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Purpose <p>The escalating demands for traditional fuels have depleted fossil fuels. Confronting global warming issues, there is a growing need to search for alternative renewable biofuel sources such as bioethanol. The bioconversion of lignocellulosic biomass has highlighted the urgency of advancing research on biofuel production. Using cellulase immobilization reduces the conversion costs and enhances bioethanol production.</p> Methods <p>The present study aimed to produce bioethanol from lignocellulosic renewable materials through cellulase immobilization and HPLC analysis. Cellulase producing bacteria were isolated from the snail gut using carboxymethyl cellulose (CMC). This isolate was subjected to ribotyping identification. The bacteria were entrapped (immobilized) and used for cellulase production on solid-state fermentation (SSF) using sugarcane bagasse as a solid substrate. <i>Saccharomyces cerevisiae</i> and cellulase were used for ethanol production under immobilized and non-immobilized bead conditions. The substrate consumption and kinetic profile of ethanol production were monitored using HPLC.</p> Results <p>The results demonstrate the central role of cellulase from cellulolytic bacteria is crucial in producing bioethanol holistically and effectively. Cellulase is predominantly produced by the culture of <i>Bacillus amyloliquefacience</i> and applied to a large amount of pretreated sugarcane bagasse. Analysis of sugarcane bagasse revealed the presence of 36.4% cellulose, 25.7% hemicellulose, 27.6% lignin, 0.2% moisture, 2.8% dry matter, 2.2% organic matter, 0.8% ash, and 2.3% protein. HPLC analysis was performed to determine sugar contents with retention time at 5.7&#xa0;min, 7.427&#xa0;min, 1.413&#xa0;min, 1.060&#xa0;min, 6.007&#xa0;min, and 5.753&#xa0;min using a mobile phase of acetonitrile: deionized water in ratio of 80:20 and 50:50. The saccharification rate was studied using immobilized cellulase and <i>S. cerevisiae</i> and compared to non-immobilized cellulase and <i>Saccharomyces cerevisiae</i>. Immobilized cellulase and <i>Saccharomyces cerevisiae</i> gave maximum saccharification (8.1% and 6.1%) compared to non-immobilized cells (3.7% and 1.95%). In addition, HPLC reported a comparative study on pure and bioethanol produced based on retention time.</p> Conclusion <p>This significant improvement in bioethanol production has been documented from the immobilized cellulase derived from <i>Bacillus amyloliquefacience</i>, which was isolated from the snail gut.</p>

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Entrapment of Cellulase of Snail Gut Bacillus amyloliquifacience for converting Sugarcane Bagasse to Bioethanol Using Saccharomyces cerevisiae

  • 期刊论文

  • 发表日期：

    2025年2月14日

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  • Sajib Podder,
  • Ranajit Kumar Shaha,
  • Ariful Haque,
  • Kahkashan Perveen,
  • Mashail Fahad Alsayed,
  • M. Murali,
  • N. Shilpa,
  • Tanzima Yeasmin,
  • Riyaz Sayyed

摘要

Purpose

The escalating demands for traditional fuels have depleted fossil fuels. Confronting global warming issues, there is a growing need to search for alternative renewable biofuel sources such as bioethanol. The bioconversion of lignocellulosic biomass has highlighted the urgency of advancing research on biofuel production. Using cellulase immobilization reduces the conversion costs and enhances bioethanol production.

Methods

The present study aimed to produce bioethanol from lignocellulosic renewable materials through cellulase immobilization and HPLC analysis. Cellulase producing bacteria were isolated from the snail gut using carboxymethyl cellulose (CMC). This isolate was subjected to ribotyping identification. The bacteria were entrapped (immobilized) and used for cellulase production on solid-state fermentation (SSF) using sugarcane bagasse as a solid substrate. Saccharomyces cerevisiae and cellulase were used for ethanol production under immobilized and non-immobilized bead conditions. The substrate consumption and kinetic profile of ethanol production were monitored using HPLC.

Results

The results demonstrate the central role of cellulase from cellulolytic bacteria is crucial in producing bioethanol holistically and effectively. Cellulase is predominantly produced by the culture of Bacillus amyloliquefacience and applied to a large amount of pretreated sugarcane bagasse. Analysis of sugarcane bagasse revealed the presence of 36.4% cellulose, 25.7% hemicellulose, 27.6% lignin, 0.2% moisture, 2.8% dry matter, 2.2% organic matter, 0.8% ash, and 2.3% protein. HPLC analysis was performed to determine sugar contents with retention time at 5.7 min, 7.427 min, 1.413 min, 1.060 min, 6.007 min, and 5.753 min using a mobile phase of acetonitrile: deionized water in ratio of 80:20 and 50:50. The saccharification rate was studied using immobilized cellulase and S. cerevisiae and compared to non-immobilized cellulase and Saccharomyces cerevisiae. Immobilized cellulase and Saccharomyces cerevisiae gave maximum saccharification (8.1% and 6.1%) compared to non-immobilized cells (3.7% and 1.95%). In addition, HPLC reported a comparative study on pure and bioethanol produced based on retention time.

Conclusion

This significant improvement in bioethanol production has been documented from the immobilized cellulase derived from Bacillus amyloliquefacience, which was isolated from the snail gut.