Identification of Non-Ribosomal Peptide Synthetase Modifications Involved in Surfactin Production and Quorum-sensing Operon of Bacillus subtilis MJ01 Isolated from Oil-Contaminated Soil

T. Rahimi, A. Niazi, S. Taghavi, E. Ebrahimie, S. Ayatollahi, Tahereh Deihimi
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Abstract

The isolation of native microorganism that produced biosurfactants in order to oil pollutants bioremediation and hydrophobic oil hydrocarbons availability inter soil texture has become important issues in bioremediation technology. Surfactin is one of the biosurfactants with more application that produced by Bacillus subtilis strains could overcome these problems. Thus in this study, we investigated operon which involved in surfactin biosynthesis and its regulator comQXPA operon due to a high level of surfactin biosynthesis by B. subtilis MJ01 isolated from oil contaminated soil based on comparative genomics approaches. Surfactin operon localized and compared among six genomes of close relative strains and MJ01 indicated that missense point mutations on genes of surfactin operon were existence. These mutations affected NPRS protein AMP-binding domain that responsible to bind amino acid to correct the situation on surfactin peptide ring. It seems that lack of hemolytic and anti-microbial function of MJ01 surfactin was due to the creation of missense mutation and modifications in the surfactin biosynthesis NPRS enzyme structure. Moreover, srf genes expression regulated by comQXPA‏ quorum sensing operon. MJ01 Quorum sensing operon rearrangement showed that part of the comQ gene was extended into comX gene and these genes had overlap region. Results suggested that in MJ01 genome has been occurred specific combination of QS genes organization. Despite high similarity of three genes comQXP‏ among MJ01 with BEST7613 and other subtilis strains group, comA gene showed high identity with spizizenii strains group‏.
从石油污染土壤中分离的枯草芽孢杆菌MJ01表面活性素产生和群体感应操作子的非核糖体肽合成酶修饰的鉴定
分离产生生物表面活性剂的天然微生物以进行石油污染物的生物修复和疏水性石油烃在土壤结构中的有效性已成为生物修复技术中的重要问题。枯草芽孢杆菌生产的表面活性剂是目前应用较多的生物表面活性剂之一。因此,本研究基于比较基因组学的方法,研究了石油污染土壤中分离的枯草芽孢杆菌MJ01高水平合成表面素所导致的参与表面素生物合成的操纵子及其调控子comQXPA。表面蛋白操纵子定位与近缘菌株6个基因组与MJ01的比较表明,表面蛋白操纵子基因存在错义点突变。这些突变影响了NPRS蛋白的amp结合结构域,该结构域负责与氨基酸结合以纠正表面蛋白肽环上的情况。MJ01表面素缺乏溶血和抗微生物功能可能是由于表面素生物合成酶NPRS结构发生错义突变和修饰所致。此外,srf基因的表达受comQXPA群体感应操纵子的调控。MJ01群体感应操纵子重排表明comQ基因部分延伸到comX基因中,这些基因有重叠区。结果表明,在MJ01基因组中已经发生了特异的QS基因组合组织。MJ01的comQXP基因与BEST7613等枯草菌群具有较高的相似性,而comA基因与spizizenii菌株群具有较高的一致性。
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