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    细叶小檗不同生长部位生物碱抑菌活性研究

    Antibacterial activity of alkaloids from different growth parts of Berberis poiretii

    • 摘要:
        目的  对细叶小檗不同生长部位(根、茎、叶、果)生物碱抑菌活性进行对比评价,旨在筛选细叶小檗抑菌有效部位和主要抑菌成分,为拓宽其全植株应用领域及研发新型天然食品防腐剂的研究提供理论依据。
        方法  以抑菌圈直径、最低抑菌质量浓度和最低杀菌质量浓度为考查指标,采用牛津杯抑菌法,研究各生长部位生物碱对4种食源性致病菌(大肠杆菌、金黄色葡萄球菌、沙门氏菌、枯草芽孢杆菌)的抑制作用,采用高效液相色谱法测定生物碱的种类和含量。
        结果  纯化前后细叶小檗4种不同生长部位生物碱对大肠杆菌、金黄色葡萄球菌、沙门氏菌、枯草芽孢杆菌这4种常见食源性致病菌均呈现正相关的剂效关系。其中,纯化前细叶小檗实生物碱的抑菌效果最佳,当其生物碱质量浓度为60 g/L时,抑菌圈直径大于20 mm。通过二倍稀释法研究表明:其对大肠杆菌、沙门氏菌、枯草芽孢杆菌的最低抑菌质量浓度(MIC)均为1.25 g/L,最低杀菌质量浓度(MBC)均为2.50 g/L,对金黄色葡萄球菌的MIC为2.50 g/L,MBC为5.00 g/L。纯化后细叶小檗根生物碱的抑菌效果最佳,当其生物碱质量浓度为40 g/L时,抑菌圈直径大于20 mm。通过二倍稀释法研究表明:其对大肠杆菌、金黄色葡萄球菌的MIC均为0.31 g/L,MBC均为0.63 g/L,对枯草芽孢杆菌、沙门氏菌的MIC为0.63 g/L,MBC为1.25 g/L。液相色谱检测结果显示:纯化后生物碱构成包括小檗碱(0.28 ~ 41.69 g)、药根碱(0.32 ~ 12.67 g)、巴马汀(0.25 ~ 17.09 g)。
        结论  细叶小檗根、茎和果这3个部位对供试菌株有较强的抑菌作用,叶抑菌效果稍弱。4个不同生长部位基本生物碱单体种类相同,但含量存在显著差异。当生物碱质量浓度为60 g/L时,纯化前不同部位抑菌作用强弱次序为果 > 根 > 茎 > 叶;纯化后不同部位抑菌作用强弱次序为根 > 茎 > 实 > 叶。

       

      Abstract:
        Objective  In this paper, the antibacterial activity of alkaloids from different parts (root, stem, leaf and fruit) of Berberis poiretii was evaluated to provide a theoretical basis for broadening the application fields of whole plant and developing new natural food preservatives.
        Method  Taking the diameter of bacteriostatic zone, the lowest bacteriostatic concentration and the lowest bacteriostatic concentration as the examination index, the Oxford Cup bacteriostatic method was used to study the inhibitory effects of alkaloids from different growth sites on four food-borne pathogens (Escherichia coli, Staphylococcus aureus, Salmonella spp., Bacillus subtilis), the types and contents of alkaloids were determined by high-performance liquid chromatography method.
        Result  Before and after purification, the alkaloids from 4 different parts of Berberis poiretii showed positive dose-effect relationship to four common food-borne pathogenic bacteria, i.e. Escherichia coli, Staphylococcus aureus, Salmonella spp., Bacillus subtilis. When the concentration of alkaloids was 60 g/L, the diameter of bacteriostatic zone was more than 20 mm. The minimum inhibitory concentration (MIC) to Escherichia coli, Salmonella spp. and Bacillus subtilis was 1.25 g/L, minimum bactericidal concentration (MBC) was 2.50 g/L, and the MIC to Staphylococcus aureus was 2.50 g/L, MBC was 5.00 g/L. When the concentration of alkaloid was 40 g/L, the diameter of bacteriostatic zone was more than 20 mm. The results showed that the MIC for Escherichia coli and Salmonella spp. was 0.31 g/L, MBC was 0.63 g/L, and the MIC for Bacillus subtilis and Salmonella spp. was 0.63 g/L, respectively, MBC was 1.25 g/L. The results of HPLC showed that the purified alkaloids were berberine (0.28–41.69 g), jatrorrhizine (0.32–12.67 g) and palmatine (0.25–17.09 g) .
        Conclusion  The root, stem and fruit of Berberis poiretii have stronger bacteriostatic effect on the tested strains, but the leaf has weaker bacteriostatic effect. The basic alkaloid monomers in 4 different growing parts are the same, but the content of the monomers is significantly different. When the concentration of alkaloid is 60 g/L, the order of bacteriostasis is fruit > root > stem > leaf, and the order of bacteriostasis is root > stem > fruit > leaf.

       

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