• 中文核心期刊
  • CSCD来源期刊
  • 中国科技核心期刊
  • CA、CABI、ZR收录期刊

植物乳杆菌LV02抑菌特性及发酵培养基优化研究

Study on Bacteriostatic Characteristics of Lactobacillus plantarum LV02 and Optimization of Fermentation Medium

  • 摘要:
      目的   优化植物乳杆菌LV02发酵培养基,研究其抑菌特性,为开发抗菌保鲜类产品提供技术参考。
      方法   以大肠杆菌YS为指示菌,抑菌圈直径及生物量OD600为评价指标。在单因素试验的基础上,采用PB试验初步确定各因素的高低水平,接着采用最陡爬坡试验进一步确定步长及方向来接近最大产值,确定CCD试验中心点。最后,采用CCD设计试验,研究各影响因素及其交互作用对植物乳杆菌LV02抑菌性及生物量OD600的影响,确定各影响因素的最佳水平。并通过牛津杯法研究LV02的抑菌特性,来确定应用环境的设定范围。
      结果   LV02的优化发酵培养基配方为:葡萄糖34.07 g·L−1、酵母浸粉18.12 g·L−1、磷酸氢二钾2 g·L−1、硫酸锰0.16 g·L−1、乙酸钠5 g·L−1、硫酸镁0.20 g·L−1、柠檬酸铵1 g·L−1、吐温80 1 mL·L−1、胡萝卜汁50 mL·L−1,蒸馏水1 L。以5%接种,37 ℃培养24 h,结果为对大肠杆菌YS的抑菌圈直径比未优化前提高了近26%,OD600提高了12%。通过抑菌特性的分析,确定了粗提植物乳杆菌LV02的细菌素所需硫酸铵饱和度为80%,证明了植物乳杆菌LV02具有热稳定性(100 ℃,120 min)、酸碱稳定性(pH 3.0~7.5)及抑菌性。
      结论   采用CCD试验优化植物乳杆菌LV02发酵培养基,可有效提高生物量OD600及对大肠杆菌YS的抑菌性。通过牛津杯法研究LV02的抑菌特性,确定了pH、温度具稳定性的设定范围。

     

    Abstract:
      Objective  Bacteriostatic property of Lactobacillus plantarum LV02 bacteriocin produced on an optimized fermentation medium was determined.
      Method  Based on a single-factor design, the PB and the steepest ascent experiments were conducted to locate the center point of the step length and direction of the influencing factors for a CCD test. Subsequently, effects of different media on the bacteriostatic property and OD600 of the cultured L. plantarum LV02 were evaluated for formulation optimization. The Oxford cup method was employed to determine the bacteriostatic capacity and stability under application conditions of the resulting LV02 bacteriocin produced on the optimized medium.
      Result  The optimized formula for the LV02 fermentation medium in 1 L of water constituted 34.07 g of glucose, 18.12 g of yeast extract, 2 g of dipotassium hydrogen phosphate, 0.16 g of manganese sulfate, 5 g of sodium acetate, 0.20 g of magnesium sulfate, 1 g of ammonium citrate, 1 mL of Tween 80, and 50 mL of carrot juice. On the optimized medium, LV02 grew to yield a 12% increase on OD600 and bacteriocin with a 26% increase on the diameter of inhibition zone against Escherichia coli YS. For the crude LV02 bacteriocin extraction, 80% saturation concentration of ammonium sulfate was used. The antibacterial LV02 bacteriocin was stable under 100 ℃ for 120 m and pH 3.0~7.5.
      Conclusion  L. plantarum LV02 cultured on the optimized fermentation medium was bacteriostatic against E. coli YS. The fermentation produced bacteriocin with desirable stabilities to heat, acid, and slight alkaline condition and was considered promising for the development of an antibacterial agent.

     

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