乳酸乳球菌革兰氏阳性菌增强基质颗粒制备方法的比较  

Comparison of the preparation methods for Gram-positive bacteria enhancer matrix particles of Lactococcus lactis

作　　者：彭双婷 许洵锴 曾旭健 王燚 苏灿文 梁晋嘉 林鸿颖 陈秋 包银莉 PENG Shuangting;XU Xunkai;ZENG Xujian;WANG Yi;SU Canwen;LIANG Jinjia;LIN Hongying;CHEN Qiu;BAO Yinli(Longyan University,Longyan 364012,China;Fujian Agriculture and Forestry University,Fuzhou 350002,China;Fujian Provincial University Engineering Research Center for Prevention and Control of Animal-Brone Zoonoses,Longyan 364012,China)

机构地区：[1]龙岩学院,福建龙岩364012 [2]福建农林大学,福州350002 [3]动物源性人兽共患病防控福建省高校工程研究中心,福建龙岩364012

出　　处：《黑龙江畜牧兽医》2025年第2期79-85,共7页Heilongjiang Animal Science And veterinary Medicine

基　　金：中央引导地方科技发展资金项目(2022L3067);福建省自然科学基金项目(2020J05202);福建省教育厅中青年教师教育科研项目(JAT190738);国家级大学生创新创业训练计划资助项目(202111312021);上杭县奇迈科技创新基金项目(2020SHQM13)。

摘　　要：为了高效制备高质量乳酸乳球菌革兰氏阳性菌增强基质(Gram-positive enhancer matrix,GEM)颗粒,试验分别采用0.625%、1.25%、2.50%、5.00%、10.00%的三氯乙酸(TCA)、盐酸(HCl)、乙酸(HAc)、硫酸(H2 SO4)加热煮沸乳酸乳球菌MG1363的方法制备了GEM颗粒,并检测了GEM颗粒得率、蛋白质去除效果、颗粒完整性,同时测定了TCA制备的GEM颗粒与锚定蛋白(protein anchor,PA)的结合能力。结果表明:TCA与HAc加热煮沸法制备的GEM颗粒平均得率相似,且极显著高于HCl和H2 SO4(P<0.01),其中5.00%HAc制备的GEM颗粒得率最高,可达99.27%;各浓度(除10%外)HAc、0.625%和1.25%TCA、0.625%HCl及0.625%H2 SO4制备的GEM颗粒蛋白质去除效果较好;各浓度TCA、HCl、HAc及0.625%H2 SO4制备的GEM颗粒与未经酸处理的乳酸乳球菌形态相似,而1.25%、2.50%、5.00%、10.00%H2 SO4制备的颗粒明显圆缩且边缘粗糙;不同浓度TCA制备的GEM颗粒均可与PA结合,且1 U 1.25%TCA制备的GEM颗粒可结合不少于142.86μg的PA。说明低浓度(0.625%、1.25%)的TCA和HCl及5.00%HAc具有更好的提取效果,且1.25%TCA制备的GEM颗粒具有较好的PA结合能力。In order to efficiently prepare high-quality Gram-positive enhancer matrix(GEM)particles of Lactococcus lactis,in the test,GEM particles were prepared by heating and boiling Lactococcus lactis MG1363 at concentrations of 0.625%,1.25%,2.50%,5.00%and 10.00%trichloroacetic acid(TCA),hydrochloric acid(HCl),acetic acid(HAc)and sulfuric acid(H2 SO4),respectively.GEM particle yields,protein removal effect,and particle integrity were detected.At the same time,the binding ability of GEM particles prepared by TCA to protein anchor(PA)was determined.The results showed that the average yields of GEM particles prepared by TCA and HAc heating and boiling methods were similar and extremely significantly higher than those prepared by HCl and H2 SO4(P<0.01),and the yield of GEM particles prepared by 5.00%HAc was the highest,reaching 99.27%.The protein removal effects of GEM particles prepared by all concentrations of HAc(except 10%),0.625%and 1.25%TCA,0.625%HCl and 0.625%H2 SO4 were better.The GEM particles prepared by TCA,HCl,HAc and 0.625%H2 SO4 were similar to those of Lactococcus lactis without acid treatment,but the particles prepared by 1.25%,2.50%,5.00%and 10.00%H2 SO4 were significantly rounded and had rough edges.GEM particles prepared by different concentrations of TCA could bind to PA,and GEM particles prepared by 1 U of 1.25%TCA could bind no less than 142.86μg of PA.These results indicated that the low concentrations of(0.625%,1.25%)TCA,HCl and 5.00%HAc had better extraction effects,and the GEM particles prepared by 1.25%TCA had better PA binding capacity.

关 键 词：革兰氏阳性菌增强基质颗粒 得率 蛋白质去除效率 颗粒完整性 结合能力 

分 类 号：S852.4[农业科学—基础兽医学]