花椒抑菌成分提取方法及抑菌机理研究

Antibacterial Effect and Extraction Method from Chinese Prickly Ash

目的:研究花椒抑菌成分的提取方法及抑菌机理,为花椒的高效利用以及新型食品生物抑菌剂的开发提供理论基础。方法:对超声、回流、闪式3种花椒提取方法进行比较,选出最佳方法,确定最佳提取条件;采用牛津杯法及倍半稀释法测定花椒提取物抑菌圈及最小抑菌浓度(MIC);测定添加提取物前后细菌的生长曲线及电导率,对抑菌机理做初步分析。结果:3种提取方法比较:闪式>回流>超声;闪式提取最佳条件:95%乙醇,6min;花椒提取物对金黄色葡萄球菌、藤黄节杆菌、大肠杆菌、铜绿假单胞菌、白色念珠菌及青霉菌的抑菌圈分别为26.09,37.89,21.58,5.71,27.47及26.01 mm,对金黄色葡萄球菌、大肠杆菌及白色念珠菌MIC分别为7.5,7.5,10 mg/m L;生长曲线和电导率试验结果表明,花椒提取物的抑菌作用主要表现在对数生长期,作用后菌悬液电导率明显增加。结论:闪式提取法为花椒提取物最佳提取方法,其可抑制多种微生物生长,主要通过改变细胞膜通透性及影响细胞壁发挥抑菌作用。

Objective： Effective extraction methods and antibacterial mechanism of Chinese prickly ash had been studied, which would provide theoretical basis for efficient utilization of Chinese prickly ash and the exploitation of new food antibacterial agent. Methods： Among three extraction methods from Chinese prickly ash： ultrasound, reflux and flash extraction method, the best extraction method was chosen out, and the best extraction conditions were determined by comparing antibacterial effects. Chinese prickly ash extract was used for antibacterial activity by oxford cup test. The minimum inhibitory concentrations（MIC） of the extract was tested by tube times dilution. The growth curve of bacteria and the conductivity of bacteria suspension were detected before and after the Chinese prickly ash extract was added to the microbial medium, which would contribute to the study on the antibacterial mechanism preliminarily. Results： Comparative analysis of the three extraction methods showed that flash extraction method was the best method and the ultrasonic extraction method was the worst one. The best conditions of flash extraction method were： 95% ethanol and 6 min.The inhibition zone diameters of the extract to Staphylococcus aureus, Gambogic coli, Escherichia coli, Pseudomonas aeruginosa, Candida albicans and penicillium were 26.09 mm, 37.89 mm, 21.58 mm, 5.71 mm, 27.47 mm and 26.01 mm.The MIC value of Staphylococcus aureus, Candida albicans and Escherichia coli were 7.5 g/L, 7.5 g/L and 10 g/L. The results of growth curve test and the conductivity test showed that Chinese prickly ash extract had a significant effect on strains during bacterial logarithmic phase and the bacteria suspension conductivity increased significantly after the extract was added. Conclusion： Flash extraction method was the best one. Chinese prickly ash extract could inhibit the growth of many microbes and its antibacterial activity was mainly due to the changes of the cell membrane permeability and the damage of the cell wall.