Abstract：
Synthetic biology is contributing to the advancement of the global net-negative carbon economy, with emphasis on formate as a member of the one-carbon substrate garnering substantial attention. In this study, we employed base editing tools to facilitate adaptive evolution, achieving a formate tolerance of Yarrowia lipolytica to 1 M within 2 months. This effort resulted in two mutant strains, designated as M25-70 and M25-14, both exhibiting significantly enhanced formate utilization capabilities. Transcriptomic analysis revealed the upregulation of nine endogenous genes encoding formate dehydrogenases when cultivated utilizing formate as the sole carbon source. Furthermore, we uncovered the pivotal role of the glyoxylate and threonine-based serine pathway in enhancing glycine supply to promote formate assimilation. The full potential of Y. lipolytica to tolerate and utilize formate establishing the foundation for pyruvate carboxylase-based carbon sequestration pathways. Importantly, this study highlights the existence of a natural formate metabolic pathway in Y. lipolytica.
摘要：
合成生物学正在为全球净负碳经济的发展做出贡献，强调甲酸盐作为一碳底物的一员引起了广泛的关注。在这项研究中,我们采用了基础编辑工具来促进自适应进化，在2个月内实现Yarrowialipolytica对1M的甲酸耐受性。这项努力导致了两个突变菌株，命名为M25-70和M25-14,两者均表现出显著提高的甲酸盐利用能力。转录组分析显示，当利用甲酸盐作为唯一碳源培养时，编码甲酸脱氢酶的9个内源基因上调。此外，我们发现了基于乙醛酸和苏氨酸的丝氨酸途径在增强甘氨酸供应以促进甲酸同化方面的关键作用。Y.Lipolytica耐受和利用甲酸的全部潜力为基于丙酮酸羧化酶的碳封存途径奠定了基础。重要的是,这项研究强调了Y.Lipolytica中天然甲酸代谢途径的存在。
