{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T22:07:23Z","timestamp":1788991643854,"version":"build-2803163510"},"reference-count":33,"publisher":"American Chemical Society (ACS)","issue":"35","license":[{"start":{"date-parts":[[2026,8,27]],"date-time":"2026-08-27T00:00:00Z","timestamp":1787788800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100006477","name":"National Taiwan University","doi-asserted-by":"publisher","award":["NTU-CC-113L894802"],"award-info":[{"award-number":["NTU-CC-113L894802"]}],"id":[{"id":"10.13039\/501100006477","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100006477","name":"National Taiwan University","doi-asserted-by":"publisher","award":["NTU-CC-114L893002"],"award-info":[{"award-number":["NTU-CC-114L893002"]}],"id":[{"id":"10.13039\/501100006477","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100006477","name":"National Taiwan University","doi-asserted-by":"publisher","award":["NTU-CC-115L891702"],"award-info":[{"award-number":["NTU-CC-115L891702"]}],"id":[{"id":"10.13039\/501100006477","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100020950","name":"National Science and Technology Council","doi-asserted-by":"publisher","award":["111-2113-M-002-015-MY3"],"award-info":[{"award-number":["111-2113-M-002-015-MY3"]}],"id":[{"id":"10.13039\/501100020950","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100020950","name":"National Science and Technology Council","doi-asserted-by":"publisher","award":["111-2311-B-002-008-MY3"],"award-info":[{"award-number":["111-2311-B-002-008-MY3"]}],"id":[{"id":"10.13039\/501100020950","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100020950","name":"National Science and Technology Council","doi-asserted-by":"publisher","award":["114-2113-M-002-009-MY3"],"award-info":[{"award-number":["114-2113-M-002-009-MY3"]}],"id":[{"id":"10.13039\/501100020950","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,9,9]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Mannans are abundant plant hemicelluloses, and endo-\u03b2-mannanases are important biocatalysts for their conversion into functional manno-oligosaccharides. Here, we report the structural and functional characterization of a glycoside hydrolase family 134 \u03b2-mannanase from Aspergillus nidulans (AnGH134) and a structure-guided engineering strategy to improve its performance on locust bean gum. The 1.75 \u00c5 crystal structure reveals the conserved lysozyme-like fold of GH134 enzymes and supports an inverting catalytic mechanism with Glu43 and Asp55 as the putative catalytic residues. Docking, mutational, and molecular dynamics analyses indicate that AnGH134 uses an extended substrate-binding groove and that groove-exit residues and the C-terminal region contribute to productive catalysis. Guided by these findings, N-terminal fusion of CBM10 enhanced catalytic efficiency and thermal stability, whereas C-terminal fusion was detrimental. These results provide a framework for engineering GH134 mannanases.<\/jats:p>","DOI":"10.1021\/acs.jafc.6c05159","type":"journal-article","created":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T18:28:32Z","timestamp":1788287312000},"page":"27687-27697","source":"Crossref","is-referenced-by-count":0,"title":["Structural Characterization\nand Engineering of a GH134\n\u03b2-Mannanase from\n                    <i>Aspergillus nidulans<\/i>\n                    for Enhancement of Activity and Stability"],"prefix":"10.1021","volume":"74","author":[{"given":"Sheng-Chia","family":"Chen","sequence":"first","affiliation":[{"name":"National Taiwan University , , ,","place":["Taipei, Taiwan, 106319"]},{"name":"National Kaohsiung University of Science and Technology , , ,","place":["Kaohsiung, Taiwan, 811213"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Po-Chih","family":"Kuo","sequence":"additional","affiliation":[{"name":"National Taiwan University , , ,","place":["Taipei, Taiwan, 106319"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wen-Ming","family":"Chen","sequence":"additional","affiliation":[{"name":"National Kaohsiung University of Science and Technology , , ,","place":["Kaohsiung, Taiwan, 811213"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shih-Yi","family":"Sheu","sequence":"additional","affiliation":[{"name":"National Kaohsiung University of Science and Technology , , ,","place":["Kaohsiung, Taiwan, 811213"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu-Bin","family":"Huang","sequence":"additional","affiliation":[{"name":"National Kaohsiung University of Science and Technology , , ,","place":["Kaohsiung, Taiwan, 811213"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cheng-Yang","family":"Huang","sequence":"additional","affiliation":[{"name":"National Taiwan University , , ,","place":["Taipei, Taiwan, 106319"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"I-Weh","family":"Chien","sequence":"additional","affiliation":[{"name":"National Taiwan University , , ,","place":["Taipei, Taiwan, 106319"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8036-7563","authenticated-orcid":true,"given":"Tzong-Huei","family":"Lee","sequence":"additional","affiliation":[{"name":"National