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<p>Starch is a vital raw material for numerous industries, but conventional processing methods are energy-intensive and environmentally detrimental. Raw starch-digesting amylases provide a sustainable alternative by directly hydrolyzing raw starch at temperatures below its gelatinization point. A novel raw starch-digesting α-amylase (Amy-38) from <i>Bacillus cereus</i> spH1 was purified to homogeneity after ammonium sulfate precipitation and gel filtration, achieving a 17.8-fold purification and 12.3% yield. The purified amylase presents a molecular weight of 68&#xa0;kDa, with optimal activity at pH 7.0 and 60&#xa0;°C. Optimal stability was observed at pH levels of 6.0–7.5 and temperatures of 40–55&#xa0;°C. The enzyme retained 90% or 85% of its activity after 30&#xa0;min incubation at pH 7.5 or 45&#xa0;°C, respectively. Moreover, Ca²⁺ was the most effective activator for Amy-38 (224%) followed by Mg²⁺ (176%). The amylase exhibited specificity for starch, with average Km and Vmax values of 18.18&#xa0;mg/mL and 72.99 µmol/min/mL, respectively. The hydrolysis of raw wheat starch was optimized using Response Surface Methodology. Using 5% native wheat starch, 52.14% reducing sugars were produced at 60&#xa0;°C for 3&#xa0;h, eliminating the need for energy-intensive gelatinization. These results demonstrate that Amy-38 is a potent catalyst for efficient hydrolysis of raw starch, positioning it as a promising and sustainable biocatalyst for industrial applications.</p> Graphical Abstract <p></p>

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Towards Sustainable Biocatalysis: A Novel Thermostable Raw Starch-Digesting Amylase from Bacillus Cereus as a Green and Eco-Friendly Alternative for Starch Processing

  • 期刊论文

  • 发表日期：

    2026年1月3日

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  • Bouthaina Ben Hadj Hmida,
  • Sameh Ben Mabrouk,
  • Monia Blibech,
  • Adel Sayari,
  • Aida Hmida-Sayari

摘要

Starch is a vital raw material for numerous industries, but conventional processing methods are energy-intensive and environmentally detrimental. Raw starch-digesting amylases provide a sustainable alternative by directly hydrolyzing raw starch at temperatures below its gelatinization point. A novel raw starch-digesting α-amylase (Amy-38) from Bacillus cereus spH1 was purified to homogeneity after ammonium sulfate precipitation and gel filtration, achieving a 17.8-fold purification and 12.3% yield. The purified amylase presents a molecular weight of 68 kDa, with optimal activity at pH 7.0 and 60 °C. Optimal stability was observed at pH levels of 6.0–7.5 and temperatures of 40–55 °C. The enzyme retained 90% or 85% of its activity after 30 min incubation at pH 7.5 or 45 °C, respectively. Moreover, Ca²⁺ was the most effective activator for Amy-38 (224%) followed by Mg²⁺ (176%). The amylase exhibited specificity for starch, with average Km and Vmax values of 18.18 mg/mL and 72.99 µmol/min/mL, respectively. The hydrolysis of raw wheat starch was optimized using Response Surface Methodology. Using 5% native wheat starch, 52.14% reducing sugars were produced at 60 °C for 3 h, eliminating the need for energy-intensive gelatinization. These results demonstrate that Amy-38 is a potent catalyst for efficient hydrolysis of raw starch, positioning it as a promising and sustainable biocatalyst for industrial applications.

Graphical Abstract