced biogas production was attributed to improved hydrolysis of complex organic compounds, resulting in greater substrate bioavailability for methanogenic microorganisms. Moreover, methane fermentation led to the complete elimination of E. coli from all supernatants, confirming the hygienization potential of the process. The results of this study indicate that enzymatic pretreatment may serve as a viable strategy to improve both the energy efficiency and hygienic safety of anaerobic digestion processes, with relevance for future optimization and full-scale wastewater treatment applications.
Keywords: biogas; methane; sewage sludge; supernatants; anaerobic digestion; E. coli biogas; methane; sewage sludge; supernatants; anaerobic digestion; E. coli

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MDPI and ACS Style

Macherzyński, B.; Wszelaka-Rylik, M.; Marszałek, A.; Popowska-Nowak, E. The Effect of Enzymatic Disintegration Using Cellulase and Lysozyme on the Efficiency of Methane Fermentation of Sewage Sludge. Energies 2025, 18, 5597. https://doi.org/10.3390/en18215597

AMA Style

Macherzyński B, Wszelaka-Rylik M, Marszałek A, Popowska-Nowak E. The Effect of Enzymatic Disintegration Using Cellulase and Lysozyme on the Efficiency of Methane Fermentation of Sewage Sludge. Energies. 2025; 18(21):5597. https://doi.org/10.3390/en18215597

Chicago/Turabian Style

Macherzyński, Bartłomiej, Małgorzata Wszelaka-Rylik, Anna Marszałek, and Elżbieta Popowska-Nowak. 2025. "The Effect of Enzymatic Disintegration Using Cellulase and Lysozyme on the Efficiency of Methane Fermentation of Sewage Sludge" Energies 18, no. 21: 5597. https://doi.org/10.3390/en18215597

APA Style

Macherzyński, B., Wszelaka-Rylik, M., Marszałek, A., & Popowska-Nowak, E. (2025). The Effect of Enzymatic Disintegration Using Cellulase and Lysozyme on the Efficiency of Methane Fermentation of Sewage Sludge. Energies, 18(21), 5597. https://doi.org/10.3390/en18215597

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