{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T12:43:21Z","timestamp":1786538601637,"version":"build-2736575974"},"reference-count":26,"publisher":"American Chemical Society (ACS)","issue":"31","license":[{"start":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T00:00:00Z","timestamp":1785283200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T00:00:00Z","timestamp":1785283200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T00:00:00Z","timestamp":1785283200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["W2533092"],"award-info":[{"award-number":["W2533092"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"\u2018111 Center\u2019, China","award":["D26021"],"award-info":[{"award-number":["D26021"]}]},{"name":"Innovation & Entrepreneurship Team Project of Guangzhou, China","award":["2024D03J0012"],"award-info":[{"award-number":["2024D03J0012"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,8,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Caffeic acid (CA) is a valuable phenylpropanoid with applications in food, pharmaceutical, and chemical industries. Microbial production of CA is often limited by the terminal hydroxylation catalyzed by 4-hydroxyphenylacetate 3-monooxygenase (HpaBC). Here, we constructed a de novo CA biosynthetic pathway in Escherichia coli and enhanced production through systematic metabolic and spatial engineering. Optimization of l-tyrosine supply and HpaBC expression increased CA production to 61.0 mg\/L. However, further enhancement of glucose uptake and precursor supply was insufficient, indicating that hydroxylation remained a major limitation. To address this, HpaBC was relocated to the periplasm via the Tat pathway, increasing CA production 4.9-fold to 299.1 mg\/L with reduced byproduct formation. Further lpp+14 mediated periplasmic remodeling and fermentation optimization increased shake-flask production to 463.7 mg\/L. Finally, fed-batch fermentation achieved 5.1 g\/L CA in a 3 L bioreactor. This study highlights periplasmic engineering as an effective strategy for improving oxidation-dependent phenylpropanoid biosynthesis.<\/jats:p>","DOI":"10.1021\/acs.jafc.6c05246","type":"journal-article","created":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T13:17:23Z","timestamp":1785849443000},"page":"24542-24552","source":"Crossref","is-referenced-by-count":0,"title":["Periplasmic Engineering\nEnhances Terminal Hydroxylation\nfor Efficient Caffeic Acid Biosynthesis in\n                    <i>Escherichia coli<\/i>"],"prefix":"10.1021","volume":"74","author":[{"given":"Shangyi","family":"Wang","sequence":"first","affiliation":[{"name":"Guangzhou University , , 230 Wai Huan Xi Road , ,","place":["Guangzhou, China, 510006"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuqi","family":"Zhuo","sequence":"additional","affiliation":[{"name":"Guangzhou University , , 230 Wai Huan Xi Road , ,","place":["Guangzhou, China, 510006"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jamila Akter","family":"Tuly","sequence":"additional","affiliation":[{"name":"Guangzhou University , , 230 Wai Huan Xi Road , ,","place":["Guangzhou, China, 510006"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6072-1289","authenticated-orcid":true,"given":"Hossain M.","family":"Zabed","sequence":"additional","affiliation":[{"name":"Guangzhou University , , 230 Wai Huan Xi Road , ,","place":["Guangzhou, China, 510006"]},{"name":"Guangzhou University , , 230 Wai Huan Xi Road , ,","place":["Guangzhou, China, 510006"]},{"name":"Guangzhou University , , 230 Wai Huan Xi Road , ,","place":["Guangzhou, China, 510006"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yufei","family":"Zhang","sequence":"additional","affiliation":[{"name":"Guangzhou University , , 230 Wai Huan Xi Road , ,","place":["Guangzhou, China, 510006"]},{"name":"Guangzhou University , , 230 Wai Huan Xi Road , ,","place":["Guangzhou, China, 510006"]},{"name":"Guangzhou University , , 230 Wai Huan Xi Road , ,","place":["Guangzhou, China, 510006"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guoyan","family":"Zhang","sequence":"additional","affiliation":[{"name":"Guangzhou University , , 230 Wai Huan Xi Road , ,","place":["Guangzhou, China, 510006"]},{"name":"Guangzhou University , , 230 Wai Huan Xi Road , ,","place":["Guangzhou, China, 510006"]},{"name":"Guangzhou University , , 230 Wai Huan Xi Road , ,","place":["Guangzhou, China, 510006"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuehui","family":"Tian","sequence":"additional","affiliation":[{"name":"Guangzhou University , , 230 Wai Huan Xi Road , ,","place":["Guangzhou, China, 510006"]},{"name":"Guangzhou University , , 230 Wai Huan Xi Road , ,","place":["Guangzhou, China, 510006"]},{"name":"Guangzhou University , , 230 Wai Huan Xi Road , ,","place":["Guangzhou, China, 510006"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jia","family":"Li","sequence":"additional","affiliation":[{"name":"Guangzhou University , , 230 Wai Huan Xi Road , ,","place":["Guangzhou, China, 510006"]},{"name":"Guangzhou University , , 230 Wai Huan Xi Road , ,","place":["Guangzhou, China, 510006"]},{"name":"Guangzhou University , , 230 Wai Huan Xi Road , ,","place":["Guangzhou, China, 510006"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junhua","family":"Yun","sequence":"additional","affiliation":[{"name":"Guangzhou University , , 230 Wai Huan Xi Road , ,","place":["Guangzhou, China, 510006"]},{"name":"Guangzhou University , , 230 Wai Huan Xi Road , ,","place":["Guangzhou, China, 510006"]},{"name":"Guangzhou University , , 230 Wai Huan Xi Road , ,","place":["Guangzhou, China, 510006"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5817-1609","authenticated-orcid":true,"given":"Xianghui","family":"Qi","sequence":"additional","affiliation":[{"name":"Guangzhou University , , 230 Wai Huan Xi Road , ,","place":["Guangzhou, China, 510006"]},{"name":"Guangzhou University , , 230 Wai Huan Xi Road , ,","place":["Guangzhou, China, 510006"]},{"name":"Guangzhou University , , 230 Wai Huan Xi Road , ,","place":["Guangzhou, China, 510006"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,7,29]]},"reference":[{"issue":"1\u20132","key":"2026081208101681000_cit1","doi-asserted-by":"publisher","first-page":"136","DOI":"10.1016\/j.ijpharm.2010.09.035","article-title":"In vitro and in vivo\nantioxidant properties of chlorogenic acid and caffeic acid","volume":"403","author":"Sato","year":"2011","journal-title":"Int. J. Pharm."},{"issue":"2","key":"2026081208101681000_cit2","doi-asserted-by":"publisher","DOI":"10.3390\/molecules25020247","article-title":"Caffeic\nacid attenuates multi-drug resistance in cancer cells by inhibiting\nefflux function of human P-glycoprotein","volume":"25","author":"Teng","year":"2020","journal-title":"Molecules"},{"key":"2026081208101681000_cit3","doi-asserted-by":"publisher","first-page":"106","DOI":"10.1016\/j.fct.2018.10.040","article-title":"Reproductive\nand developmental toxicity study of caffeic acid in mice","volume":"123","author":"Liu","year":"2019","journal-title":"Food Chem. Toxicol."},{"key":"2026081208101681000_cit4","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.ymben.2019.03.002","article-title":"Balancing the non-linear rosmarinic\nacid biosynthetic pathway by modular co-culture engineering","volume":"54","author":"Li","year":"2019","journal-title":"Metab. Eng."},{"issue":"9","key":"2026081208101681000_cit5","doi-asserted-by":"publisher","first-page":"2816","DOI":"10.1021\/acs.jafc.0c07588","article-title":"De novo biosynthesis of chlorogenic acid using an artificial\nmicrobial community","volume":"69","author":"Li","year":"2021","journal-title":"J. Agric. Food Chem."