A proteomic, reverse-genetic and sequencing approach to reveal genes and enzymes for LAS-surfactant degradation in bacterial communities

一种蛋白质组学、反向遗传学和测序方法，用于揭示细菌群落中 LAS 表面活性剂降解的基因和酶

• 批准号: 187067634
• 负责人: Professor Dr. David Schleheck, Ph.D.
• 金额: --
• 依托单位: Arbeitsgruppe Limnische Mikrobiologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2010
• 资助国家: 德国
• 起止时间: 2009-12-31 至 2014-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/187067634?language=en
• 关键词: proteomic; reverse; genetic; sequencing; approach

The complete biodegradation of industrial chemicals can impose great challenges to environmental microbial communities. One prominent example is the mineralization of xenobiotic laundry surfactant LAS (linear alkylbenzene sulfonate; sec. 4-sulfophenyl alkane). Commercial LAS is a complex mixture of isomers, and its mineralization is accomplished by heterotrophic bacterial communities (LAS-community) in two tiers: bacteria in the first tier utilize the LAS-alkyl chains for growth and convert LAS to short-chain SPCs (sulfophenyl carboxylates); bacteria in the second tier mineralize these SPCs. To date, the complete LAS and SPC pathways and the genes and enzymes involved remain unknown, as well as the phylogenetic composition of the LAS-community, e.g. in sewage treatment plants. Now, a three-member, genome-sequenced ‘model LAS-community’ is available, Parvibaculum lavamentivorans DS-1 (first tier) with Comamonas testosteroni KF-1 and Delftia acidovorans SPH-1 (second tier). This allows for renewed attack on understanding LAS- and SPC-degradation, through a proteomic and reverse-genetic approach. The exploration is expected to deliver insight into the evolution of these important, novel degradation pathways, and, e.g., discover novel P450 monooxygenases and Baeyer-Villiger-type monooxygenases. However, the ‘model LAS-community’ mineralizes only a subset of the many SPCs generated from commercial LAS. Hence, the environmental community needed to catalyze the complete degradation of commercial LAS is expected to be much more complex, but this has never been demonstrated. Therefore, also the environmental LAS-community will be explored, firstly, for its phylogenetic complexity, and secondly, for the abundance and diversity of key genes for LAS- and SPC-degradation, through a massively-parallel sequencing approach.

工业化学品的完全生物降解会给环境微生物群落带来巨大挑战。一个突出的例子是外源洗衣表面活性剂 LAS（直链烷基苯磺酸盐；仲 4-磺基苯基烷烃）的矿化。商业LAS是异构体的复杂混合物，其矿化是通过两层异养细菌群落（LAS群落）完成的：第一层细菌利用LAS烷基链生长并将LAS转化为短链SPC（磺基苯基羧酸盐）；第二层的细菌使这些 SPC 矿化。迄今为止，完整的 LAS 和 SPC 途径以及所涉及的基因和酶以及 LAS 群落的系统发育组成仍然未知。在污水处理厂。现在，一个由三名成员组成的基因组测序“模型 LAS 群落”现已推出，Parvibaculum lavamentivorans DS-1（第一层）、睾酮丛毛单胞菌 KF-1 和 Delftia Acidovorans SPH-1（第二层）。这使得通过蛋白质组学和反向遗传学方法重新了解 LAS 和 SPC 降解成为可能。这项探索预计将深入了解这些重要的新型降解途径的演变，例如发现新型 P450 单加氧酶和 Baeyer-Villiger 型单加氧酶。然而，“模型 LAS 社区”仅矿化了商业 LAS 生成的众多 SPC 的一个子集。因此，催化商业 LAS 完全降解所需的环境界预计要复杂得多，但这从未得到证实。因此，我们还将通过大规模并行测序方法，首先探索环境 LAS 群落的系统发育复杂性，其次探索 LAS 和 SPC 降解关键基因的丰度和多样性。