Combinatorial mutagenesis of high-yielding esterase bacteria and optimization of solid-state fermentation
ZHAO Zhijun1, ZHANG Yanzhu1,3, LIU Yanbo1, ZHOU Pingping1, GE Shaohua2, SUN Xiyu1*
1(Liquor College,Henan University of Animal Husbandry and Economy,Zhengzhou 450046,China) 2(Baofeng Liquor Industry Co.Ltd,Pingdingshan 467500,China) 3(College of Food and Pharmaceutical Science,Ningbo University,Ningbo 315211,China)
Abstract: In order to obtain bacteria with high esterase production,combinatorial mutagenesis using ultraviolet (UV) and diethyl sulfate (DES) technology was applied to previous isolated strain Bacillus velezensis Nz 1.201.The high-yielding mutant strain D27 was selected by the transparent circle method for primary screening and the esterase activity measurement for rescreening.After five consecutive subcultures,its genetic performance was relatively stable.The enzyme activity reached 62.45 U/100 g,which was 66% higher than that of the original strain.Mutant strain D27 was further used in solid-state fermentation.Based on single factor experiment,Box-Behnken design-response surface methodology was used to optimize culture conditions for mutant D27 with esterase activity as the response value.The results showed that the optimal culture conditions were:33℃,6 days,strain inoculation volume of 3%,and raw material moisture content of 73%.Under the optimal condition,the highest esterase activity was 89.52 U /100 g,an increase of 137% compared with the initial strain.Studies have shown that the mutant strain D27 has a higher ability to produce esterase under solid-state fermentation,and has certain application potential in liquor production.
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