不同稀酸制备革兰氏阳性增强基质(GEM)颗粒的比较  

Comparison on the preparation of gram-positive enhancer matrix(GEM) particles by different dilute acids

作　　者：李鹏成[1] 张晓燕 乔绪稳[1] 郑其升[1] 侯继波[1] LI Peng-cheng;ZHANG Xiao-yan;QIAO Xu-wen;ZHENG Qi-sheng;HOU Ji-bo(Institute of Veterinary Immunology ＆ Engineering,Jiangsu Academy of Agricultural Sciences,National Research Center of Engineering and Technology for Veterinary Biologicals,Nanjing 210014,China;Yuneheng Agricultural Vocation-technical College of Shanxi,Yuneheng 044000,China)

机构地区：[1]江苏省农业科学院动物免疫工程研究所,国家兽用生物制品工程研究中心,江苏南京210014 [2]山西运城农业职业技术学院,山西运城044000

出　　处：《江苏农业学报》2018年第3期605-611,共7页Jiangsu Journal of Agricultural Sciences

基　　金：国家自然科学基金项目(31502105); 江苏省农业科学院基本科研业务专项项目[ZX(16)2030]

摘　　要：本研究旨在比较不同稀酸制备革兰氏阳性增强基质(GEM)颗粒的效果。首先将3种不同的稀酸[硫酸(H2SO4)、盐酸(HCl)、三氯乙酸(TCA)]煮沸,制备GEM颗粒,分别比较得率(细菌计数)、蛋白去除率(SDSPAGE和SDS解离蛋白定量分析)以及GEM颗粒与锚钩蛋白(PA)的亲和活性等,最后进行4℃以及冷冻干燥后4℃下GEM的保存期研究。结果显示,0.025 mol/L的H2SO4制备GEM颗粒得率为89.8%,蛋白去除效果和锚定活性均较好。0.050 mol/L HCl与0.100 mol/L HCl制备的GEM颗粒蛋白质去除效率无显著差异,0.050 mol/L HCl制备GEM的得率(99.7%)高于0.100 mol/L HCl(86.5%),但0.050 mol/L HCl制备的GEM颗粒与PA的锚定活性不及0.100 mol/L HCl制备的GEM颗粒。不同浓度TCA制备的GEM颗粒得率、锚定活性均较好,但蛋白质去除效果较差。GEM颗粒4℃保存18个月,其活性不受影响,且冷冻干燥对GEM颗粒活性无影响。0.025 mol/L的H2SO4和0.100 mol/L的HCl可制备高质量的GEM。The aim of this study was to compare the effects of gram-positive enhancer matrix（ GEM） prepared with different dilute acids. Three different dilute acids,H2SO4,HCl and TCA were applied to prepare the GEM particles by boiling. The optimal preparation conditions were determined by comparing the yield（ bacterial count）,protein removal rate（ SDS-PAGE and protein quantification） and the affinity activity of GEM particles and protein anchor（ PA）. Furthermore,the shelf life of GEM was researched at 4 ℃ and after freeze-drying. The results showed that the yield of the GEM particles was 89.8% under the treatment of 0.025 mol/L H2SO4,and the protein removal effect and anchoring activity were ideal.There was no difference in the protein removal rate of GEM under the treatment of 0.050 mol/L HCl and 0.100 mol/L HCl,and the yield under the treatment of 0. 050 mol/L HCl（ 99. 7%） was higher than that under the treatment of0. 100 mol/L HCl（ 86. 5%）. However,the anchoring activity of GEM particles with PA under the treatment of0. 050 mol/L HCl was less than that under the treatment of0. 100 mol/L HCl. The yield and anchoring activity of GEM particles obtained by different concentration of TCA were ideal,but the protein removal rate was poor. GEM particles were kept 18 months at 4 ℃. Freeze drying couldn＇t influence the activity of GEM particles. The results indicated that the treatments of 0.025 mol/L H2SO4 and 0.100 mol/L HCl could produce high-quality GEM particles.

关 键 词：乳酸乳球菌 革兰氏阳性增强基质(GEM) 稀酸 

分 类 号：S852.43[农业科学—基础兽医学]