},{"key":"2026081208101681000_cit6","doi-asserted-by":"publisher","first-page":"89","DOI":"10.1016\/j.ymben.2017.09.011","article-title":"Engineering a bacterial\nplatform for total biosynthesis of caffeic acid derived phenethyl\nesters and amides","volume":"44","author":"Wang","year":"2017","journal-title":"Metab. Eng."},{"issue":"24","key":"2026081208101681000_cit7","doi-asserted-by":"publisher","first-page":"6553","DOI":"10.1016\/j.bmcl.2008.10.046","article-title":"Synthesis of trans-caffeate\nanalogues and their bioactivities against HIV-1 integrase and cancer\ncell lines","volume":"18","author":"Xia","year":"2008","journal-title":"Bioorg. Med. Chem. Lett."},{"issue":"1","key":"2026081208101681000_cit8","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-019-09848-w","article-title":"Synthetic biology strategies for\nmicrobial biosynthesis of plant natural products","volume":"10","author":"Cravens","year":"2019","journal-title":"Nat. Commun."},{"key":"2026081208101681000_cit9","doi-asserted-by":"publisher","DOI":"10.1186\/1475-2859-11-42","article-title":"Biosynthesis of caffeic\nacid in Escherichia coli using its\nendogenous hydroxylase complex","volume":"11","author":"Lin","year":"2012","journal-title":"Microb. Cell\nFact."},{"issue":"2","key":"2026081208101681000_cit10","doi-asserted-by":"publisher","first-page":"287","DOI":"10.1016\/j.eng.2018.11.029","article-title":"Engineering the biosynthesis\nof caffeic acid in Saccharomyces cerevisiae with heterologous enzyme combinations","volume":"5","author":"Liu","year":"2019","journal-title":"Engineering"},{"key":"2026081208101681000_cit11","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2025.1570406","article-title":"Heterologous\nproduction of caffeic acid in microbial hosts: current status and\nperspectives","volume":"16","author":"Li","year":"2025","journal-title":"Front. Microbiol."},{"issue":"12","key":"2026081208101681000_cit12","doi-asserted-by":"publisher","first-page":"3188","DOI":"10.1002\/bit.24988","article-title":"Caffeic acid production\nenhancement by engineering\na phenylalanine over-producing Escherichia coli strain","volume":"110","author":"Huang","year":"2013","journal-title":"Biotechnol. Bioeng."},{"issue":"1","key":"2026081208101681000_cit13","doi-asserted-by":"publisher","DOI":"10.1186\/s12934-020-01300-9","article-title":"Heterologous\ncaffeic acid biosynthesis in Escherichia coli is affected by choice of tyrosine\nammonia lyase and redox partners for bacterial cytochrome P450","volume":"19","author":"Haslinger","year":"2020","journal-title":"Microb. Cell Fact."},{"issue":"5","key":"2026081208101681000_cit14","doi-asserted-by":"publisher","first-page":"520","DOI":"10.1038\/s41589-022-01014-6","article-title":"Engineering\ncofactor supply and recycling\nto drive phenolic acid biosynthesis in yeast","volume":"18","author":"Chen","year":"2022","journal-title":"Nat. Chem. Biol."},{"issue":"3","key":"2026081208101681000_cit15","first-page":"794","article-title":"Cofactor manipulation\nto drive biosynthesis of natural products in microbial cell factories","volume":"7","author":"Gao","year":"2022","journal-title":"Synth. Syst. Biotechnol."},{"issue":"10","key":"2026081208101681000_cit16","doi-asserted-by":"publisher","first-page":"2554","DOI":"10.1002\/bit.26797","article-title":"Metabolic engineering\nof Escherichia coli for the enhanced\nproduction of L-tyrosine","volume":"115","author":"Kim","year":"2018","journal-title":"Biotechnol. Bioeng."},{"issue":"1","key":"2026081208101681000_cit17","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.2004935","article-title":"The gram-negative\nbacterial periplasm: size matters","volume":"16","author":"Miller","year":"2018","journal-title":"PLoS Biol."