Bacilysin, as the simplest peptide antibiotic made up of only L-alanine and L-anticapsin, is produced and excreted by Bacillus subtilis under the control of quorum sensing. We analyzed bacilysin-nonproducing strain OGU1 which was obtained by bacA-targeted pMutin T3 insertion into the parental strain genome resulting in a genomic organization (bacA′::lacZ::erm::bacABCDEF) to form an IPTG-inducible bac operon. Although IPTG induction provided 3- to 5-fold increment in the transcription of bac operon genes, no bacilysin activity was detectable in bioassays and inability of the OGU1 to form bacilysin was confirmed by UPLC-mass spectrometry analysis. Phenotypic analyses revealed the deficiencies in OGU1 with respect to colony pigmentation, spore coat proteins, spore resistance and germination, which could be rescued by external addition of bacilysin concentrate into its cultures. 2DE MALDI-TOF/MS and nanoLC-MS/MS were used as complementary approaches to compare cytosolic proteomes of OGU1. 2-DE identified 159 differentially expressed proteins corresponding to 121 distinct ORFs. In nanoLC-MS/MS, 76 proteins were differentially expressed in OGU1. Quantitative transcript analyses of selected genes validated the proteomic findings. Overall, the results pointed to the impact of bacilysin on expression of certain proteins of sporulation and morphogenesis; the members of mother cell compartment-specific σ^E and σ^K regulons in particular, quorum sensing and two component-global regulatory systems, peptide transport, stress response as well as CodY- and ScoC-regulated proteins.

中文翻译：

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对bac操纵子沉默的菌株的分析表明，杆菌素对枯草芽孢杆菌具有多效性

枯草杆菌蛋白酶是仅由L-丙氨酸和L-抗辣椒素组成的最简单的肽抗生素，由枯草芽孢杆菌在群体感应的控制下产生和排泄。我们分析了不产生杆菌素的菌株OGU1，该菌株是通过将bacA靶向的pMutin T3插入亲本菌株基因组而获得的，从而导致了基因组组织（bacA':: lacZ :: erm :: bacABCDEF）形成了IPTG诱导型bac操纵子。尽管IPTG诱导在bac的转录中提供了3至5倍的增量操纵子基因，在生物测定中未检测到杆菌素活性，并且通过UPLC-质谱分析法确认了OGU1不能形成杆菌素。表型分析显示，OGU1在菌落色素沉着，孢子外壳蛋白，孢子抗性和萌发方面存在缺陷，可以通过在培养物中外加浓缩杆菌素来挽救这种缺陷。2DE MALDI-TOF / MS和nanoLC-MS / MS被用作比较OGU1胞质蛋白质组的补充方法。2-DE鉴定了159个差异表达的蛋白质，对应于121个不同的ORF。在nanoLC-MS / MS中，OGU1中差异表达了76种蛋白质。所选基因的转录本定量分析证实了蛋白质组学发现。总体，结果表明，杆菌素对某些孢子形成和形态发生蛋白的表达有影响。母细胞室特异性成员^È和σ ^ķ尤其，群体感应调节子和双组份全球监管系统，肽转运，应激反应以及CodY-和ScoC调节的蛋白质。