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<p>The suboptimal ethanol yield in “Burukutu,” a traditional sorghum-based alcoholic beverage, stems from the recalcitrance of the grain’s starch–protein matrix to enzymatic hydrolysis. This study compared the performance of free and alginate-entrapped (2% w/v) <i>α</i>-amylase, glucoamylase, and protease, applied in various combinations, to optimize saccharification kinetics and bioethanol productivity in red and white sorghum cultivars. Results indicated that the triple-enzyme cocktail (<i>α</i>-amylase + glucoamylase + protease) was significantly superior to individual or dual-enzyme systems (<i>F</i> = 3740.50, <i>p</i> &lt; 0.001), emphasizing the critical role of protease in deconstructing the protein barrier to enhance starch accessibility. Red sorghum exhibited significantly higher saccharification depth and ethanol titers than the white cultivar (<i>p</i> &lt; 0.001), likely due to cultivar-specific differences in grain architecture. Peak ethanol concentrations reached 35.3 ± 0.2% v/v in red sorghum using the free triple-enzyme system, while the immobilized system achieved 29.70 ± 0.36% v/v at 72&#xa0;h. Notably, the immobilized biocatalysts demonstrated robust operational stability, maintaining high sugar utilization efficiencies (&gt; 85%) through six consecutive fermentation cycles. While a gradual decline in efficiency was observed (<i>F</i> = 151.95, <i>p</i> &lt; 0.001), the immobilized multi-enzyme framework retained substantial activity, particularly in the red sorghum matrix. These findings demonstrate that synergistic multi-enzyme immobilization substantially improves starch-to-ethanol conversion and process sustainability, offering a scalable model for intensifying sorghum-based bioethanol production.</p>

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Integration of Enzyme Immobilization and Multi-Enzyme Hydrolysis to Enhance Saccharification Efficiency and Ethanol Yield in Sorghum Fermentation Systems

  • 期刊论文

  • 发表日期：

    2026年4月28日

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  • Muinat M. Adeyanju,
  • Adedeji N. Ademakinwa,
  • Kazeem O. Alli,
  • Adefemi O. Adefuye,
  • Emmanuel Sunday Omirin,
  • Olalekan O. Bakare,
  • Samuel O. Olalekan,
  • Enitan O. Adesanya,
  • Odunayo C. Atewolara-Odule,
  • Kuburat T. Odufuwa,
  • Oseyemi O. Olubomehin,
  • Bamidele S. Fagbohunka

摘要

The suboptimal ethanol yield in “Burukutu,” a traditional sorghum-based alcoholic beverage, stems from the recalcitrance of the grain’s starch–protein matrix to enzymatic hydrolysis. This study compared the performance of free and alginate-entrapped (2% w/v) α-amylase, glucoamylase, and protease, applied in various combinations, to optimize saccharification kinetics and bioethanol productivity in red and white sorghum cultivars. Results indicated that the triple-enzyme cocktail (α-amylase + glucoamylase + protease) was significantly superior to individual or dual-enzyme systems (F = 3740.50, p < 0.001), emphasizing the critical role of protease in deconstructing the protein barrier to enhance starch accessibility. Red sorghum exhibited significantly higher saccharification depth and ethanol titers than the white cultivar (p < 0.001), likely due to cultivar-specific differences in grain architecture. Peak ethanol concentrations reached 35.3 ± 0.2% v/v in red sorghum using the free triple-enzyme system, while the immobilized system achieved 29.70 ± 0.36% v/v at 72 h. Notably, the immobilized biocatalysts demonstrated robust operational stability, maintaining high sugar utilization efficiencies (> 85%) through six consecutive fermentation cycles. While a gradual decline in efficiency was observed (F = 151.95, p < 0.001), the immobilized multi-enzyme framework retained substantial activity, particularly in the red sorghum matrix. These findings demonstrate that synergistic multi-enzyme immobilization substantially improves starch-to-ethanol conversion and process sustainability, offering a scalable model for intensifying sorghum-based bioethanol production.