A strain with high nattokinase activity was screened from naturally fermented pastes with casein plate selecting and shaking culturing, the strain SH21 was identified as Bacillus subtilis by 16S rRNA gene sequencing. Using nattokinase activity (NK) and purine as indicators, the liquid fermentation conditions of Bacillus subtilis SH21 were optimized by single factor experiments and response surface methodology. The single factor tests were designed to investigate the influence of carbon sources, nitrogen sources, pH, inoculation, fermentation temperature and fermentation time on nattokinase activity and purine content. Then the Plackett-Burman test was designed, and three significant factors, pH, inoculation and fermentation temperature were obtained. Finally, the Box-Behnken design was used and the response surface analysis was performed. The optimized fermentation condition was pH 6.0, with the inoculation amount 3%, and fermentation temperature 38℃. The enzyme activity increased from 166.99 U/mL to 204.52 U/mL, which was improved by 1.22 folds. While the purine content decreased from 75.4 mg/L to 51.27 mg/L, reduced by 32% under the optimized condition. This study lays the foundation for the further development of B. subtilis fermentation products with high nattokinase activity and low purine production.
PANG Yuanxiang
,
XIE Yuanhong
,
JIN Junhua
,
LIU Hui
,
ZHANG Hongxing
. Isolation and optimization of Bacillus subtilis SH21 for low purine and high nattokinase activity[J]. Food and Fermentation Industries, 2021
, 47(11)
: 194
-199
.
DOI: 10.13995/j.cnki.11-1802/ts.026039
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