不动杆菌属中aidE基因编码高丝氨酸内酯酶
发布时间：2018-11-03 16:16
【摘要】：群体感应(Quorum sensing,QS)是细菌在进化过程中形成的依赖于群体密度的细菌间交流方式。许多革兰氏阴性细菌以N-酰基高丝氨酸内酯(AHL)为信号分子,感应自身群体密度并调控致病基因表达。因此,淬灭AHLs信号分子可防治此类细菌引起的植物病害。本实验室前期已筛选得到了一株具有AHLs信号降解能力的不动杆菌菌株Acinetobacter sp.77,本研究通过基因组文库筛选,自菌株77中克隆得到具有AHLs降解活性的基因aidE。该基因编码268个氨基酸。序列一致性比较发现aidE的氨基酸序列与吉伦伯不动杆菌Acinetobacter gyllenbergii CIP110306中β-内酰胺酶一致性高达95%,但与已知的AHLs降解酶序列一致性较低,最高为缓黄分支杆菌Mycobacterium lentiflavum中AHL内酯酶Att M/Aii B家族蛋白(CQD23908.1),一致性仅为33%。通过高压液相色谱(HPLC)分析Aid E蛋白处理N-己酰基高丝氨酸内酯(C6-HSL)的反应产物,证明aidE为AHL内酯酶。序列比对研究发现,aidE基因在不动杆菌属中并不保守,其在菌株77基因组中的上下游的基因排列存在菌株水平的特异性,且aidE基因下游存在疑似IS插入序列,上述证据表明aidE基因有可能是通过水平转移进入Acinetobacter sp.77基因组中,或其在基因组中的位置发生过重排。表达aidE的软腐果胶杆菌Z3-3中完全检测不到AHLs信号产生,且致病力明显降低。综上所述,aidE为新发现的AHL内酯酶。在防治依赖QS系统表达致病性的细菌病害中具有应用潜力。
[Abstract]:Group sensing (Quorum sensing,QS) is a kind of communication between bacteria which depends on population density during the evolution of bacteria. Many Gram-negative bacteria use N-acyl high-serine lactone (AHL) as signal molecule, which induces their own population density and regulates the expression of pathogenic genes. Therefore, quenched AHLs signaling molecules can prevent and cure plant diseases caused by these bacteria. A strain of Acinetobacter sp.77, with the ability of AHLs signal degradation was screened in our laboratory. The gene aidE. with AHLs degradation activity was cloned from strain 77 by genomic library screening. The gene encodes 268 amino acids. The results showed that the amino acid sequence of aidE was as high as 95% of 尾 -lactamase in Acinetobacter cereus Acinetobacter gyllenbergii CIP110306, but it was not consistent with the known AHLs degrading enzyme sequence. The highest was Att M/Aii B family protein (CQD23908.1) in AHL of Mycobacterium bradyloides Mycobacterium lentiflavum, and the consistency was only 33%. The reaction product of N-caproyl homoserine lactone (C6-HSL) treated with Aid E protein was analyzed by high performance liquid chromatography (HPLC). It was proved that aidE was an esterase in AHL. Sequence alignment showed that the aidE gene was not conserved in Acinetobacter genus, and the upstream and downstream genes in the genome of strain 77 had the specificity of strain level, and there was a suspected IS insertion sequence in the downstream of the aidE gene. The above evidences suggest that the aidE gene may be transferred horizontally into the Acinetobacter sp.77 genome or its position in the genome has been rearranged. AHLs signal was not detected in Pectinobacterium soft Pectinobacterium Z3-3, which expressed aidE, and its pathogenicity was significantly decreased. In conclusion, aidE is a newly discovered esterase in AHL. It has the potential to prevent and cure bacterial diseases which depend on the expression of QS system.
【作者单位】： 中国农业大学植物病理系;
【基金】：国家自然科学基金(Nos.31272082,31572045) 国家重点基础研究发展计划(973计划)(No.2015CB150605) 国家重点研发计划重点专项(No.2017YFD0201108)资助~~
【分类号】：Q933
[Abstract]:Group sensing (Quorum sensing,QS) is a kind of communication between bacteria which depends on population density during the evolution of bacteria. Many Gram-negative bacteria use N-acyl high-serine lactone (AHL) as signal molecule, which induces their own population density and regulates the expression of pathogenic genes. Therefore, quenched AHLs signaling molecules can prevent and cure plant diseases caused by these bacteria. A strain of Acinetobacter sp.77, with the ability of AHLs signal degradation was screened in our laboratory. The gene aidE. with AHLs degradation activity was cloned from strain 77 by genomic library screening. The gene encodes 268 amino acids. The results showed that the amino acid sequence of aidE was as high as 95% of 尾 -lactamase in Acinetobacter cereus Acinetobacter gyllenbergii CIP110306, but it was not consistent with the known AHLs degrading enzyme sequence. The highest was Att M/Aii B family protein (CQD23908.1) in AHL of Mycobacterium bradyloides Mycobacterium lentiflavum, and the consistency was only 33%. The reaction product of N-caproyl homoserine lactone (C6-HSL) treated with Aid E protein was analyzed by high performance liquid chromatography (HPLC). It was proved that aidE was an esterase in AHL. Sequence alignment showed that the aidE gene was not conserved in Acinetobacter genus, and the upstream and downstream genes in the genome of strain 77 had the specificity of strain level, and there was a suspected IS insertion sequence in the downstream of the aidE gene. The above evidences suggest that the aidE gene may be transferred horizontally into the Acinetobacter sp.77 genome or its position in the genome has been rearranged. AHLs signal was not detected in Pectinobacterium soft Pectinobacterium Z3-3, which expressed aidE, and its pathogenicity was significantly decreased. In conclusion, aidE is a newly discovered esterase in AHL. It has the potential to prevent and cure bacterial diseases which depend on the expression of QS system.
【作者单位】： 中国农业大学植物病理系;
【基金】：国家自然科学基金(Nos.31272082,31572045) 国家重点基础研究发展计划(973计划)(No.2015CB150605) 国家重点研发计划重点专项(No.2017YFD0201108)资助~~
【分类号】：Q933
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