{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,13]],"date-time":"2026-11-13T04:36:00Z","timestamp":1794544560425,"version":"4.9.1"},"reference-count":39,"publisher":"American Chemical Society (ACS)","issue":"10","license":[{"start":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T00:00:00Z","timestamp":1790294400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T00:00:00Z","timestamp":1790294400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T00:00:00Z","timestamp":1790294400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/501100003399","name":"Science and Technology Commission of Shanghai Municipality","doi-asserted-by":"publisher","award":["24ZR1451000"],"award-info":[{"award-number":["24ZR1451000"]}],"id":[{"id":"10.13039\/501100003399","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100014900","name":"School of Physical Science and Technology, ShanghaiTech University","doi-asserted-by":"publisher","award":["SPST-YSFZ-2024-01"],"award-info":[{"award-number":["SPST-YSFZ-2024-01"]}],"id":[{"id":"10.13039\/501100014900","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32571664"],"award-info":[{"award-number":["32571664"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,10,16]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Raspberry ketone (RK) is a high-value flavor and fragrance compound widely used in the food, beverage, and cosmetic industries. However, existing biosynthetic routes largely rely on native metabolic pathways that involve multiple enzymatic steps and exhibit limited production efficiency. Here, we designed and constructed a retrosynthesis-guided artificial biosynthetic pathway for cell-free RK production. The pathway combines an artificial aldolase (ALD) with the native benzalacetone reductase (BAR) to convert 4-hydroxybenzaldehyde and acetone into RK through only two enzymatic steps. Following optimization of enzyme architecture and reaction conditions, the system produced 1.33 mM RK with a 66% conversion yield. We further extended this platform to the synthesis of the cardiotonic drug dobutamine through a non-natural reductive amination cascade using an engineered short-chain dehydrogenase\/reductase (SDR). Rational protein engineering and byproduct recycling increased dobutamine production by approximately 104-fold relative to the wild-type enzyme. Both biosynthetic modules remained robust upon scale-up from microliter to milliliter volumes, and preparative-scale synthesis enabled isolation of 19.2 mg of RK. Overall, this work establishes artificial biosynthetic pathways for the production of a plant-derived natural product and a clinically relevant drug molecule, highlighting the potential of cell-free biomanufacturing as a sustainable platform for the synthesis of value-added chemicals and pharmaceuticals.<\/jats:p>","DOI":"10.1021\/acssynbio.6c00642","type":"journal-article","created":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T06:29:57Z","timestamp":1790317797000},"page":"4371-4379","source":"Crossref","is-referenced-by-count":1,"title":["Artificial Biosynthetic Pathways for Cell-Free Production of Raspberry Ketone and the Cardiotonic Drug Dobutamine"],"prefix":"10.1021","volume":"15","author":[{"given":"Yuhao","family":"Zhang","sequence":"first","affiliation":[{"name":"ShanghaiTech University , , ,","place":["Shanghai, China, 201210"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wan-Qiu","family":"Liu","sequence":"additional","affiliation":[{"name":"ShanghaiTech University , , ,","place":["Shanghai, China, 201210"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2359-238X","authenticated-orcid":true,"given":"Jian","family":"Li","sequence":"additional","affiliation":[{"name":"ShanghaiTech University , , ,","place":["Shanghai, China, 201210"]},{"name":"ShanghaiTech University , , ,","place":["Shanghai, China, 201210"]},{"name":"ShanghaiTech University , , ,","place":["Shanghai, China, 201210"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,9,25]]},"reference":[{"key":"2026101610062598300_cit1","doi-asserted-by":"publisher","first-page":"100606","DOI":"10.1016\/j.fufo.2025.100606","article-title":"Advancements and challenges in the bioproduction of raspberry ketone by precision fermentation","volume":"11","author":"Gao","year":"2025","journal-title":"Future Foods"},{"key":"2026101610062598300_cit2","doi-asserted-by":"publisher","DOI":"10.1111\/jfbc.14018","article-title":"Potential metabolic activities of raspberry ketone","volume":"46","author":"Li","year":"2022","journal-title":"J. Food Biochem."},{"key":"2026101610062598300_cit3","doi-asserted-by":"publisher","first-page":"926","DOI":"10.1007\/s10886-024-01499-6","article-title":"A review on natural phenylbutanoid attractants: Occurrence, distribution, and role in nature, especially in relation to Dacini fruit fly behavior and pollination","volume":"50","author":"Tan","year":"2024","journal-title":"J. Chem. Ecol."},{"key":"2026101610062598300_cit4","doi-asserted-by":"publisher","first-page":"615","DOI":"10.1002\/ffj.3678","article-title":"Advances in the synthesis and applications of raspberry ketone: A review","volume":"36","author":"Guo","year":"2021","journal-title":"Flavour. Fragr. J."},{"key":"2026101610062598300_cit5","doi-asserted-by":"publisher","first-page":"1323","DOI":"10.1002\/jctb.722","article-title":"Bioflavours and fragrances via fermentation and biocatalysis","volume":"77","author":"Vandamme","year":"2002","journal-title":"J. Chem. Technol. Biotechnol."},{"issue":"6673","key":"2026101610062598300_cit6","doi-asserted-by":"publisher","DOI":"10.1126\/science.adh8615","article-title":"From nature to industry: Harnessing enzymes for biocatalysis","volume":"382","author":"Buller","year":"2023","journal-title":"Science"},{"key":"2026101610062598300_cit7","doi-asserted-by":"publisher","first-page":"3269","DOI":"10.1038\/s41467-026-69968-y","article-title":"Engineering LmrR protein for L-proline-based asymmetric aldol biocatalysis","volume":"17","author":"Lu","year":"2026","journal-title":"Nat. Commun."},{"key":"2026101610062598300_cit8","doi-asserted-by":"publisher","first-page":"1270","DOI":"10.1002\/biot.200700076","article-title":"Microbial production of natural raspberry ketone","volume":"2","author":"Beekwilder","year":"2007","journal-title":"Biotechnol. 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Food Chem."},{"issue":"1","key":"2026101610062598300_cit13","doi-asserted-by":"publisher","first-page":"92","DOI":"10.1186\/s12934-020-01351-y","article-title":"Synthesis of the character impact compound raspberry ketone and additional flavoring phenylbutanoids of biotechnological interest with Corynebacterium glutamicum","volume":"19","author":"Milke","year":"2020","journal-title":"Microb. Cell Fact."},{"key":"2026101610062598300_cit14","doi-asserted-by":"publisher","first-page":"ysab021","DOI":"10.1093\/synbio\/ysab021","article-title":"High-yield \u2018one-pot\u2019 biosynthesis of raspberry ketone, a high-value fine chemical","volume":"6","author":"Moore","year":"2021","journal-title":"Synth. Biol."},{"key":"2026101610062598300_cit15","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1007\/s00897960034a","article-title":"Rheosmin (\u201craspberry ketone\u201d) and zingerone, and their preparation by crossed aldol-catalytic hydrogenation sequences","volume":"1","author":"Smith","year":"1996","journal-title":"Chem. 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