},{"key":"2026081208101681000_cit18","doi-asserted-by":"publisher","first-page":"415","DOI":"10.1016\/j.ymben.2025.06.006","article-title":"Engineering of Escherichia coli cytoplasm and periplasm for efficient synthesis\nof salvianic acid A","volume":"91","author":"Tian","year":"2025","journal-title":"Metab. Eng."},{"issue":"24","key":"2026081208101681000_cit19","doi-asserted-by":"publisher","first-page":"8460","DOI":"10.1073\/pnas.0500737102","article-title":"Signal peptide\u2013chaperone\ninteractions\non the twin-arginine protein transport pathway","volume":"102","author":"Hatzixanthis","year":"2005","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"issue":"8","key":"2026081208101681000_cit20","doi-asserted-by":"publisher","first-page":"1678","DOI":"10.1021\/bi201852d","article-title":"Conserved signal peptide\nrecognition systems across the prokaryotic domains","volume":"51","author":"Coulthurst","year":"2012","journal-title":"Biochemistry"},{"issue":"7","key":"2026081208101681000_cit21","doi-asserted-by":"publisher","first-page":"2506","DOI":"10.1128\/AEM.04023-14","article-title":"Multigene editing in the Escherichia coli genome via the CRISPR-Cas9 system","volume":"81","author":"Jiang","year":"2015","journal-title":"Appl. Environ.\nMicrobiol."},{"key":"2026081208101681000_cit22","doi-asserted-by":"publisher","DOI":"10.1016\/j.biortech.2022.128320","article-title":"Enhancing caffeic acid production\nin Escherichia coli by engineering\nthe biosynthesis pathway and transporter","volume":"368","author":"Wang","year":"2023","journal-title":"Bioresour.\nTechnol."},{"key":"2026081208101681000_cit23","doi-asserted-by":"publisher","DOI":"10.1016\/j.enzmictec.2023.110193","article-title":"Caffeic acid production from glucose using metabolically engineered Escherichia coli","volume":"164","author":"Sakae","year":"2023","journal-title":"Enzyme Microb.\nTechnol."},{"key":"2026081208101681000_cit24","doi-asserted-by":"publisher","DOI":"10.1186\/s12934-014-0126-z","article-title":"Engineering Escherichia coli to overproduce aromatic amino acids and derived\ncompounds","volume":"13","author":"Rodriguez","year":"2014","journal-title":"Microb. Cell Fact."},{"issue":"12","key":"2026081208101681000_cit25","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.2004303","article-title":"Communication\nacross the bacterial\ncell envelope depends on the size of the periplasm","volume":"15","author":"Asmar","year":"2017","journal-title":"PLoS Biol."},{"issue":"8","key":"2026081208101681000_cit26","first-page":"1138","article-title":"Structural and functional characterization\nof 4-hydroxyphenylacetate\n3-monooxygenase from Escherichia coli","volume":"21","author":"Deng","year":"2020","journal-title":"ChemBioChem"}],"container-title":["Journal of Agricultural\nand Food Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.jafc.6c05246","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/31\/24542\/66330459\/acs.jafc.6c05246.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/31\/24542\/66330459\/acs.jafc.6c05246.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T12:21:36Z","timestamp":1786537296000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jafcau\/article\/74\/31\/24542\/5236747\/Periplasmic-Engineering-Enhances-Terminal"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,29]]},"references-count":26,"journal-issue":{"issue":"31","published-online":{"date-parts":[[2026,7,29]]},"published-print":{"date-parts":[[2026,8,12]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.jafc.6c05246","relation":{},"ISSN":["0021-8561","1520-5118"],"issn-type":[{"value":"0021-8561","type":"print"},{"value":"1520-5118","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,29]]}}}