Thermophilic fermentation enhanced by eggshell-derived hydroxyapatite for sustainable hydrogen production

Shishita Zahan Zisha , Hasfalina Che Man , Rozita Omar , Suraya Abdul Rashid , Tan Jian Ping , Shareena Fairuz , Abdul Manaf , Tabassum Mumtaz , Nur Syakina Jamali

Bioresources and Bioprocessing ›› 2025, Vol. 12 ›› Issue (1) : 136

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Bioresources and Bioprocessing ›› 2025, Vol. 12 ›› Issue (1) :136 DOI: 10.1186/s40643-025-00976-4
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Thermophilic fermentation enhanced by eggshell-derived hydroxyapatite for sustainable hydrogen production

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Abstract

This study presents the eco-friendly synthesis of Hydroxyapatite (HAP) using chicken eggshell biowaste as a sustainable calcium source to enhance biohydrogen production. The stoichiometric ratio of Ca/P was determined to be 1.67 by evaluating pH, temperature, and calcination time, revealing that pH 10.5, temperature 950ᵒC, and calcination time of 2 h were optimal. Structural and morphological characterization confirmed the successful formation of crystalline HAP with 78% crystallinity (XRD), accurate Ca/P ratio (EDS), and phosphate group presence (FTIR). The synthesized HAP was applied as a bio-additive in thermophilic dark fermentation using a Thermoanaerobacterium-enriched sludge. The optimal HAP dosage (560 mg/L) yielded the highest hydrogen yield of 1.085 mol H2/mol sugar and a hydrogen production rate of 27.969 mL H2/(L h), representing a 59.73% increase over the control. Volatile fatty acid analysis and microbial profiling confirmed enhanced butyrate metabolism and a dominant presence of Thermoanaerobacterium thermosaccharolyticum. This work demonstrates a novel waste-to-value approach by integrating green-synthesized HAP with thermophilic fermentation, contributing to both waste valorization and renewable hydrogen production.

Keywords

Green synthesis / Hydroxyapatite from eggshells / Optimization using response surface methodology / Stoichiometric ratio / Biohydrogen production

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Shishita Zahan Zisha, Hasfalina Che Man, Rozita Omar, Suraya Abdul Rashid, Tan Jian Ping, Shareena Fairuz, Abdul Manaf, Tabassum Mumtaz, Nur Syakina Jamali. Thermophilic fermentation enhanced by eggshell-derived hydroxyapatite for sustainable hydrogen production. Bioresources and Bioprocessing, 2025, 12(1): 136 DOI:10.1186/s40643-025-00976-4

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Funding

Organization for Women in Science for the Developing World(Organization for Women in Science for the Developing World)

Ministry of Higher Education and Scientific Research(FRGS/1/2023/TK08/UPM/02/3)

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