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Purpose <p>This study aimed to effectively valorize starchy agro-residues (e.g., oat, rice, and wheat brans) for biohydrogen production through Clostridial fermentation using both separate hydrolysis and fermentation (SHF) and consolidated bioprocessing (CBP) strategies.</p> Methods <p>Four <i>Clostridium</i> strains (BOH3, G117, NCIM 2337, and NCIM 2825) were screened for amylase activity and hydrogen production in minimal media containing starch and mixed sugars. Mild acid–moist heat pretreatment (0.5&#xa0;N HCl, 121&#xa0;°C, 30&#xa0;min) of starchy brans (50&#xa0;g/L) was used to generate fermentable hydrolysates, which were evaluated alongside direct fermentation of native brans.</p> Results <p><i>Clostridium beijerinckii</i> G117 exhibited the highest amylase activity (14.37 U/mg) and hydrogen yield (257 mL/g). Pretreated brans yielded hydrolysates containing 14–17&#xa0;g/L starch, 6–16&#xa0;g/L reducing sugars, and 1.25–2.80&#xa0;g/L protein, with low phenolic and mineral content. Fermentation of hydrolysates (12.5&#xa0;g/L total soluble sugars (TSS)) produced 3,280 mL/L biohydrogen, with yields of 340 mL/g TSS and 2.76&#xa0;mol H<sub>2</sub>/mol glucose. In contrast, direct fermentation of native brans (25&#xa0;g/L) enabled the secretion of extracellular amylase (6.50–11.50 U/mg) and xylanase (0.9–2.0 U/mg), resulting in higher biohydrogen production (3,960 mL/L) with comparable yields. Mass and electron balance analyses showed high CBP efficiencies (83–93%), comparable to separate hydrolysis and fermentation (72–96%).</p> Conclusion <p>This study demonstrates the efficient valorization of agro-starchy brans for biohydrogen production. Oat and wheat brans supported higher biohydrogen yields than rice bran through direct consolidated bioprocessing by <i>C. beijerinckii</i> G117, representing the first report of this process.</p> Graphical Abstract <p></p> <p><b>Statement of Novelty</b></p> <p>This study reports the first successful use of native and pretreated rice, wheat, and oat brans as starchy agro-residues for Clostridial biohydrogen production. Among four strains evaluated, <i>Clostridium beijerinckii</i> G117 showed the highest amylolytic and hydrogenogenic potential, with amylase activity of 16.80 U/mg and hydrogen yields exceeding 250 mL/g substrate. The strain efficiently converted bran hydrolysates (~ 12.5&#xa0;g/L total soluble sugars) to 3060 mL/L hydrogen, achieving yields of 340 mL/g substrate and 2.76&#xa0;mol H<sub>2</sub>/mol glucose. Notably, G117 directly fermented native brans without external enzymes due to its inherent amylase and xylanase activities, producing up to 3960 mL/L hydrogen. These findings demonstrate the feasibility of consolidated bioprocessing for cost-effective and sustainable biohydrogen production.</p>

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Biohydrogen Poduction Using Pretreated and Native Oat, Rice, and Wheat Brans by Clostridium beijerinckii G117

  • 期刊论文

  • 发表日期：

    2026年8月4日

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  • Akanksha Jain,
  • Md. Ebrahim Khalil,
  • Phuntsho Yenten,
  • Sidharth UC,
  • Venkata Giridhar Poosarla,
  • Aruliah Rajasekar,
  • Vignesh Natarajan,
  • Chandraraj Krishnan,
  • Kun Lin Yang,
  • Gobinath Rajagopalan

摘要

Purpose

This study aimed to effectively valorize starchy agro-residues (e.g., oat, rice, and wheat brans) for biohydrogen production through Clostridial fermentation using both separate hydrolysis and fermentation (SHF) and consolidated bioprocessing (CBP) strategies.

Methods

Four Clostridium strains (BOH3, G117, NCIM 2337, and NCIM 2825) were screened for amylase activity and hydrogen production in minimal media containing starch and mixed sugars. Mild acid–moist heat pretreatment (0.5 N HCl, 121 °C, 30 min) of starchy brans (50 g/L) was used to generate fermentable hydrolysates, which were evaluated alongside direct fermentation of native brans.

Results

Clostridium beijerinckii G117 exhibited the highest amylase activity (14.37 U/mg) and hydrogen yield (257 mL/g). Pretreated brans yielded hydrolysates containing 14–17 g/L starch, 6–16 g/L reducing sugars, and 1.25–2.80 g/L protein, with low phenolic and mineral content. Fermentation of hydrolysates (12.5 g/L total soluble sugars (TSS)) produced 3,280 mL/L biohydrogen, with yields of 340 mL/g TSS and 2.76 mol H2/mol glucose. In contrast, direct fermentation of native brans (25 g/L) enabled the secretion of extracellular amylase (6.50–11.50 U/mg) and xylanase (0.9–2.0 U/mg), resulting in higher biohydrogen production (3,960 mL/L) with comparable yields. Mass and electron balance analyses showed high CBP efficiencies (83–93%), comparable to separate hydrolysis and fermentation (72–96%).

Conclusion

This study demonstrates the efficient valorization of agro-starchy brans for biohydrogen production. Oat and wheat brans supported higher biohydrogen yields than rice bran through direct consolidated bioprocessing by C. beijerinckii G117, representing the first report of this process.

Graphical Abstract

Statement of Novelty

This study reports the first successful use of native and pretreated rice, wheat, and oat brans as starchy agro-residues for Clostridial biohydrogen production. Among four strains evaluated, Clostridium beijerinckii G117 showed the highest amylolytic and hydrogenogenic potential, with amylase activity of 16.80 U/mg and hydrogen yields exceeding 250 mL/g substrate. The strain efficiently converted bran hydrolysates (~ 12.5 g/L total soluble sugars) to 3060 mL/L hydrogen, achieving yields of 340 mL/g substrate and 2.76 mol H2/mol glucose. Notably, G117 directly fermented native brans without external enzymes due to its inherent amylase and xylanase activities, producing up to 3960 mL/L hydrogen. These findings demonstrate the feasibility of consolidated bioprocessing for cost-effective and sustainable biohydrogen production.