{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,27]],"date-time":"2026-07-27T06:59:04Z","timestamp":1785135544783,"version":"3.55.0"},"reference-count":66,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2026,7,26]],"date-time":"2026-07-26T00:00:00Z","timestamp":1785024000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"Natural Science Foundation of China","doi-asserted-by":"publisher","award":["31972176"],"award-info":[{"award-number":["31972176"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>In this study, a previously uncharacterized zearalenone lactone hydrolase, ZHD30L, from Knufia peltigerae was identified and characterized, and semi-rational design was employed to enhance its thermostability. The results indicate that the optimal reaction conditions for this enzyme are 40 \u00b0C and pH 9.0, under which it catalyzes the ring-opening of the ZEN lactone to generate the non-toxic product HZEN. The mutant with five combined sites Mut5, (E11T-A127V-I180K-H235P-V156I), was obtained through semi-rational design combined with multiple rounds of iterative superposition and negative elimination strategies. Thermodynamic characterization revealed that, compared to the wild type, the t1\/2 of Mut5 at 48 \u00b0C was significantly extended from 1.67 min to 796.8 min, and the Tm value increased from 39.56 \u00b0C to 50.04 \u00b0C. The markedly prolonged thermal half-life and increased apparent melting temperature may improve the operational robustness of Mut5 during moderate-temperature feed and food processing or post-processing applications, potentially reducing activity loss and the need for repeated enzyme additions. Concurrently, the kcat\/KM improved from 19.08 \u03bcM\u22121\u00b7s\u22121 (95% Cl = 12.80 to 25.36 \u03bcM\u22121\u00b7s\u22121) to 33.29 \u03bcM\u22121\u00b7s\u22121 (95% Cl = 23.20 to 43.38 \u03bcM\u22121\u00b7s\u22121). Under optimal reaction conditions, WT and Mut5 degraded 76.51% \u00b1 1.65% and 99.13% \u00b1 0.71% of ZEN (10 \u03bcg\/mL), respectively, within 3 min at equal enzyme concentrations. Molecular dynamics simulations indicate that the H235P and E11T mutations reduce local conformational entropy, the A127V and V156I mutations optimize internal hydrophobic stacking, and the I180K mutation reshapes the local charge microenvironment. Furthermore, Mut5 exhibited an overall RMSD at high temperatures that was 0.026 \u00c5 lower than that of the wild type, and a global free energy minimum that was 0.36 kcal\/mol lower. These computational results offer possible structural interpretations, providing a theoretical reference for developing highly efficient enzyme preparations for the feed and food industries.<\/jats:p>","DOI":"10.3390\/toxins18080324","type":"journal-article","created":{"date-parts":[[2026,7,27]],"date-time":"2026-07-27T05:49:27Z","timestamp":1785131367000},"page":"324","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Identification and Thermal Stability Modification of a Zearalenone Lactone Hydrolase ZHD30L"],"prefix":"10.3390","volume":"18","author":[{"given":"Xingsai","family":"Liu","sequence":"first","affiliation":[{"name":"Food Laboratory of Zhongyuan, Luohe 462300, China"},{"name":"College of Biological Engineering, Henan University of Technology, Zhengzhou 450001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fengguang","family":"Zhao","sequence":"additional","affiliation":[{"name":"College of Biological Engineering, Henan University of Technology, Zhengzhou 450001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-9323-0255","authenticated-orcid":false,"given":"Kashif Iqbal","family":"Sahibzada","sequence":"additional","affiliation":[{"name":"Department of Health Professional Technologies, Faculty of Allied Health Sciences, The University of Lahore, Lahore 54570, Pakistan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yaping","family":"Zhang","sequence":"additional","affiliation":[{"name":"College of Biological Engineering, Henan University of Technology, Zhengzhou 450001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shan","family":"Wei","sequence":"additional","affiliation":[{"name":"College of Biological Engineering, Henan University of Technology, Zhengzhou 450001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pingping","family":"Tian","sequence":"additional","affiliation":[{"name":"College of Food and Bioengineering, Henan University of Science and Technology, Luoyang 471023, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yingying","family":"Wang","sequence":"additional","affiliation":[{"name":"College of Biological Engineering, Henan University of Technology, Zhengzhou 450001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuansen","family":"Hu","sequence":"additional","affiliation":[{"name":"College of Biological Engineering, Henan University of Technology, Zhengzhou 450001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5342-0656","authenticated-orcid":false,"given":"Yangyong","family":"Lv","sequence":"additional","affiliation":[{"name":"Food Laboratory of Zhongyuan, Luohe 462300, China"},{"name":"College of Biological Engineering, Henan University of Technology, Zhengzhou 450001, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2773","DOI":"10.1080\/10408398.2019.1658570","article-title":"Worldwide contamination of food-crops with mycotoxins: Validity of the widely cited \u2018FAO estimate\u2019 of 25","volume":"60","author":"Eskola","year":"2020","journal-title":"Crit. 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