广西红树林土壤产淀粉酶菌株的筛选及产酶条件优化

Screening of amylase-producing strains in Guangxi mangrove soil and optimization of enzyme-producing conditions

  • 摘要: 目的:从广西石角红树林自然保护区红树林土壤中筛选产淀粉酶菌株,通过Plackett-Burman实验和响应面优化其产酶条件,以期建立高产淀粉酶的发酵工艺。方法:通过平板筛选,从红树林土壤中获得产淀粉酶菌株;通过生理生化与16S rDNA测序确定菌株种类;通过单因素优化,Plackett-Burman实验和响应面优化得出最适产酶条件。结果:从红树林土壤中获得产淀粉酶菌株;鉴定为贝莱斯芽孢杆菌(Bacillus velezensis),命名为Bacillus velezensis MG5-8;获得淀粉酶的最适温度为70℃,最适pH为6.0;经优化获得该菌株产淀粉酶最适温度为30℃,最适产酶碳源为玉米浆粉,最适产酶氮源为黄豆饼粉与牛肉膏;最适产酶条件为黄豆饼粉0.51%、牛肉膏0.55%、玉米浆粉0.34%,在此条件下该菌株的理论酶活性最大值为134.03 U/mL,模型经验证测得B. velezensis MG5-8淀粉酶活性为130.28 U/mL,为理论值的97.2%,是优化前酶活的5.63倍。结论:成功分离出一株产淀粉酶的菌株(B. velezensis MG5-8),在优化的条件下酶活最高为130.28 U/mL。

     

    Abstract: Objective: An amylase-producing strain was screened and identified from mangrove soil in Shijiao Mangrove Nature Reserve of Guangxi. The amylase-producing process was optimized by Plackett-Burman design and response surface optimization, in order to establish the fermentation process of high amylase production.Methods: The amylase-producing strain was obtained from mangrove soil by plate screening. The strain was identified by physiological, biochemical and 16S rDNA sequencing. The optimized condition for enzyme production was obtained by single factor optimization, Plackett-Burman design and response surface optimization. Results: The amylase-producing strain was obtained from mangrove soil. Bacillus velezensis was identified and named Bacillus velezensis MG5-8. The optimum temperature and pH of the amylase were 70 ℃ and 6.0, respectively. The optimum temperature for amylase production was 30 ℃, the optimum carbon source for amylase production was corn steep liquor powder, and the optimum nitrogen source was soybean powder and beef extract by single factor optimization. The optimum enzyme production conditions were as following: soybean powder 0.51%, beef extract 0.55%, corn steep liquor powder 0.34%. Under these conditions, the theoretical maximum enzyme activity of this strain was 134.03 U/mL. The activity of B. velezensis MG5-8 amylase was 130.28 U/mL,which was 97.2% of the theoretical value and 5.63 folds of the enzyme activity before optimization. Conclusion: A strain producing amylase(B. velezensis MG5-8) is successfully isolated and identified. The maximum enzyme activity is 130.28 U/mL after optimization.

     

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