Huang Jean J, Han Jong-In, Zhang Lian-Hui, Leadbetter Jared R
Department of Biology, California Institute of Technology, Pasadena, California 91125, USA.
Appl Environ Microbiol. 2003 Oct;69(10):5941-9. doi: 10.1128/AEM.69.10.5941-5949.2003.
Acyl-homoserine lactones (AHLs) are employed by several Proteobacteria as quorum-sensing signals. Past studies have established that these compounds are subject to biochemical decay and can be used as growth nutrients. Here we describe the isolation of a soil bacterium, Pseudomonas strain PAI-A, that degrades 3-oxododecanoyl-homoserine lactone (3OC12HSL) and other long-acyl, but not short-acyl, AHLs as sole energy sources for growth. The small-subunit rRNA gene from strain PAI-A was 98.4% identical to that of Pseudomonas aeruginosa, but the soil isolate did not produce obvious pigments or AHLs or grow under denitrifying conditions or at 42 degrees C. The quorum-sensing bacterium P. aeruginosa, which produces both 3OC12HSL and C4HSL, was examined for the ability to utilize AHLs for growth. It did so with a specificity similar to that of strain PAI-A, i.e., degrading long-acyl but not short-acyl AHLs. In contrast to the growth observed with strain PAI-A, P. aeruginosa strain PAO1 growth on AHLs commenced only after extremely long lag phases. Liquid-chromatography-atmospheric pressure chemical ionization-mass spectrometry analyses indicate that strain PAO1 degrades long-acyl AHLs via an AHL acylase and a homoserine-generating HSL lactonase. A P. aeruginosa gene, pvdQ (PA2385), has previously been identified as being a homologue of the AHL acylase described as occurring in a Ralstonia species. Escherichia coli expressing pvdQ catalyzed the rapid inactivation of long-acyl AHLs and the release of HSL. P. aeruginosa engineered to constitutively express pvdQ did not accumulate its 3OC12HSL quorum signal when grown in rich media. However, pvdQ knockout mutants of P. aeruginosa were still able to grow by utilizing 3OC12HSL. To our knowledge, this is the first report of the degradation of AHLs by pseudomonads or other gamma-Proteobacteria, of AHL acylase activity in a quorum-sensing bacterium, of HSL lactonase activity in any bacterium, and of AHL degradation with specificity only towards AHLs with long side chains.
酰基高丝氨酸内酯(AHLs)被多种变形菌用作群体感应信号。过去的研究表明,这些化合物会发生生化降解,并且可以用作生长营养物质。在此,我们描述了从土壤中分离出的一种细菌——假单胞菌PAI - A菌株,它能够降解3 - 氧代十二烷酰高丝氨酸内酯(3OC12HSL)和其他长链酰基而非短链酰基的AHLs,并将其作为唯一的生长能源。PAI - A菌株的小亚基rRNA基因与铜绿假单胞菌的该基因有98.4%的同源性,但这种土壤分离株不产生明显色素或AHLs,在反硝化条件下或42℃时也不能生长。对能产生3OC12HSL和C4HSL的群体感应细菌铜绿假单胞菌利用AHLs进行生长的能力进行了检测。其表现出与PAI - A菌株相似的特异性,即降解长链酰基而非短链酰基的AHLs。与PAI - A菌株的生长情况不同,铜绿假单胞菌PAO1菌株在AHLs上生长仅在极长的延迟期后才开始。液相色谱 - 大气压化学电离 - 质谱分析表明,PAO1菌株通过一种AHL酰基转移酶和一种产生高丝氨酸的HSL内酯酶降解长链酰基AHLs。铜绿假单胞菌的一个基因pvdQ(PA2385)先前已被鉴定为与在一种罗尔斯通氏菌中发现的AHL酰基转移酶同源。表达pvdQ的大肠杆菌能催化长链酰基AHLs的快速失活并释放出HSL。经基因工程改造后能组成型表达pvdQ的铜绿假单胞菌在丰富培养基中生长时不会积累其3OC12HSL群体感应信号。然而,铜绿假单胞菌的pvdQ基因敲除突变体仍能够利用3OC12HSL生长。据我们所知,这是关于假单胞菌或其他γ - 变形菌降解AHLs、群体感应细菌中AHL酰基转移酶活性、任何细菌中HSL内酯酶活性以及仅对具有长侧链的AHLs具有特异性的AHL降解的首次报道。