Pregnon Guillaume, Minton Nigel P, Soucaille Philippe
INSA, UPS, INP, Toulouse Biotechnology Institute (TBI), Université de Toulouse, 31400 Toulouse, France.
BBSRC/EPSRC Synthetic Biology Research Centre (SBRC), School of Life Sciences, University Park, The University of Nottingham, Nottingham NG7 2RD, UK.
Microorganisms. 2022 Sep 5;10(9):1790. doi: 10.3390/microorganisms10091790.
is an acetogen that can produce butyrate along with acetate as the main fermentation end-product from methanol, a promising C1 feedstock. Although physiological characterization of B2 during methylotrophy was previously performed, the strain was cultured in a semi-defined medium, limiting the scope for further metabolic insights. Here, we sequenced the complete genome of the native strain and performed adaptive laboratory evolution to sustain growth on methanol mineral medium. The evolved population significantly improved its maximal growth rate by 3.45-fold. Furthermore, three clones from the evolved population were isolated on methanol mineral medium without cysteine by the addition of sodium thiosulfate. To identify mutations related to growth improvement, the whole genomes of wild-type B2, the 10th, 25th, 50th, and 75th generations, and the three clones were sequenced. We explored the total proteomes of the native and the best evolved clone (n°2) and noticed significant differences in proteins involved in gluconeogenesis, anaplerotic reactions, and sulphate metabolism. Furthermore, a homologous recombination was found in subunit S of the type I restriction-modification system between both strains, changing the structure of the subunit, its sequence recognition and the methylome of the evolved clone. Taken together, the genomic, proteomic and methylomic data suggest a possible epigenetic mechanism of metabolic regulation.
是一种产乙酸菌,它可以将甲醇作为有前景的C1原料,产生丁酸以及乙酸作为主要发酵终产物。尽管之前对B2在甲基营养过程中的生理特性进行了研究,但该菌株是在半限定培养基中培养的,这限制了进一步进行代谢研究的范围。在此,我们对该天然菌株的完整基因组进行了测序,并进行了适应性实验室进化,以维持其在甲醇矿物培养基上的生长。进化后的菌群将其最大生长速率显著提高了3.45倍。此外,通过添加硫代硫酸钠,从进化后的菌群中分离出三个克隆,它们能在不含半胱氨酸的甲醇矿物培养基上生长。为了鉴定与生长改善相关的突变,对野生型B2、第10代、第25代、第50代和第75代以及这三个克隆的全基因组进行了测序。我们探索了天然菌株和进化程度最佳的克隆(编号2)的总蛋白质组,发现参与糖异生、回补反应和硫酸盐代谢的蛋白质存在显著差异。此外,在两种菌株之间的I型限制修饰系统的S亚基中发现了同源重组,这改变了该亚基的结构、其序列识别以及进化后克隆的甲基化组。综合来看,基因组、蛋白质组和甲基化组数据表明存在一种可能的代谢调控表观遗传机制。