BBSRC/EPSRC Synthetic Biology Research Centre (SBRC), School of Life Sciences, University Park, The University of Nottingham, Nottingham, UK.
Present address: Independent Commodity Intelligence Service, Bishopsgate, London, UK.
Microbiology (Reading). 2020 Jun;166(6):579-592. doi: 10.1099/mic.0.000916.
The strictly anaerobic bacterium is well known for its ability to convert sugars into organic acids and solvents, most notably the potential biofuel butanol. However, the regulation of its fermentation metabolism, in particular the shift from acid to solvent production, remains poorly understood. The aim of this study was to investigate whether cell-cell communication plays a role in controlling the timing of this shift or the extent of solvent formation. Analysis of the available genome sequences revealed the presence of eight putative RRNPP-type quorum-sensing systems, here designated to , each consisting of an RRNPP-type regulator gene followed by a small open reading frame encoding a putative signalling peptide precursor. The identified regulator and signal peptide precursor genes were designated to and to , respectively. Triplicate regulator mutants were generated in strain ATCC 824 for each of the eight systems and screened for phenotypic changes. The mutants showed increased solvent formation during early solventogenesis and hence the QssB system was selected for further characterization. Overexpression of severely reduced solvent and endospore formation and this effect could be overcome by adding short synthetic peptides to the culture medium representing a specific region of the QspB signalling peptide precursor. In addition, overexpression of increased the production of acetone and butanol and the initial (48 h) titre of heat-resistant endospores. Together, these findings establish a role for QssB quorum sensing in the regulation of early solventogenesis and sporulation in .
严格厌氧菌以将糖转化为有机酸和溶剂而闻名,特别是有潜力的生物燃料丁醇。然而,其发酵代谢的调控,特别是从酸到溶剂生产的转变,仍然知之甚少。本研究旨在探讨细胞间通讯是否在控制这种转变的时机或溶剂形成的程度方面发挥作用。对现有基因组序列的分析揭示了存在八个推定的 RRNPP 型群体感应系统,这里指定为 至 ,每个系统都由 RRNPP 型调节剂基因后面跟着一个编码假定信号肽前体的小开放阅读框组成。鉴定的调节剂和信号肽前体基因分别指定为 至 。针对每个系统在 ATCC 824 菌株中生成了三倍的调节剂突变体,并对其表型变化进行了筛选。 突变体在早期溶剂生成过程中表现出增加的溶剂形成,因此选择 QssB 系统进行进一步表征。 过表达严重减少了溶剂和内生孢子的形成,并且可以通过向培养基中添加短的合成肽来克服该效应,该合成肽代表 QspB 信号肽前体的特定区域。此外, 过表达增加了丙酮和丁醇的产量以及耐热内生孢子的初始(48 h)产量。总之,这些发现确立了 QssB 群体感应在 中早期溶剂生成和孢子形成调控中的作用。