{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T09:18:11Z","timestamp":1788254291124,"version":"build-2803163510"},"reference-count":47,"publisher":"American Chemical Society (ACS)","issue":"33","license":[{"start":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T00:00:00Z","timestamp":1786492800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T00:00:00Z","timestamp":1786492800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T00:00:00Z","timestamp":1786492800000},"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":["32371493"],"award-info":[{"award-number":["32371493"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32501309"],"award-info":[{"award-number":["32501309"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,8,26]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Spermidine, a natural polyamine, is commonly used as a dietary supplement. Microbial synthesis of spermidine is a sustainable and effective alternative to traditional extraction methods but is constrained by low yields. In this study, enzyme engineering strategies were applied to increase the spermidine yield in Bacillus amyloliquefaciens. Alanine scanning mutagenesis of agmatinase SpeB increased spermidine yield by 78.3%. Enzyme kinetic analysis and molecular dynamics (MD) simulations further revealed that the increase in yield was attributed to the enhanced substrate affinity of SpeB. Additionally, molecular docking, negative feedback regulation, and substrate channel engineering were used to engineer spermidine synthase SpeE1, resulting in a 178.2% increase in spermidine yield. Through fed-batch fermentation in shake flasks, the coexpression-engineered strain PM1\/SpeE1E159P\/S157L\/Q32N-SpeBH151A\/Y144A exhibited a 5.6-fold increase in spermidine yield, reaching up to 851.15 mg\/L, compared to the control strain PM1\/SpeE-SpeB. This work provides an effective enzyme engineering approach for the high-level microbial synthesis of spermidine.<\/jats:p>","DOI":"10.1021\/acs.jafc.6c01482","type":"journal-article","created":{"date-parts":[[2026,8,12]],"date-time":"2026-08-12T07:02:25Z","timestamp":1786518145000},"page":"26473-26483","source":"Crossref","is-referenced-by-count":0,"title":["Rational Design\nof Agmatinase SpeB and Spermidine\nSynthase SpeE Enhances Spermidine Production in\n                    <i>Bacillus amyloliquefaciens<\/i>"],"prefix":"10.1021","volume":"74","author":[{"given":"Yingchao","family":"Wu","sequence":"first","affiliation":[{"name":"Huazhong Agricultural University , , ,","place":["Wuhan, China, 430070"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ziyue","family":"Zhao","sequence":"additional","affiliation":[{"name":"Huazhong Agricultural University , , ,","place":["Wuhan, China, 430070"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meiye","family":"Li","sequence":"additional","affiliation":[{"name":"Huazhong Agricultural University , , ,","place":["Wuhan, China, 430070"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Keyao","family":"An","sequence":"additional","affiliation":[{"name":"Huazhong Agricultural University , , ,","place":["Wuhan, China, 430070"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dian","family":"Zou","sequence":"additional","affiliation":[{"name":"Huazhong Agricultural University , , ,","place":["Wuhan, China, 430070"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5731-7824","authenticated-orcid":true,"given":"Xuetuan","family":"Wei","sequence":"additional","affiliation":[{"name":"Huazhong Agricultural University , , ,","place":["Wuhan, China, 430070"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,12]]},"reference":[{"issue":"7","key":"2026090104471420500_cit1","doi-asserted-by":"publisher","first-page":"6401","DOI":"10.18632\/aging.103035","article-title":"Spermidine and spermine delay brain\naging by inducing autophagy in SAMP8 mice","volume":"12","author":"Xu","year":"2020","journal-title":"Aging"},{"key":"2026090104471420500_cit2","doi-asserted-by":"publisher","DOI":"10.1016\/j.phrs.2020.104943","article-title":"Spermidine as a target for cancer\ntherapy","volume":"159","author":"Fan","year":"2020","journal-title":"Pharmacol. 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Sci."