Genetic and functional characterization of the gene cluster directing the biosynthesis of putisolvin I and II in Pseudomonas putida strain PCL1445
Genetic and functional characterization of the gene cluster directing the biosynthesis of putisolvin I and II in Pseudomonas putida strain PCL1445
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DOI：
10.1099/mic.0.2008/016444-0
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发表时间：
2008-07-01
期刊：
影响因子：
2.8
通讯作者：
Bloemberg, Guido V.
中科院分区：
文献类型：
--
作者：
Dubern, Jean-Frederic;Coppoolse, Eric R.;Bloemberg, Guido V.
恶臭假单胞菌PCL 1445分泌两种环脂肽，putisolvin I和putisolvin II，其具有降低表面张力的能力，并且能够抑制生物膜形成并破坏包括铜绿假单胞菌的假单胞菌物种的生物膜。分离、测序并鉴定了putisolvin合成酶基因簇（pso）及其周围区域。三个基因，称为psoA，psoB和psoC，被确定和参与putisolvin的生物合成。基因产物编码负责结合putisolvin肽部分的12个氨基酸的12个模块。序列数据表明，第11个模块的腺苷酸化结构域优先识别瓦尔而不是Leu或Ile，因此有利于putisolvin I的生产超过putisolvin II。巯基化结构域的详细分析表明，前九个模块识别D形式的氨基酸残基，而随后的两个模块识别L形式，最后一个模块识别L或D形式，无差别。位于psoA上游的psoR基因与luxR型调控基因具有高度的相似性，是pso簇表达所必需的。此外，两个基因，macA和macB，位于下游的psoC被确定，并显示参与putisolvin的生产或出口。
Pseudomonas putida PCL1445 secretes two cyclic lipopeptides, putisolvin I and putisolvin II, which possess a surface-tension-reducing ability, and are able to inhibit biofilm formation and to break down biofilms of Pseudomonas species including Pseudomonas aeruginosa. The putisolvin synthetase gene cluster (pso) and its surrounding region were isolated, sequenced and characterized. Three genes, termed psoA, psoB and psoC, were identified and shown to be involved in putisolvin biosynthesis. The gene products encode the 12 modules responsible for the binding of the 12 amino acids of the putisolvin peptide moiety. Sequence data indicate that the adenylation domain of the 11th module prioritizes the recognition of Val instead of Leu or Ile and consequently favours putisolvin I production over putisolvin II. Detailed analysis of the thiolation domains suggests that the first nine modules recognize the D form of the amino acid residues while the two following modules recognize the L form and the last module the L or D form, indifferently. The psoR gene, which is located upstream of psoA, shows high similarity to luxR-type regulatory genes and is required for the expression of the pso cluster. In addition, two genes, macA and macB, located downstream of psoC were identified and shown to be involved in putisolvin production or export.