Characterization of EmpA protease in Vibrio anguillarum M3
Characterization of EmpA protease in Vibrio anguillarum M3
复制标题
鳗弧菌 M3 中 EmpA 蛋白酶的表征
DOI：
10.1007/s11802-011-1781-x
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发表时间：
2011-11
期刊：
影响因子：
--
通讯作者：
LI Jie
中科院分区：
文献类型：
--
作者：
YANG Guanpin;HAN Yifan;XIAO Peng;HAO Bin;MO Zhaolan;LI Jie
EmpA是鳗弧菌分泌的一种胞外金属蛋白酶。为了更好地了解empA在鳗弧菌M3菌株致病性中的作用，构建了empA插入突变体。在突变体中，其胞外蛋白水解活性、群集运动性、溶血活性和对大菱鲆的毒力降低。突变株和野生型菌株的胞外蛋白水解活性显著下降（5倍），生长曲线相似，表明EmpA是M3的主要胞外蛋白酶。突变体的LD_（50）比野生型增加了38倍。在培养上清中未检测到pro-EmpA，表明EmpA在24 h后自溶为成熟蛋白。M3株的EmpA分泌与NB 10株相似。突变株的减毒力与M93 Sm株相似。结果表明，EmpA对鳗弧菌M3的特异性作用不同于以往的研究，EmpA对鳗弧菌M3的群集运动和溶血活性有促进作用。这些结果为理解EmpA的功能及其在疫苗开发中的潜在应用提供了见解。
EmpA is an extracellular metalloprotease secreted by Vibrio anguillarum. For better understanding its role in the pathogenicity of V. anguillarum strain M3, empA insertion mutant was constructed. In the mutant it decreased in extracellular proteolytic activity, swarming motility, hemolytic activity and virulence on turbot (Scophthalmus maximus). Significant decline (by 5-fold) of extracellular proteolytic activity and similar growth curve between mutant and wild type strains indicated that EmpA was the major extracellular protease of M3. LD50 of mutant increased by 38-fold compared with wild type. No pro-EmpA was detected in the supernatant of culture, indicating that EmpA autolyzed to mature protein after 24 h. Secretion of EmpA in M3 was similar to that in NB10 strain. Attenuated virulence of mutant was similar to that of M93Sm strain. It was demonstrated that specific operation of EmpA was different from that in previous studies and EmpA contributed to the swarming motility and hemolytic activity in V. anguillarum strain M3. The results provides insight into understanding the function of EmpA and its potential application in vaccine development.
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