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To facilitate the engineering of formaldehyde\u2010producing enzymes, the development of sensitive, user\u2010friendly, and cost\u2010effective detection methods is required. In this study, we engineered\n                    <jats:italic>Escherichia coli<\/jats:italic>\n                    to serve as a cellular biosensor capable of detecting a broad range of formaldehyde concentrations. Using both natural and promiscuous formaldehyde assimilation enzymes, we designed three distinct\n                    <jats:italic>E. coli<\/jats:italic>\n                    growth biosensor strains that depend on formaldehyde for cell growth. These strains were engineered to be auxotrophic for one or several essential metabolites that could be produced through formaldehyde assimilation. The respective assimilating enzyme was expressed from the genome to compensate the auxotrophy in the presence of formaldehyde. We first predicted the formaldehyde dependency of the biosensors by flux balance analysis and then analysed it experimentally. Subsequent to strain engineering, we enhanced the formaldehyde sensitivity of two biosensors either through adaptive laboratory evolution or modifications at metabolic branch points. The final set of biosensors demonstrated the ability to detect formaldehyde concentrations ranging approximately from 30\u2009\u03bcM to 13\u2009mM. We demonstrated the application of the biosensors by assaying the in\u00a0vivo activity of different methanol dehydrogenases in the most sensitive strain. The fully genomic nature of the biosensors allows them to be deployed as \u201cplug\u2010and\u2010play\u201d devices for high\u2010throughput screenings of extensive enzyme libraries. The formaldehyde growth biosensors developed in this study hold significant promise for advancing the field of enzyme engineering, thereby supporting the establishment of a sustainable one\u2010carbon bioeconomy.\n                  <\/jats:p>","DOI":"10.1111\/1751-7915.14527","type":"journal-article","created":{"date-parts":[[2024,7,19]],"date-time":"2024-07-19T11:10:34Z","timestamp":1721387434000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":21,"title":["Design, construction and optimization of formaldehyde growth biosensors with broad application in biotechnology"],"prefix":"10.1111","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1548-7643","authenticated-orcid":false,"given":"Karin","family":"Schann","sequence":"first","affiliation":[{"name":"Max Planck Institute of Molecular Plant Physiology  Potsdam\u2010Golm 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