紫叶小檗果小檗碱的提取及抑菌活性研究

谭婷; 朱斌; 杨倩; 吴娟娟; 史娟

doi:10.13386/j.issn1002-0306.2019120242

紫叶小檗果小檗碱的提取及抑菌活性研究

Extraction and Antibacterial Activity of Berberine from the Fruit of Berberis thunbergii var atropurpurea

摘要

摘要: 研究紫叶小檗果小檗碱的提取条件及其抑菌活性。在单因素实验和正交实验基础上,得到小檗碱的最佳提取工艺。通过平板抑菌实验测定紫叶小檗果小檗碱对马铃薯干腐病菌(Fusarium solani,F.s.)、番茄灰霉病菌(Botrytis cinerea,B.c.)、尖孢镰刀病菌(Fusarium oxysporum,F.o.)、苹果炭疽病菌(Colletotrichum gloeosporioides,C.g.)、水稻稻瘟病菌(Magnaporthe grisea,M.g.)、苹果腐烂病菌(Cytospora sp,C.s.)六种植物病菌的抑制作用;采用滤纸片法评估紫叶小檗果及葛根、紫叶小檗果配伍制剂对金黄色葡萄球菌和大肠杆菌的抑菌活性。实验结果表明,小檗碱最佳提取条件为:纤维素酶用量120 mg、酶解时间120 min、料液比1:12 g/mL、浸提时间150 min,得率达到0.198%。在小檗碱浓度为4 mg/mL时,对六种植物病菌抑菌效果由大到小排序如下:尖孢镰刀病菌(F.o.) > 苹果腐烂病菌(C.s.) > 番茄灰霉病菌(B.c.) > 水稻稻瘟病菌(M.g.) > 马铃薯干腐病菌(F.s.) > 苹果炭疽病菌(C.g.),小檗碱对尖孢镰刀病菌具有较好的抑菌活性,抑菌率在小檗碱浓度为4 mg/mL时可达到89.28%;葛根与紫叶小檗果配伍对金黄色葡萄球菌的抑菌活性较强,对大肠杆菌的抑菌活性存在最佳配伍,为葛根:紫叶小檗果质量比6:44。综上所述,本实验所得提取工艺稳定、可行,可用于紫叶小檗果中小檗碱的提取,并且紫叶小檗果中小檗碱具有较强的抑菌活性,可用于食品防腐、农业生物防治等领域。

Abstract: To optimize conditions for extracting and antibacterial activity of berberine from the fruit of Berberis thunbergii var atropurpurea, the single factor and orthogonal test were used. The inhibitory effects of berberine on six plant bacteria (Fusarium solani, Botrytis cinerea, Fusarium oxysporum, Colletotrichum gloeosporioides, Magnaporthe grisea and Cytospora sp) were determined by plate bacteriostatic tests. The antibacterial activities of the compatibility of Radix Puerariae and Berberis thunbergii var atropurpurea, and pure Berberis thunbergii var atropurpurea against Escherichia coli and Staphylococcus aureus were evaluated via the filter paper disk. The optimal extraction conditions for berberine were 120 mg of cellulase, 120 min enzymolysis time, material-liquid ratio 1:12 g·mL^-1, extraction time 150 min. The extraction rate reached 0.198%. The order of antibacterial activity of six plant pathogens was as follows:Fusarium oxysporum > Cytospora sp > Botrytis cinerea > Magnaporthe grisea > Fusarium solani > Colletotrichum gloeosporioides. Berberine had better antibacterial effect on Fusarium oxysporum. Bacteriostatic rate reached 89.28% when the content of berberine was 4 mg/mL. The combination of Berberis purpurea and Pueraria lobata had strong antibacterial activity against Staphylococcus aureus, and the best compatibility for inhibiting Escherichia coli was Pueraria lobata:Purple Berberis purpurea mass ratio 6:44. In summary, the extraction process obtained in this experiment is stable and feasible, and it can be used for the extraction of berberine from fruit of Berberis thunbergii var atropurpurea, and the berberine from purple leaf berberis has strong antibacterial activity and can be used in food preservation, agricultural biological control and other fields.