文章摘要
李淼,侯灵芝,徐兴浩,王肖肖,郑明灿,刘昂.日照海域分泌肽聚糖水解酶菌株的筛选、初步鉴定及抑菌活性测定[J].济宁医学院学报,2020,43(4):233-237
日照海域分泌肽聚糖水解酶菌株的筛选、初步鉴定及抑菌活性测定
Screening,preliminary identification of peptidoglycan hydrolase-producing bacteria from Rizhao Sea area and determination of antibacterial activity of the obtained peptidoglycan hydrolase
投稿时间:2020-06-23  
DOI:10.3969/j.issn.1000-9760.2020.04.002
中文关键词: 肽聚糖水解酶;溶菌酶;金黄色葡萄球菌;芽孢杆菌
英文关键词: Peptidoglycan hydrolase;Lysozyme;Staphylococcus aureus;Bacillus
基金项目:济宁医学院2018年校级大学生创新创业训练计划项目(cx2018040);济宁医学院2018年国家级大学生创新创业训练计划项目(20180443012)
作者单位E-mail
李淼 济宁医学院基础医学院, 济宁 272067  
侯灵芝 济宁医学院基础医学院, 济宁 272067  
徐兴浩 济宁医学院基础医学院, 济宁 272067  
王肖肖 济宁医学院基础医学院, 济宁 272067  
郑明灿 济宁医学院基础医学院, 济宁 272067  
刘昂 济宁医学院基础医学院, 济宁 272067 liuang08@163.com 
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中文摘要:
      目的 筛选能分泌肽聚糖水解酶的菌株并测定其抑菌活性,为研发新型抗菌药物提供素材。方法 从日照海域潮间带沉积物中筛选能水解肽聚糖,并且可以在以金黄色葡萄球菌为唯一营养物的液体培养基中生长的菌株,16S rRNA基因序列测定并同源性比对,确定其分类地位;诱导产酶,离心培养液得到的上清作为粗酶液,观测粗酶液对金黄色葡萄球菌的抑菌活性。结果 筛选出8株分泌肽聚糖水解酶且能够在以金黄色葡萄球菌为唯一营养物的液体培养基中生长的菌株。16S rRNA基因序列同源性比对分析表明,这8株菌均属于芽孢杆菌属。其粗酶液菌均对金黄色葡萄球菌有明显的抑制作用。与对照组相比,粗酶液处理后菌落形成单位下降比率在(61.9±7.7)%~(95.0±2.4)%。显微镜下观察,粗酶液处理后的葡萄球菌轮廓变得不清晰,数量减少。结论 海洋潮间带中存在能够分泌肽聚糖水解酶的菌株,本实验筛选的8株菌粗酶液均表现出较强的抑制金黄色葡萄球菌的活性,且粗酶液对金黄色葡萄球菌有裂解活性。
英文摘要:
      Objective To screen bacteria strains capable of secreting peptidoglycan hydrolase and determine their antibacterial activity,which provide materials for the development of new antibacterial drugs. Methods The strains were screened capable of hydrolyzing peptidoglycan from the intertidal sediments in Rizhao sea area.The strains were inoculated in a liquid medium with Staphylococcus.aureus as the sole nutrient and those strains that could grow in this medium were selected for the following experiments.First of all,the screened strains were subjected to the 16S rRNA gene sequence determination and homology comparison to determine its taxonomic status.Secondly,the selected strains were inoculated in a liquid medium with S.aureus as the sole nutrient to induce enzyme production,and the supernatant after centrifuging the culture was used as a crude enzyme.The antibacterial activity of crude enzyme against S.aureus was measured and the effect of crude enzyme solution on S.aureus was observed under a microscope. Results Eight strains that could secret peptidoglycan hydrolase and were able to grow in liquid medium with S.aureus as the sole nutrient were selected.The 16S rRNA gene sequence homology analysis showed that the eight strains belonged to the genus Bacillus.All of the crude enzymes of the selected eight strains had obvious inhibitory effects on S.aureus.Compared with the blank control,the reduction rate of colony forming units after the action of crude enzyme solution was between 61.9±7.7%~95.0±2.4%.Observed under a microscope,the outline of cells of S.aureus became unclear and the number decreased. Conclusion There are strains capable of secreting peptidoglycan hydrolase in the marine intertidal zone.The eight strains capable of secreting peptidoglycan hydrolase in this experiment belong to the genus Bacillus.The crude enzyme solutions of the eight strains screened showed strong activity against S.aureus,and microscopic examination showed that the crude enzyme solution had lytic activity against S.aureus.
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