biostudies-literatureMo XNational Natural Science Foundation of ChinaNational Key Research and Development Program of Chinae0256024https://www.ebi.ac.uk/biostudies/studies/S-EPMC12445348biostudies-literatureUnknown91(9)<i>Methylorubrum extorquens</i> AM1, a native formate-utilizing bacterium, has exhibited limited capacity to tolerate formate. In this study, we employed an adaptive laboratory evolution (ALE) strategy to develop an evolved strain FT3 derived from <i>M. extorquens</i> AM1, with enhanced formate tolerance. When cultivated with a mixture of carbon sources containing 90 mM formate and 30 mM methanol, the FT3 strain exhibited 5.3 times higher optical density (OD<sub>600</sub>) compared to the parental strain. FT3 strain was shown to efficiently utilize both methanol and formate in experiments using <sup>13</sup>C-labeled carbon sources. Furthermore, the mechanism underlying the enhanced formate tolerance in FT3 strain was investigated through a combination of DNA re-sequencing, transcriptome aApplied and environmental microbiologyAdaptively evolved <i>Methylorubrum extorquens</i> with enhanced formate tolerance and its application in 3-hydroxypropionic acid production.PMC124453482021YFC21035002247821722078169Liu ZBao KZhu LMa ZZhang CYang SMo XZhao YfalseAdaptively evolved <i>Methylorubrum extorquens</i> with enhanced formate tolerance and its application in 3-hydroxypropionic acid production.<i>Methylorubrum extorquens</i> AM1, a native formate-utilizing bacterium, has exhibited limited capacity to tolerate formate. In this study, we employed an adaptive laboratory evolution (ALE) strategy to develop an evolved strain FT3 derived from <i>M. extorquens</i> AM1, with enhanced formate tolerance. When cultivated with a mixture of carbon sources containing 90 mM formate and 30 mM methanol, the FT3 strain exhibited 5.3 times higher optical density (OD<sub>600</sub>) compared to the parental strain. FT3 strain was shown to efficiently utilize both methanol and formate in experiments using <sup>13</sup>C-labeled carbon sources. Furthermore, the mechanism underlying the enhanced formate tolerance in FT3 strain was investigated through a combination of DNA re-sequencing, transcriptome a2025-01-01T00:00:00Z2025 Sep2026-06-03T14:30:39.675Z2026-04-28T03:11:44.079ZS-EPMC124453484080153410.1128/aem.02560-24