Ученые создали штамм бактерии, способный эффективно перерабатывать формат в биотопливо

Abstract Formate is a promising, sustainable feedstock for microbial conversion into value-added products via the Wood–Ljungdahl pathway. Based on the identification of three distinct formate dehydrogenases in Clostridium sp. AWRP, this study investigated the potential of this acetogen for formate-based growth. Although the wild-type strain exhibited an extended lag phase due to formate toxicity, adaptive laboratory evolution yielded the adapted strain F30T, which demonstrated a significantly enhanced growth rate and tolerance to formate concentrations up to 300 mM. Whole-genome and transcriptomic analyses of the F30T strain revealed a sophisticated, multi-layered strategy for mitigating organic acid stress and optimizing energy conservation. Adaptive laboratory evolution resulted in six key non-synonymous mutations, including those in ackA and adhE1, and triggered the differential expression of 948 genes out of 4199 genes under the formate-supplemented condition. Specifically, F30T achieved intracellular pH buffering and supplemental ATP generation by upregulating the arginine deiminase pathway, histidine biosynthesis, and ethanolamine utilization. To maintain growth near the thermodynamic limit, the strain implemented a stringent energy-saving program by downregulating the F₀F₁-type ATP synthase and the methyl-branch of the Wood–Ljungdahl pathway, while simultaneously enhancing stress resilience through molecular chaperone upregulation. In a pH-stat fed-batch system using formic acid, the adapted F30T strain showed a higher biomass yield while achieving a similar acetate yield from formate as the wild-type strain. This study demonstrated that adaptive laboratory evolution is a highly effective strategy for enhancing the formate utilization and stress tolerance of Clostridium sp. AWRP. Through integrated genomic and transcriptomic analyses, the molecular basis of the F30T strain’s improved performance was elucidated, identifying key mutations and coordinated metabolic and regulatory mechanisms that maintain cellular homeostasis and maximize energy efficiency. These findings provide valuable molecular insights to aid the targeted design and optimization of formate-utilizing acetogens for use in microbial cell factories aimed at sustainable C1-based bioproduction. Funding This work was supported by the Korea Institute of Ocean Science and Technology (KIOST) in-house programs (EA0411 and EA0413). Author information Authors and Affiliations Marine Biotechnology & Bioresource Research Department, Korea Institute of Ocean Science and Technology, Busan, 49111, Republic of Korea Soo Jae Kwon, Seongeun Yang, Sung-Mok Lee, Joungmin Lee, Sung Gyun Kang & Hyun Sook Lee Department of Marine Biotechnology, University of Science and Technology, Daejeon, 34129, Republic of Korea Joungmin Lee, Sung Gyun Kang & Hyun Sook Lee Authors Soo Jae Kwon Search author on: PubMed Google Scholar Seongeun Yang Search author on: PubMed Google Scholar Sung-Mok Lee Search author on: PubMed Google Scholar Joungmin Lee Search author on: PubMed Google Scholar Sung Gyun Kang Search author on: PubMed Google Scholar Hyun Sook Lee Search author on: PubMed Google Scholar Corresponding author Correspondence to Hyun Sook Lee. Ethics declarations Ethics approval and consent to participate Not applicable. Consent for publication Not applicable. Competing interests The authors declare that there are no competing interests. Additional information Publisher's Note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Supplementary Information Rights and permissions Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. Reprints and permissions Profiles Hyun Sook Lee View author profile