{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T19:45:44Z","timestamp":1792784744731,"version":"4.5.0"},"reference-count":42,"publisher":"American Chemical Society (ACS)","issue":"10","license":[{"start":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T00:00:00Z","timestamp":1790121600000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T00:00:00Z","timestamp":1790121600000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T00:00:00Z","timestamp":1790121600000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"name":"National Basic Research Program of China (973 Program)","award":["2023YFA0914700"],"award-info":[{"award-number":["2023YFA0914700"]}]},{"name":"National Natural Science Foundation of China (NSFC)","award":["22322802"],"award-info":[{"award-number":["22322802"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"crossref"}]},{"name":"National Natural Science Foundation of China (NSFC)","award":["32271488"],"award-info":[{"award-number":["32271488"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"crossref"}]}],"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>Benzylisoquinoline alkaloids (BIAs) are pharmacologically valuable natural products, yet their microbial production in Escherichia coli is hindered by two major bottlenecks: (1) imbalanced metabolic flux between the two upstream precursors, 3,4-dihydroxyphenylacetaldehyde (3,4-DHPAA) and dopamine, and (2) poor soluble expression of plant-derived enzymes, particularly norcoclaurine synthase (NCS) and the methyltransferase Ps4\u2032OMT2. To address these challenges, we systematically engineered E. coli for efficient synthesis of (S)-reticuline, the universal precursor to diverse BIAs. First, to resolve the flux imbalance, we introduced a heterologous \u03c9-transaminase (\u03c9-TA) that dynamically interconverts 3,4-DHPAA and dopamine, thereby coordinating precursor supply without tedious tuning of branch pathways. Second, to overcome enzyme insolubility, we combined solubility-enhancing strategies\ue5f8including SUMO fusion, ProteinMPNN-guided design, and N-terminal truncation\ue5f8which markedly improved the soluble expression of CjNCS and Ps4\u2032OMT2. Furthermore, strengthening the S-adenosylmethionine (SAM) regeneration cycle through integration of luxS, mtn, and metF genes enhanced the efficiency of the three sequential methylation reactions. The final engineered strain achieved a (S)-reticuline titer of 347 mg\/L in shake-flask fermentation. Overall, this work establishes an efficient E. coli chassis for sustainable BIAs production by systematically tackling flux imbalance and enzyme solubility.<\/jats:p>","DOI":"10.1021\/acssynbio.6c00417","type":"journal-article","created":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T16:02:20Z","timestamp":1790179340000},"page":"4229-4239","source":"Crossref","is-referenced-by-count":0,"title":["Balancing Precursor Supply and Enhancing Enzyme Solubility for Efficient Production of (\n                    <i>S<\/i>\n                    )-Reticuline in\n                    <i>Escherichia coli<\/i>"],"prefix":"10.1021","volume":"15","author":[{"given":"Xiaxia","family":"Zhang","sequence":"first","affiliation":[{"name":"Beijing University of Chemical Technology , , ,","place":["Beijing, China, 100029"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Muhammad Hassan","family":"Butt","sequence":"additional","affiliation":[{"name":"Beijing University of Chemical Technology , , ,","place":["Beijing, China, 100029"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wen","family":"Zhang","sequence":"additional","affiliation":[{"name":"Beijing University of Chemical Technology , , ,","place":["Beijing, China, 100029"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuan","family":"Gao","sequence":"additional","affiliation":[{"name":"Beijing University of Chemical Technology , , ,","place":["Beijing, China, 100029"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shibo","family":"Guo","sequence":"additional","affiliation":[{"name":"Beijing University of Chemical Technology , , ,","place":["Beijing, China, 100029"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianfeng","family":"Fang","sequence":"additional","affiliation":[{"name":"Beijing University of Chemical Technology , , ,","place":["Beijing, China, 100029"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6969-2952","authenticated-orcid":true,"given":"Xiaolin","family":"Shen","sequence":"additional","affiliation":[{"name":"Beijing University of Chemical Technology , , ,","place":["Beijing, China, 100029"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7218-2224","authenticated-orcid":true,"given":"Jia","family":"Wang","sequence":"additional","affiliation":[{"name":"Beijing University of Chemical Technology , , ,","place":["Beijing, China, 100029"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5590-6473","authenticated-orcid":true,"given":"Xinxiao","family":"Sun","sequence":"additional","affiliation":[{"name":"Beijing University of Chemical Technology , , ,","place":["Beijing, China, 100029"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qipeng","family":"Yuan","sequence":"additional","affiliation":[{"name":"Beijing University of Chemical Technology , , ,","place":["Beijing, China, 100029"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,9,23]]},"reference":[{"issue":"5","key":"2026101610072798400_cit1","doi-asserted-by":"publisher","first-page":"647","DOI":"10.1093\/pcp\/pct020","article-title":"Benzylisoquinoline alkaloid metabolism: A century of discovery and a brave new world","volume":"54","author":"Hagel","year":"2013","journal-title":"Plant Cell Physiol."},{"issue":"6","key":"2026101610072798400_cit2","doi-asserted-by":"publisher","first-page":"1779","DOI":"10.3390\/pharmaceutics15061779","article-title":"Synthesis and modification of morphine and codeine, leading to diverse libraries with improved pain relief properties","volume":"15","author":"Zarin","year":"2023","journal-title":"Pharmaceutics"},{"issue":"5","key":"2026101610072798400_cit3","doi-asserted-by":"publisher","first-page":"712","DOI":"10.1016\/j.metabol.2008.01.013","article-title":"Efficacy of berberine in patients with type 2 diabetes mellitus","volume":"57","author":"Yin","year":"2008","journal-title":"Metabolism"},{"issue":"8","key":"2026101610072798400_cit4","doi-asserted-by":"publisher","first-page":"12349","DOI":"10.3390\/molecules190812349","article-title":"Berberine an epiphany against cancer","volume":"19","author":"Guama\u0301n Ortiz","year":"2014","journal-title":"Molecules"},{"issue":"15","key":"2026101610072798400_cit5","doi-asserted-by":"publisher","first-page":"5699","DOI":"10.1021\/jm501180v","article-title":"Progress toward the development of noscapine and derivatives as anticancer agents","volume":"58","author":"DeBono","year":"2015","journal-title":"J. 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