培养条件及人参对嗜酸乳杆菌外膜囊泡的影响  

Effects of culture conditions and ginseng on outer membrane vesicles of Lactobacillus acidophilus

作　　者：李星枚 李勇敏[2] 罗吉[2] 吕元[2] 刘芳[2] 谭小宁[2] LI Xing-mei;LI Yong-min;LUO Ji;Lü Yuan;LIU Fang;TAN Xiao-ning(Hunan University of Chinese Medicine,Changsha 410000,China;Hunan Academy of Traditional Chinese Medicine,Affiliated Hospital,Changsha 410000,China)

机构地区：[1]湖南中医药大学,湖南长沙410000 [2]湖南省中医药研究院附属医院,湖南长沙410000

出　　处：《时珍国医国药》2022年第4期824-826,共3页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金面上项目(81774163);湖南省科技创新计划项目(2017TP1033)。

摘　　要：目的探究在不同培养条件及人参对嗜酸乳杆菌分泌的外膜囊泡(OMVs)的影响。方法在厌氧培养、需氧培养环境及海博、OXOID两种不同厂家培养基的培养条件下,及人参与嗜酸乳杆菌共培养中,提取其分泌的OMVs。用透射电镜观察OMVs的大小、形态;用BCA蛋白含量测定法测定其蛋白含量;SDS-PAGE测定其蛋白分布。结果经电镜鉴定,嗜酸乳杆菌OMVs为20~200nm的双层囊泡状结构;在厌氧、需氧培养条件下,嗜酸乳杆菌OMVs的蛋白含量分别为0.63mg/mL、0.45mg/mL;在海博、OXOID两种不同厂家培养基培养的嗜酸乳杆菌OMVs蛋白含量分别为0.92mg/mL、0.45mg/mL;在有无人参影响下OMVs、Gin-OMVs蛋白含量分别为0.45mg/mL、0.28mg/mL;厌氧、需氧培养的嗜酸乳杆菌OMVs在40~55KD均有两条条带,而厌氧培养的OMVs在30~40KD另有一条条带;海博、OXOID两种不同厂家培养基培养的嗜酸乳杆菌OMVs蛋白条带无明显差别;OMVs与Gin-OMVs相比Gin-OMVs的条带较浅,且40~55KD之间只有一条条带。结论嗜酸乳杆菌在不同培养条件下所分泌的OMVs是有区别的,可选出最佳提取参数;人参可能能够抑制嗜酸乳杆菌分泌OMVs及改变其内含蛋白,此实验可供后续嗜酸乳杆菌OMVs功能研究提供一定的研究价值。Objective To investigate the effect of ginseng on OMVs secreted by Lactobacillus acidophilus under different culture conditions.Methods The OMVs secreted by Lactobacillus acidophilus were extracted under the conditions of anaerobic culture,aerobic culture and two different media of Haibo and OXOID,and human participation.The size and morphology of OMVs were observed by tem.The protein content was determined by BCA protein content assay.The protein distribution was determined by SDS-PAGE.Results The OMVs of Lactobacillus acidophilus were identified by electron microscope as 20-200 nm double-layer vesicles.Under anaerobic and aerobic culture conditions,the protein content of Latobacillus acidophilus OMVs was 0.63 mg/mL and 0.45 mg/mL,respectively.The OMVs protein content of Latobacillus acidophilus cultured in haibo and OXOID medium was 0.92 mg/mL and 0.45 mg/mL,respectively.The OMVs protein contents of OMVs and Gin-OMVs were 0.45 mg/mL and 0.28 mg/mL under the influence of ginseng or not,respectively.Both anaerobic and aerobic cultured Lactobacillus acidophilus OMVs had two bands at 40-55 kD,while anaerobic cultured Lactobaciulls acidophilus OMVs had another band at 30-40 kD.There was no Significant difference in the OMVs protein bands of Lactobacillus acidophilus cultured in the medium of haibo and OXOID.Compared with Gin-OMVs,the strips of Gin-OMVs are shallows,and there are only strips between 40 and 55 KD.Conclusion The OMVs secreted by Lactobacillus acidophilus under different culture conditions are different,and the optimal extraction conditions can be selected.Ginseng may inhibit the secretion of OMVs by Lactobacillus acidophilus and change its contained proteins.This experiment provides certain research value for the subsequent study on the OMVs function of Lactobacillus acidophilus.

关 键 词：嗜酸乳杆菌 外膜囊泡 人参 提取 鉴定 

分 类 号：R285.5[医药卫生—中药学]