{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T13:54:14Z","timestamp":1791467654676,"version":"4.3.4"},"reference-count":22,"publisher":"American Society for Microbiology","license":[{"start":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T00:00:00Z","timestamp":1791417600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T00:00:00Z","timestamp":1791417600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"DOI":"10.13039\/501100003052","name":"Ministry of Trade, Industry and Energy","doi-asserted-by":"publisher","award":["RS-2024-00488503"],"award-info":[{"award-number":["RS-2024-00488503"]}],"id":[{"id":"10.13039\/501100003052","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003725","name":"National Research Foundation of Korea","doi-asserted-by":"publisher","award":["RS-2020-NR049543"],"award-info":[{"award-number":["RS-2020-NR049543"]}],"id":[{"id":"10.13039\/501100003725","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Appl Environ Microbiol"],"abstract":"ABSTRACT<\/jats:title>\n \n \n \n 1,3-Butanediol (1,3-BDO) exists as (R)- and (S)-enantiomers, yet how aerobic bacteria catabolize the (S)-form has been unknown. Here, we define (S)-1,3-BDO degradation pathway in\n Pseudomonas putida<\/jats:italic>\n KT2440 and uncover an enantioselective enzyme that both controls the pathway and enables a chiral resolution. Transcriptomics, quantitative RT-PCR, and systematic gene deletions show that the\n ped<\/jats:italic>\n cluster oxidizes (S)-1,3-BDO to (S)-3-hydroxybutyrate [(S)-3-HB] using the same upstream machinery as (R)-1,3-BDO and 1,4-BDO, but that the downstream route then diverges sharply. Rather than the free-acid dehydrogenase route used for (R)-3-HB, (S)-3-HB is catabolized through an acyl-CoA synthetase, Acs (PP_3458): a \u0394\n acs<\/jats:italic>\n mutant cannot grow on (S)-3-HB, and complementation restores growth, establishing that PP_3458 activates (S)-3-HB to (S)-3-hydroxybutyryl-CoA for \u03b2-oxidation. This ATP-dependent CoA-activation route is mechanistically distinct from the ATP-independent (S)-3-hydroxybutyrate dehydrogenase (3SHBDH) pathway of anaerobic\n Clostridia<\/jats:italic>\n , identifying a different enzymatic solution to (S)-3-HB assimilation in aerobes. We further show that PP_3458 strongly prefers (S)- over (R)-3-HB and is inhibited by (R)-3-HB, a crosstalk that explains why a (R)-3-HB-accumulating mutant arrests on racemic 3-HB. Two applications follow directly: whole-cell kinetic resolution that converts racemic 3-HB to (S)-3-HB at high enantiomeric excess (ee > 0.99), and, in a strain unable to degrade (S)-3-HB, near-quantitative recovery\u2014providing a chiral pharmaceutical building block; and\n ped<\/jats:italic>\n -based biosensors that report 1,3-BDO across both enantiomers and function in\n P. putida<\/jats:italic>\n and\n Escherichia coli<\/jats:italic>\n . Some mechanistic details\u2014the inhibition mode and the individual downstream \u03b2-oxidation steps\u2014remain to be fully resolved.\n <\/jats:p>\n \n IMPORTANCE<\/jats:title>\n \n 1,3-Butanediol is a commercially important C\n 4<\/jats:sub>\n diol, but the microbial fate of its (S)-enantiomer in aerobic bacteria has been a gap in our understanding of diol metabolism. We show that\n Pseudomonas putida<\/jats:italic>\n KT2440 degrades (S)-1,3-BDO through a route that, after a shared\n ped<\/jats:italic>\n -dependent oxidation to (S)-3-hydroxybutyrate, depends on an acyl-CoA synthetase (PP_3458) to activate the (S)-acid as its CoA thioester. This ATP-dependent CoA-activation step is a distinct enzymatic strategy from the ATP-independent dehydrogenase route that anaerobic bacteria use for the same substrate, showing that aerobes and anaerobes solve (S)-3-HB assimilation differently. The work also turns this enzymology into utility: because PP_3458 is enantioselective and is inhibited by (R)-3-hydroxybutyrate, the strain can be used for whole-cell kinetic resolution that yields enantiopure (S)-3-hydroxybutyrate\u2014a chiral pharmaceutical building block\u2014from racemic 3-hydroxybutyrate, and the\n ped<\/jats:italic>\n -based biosensors reported here detect 1,3-BDO in both\n P. putida<\/jats:italic>\n and\n Escherichia coli<\/jats:italic>\n , providing a portable tool for strain screening.\n <\/jats:p>\n <\/jats:sec>\n <\/jats:sec>","DOI":"10.1128\/aem.01295-26","type":"journal-article","created":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T13:00:41Z","timestamp":1791464441000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["Degradation of (S)-1,3-butanediol in\n Pseudomonas putida<\/i>\n KT2440 reveals a CoA-activation route and enantiomeric crosstalk between (R)- and (S)-3-hydroxybutyrate catabolism"],"prefix":"10.1128","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0496-0917","authenticated-orcid":true,"given":"Nandakumar","family":"Arumugam","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/017cjz748","id-type":"ROR","asserted-by":"publisher"}],"name":"School of Energy and Chemical Engineering, Ulsan National Institute of Science and Technology","place":["Ulsan, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-3751-6877","authenticated-orcid":true,"given":"Tayyab","family":"Islam","sequence":"additional","affiliation":[{"name":"R&D Center, ACTIVON Co., Ltd.","place":["Cheongju, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2059-2387","authenticated-orcid":true,"given":"Joon Young","family":"Park","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/017cjz748","id-type":"ROR","asserted-by":"publisher"}],"name":"Center for Bio-based Chemistry, Korea Research Institute of Chemical Technology","place":["Ulsan, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Minchang","family":"Jang","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/05apxxy63","id-type":"ROR","asserted-by":"publisher"}],"name":"Graduate School of Engineering Biology, Korea Advanced Institute of Science and Technology","place":["Daejeon, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5934-2633","authenticated-orcid":true,"given":"Mugesh","family":"Sankaranarayanan","sequence":"additional","affiliation":[{"name":"Center for Metabolic Engineering & Synthetic Biology, Department of Biotechnology, Vel Tech Rangarajan Dr. Sagunthala R&D Institute of Science and Technology","place":["Chennai, India"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6489-032X","authenticated-orcid":true,"given":"Donghyuk","family":"Kim","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/05apxxy63","id-type":"ROR","asserted-by":"publisher"}],"name":"Graduate School of Engineering Biology, Korea Advanced Institute of Science and Technology","place":["Daejeon, Republic of Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1711-4760","authenticated-orcid":true,"given":"Sung Kuk","family":"Lee","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/017cjz748","id-type":"ROR","asserted-by":"publisher"}],"name":"School of Energy and Chemical Engineering, Ulsan National Institute of Science and Technology","place":["Ulsan, Republic of 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