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

Appl Biochem Biotechnol. 2023 Jan;195(1):68-85. doi: 10.1007/s12010-022-04097-y. Epub 2022 Aug 15.

Abstract

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.

Keywords: Bacillus subtilis; Fermentation temperature; Heavy ion mutagenesis; α-Amylase.

MeSH terms

  • Bacillus subtilis / genetics
  • Bacillus subtilis / metabolism
  • Fermentation
  • Heavy Ions*
  • Mutagenesis
  • alpha-Amylases* / genetics
  • alpha-Amylases* / metabolism

Substances

  • alpha-Amylases