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<p>Glucoamylases capable of hydrolyzing raw starch at moderate temperatures exhibit significant potential for the direct and efficient hydrolysis of raw starch. Using bioinformatics-assisted mining strategies, four candidate glucoamylases were initially identified from the GenBank database. Among these candidates, SeGA demonstrated high specific activities of 412.94&#xa0;U/mg toward soluble starch and 143.51&#xa0;U/mg toward raw corn starch. The half-life of SeGA was 192 h at 40&#xa0;°C. Using SeGA as the initial enzyme, FuncLib design was performed to enhance its activity. The mutant SeGA-21 exhibited higher specific activity of 466.22 U/mg toward soluble starch and 173.98&#xa0;U/mg toward raw corn starch, representing increases of 1.13-fold and 1.21-fold compared to SeGA, respectively. The half-life of SeGA-21 at 40&#xa0;°C increased to 240 h, representing a 25% increase compared with SeGA. SeGA-21 can efficiently hydrolyze 30% raw corn starch, achieving a hydrolysis rate of approximately 31% after a 9-h reaction. When working synergistically with amylolytic enzymes, a hydrolysis rate of 71.5% was achieved for 30% raw corn starch at 40&#xa0;°C within 15 h of reaction. Due to its ability to hydrolyze raw starch, moderate reaction temperature requirements, and excellent thermostability, SeGA-21 can be regarded as an efficient enzyme for the cold hydrolysis of raw starch and starch modification.</p>

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Discovery and design of novel glucoamylases suitable for raw starch hydrolysis at moderate temperatures via an integrated bioinformatics-assisted strategy

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年1月14日

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  • Shuangshuang Ge,
  • Yimeng Song,
  • Tong Ye,
  • Xueting Zhang,
  • Xuecheng Zhang,
  • Yinliang Zhang,
  • Yazhong Xiao,
  • Wei Fang

摘要

Glucoamylases capable of hydrolyzing raw starch at moderate temperatures exhibit significant potential for the direct and efficient hydrolysis of raw starch. Using bioinformatics-assisted mining strategies, four candidate glucoamylases were initially identified from the GenBank database. Among these candidates, SeGA demonstrated high specific activities of 412.94 U/mg toward soluble starch and 143.51 U/mg toward raw corn starch. The half-life of SeGA was 192 h at 40 °C. Using SeGA as the initial enzyme, FuncLib design was performed to enhance its activity. The mutant SeGA-21 exhibited higher specific activity of 466.22 U/mg toward soluble starch and 173.98 U/mg toward raw corn starch, representing increases of 1.13-fold and 1.21-fold compared to SeGA, respectively. The half-life of SeGA-21 at 40 °C increased to 240 h, representing a 25% increase compared with SeGA. SeGA-21 can efficiently hydrolyze 30% raw corn starch, achieving a hydrolysis rate of approximately 31% after a 9-h reaction. When working synergistically with amylolytic enzymes, a hydrolysis rate of 71.5% was achieved for 30% raw corn starch at 40 °C within 15 h of reaction. Due to its ability to hydrolyze raw starch, moderate reaction temperature requirements, and excellent thermostability, SeGA-21 can be regarded as an efficient enzyme for the cold hydrolysis of raw starch and starch modification.