{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T11:17:41Z","timestamp":1788434261966,"version":"build-2803163510"},"reference-count":39,"publisher":"American Chemical Society (ACS)","issue":"17","license":[{"start":{"date-parts":[[2026,8,15]],"date-time":"2026-08-15T00:00:00Z","timestamp":1786752000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"publisher","award":["2R15GM120704-03"],"award-info":[{"award-number":["2R15GM120704-03"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"publisher","award":["2R15GM12704-01"],"award-info":[{"award-number":["2R15GM12704-01"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100019684","name":"Haverford College","doi-asserted-by":"publisher","award":["NA"],"award-info":[{"award-number":["NA"]}],"id":[{"id":"10.13039\/100019684","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,9,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Strategic engineering of natural product biosynthetic pathways through the incorporation of alternative, tunable carbon-based building blocks represents a promising approach for accessing medicinally relevant molecules. However, efforts toward this goal have been hindered by the substrate specificity of component enzymes. In type I and type II fatty acid synthases (FASs) and polyketide synthases (PKSs), the acyltransferase (AT) selects a specific malonyl-based coenzyme A (CoA) building block and transfers it onto the acyl carrier protein (ACP) for subsequent processing. Inspired by the observation that some ACPs can bypass the AT and \u201cself-acylate\u201d, we herein explored the tolerance of FAS and PKS ACPs to load both a variety of CoA substrates and ethane thioester (ET) analogs serving as truncated CoA building blocks. We observe that the Escherichia coli (E. coli) AT, FabD, can load and transfer methylmalonyl-CoA (mm-CoA) and malonyl-CoA (m-CoA) onto three ACPs: the type II Streptomyces coelicolor actinorhodin PKS ACP (ActACP), the E. coli type II FAS ACP (AcpP), and the type I Saccharopolyspora erythraea 6-deoxyerythronolide B PKS ACP6 (DEBS ACP6). Synthesized ET analogs of mm-CoA and m-CoA were loaded onto all three ACPs through FabD-assisted acylation. Additionally, both in the presence and absence of FabD, ACPs could be acylated with ET analogs of fluoromalonyl-, succinyl-, and glutaryl- building blocks. Overall, this work pushes the limits of ACP substrate loading, revealing new complexity in carbon-based building block selection and establishing foundations for novel routes toward diverse functional group incorporation in FAS\/PKS biosynthetic pathways.<\/jats:p>","DOI":"10.1021\/acs.biochem.6c00474","type":"journal-article","created":{"date-parts":[[2026,8,15]],"date-time":"2026-08-15T16:01:21Z","timestamp":1786809681000},"page":"2656-2660","source":"Crossref","is-referenced-by-count":0,"title":["Creating New\nAccess Points for Strategic Synthase\nEngineering: Ethane Thioester Analogs as Truncated Coenzyme A Building\nBlocks Unlock Expanded Acyl Carrier Protein Loading Capabilities"],"prefix":"10.1021","volume":"65","author":[{"given":"Jana C.","family":"Bowler","sequence":"first","affiliation":[{"name":"Haverford College , , , ,","place":["Haverford, Pennsylvania, United States, 19041"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna-Lee C.","family":"Thompson","sequence":"additional","affiliation":[{"name":"Haverford College , , , ,","place":["Haverford, Pennsylvania, United States, 19041"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Colby M.","family":"Freeman","sequence":"additional","affiliation":[{"name":"Haverford College , , , ,","place":["Haverford, Pennsylvania, United States, 19041"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yarra L.","family":"Ellett","sequence":"additional","affiliation":[{"name":"Haverford College , , , ,","place":["Haverford, Pennsylvania, United States, 19041"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8890-8985","authenticated-orcid":true,"given":"Leah M.","family":"Seebald","sequence":"additional","affiliation":[{"name":"Haverford College , , , ,","place":["Haverford, Pennsylvania, United States, 19041"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4342-2815","authenticated-orcid":true,"given":"Louise K.","family":"Charkoudian","sequence":"additional","affiliation":[{"name":"Haverford College , , , ,","place":["Haverford, Pennsylvania, United States, 19041"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,15]]},"reference":[{"key":"2026090306413567900_cit1","doi-asserted-by":"publisher","first-page":"129377","DOI":"10.1016\/j.bmcl.2023.129377","volume":"91","author":"West","year":"2023","journal-title":"Bioorg. Med. Chem. Lett."},{"issue":"2","key":"2026090306413567900_cit2","doi-asserted-by":"publisher","first-page":"147","DOI":"10.1016\/S0969-2126(03)00004-2","volume":"11","author":"Keatinge-Clay","year":"2003","journal-title":"Structure"},{"issue":"13","key":"2026090306413567900_cit3","doi-asserted-by":"publisher","first-page":"1991","DOI":"10.1002\/cbic.201402206","volume":"15","author":"Bravo-Rodriguez","year":"2014","journal-title":"ChemBioChem."},{"issue":"85","key":"2026090306413567900_cit4","doi-asserted-by":"publisher","first-page":"20130297","DOI":"10.1098\/rsif.2013.0297","volume":"10","author":"Dunn","year":"2013","journal-title":"J. R. Soc. Interface"},{"issue":"5","key":"2026090306413567900_cit5","doi-asserted-by":"publisher","first-page":"1961","DOI":"10.1021\/jacs.8b10521","volume":"141","author":"Kalkreuter","year":"2019","journal-title":"J. Am. Chem. 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