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Isolation and Screening of High-Yielding α-Amylase Mutants of Bacillus subtilis by Heavy Ion Mutagenesis


To improve fermentative production of α-amylase, heavy-ion mutagenesis technology was used to irradiate Bacillus subtilis (B. subtilis) to obtain the high yielding mutants in this study. After continuous cultivation for 12 generations, eight mutants exhibited positive mutation rate with greater H/C. The α-amylase production was stable and obviously exceeded that by the parent strain, which shows that the mutants have a good genetic stability. Among the mutants, the α-amylase activity of B. subtilis KC-180–2 was 72.26 U·mL−1, which was 82.34% higher than that of the original strain. After optimization of fermentation conditions and media, the α-amylase activity of B. subtilis KC-180–2 reached a maximum of 156.83 U·mL−1 at 36 h in a bioreactor. In addition, the optimized fermentation temperature of B. subtilis KC-180–2 was increased to 49℃, indicating B. subtilis KC-180–2 possesses high-temperature resistance, which has great application prospects for industrial fermentation for α-amylase production.

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Data Availability

All data generated or analyzed during this study are included in this published article.


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We are very grateful to researcher Bo Yu from the Institute of Microbiology, Chinese Academy of Sciences for providing the original strains. In addition, we thank the Institute of Modern Physics, Chinese Academy of Sciences for providing the heavy ion beam.


The work was supported by Inner Mongolia Major Science and Technology Projects (2019ZD021).

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Jin-na Cui and Zhan-ying Liu conceived and designed research. Jin-na Cui and Yan-xin Liu collected data and conducted experiments. Zhan-ying Liu provides research guidance, research funding, laboratories, and other material resources related to experiments. Jin-na Cui and Wei Hu wrote the draft paper, Jian-hua Hu, Yong-li Li revised the paper. Jin-na Cui had primary responsibility for final content. All authors read and approved the manuscript.

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Correspondence to Zhan-Ying Liu.

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Cui, JN., Hu, W., Liu, YX. et al. Isolation and Screening of High-Yielding α-Amylase Mutants of Bacillus subtilis by Heavy Ion Mutagenesis. Appl Biochem Biotechnol (2022).

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  • α-Amylase
  • Bacillus subtilis
  • Heavy ion mutagenesis
  • Fermentation temperature