{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T19:44:45Z","timestamp":1792784685117,"version":"4.5.0"},"reference-count":31,"publisher":"American Chemical Society (ACS)","issue":"39","license":[{"start":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T00:00:00Z","timestamp":1789948800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T00:00:00Z","timestamp":1789948800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T00:00:00Z","timestamp":1789948800000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"name":"National Natural Science Foundation of China for Cultivation Fund of Nanyang Normal University","award":["2025PY005"],"award-info":[{"award-number":["2025PY005"]}],"id":[{"id":"https:\/\/ror.org\/01f7yer47","id-type":"ROR","asserted-by":"crossref"}]},{"name":"National Natural Science Foundation of China for Cultivation Fund of Nanyang Normal University","award":["2025PY031"],"award-info":[{"award-number":["2025PY031"]}],"id":[{"id":"https:\/\/ror.org\/01f7yer47","id-type":"ROR","asserted-by":"crossref"}]},{"name":"Natural Science Foundation Project of Henan Province","award":["252300423631"],"award-info":[{"award-number":["252300423631"]}]},{"name":"Henan Provincial Joint Fund for Science and Technology","award":["235101610003"],"award-info":[{"award-number":["235101610003"]}]},{"name":"Henan Provincial Joint Fund for Science and Technology","award":["235101610044"],"award-info":[{"award-number":["235101610044"]}]},{"name":"Project of key scientific research of universities in Henan Province","award":["25B416003"],"award-info":[{"award-number":["25B416003"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,10,7]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>2-Deoxy-D-ribose-5-phosphate aldolase (DERA) is a useful biocatalyst, but its application is limited by poor thermostability. Here, a flexibility-guided engineering strategy was applied to Escherichia coli DERA (EcDERA) to improve thermostability while retaining catalytic performance. Molecular dynamics simulations were used to identify flexible regions for mutation design. Among the variants tested, D26P\/Y233F showed the best overall performance. Its half-life at 60 \u00b0C increased from 103 to 363 min (3.5-fold), and its specific activity increased by 60.9% (17.28 vs 10.74 U\u00b7mg\u20131) compared with the wild type. Structural and molecular dynamics analyses were consistent with reduced N-terminal fluctuations for D26P and changes in the local hydrophobic environment and substrate-binding interactions associated with Y233F. The double mutant also retained a catalytic efficiency comparable to that of the wild type. Targeted modification of flexible regions therefore offers a practical route to improving EcDERA thermostability while preserving catalytic performance.<\/jats:p>","DOI":"10.1021\/acs.jafc.6c08519","type":"journal-article","created":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T23:02:05Z","timestamp":1790031725000},"page":"30987-30998","source":"Crossref","is-referenced-by-count":0,"title":["Local Flexible-Region\nEngineering Improves the Thermostability\nand Catalytic Activity of 2-Deoxy-D-Ribose-5-Phosphate Aldolase"],"prefix":"10.1021","volume":"74","author":[{"given":"Xueyang","family":"Bai","sequence":"first","affiliation":[{"name":"Nanyang Normal University , , 1638 Wolong Road , , ,","place":["Nanyang, Henan, People\u2019s Republic of China, 473061"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yangwan","family":"Li","sequence":"additional","affiliation":[{"name":"Nanyang Normal University , , 1638 Wolong Road , , ,","place":["Nanyang, Henan, People\u2019s Republic of China, 473061"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fangyuan","family":"Tian","sequence":"additional","affiliation":[{"name":"Nanyang Normal University , , 1638 Wolong Road , , ,","place":["Nanyang, Henan, People\u2019s Republic of China, 473061"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mingzhu","family":"Zhang","sequence":"additional","affiliation":[{"name":"Nanyang Normal University , , 1638 Wolong Road , , ,","place":["Nanyang, Henan, People\u2019s Republic of China, 473061"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiang","family":"Zhang","sequence":"additional","affiliation":[{"name":"Nanyang Normal University , , 1638 Wolong Road , , ,","place":["Nanyang, Henan, People\u2019s Republic of China, 473061"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yunchao","family":"Kan","sequence":"additional","affiliation":[{"name":"Nanyang Normal University , , 1638 Wolong Road , , ,","place":["Nanyang, Henan, People\u2019s Republic of China, 473061"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2100-4145","authenticated-orcid":true,"given":"Dandan","family":"Li","sequence":"additional","affiliation":[{"name":"Nanyang Normal University , , 1638 Wolong Road , , ,","place":["Nanyang, Henan, People\u2019s Republic of China, 473061"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongling","family":"Shi","sequence":"additional","affiliation":[{"name":"Nanyang Normal University , , 1638 Wolong Road , , ,","place":["Nanyang, Henan, People\u2019s Republic of China, 473061"]},{"name":"Dalian University of Technology , , 2 Linggong Road , , ,","place":["Dalian, Liaoning, People\u2019s Republic of China, 116024"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lunguang","family":"Yao","sequence":"additional","affiliation":[{"name":"Nanyang Normal University , , 1638 Wolong Road , , ,","place":["Nanyang, Henan, People\u2019s Republic of China, 473061"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0352-0805","authenticated-orcid":true,"given":"Cunduo","family":"Tang","sequence":"additional","affiliation":[{"name":"Nanyang Normal University , , 1638 Wolong Road , , ,","place":["Nanyang, Henan, People\u2019s Republic of China, 473061"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,9,21]]},"reference":[{"issue":"21","key":"2026100711064373400_cit1","doi-asserted-by":"publisher","first-page":"13236","DOI":"10.1021\/acscatal.1c03911","article-title":"Unlocking Asymmetric Michael Additions\nin an Archetypical Class I Aldolase by Directed Evolution","volume":"11","author":"Kunzendorf","year":"2021","journal-title":"ACS Catal."},{"issue":"5541","key":"2026100711064373400_cit2","doi-asserted-by":"publisher","first-page":"369","DOI":"10.1126\/science.1063601","article-title":"Observation of Covalent\nIntermediates in an Enzyme\nMechanism at Atomic Resolution","volume":"294","author":"Heine","year":"2001","journal-title":"Science"},{"issue":"4","key":"2026100711064373400_cit3","doi-asserted-by":"publisher","first-page":"1019","DOI":"10.1016\/j.jmb.2004.08.066","article-title":"Analysis of the\nClass I Aldolase Binding Site Architecture Based on the Crystal Structure\nof 2-Deoxyribose-5-phosphate Aldolase at 0.99\u00c5 Resolution","volume":"343","author":"Heine","year":"2004","journal-title":"J. Mol. Biol."},{"issue":"16","key":"2026100711064373400_cit4","doi-asserted-by":"publisher","first-page":"5788","DOI":"10.1073\/pnas.0307563101","article-title":"Development\nof an efficient, scalable,\naldolase-catalyzed process for enantioselective synthesis of statin\nintermediates","volume":"101","author":"Greenberg","year":"2004","journal-title":"Proc. Natl. Acad. Sci. U. S.\nA."},{"issue":"5","key":"2026100711064373400_cit5","doi-asserted-by":"publisher","first-page":"537","DOI":"10.1002\/biot.200600020","article-title":"Directed evolution of an industrial\nbiocatalyst: 2-deoxy-D-ribose 5-phosphate aldolase","volume":"1","author":"Jennewein","year":"2006","journal-title":"Biotechnol. 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J. Biol. Macromol."},{"issue":"8","key":"2026100711064373400_cit22","doi-asserted-by":"publisher","first-page":"883","DOI":"10.3390\/catal10080883","article-title":"Discovery and Engineering\nof an Aldehyde Tolerant 2-deoxy-D-ribose 5-phosphate Aldolase (DERA)\nfrom Pectobacterium atrosepticum","volume":"10","author":"Haridas","year":"2020","journal-title":"Catalysts"},{"issue":"1","key":"2026100711064373400_cit23","doi-asserted-by":"publisher","first-page":"29","DOI":"10.1007\/s10295-012-1213-y","article-title":"Characterization\nand application of a newly synthesized 2-deoxyribose-5-phosphate aldolase","volume":"40","author":"You","year":"2013","journal-title":"J. Ind. Microbiol. Biotechnol."},{"issue":"2","key":"2026100711064373400_cit24","doi-asserted-by":"publisher","first-page":"119","DOI":"10.1016\/j.bbapap.2005.06.005","article-title":"How to\nstudy proteins by circular\ndichroism","volume":"1751","author":"Kelly","year":"2005","journal-title":"Biochim. Biophys. 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Food Chem."},{"issue":"34","key":"2026100711064373400_cit30","doi-asserted-by":"publisher","first-page":"12785","DOI":"10.1021\/acs.jafc.3c03871","article-title":"Simultaneously Enhanced Thermostability\nand Catalytic Activity of Xylanase from Streptomyces rameus L2001\nby Rigidifying Flexible Regions in Loop Regions of the N-Terminus","volume":"71","author":"Wu","year":"2023","journal-title":"J. Agric. Food Chem."},{"issue":"1","key":"2026100711064373400_cit31","doi-asserted-by":"publisher","first-page":"466","DOI":"10.1021\/acscatal.8b03561","article-title":"Switching Cofactor\nDependence of 7\u03b2-Hydroxysteroid Dehydrogenase for Cost-Effective\nProduction of Ursodeoxycholic Acid","volume":"9","author":"You","year":"2019","journal-title":"ACS Catal."}],"container-title":["Journal of Agricultural\nand Food Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.jafc.6c08519","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/39\/30987\/68588414\/acs.jafc.6c08519.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/39\/30987\/68588414\/acs.jafc.6c08519.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,7]],"date-time":"2026-10-07T15:37:56Z","timestamp":1791387476000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jafcau\/article\/74\/39\/30987\/5434769\/Local-Flexible-Region-Engineering-Improves-the"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,21]]},"references-count":31,"journal-issue":{"issue":"39","published-online":{"date-parts":[[2026,9,21]]},"published-print":{"date-parts":[[2026,10,7]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.jafc.6c08519","relation":{},"ISSN":["0021-8561","1520-5118"],"issn-type":[{"value":"0021-8561","type":"print"},{"value":"1520-5118","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,9,21]]}}}