Data_Sheet_1_Genomic potential and physiological characteristics of C1 metabolism in novel acetogenic bacteria.pdf

Acetogenic bacteria can utilize C1 compounds, such as carbon monoxide (CO), formate, and methanol, via the Wood-Ljungdahl pathway (WLP) to produce biofuels and biochemicals. Two novel acetogenic bacteria of the family Eubacteriaceae ES2 and ES3 were isolated from Eulsukdo, a delta island in South Korea. We conducted whole genome sequencing of the ES strains and comparative genome analysis on the core clusters of WLP with Acetobacterium woodii DSM1030T and Eubacterium limosum ATCC8486T. The methyl-branch cluster included a formate transporter and duplicates or triplicates copies of the fhs gene, which encodes formyl-tetrahydrofolate synthetase. The formate dehydrogenase cluster did not include the hydrogenase gene, which might be replaced by a functional complex with a separate electron bifurcating hydrogenase (HytABCDE). Additionally, duplicated copies of the acsB gene, encoding acetyl-CoA synthase, are located within or close to the carbonyl-branch cluster. The serum bottle culture showed that ES strains can utilize a diverse range of C1 compounds, including CO, formate, and methanol, as well as CO2. Notably, ES2 exhibited remarkable resistance to high concentrations of C1 substrates, such as 100% CO (200 kPa), 700 mM formate, and 500 mM methanol. Moreover, ES2 demonstrated remarkable growth rates under 50% CO (0.45 h−1) and 200 mM formate (0.34 h−1). These growth rates are comparable to or surpassing those previously reported in other acetogenic bacteria. Our study introduces novel acetogenic ES strains and describes their genetic and physiological characteristics, which can be utilized in C1-based biomanufacturing.

乙酸发酵细菌能够通过伍德-吕恩达尔途径（WLP）利用C1化合物，如一氧化碳（CO）、甲酸盐和甲醇，以生产生物燃料和生物化学品。从韩国南部的Eulsukdo三角洲岛屿中分离出了两种新型的乙酸发酵细菌，分别属于欧巴氏菌科（Eubacteriaceae）的ES2和ES3。我们对ES菌株进行了全基因组测序，并对比分析了伍德（Acetobacterium woodii DSM1030T）和欧巴氏菌（Eubacterium limosum ATCC8486T）的WLP核心群。甲基分支群包含甲酸盐转运蛋白以及fhs基因的重复或三重拷贝，该基因编码甲酰四氢叶酸合酶。甲酸盐脱氢酶群不包含氢酶基因，可能被一个具有独立电子分叉氢酶（HytABCDE）的功能性复合物所替代。此外，编码乙酰辅酶A合酶的acsB基因的重复拷贝位于羰基分支群内或附近。血清瓶培养结果显示，ES菌株能够利用包括CO、甲酸盐和甲醇在内的多种C1化合物，以及二氧化碳。值得注意的是，ES2对高浓度的C1底物表现出显著的抗性，例如100% CO（200 kPa）、700 mM甲酸盐和500 mM甲醇。此外，ES2在50% CO（0.45 h−1）和200 mM甲酸盐（0.34 h−1）下的生长速度与之前报道的其他乙酸发酵细菌相当或更高。本研究引入了新的乙酸发酵ES菌株，并描述了它们的遗传和生理特性，这些特性可用于基于C1的生物制造。