},{"issue":"7","key":"2026090104471420500_cit32","doi-asserted-by":"publisher","DOI":"10.3390\/microorganisms11071689","article-title":"Enhancing the catalytic activity\nof glycolate oxidase\nfrom Chlamydomonas reinhardtii through semi-rational\ndesign","volume":"11","author":"Feng","year":"2023","journal-title":"Microorganisms"},{"issue":"39","key":"2026090104471420500_cit33","doi-asserted-by":"publisher","first-page":"12672","DOI":"10.1021\/acs.jafc.2c04733","article-title":"Improving\nthe catalytic efficiency of Aspergillus fumigatus glucoamylase toward raw starch by engineering its N-glycosylation\nsites and saturation mutation","volume":"70","author":"Song","year":"2022","journal-title":"J. Agric. 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Food Chem."},{"key":"2026090104471420500_cit36","doi-asserted-by":"publisher","first-page":"30","DOI":"10.1016\/j.foodchem.2018.01.108","article-title":"Enhancement of Z-aspartame\nsynthesis by rational engineering of metalloprotease","volume":"253","author":"Zhu","year":"2018","journal-title":"Food Chem."},{"issue":"2","key":"2026090104471420500_cit37","doi-asserted-by":"publisher","first-page":"871","DOI":"10.1021\/acs.jcim.8b00926","article-title":"Alanine scanning\neffects on the biochemical and biophysical\nproperties of intrinsically disordered proteins: A case study of the\nhistidine to alanine mutations in amyloid-\u03b242","volume":"59","author":"Coskuner-Weber","year":"2019","journal-title":"J. Chem. Inf. 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Macromol."},{"issue":"1","key":"2026090104471420500_cit40","doi-asserted-by":"publisher","first-page":"267","DOI":"10.1021\/acs.jafc.0c07143","article-title":"Biosynthesis of a novel bioactive\nmetabolite of spermidine from Bacillus amyloliquefaciens\u202f: Gene mining, sequence analysis, and combined expression","volume":"69","author":"Zou","year":"2021","journal-title":"J. Agric. Food Chem."},{"issue":"1","key":"2026090104471420500_cit41","doi-asserted-by":"publisher","DOI":"10.1038\/s42004-023-00946-x","article-title":"Mechanistic\ninvestigation of a D to N mutation in DAHP synthase that dictates\ncarbon flux into the shikimate pathway in yeast","volume":"6","author":"Liu","year":"2023","journal-title":"Commun. Chem."},{"issue":"4","key":"2026090104471420500_cit42","doi-asserted-by":"publisher","first-page":"299","DOI":"10.1038\/nchem.2459","article-title":"Substrate channelling\nas an approach to cascade reactions","volume":"8","author":"Wheeldon","year":"2016","journal-title":"Nat. Chem."},{"issue":"9","key":"2026090104471420500_cit43","doi-asserted-by":"publisher","first-page":"5506","DOI":"10.1021\/jacs.3c00195","article-title":"Enabling peroxygenase\nactivity in cytochrome P450 monooxygenases\nby engineering hydrogen peroxide tunnels","volume":"145","author":"Zhao","year":"2023","journal-title":"J.\nAm. Chem. Soc."},{"issue":"14","key":"2026090104471420500_cit44","doi-asserted-by":"publisher","first-page":"5332","DOI":"10.1021\/acssuschemeng.5c00435","article-title":"Enhancing the substrate\nspecificity\nof nitrilase toward aliphatic nitriles based on substrate channel\ndesign","volume":"13","author":"Bian","year":"2025","journal-title":"ACS Sustainable Chem. 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Eng."}],"container-title":["Journal of Agricultural\nand Food Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.jafc.6c01482","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/33\/26473\/66506771\/acs.jafc.6c01482.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/33\/26473\/66506771\/acs.jafc.6c01482.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T08:47:32Z","timestamp":1788252452000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jafcau\/article\/74\/33\/26473\/5253186\/Rational-Design-of-Agmatinase-SpeB-and-Spermidine"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,12]]},"references-count":47,"journal-issue":{"issue":"33","published-online":{"date-parts":[[2026,8,12]]},"published-print":{"date-parts":[[2026,8,26]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.jafc.6c01482","relation":{},"ISSN":["0021-8561","1520-5118"],"issn-type":[{"value":"0021-8561","type":"print"},{"value":"1520-5118","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,12]]}}}