{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,29]],"date-time":"2025-12-29T05:54:53Z","timestamp":1766987693782,"version":"3.48.0"},"reference-count":52,"publisher":"Wiley","issue":"12","license":[{"start":{"date-parts":[[2025,12,25]],"date-time":"2025-12-25T00:00:00Z","timestamp":1766620800000},"content-version":"vor","delay-in-days":24,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000288","name":"Royal Society","doi-asserted-by":"publisher","award":["NAF\\R2\\180721"],"award-info":[{"award-number":["NAF\\R2\\180721"]}],"id":[{"id":"10.13039\/501100000288","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100014013","name":"UK Research and Innovation","doi-asserted-by":"publisher","award":["EP\/X032078\/1"],"award-info":[{"award-number":["EP\/X032078\/1"]}],"id":[{"id":"10.13039\/100014013","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["enviromicro-journals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Microbial Biotechnology"],"published-print":{"date-parts":[[2025,12]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Isobutanol, a promising biofuel with higher energy content than ethanol, presents a sustainable alternative through biosynthesis. However, enhancing yield remains challenging due to the inefficiencies in microbial synthesis. This study introduces a transcription factor\u2010based biosensor using the AlkS\u2010PalkB system in\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>Escherichia coli<\/jats:italic>\n                    <\/jats:styled-content>\n                    , which correlates green fluorescence with isobutanol concentration. Employing directed evolution, we modified AlkS to detect isobutanol, significantly improving biosensor specificity. Initial modifications increased the dynamic response from non\u2010detectable to a 2.60\u2010fold change. Subsequent optimisations through site\u2010directed mutagenesis and promoter engineering further enhanced this response to a 5.56\u2010fold change, equivalent to a 114% increase. Although engineered for isobutanol detection with high sensitivity, the engineered biosensor retains responsiveness to several short\u2010chain alcohols. This biosensor provides a foundation for high\u2010throughput screening of isobutanol and other short\u2010chain alcohol\u2010producing strains, though additional improvements in selectivity and operating range may be required for efficient implementation.\n                  <\/jats:p>","DOI":"10.1111\/1751-7915.70288","type":"journal-article","created":{"date-parts":[[2025,12,26]],"date-time":"2025-12-26T01:58:26Z","timestamp":1766714306000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Engineering an\n                    <scp>AlkS<\/scp>\n                    \u2010\n                    <scp>PalkB<\/scp>\n                    Transcription Factor\u2010Based Biosensor With Improved Sensitivity to Isobutanol and Other Short\u2010Chain Alcohols"],"prefix":"10.1111","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0009-0004-1884-7223","authenticated-orcid":false,"given":"Jiaming","family":"Chen","sequence":"first","affiliation":[{"name":"Department of Biochemical Engineering University College London  London UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-4758-8772","authenticated-orcid":false,"given":"Xizi","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Biochemical Engineering University College London  London UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-1110-8257","authenticated-orcid":false,"given":"Can","family":"Zhu","sequence":"additional","affiliation":[{"name":"Department of Biochemical Engineering University College London  London 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