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<p>The demand for sustainable and eco-friendly processes in the paper industry has driven the exploration of efficient enzymatic systems. In this study, cellulase production from thermophilic <i>Bacillus licheniformis</i> (NCBI Accession No.: KR340466) was optimized using Response Surface Methodology (RSM), resulting in an 8.4-fold enhancement in enzyme activity (0.15 to 1.26 U/ml) under optimal conditions (50&#xa0;°C, 36&#xa0;h, pH 6.0, and 2%, v/v inoculum of 42&#xa0;h old culture). The enzyme showed maximum activity at 50&#xa0;°C in 0.075&#xa0;M phosphate buffer (pH 7.5) with a reaction time of 20&#xa0;min. Kinetic parameters indicated strong substrate affinity toward carboxymethylcellulose, with a <i>K</i><sub><i>m</i></sub> of 0.472&#xa0;mg/ml and <i>V</i><sub><i>max</i></sub> of 1.11 µmol/ml/min. Metal ions analysis revealed activation by Na⁺ and inhibition by Hg²⁺ and Mg²⁺, while protein inhibitors significantly reduced enzyme activity. Importantly, the study establishes a direct link between optimized enzyme production and its industrial application. Treatment of recycled paper pulp with the optimized cellulase led to fiber defibrillation, fragmentation, and surface smoothening, as confirmed by AFM. FTIR analysis demonstrated reduction in cellulose and hemicellulose content along with removal of chromophoric and hydrophobic compounds, resulting in improved pulp characteristics, including increased opacity of deinked paper. This study presents a novel integrated approach combining statistical optimization, enzymatic characterization, and functional application in pulp bioprocessing, highlighting the potential of <i>Bacillus licheniformis</i> cellulase as a sustainable alternative to chemical treatments in paper recycling.</p>

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Production optimization of cellulase from Bacillus licheniformis and its application in paper pulp modification and biobleaching

  • 期刊论文

  • 发表日期：

    2026年7月4日

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  • Pooja Thakur,
  • Kamal Kumar Bhardwaj,
  • Savitri,
  • Reena Gupta

摘要

The demand for sustainable and eco-friendly processes in the paper industry has driven the exploration of efficient enzymatic systems. In this study, cellulase production from thermophilic Bacillus licheniformis (NCBI Accession No.: KR340466) was optimized using Response Surface Methodology (RSM), resulting in an 8.4-fold enhancement in enzyme activity (0.15 to 1.26 U/ml) under optimal conditions (50 °C, 36 h, pH 6.0, and 2%, v/v inoculum of 42 h old culture). The enzyme showed maximum activity at 50 °C in 0.075 M phosphate buffer (pH 7.5) with a reaction time of 20 min. Kinetic parameters indicated strong substrate affinity toward carboxymethylcellulose, with a Km of 0.472 mg/ml and Vmax of 1.11 µmol/ml/min. Metal ions analysis revealed activation by Na⁺ and inhibition by Hg²⁺ and Mg²⁺, while protein inhibitors significantly reduced enzyme activity. Importantly, the study establishes a direct link between optimized enzyme production and its industrial application. Treatment of recycled paper pulp with the optimized cellulase led to fiber defibrillation, fragmentation, and surface smoothening, as confirmed by AFM. FTIR analysis demonstrated reduction in cellulose and hemicellulose content along with removal of chromophoric and hydrophobic compounds, resulting in improved pulp characteristics, including increased opacity of deinked paper. This study presents a novel integrated approach combining statistical optimization, enzymatic characterization, and functional application in pulp bioprocessing, highlighting the potential of Bacillus licheniformis cellulase as a sustainable alternative to chemical treatments in paper recycling.