不動桿菌屬中aidE基因編碼高絲氨酸內(nèi)酯酶
發(fā)布時間：2018-11-03 16:16
【摘要】：群體感應(yīng)(Quorum sensing,QS)是細菌在進化過程中形成的依賴于群體密度的細菌間交流方式。許多革蘭氏陰性細菌以N-酰基高絲氨酸內(nèi)酯(AHL)為信號分子,感應(yīng)自身群體密度并調(diào)控致病基因表達。因此,淬滅AHLs信號分子可防治此類細菌引起的植物病害。本實驗室前期已篩選得到了一株具有AHLs信號降解能力的不動桿菌菌株Acinetobacter sp.77,本研究通過基因組文庫篩選,自菌株77中克隆得到具有AHLs降解活性的基因aidE。該基因編碼268個氨基酸。序列一致性比較發(fā)現(xiàn)aidE的氨基酸序列與吉倫伯不動桿菌Acinetobacter gyllenbergii CIP110306中β-內(nèi)酰胺酶一致性高達95%,但與已知的AHLs降解酶序列一致性較低,最高為緩黃分支桿菌Mycobacterium lentiflavum中AHL內(nèi)酯酶Att M/Aii B家族蛋白(CQD23908.1),一致性僅為33%。通過高壓液相色譜(HPLC)分析Aid E蛋白處理N-己酰基高絲氨酸內(nèi)酯(C6-HSL)的反應(yīng)產(chǎn)物,證明aidE為AHL內(nèi)酯酶。序列比對研究發(fā)現(xiàn),aidE基因在不動桿菌屬中并不保守,其在菌株77基因組中的上下游的基因排列存在菌株水平的特異性,且aidE基因下游存在疑似IS插入序列,上述證據(jù)表明aidE基因有可能是通過水平轉(zhuǎn)移進入Acinetobacter sp.77基因組中,或其在基因組中的位置發(fā)生過重排。表達aidE的軟腐果膠桿菌Z3-3中完全檢測不到AHLs信號產(chǎn)生,且致病力明顯降低。綜上所述,aidE為新發(fā)現(xiàn)的AHL內(nèi)酯酶。在防治依賴QS系統(tǒng)表達致病性的細菌病害中具有應(yīng)用潛力。
[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.
【作者單位】： 中國農(nóng)業(yè)大學植物病理系;
【基金】：國家自然科學基金(Nos.31272082,31572045) 國家重點基礎(chǔ)研究發(fā)展計劃(973計劃)(No.2015CB150605) 國家重點研發(fā)計劃重點專項(No.2017YFD0201108)資助~~
【分類號】：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.
【作者單位】： 中國農(nóng)業(yè)大學植物病理系;
【基金】：國家自然科學基金(Nos.31272082,31572045) 國家重點基礎(chǔ)研究發(fā)展計劃(973計劃)(No.2015CB150605) 國家重點研發(fā)計劃重點專項(No.2017YFD0201108)資助~~
【分類號】：Q933
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