The biocontrol strain Pseudomonas fluorescens F113 produces the Rhizobium small bacteriocin, N-(3-hydroxy-7-cis-tetradecenoyl)homoserine lactone, via HdtS, a putative novel N-acylhomoserine lactone synthase
The biocontrol strain Pseudomonas fluorescens F113 produces the Rhizobium small bacteriocin, N-(3-hydroxy-7-cis-tetradecenoyl)homoserine lactone, via HdtS, a putative novel N-acylhomoserine lactone synthase
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DOI：
10.1099/00221287-146-10-2469
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发表时间：
2000-10-01
期刊：
影响因子：
--
通讯作者：
Williams, P
中科院分区：
文献类型：
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作者：
Laue, RE;Jiang, Y;Williams, P
几种不同的假单胞菌产生N-酰基高丝氨酸内酯（AHLs），群体感应信号分子，参与细胞密度依赖性控制次级代谢产物和毒力基因表达。当荧光假单胞菌F113与能够灵敏地检测短（C-4-C-8）或长（C-10-C-14）酰基链AHL的阿勒生物传感器交叉划线时，没有检测到活性。然而，通过用二氯甲烷提取无细胞的固定相培养上清液，然后通过反相HPLC，得到三个不同的馏分能够激活阿勒生物传感器。随后，使用高分辨率MS和化学合成对三种AHLs进行了表征。它们是（i）N-（3-羟基-7-顺式-十四烯酰基）高丝氨酸内酯（30 H，C-14：1-HSL），一种由于其生长抑制特性而先前被称为根瘤菌小细菌素的分子，（ii）N-十氨酰高丝氨酸内酯（C-10-HSL）和（iii）N-己酰高丝氨酸内酯（C-6-HSL）。克隆了一个能指导大肠杆菌中所有三种荧光假单胞菌AHLs合成的基因（hdtS）并测序。hdtS的体外转录/翻译产生了一个约33 kDa的蛋白质，该蛋白质能够指导3OH、C-14：1-HSL、C-10-HSL和C-6-HSL在E.在大肠杆菌中，HdtS不属于已知的阿勒合酶家族（LuxI或LuxM）中的任一个，并且与溶血磷脂酸酰基转移酶家族相关。因此，HdtS可能构成能够进行阿勒生物合成的第三蛋白质家族的成员。
Several different species of Pseudomonas produce N-acylhomoserine lactones (AHLs), quorum-sensing signal molecules which are involved in the cell-density-dependent control of secondary metabolite and virulence gene expression. When Pseudomonas fluorescens F113 was cross-streaked against AHL biosensors capable of sensitively detecting either short (C-4-C-8) or long (C-10-C-14) acyl chain AHLs, no activity was detectable. However, by extracting cell-free stationary-phase culture supernatants with dichloromethane followed by reverse-phase HPLC, three distinct fractions were obtained capable of activating the AHL biosensors. Three AHLs were subsequently characterized using high-resolution MS and chemical synthesis. These were (i) N-(3-hydroxy-7-cis-tetradecenoyl)homoserine lactone (30H,C-14:1-HSL), a molecule previously known as the Rhizobium leguminosarum small bacteriocin as a consequence of its growth inhibitory properties, (ii) N-decamoylhomoserine lactone (C-10-HSL) and (iii) N-hexanoylhomoserine lactone (C-6-HSL). A gene (hdtS) capable of directing synthesis of all three P, fluorescens AHLs in Escherichia coli was cloned and sequenced. In vitro transcription/translation of hdtS yielded a protein of approximately 33 kDa capable of directing the synthesis of 3OH,C-14:1-HSL, C-10-HSL and C-6-HSL in E. coli, HdtS does not belong to either of the known AHL synthase families (LuxI or LuxM) and is related to the lysophosphatidic acid acyltransferase family. HdtS may therefore constitute a member of a third protein family capable of AHL biosynthesis.