Taiwan University , , ,","place":["Taipei, Taiwan, 106319"]},{"name":"National Taiwan University , , ,","place":["Taipei, Taiwan, 106319"]},{"name":"National Taiwan University , , ,","place":["Taipei, Taiwan, 106319"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0008-7383","authenticated-orcid":true,"given":"Chun-Hua","family":"Hsu","sequence":"additional","affiliation":[{"name":"National Taiwan University , , ,","place":["Taipei, Taiwan, 106319"]},{"name":"National Taiwan University , , ,","place":["Taipei, Taiwan, 106319"]},{"name":"National Taiwan University and Academia Sinica , , ,","place":["Taipei, Taiwan, 106319"]},{"name":"National Taiwan University , , ,","place":["Taipei, Taiwan, 106319"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,27]]},"reference":[{"issue":"13","key":"2026090917072228500_cit1","doi-asserted-by":"publisher","first-page":"3189","DOI":"10.3390\/ijms20133189","article-title":"Glucan and Mannan-Two Peas in a Pod","volume":"20","author":"Korolenko","year":"2019","journal-title":"Int. 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Biotechnol."},{"issue":"40","key":"2026090917072228500_cit7","doi-asserted-by":"publisher","first-page":"25479","DOI":"10.1021\/acs.jafc.5c03141","article-title":"Unveiling the Structural\nand Biochemical Characteristics\nof an Acidophilic \u03b2-Mannanase from Soybean (Glycine max )","volume":"73","author":"Lin","year":"2025","journal-title":"J. Agric. Food Chem."},{"issue":"22","key":"2026090917072228500_cit8","doi-asserted-by":"publisher","first-page":"13781","DOI":"10.1021\/acs.jafc.5c04047","article-title":"Enzymatic Characterization\nof a Rumen Microorganism-Derived\nMultifunctional Glycoside Hydrolase and Its GH26 Domain with Mannanase\nActivity","volume":"73","author":"Zhao","year":"2025","journal-title":"J. Agric. Food Chem."},{"key":"2026090917072228500_cit9","doi-asserted-by":"publisher","first-page":"899","DOI":"10.1016\/j.ijbiomac.2021.04.075","article-title":"Functional\nand structural investigation of a novel beta-mannanase BaMan113A from\nBacillus sp. N16\u20135","volume":"182","author":"Liu","year":"2021","journal-title":"Int. J. Biol. Macromol."},{"issue":"12","key":"2026090917072228500_cit10","doi-asserted-by":"publisher","first-page":"896","DOI":"10.1021\/acscentsci.6b00232","article-title":"A \u03b2-Mannanase with a Lysozyme-like\nFold and a Novel Molecular Catalytic Mechanism","volume":"2","author":"Jin","year":"2016","journal-title":"ACS Cent. Sci."},{"issue":"6","key":"2026090917072228500_cit11","doi-asserted-by":"publisher","first-page":"1376","DOI":"10.1016\/j.bbagen.2018.03.016","article-title":"Structural\nand biochemical insights into the substrate-binding mechanism of a\nnovel glycoside hydrolase family 134 beta-mannanase","volume":"1862","author":"You","year":"2018","journal-title":"Biochim. Biophys. Acta, Gen. 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Biol."},{"issue":"Pt 7","key":"2026090917072228500_cit24","doi-asserted-by":"publisher","first-page":"861","DOI":"10.1107\/S0907444912015636","article-title":"Improved crystallographic models\nthrough iterated local density-guided model deformation and reciprocal-space\nrefinement","volume":"68","author":"Terwilliger","year":"2012","journal-title":"Acta Crystallogr., Sect. D"},{"issue":"Pt 12","key":"2026090917072228500_cit25","doi-asserted-by":"publisher","first-page":"2126","DOI":"10.1107\/S0907444904019158","article-title":"Coot: model-building\ntools for molecular graphics","volume":"60","author":"Emsley","year":"2004","journal-title":"Acta Crystallogr.,\nSect. 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Chem."},{"issue":"9","key":"2026090917072228500_cit32","doi-asserted-by":"publisher","first-page":"2371","DOI":"10.1021\/ct900275y","article-title":"An Implementation\nof the Smooth Particle Mesh Ewald\nMethod on GPU Hardware","volume":"5","author":"Harvey","year":"2009","journal-title":"J. Chem. Theory Comput."},{"issue":"4","key":"2026090917072228500_cit33","doi-asserted-by":"publisher","first-page":"420","DOI":"10.1038\/s41587-019-0036-z","article-title":"SignalP 5.0 improves signal peptide predictions using\ndeep neural networks","volume":"37","author":"Almagro Armenteros","year":"2019","journal-title":"Nat. Biotechnol."}],"container-title":["Journal of Agricultural\nand Food Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.jafc.6c05159","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/35\/27687\/67492926\/acs.jafc.6c05159.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/jafcau\/article-pdf\/74\/35\/27687\/67492926\/acs.jafc.6c05159.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T21:15:30Z","timestamp":1788988530000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/jafcau\/article\/74\/35\/27687\/5337612\/Structural-Characterization-and-Engineering-of-a"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,27]]},"references-count":33,"journal-issue":{"issue":"35","published-online":{"date-parts":[[2026,8,27]]},"published-print":{"date-parts":[[2026,9,9]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.jafc.6c05159","relation":{},"ISSN":["0021-8561","1520-5118"],"issn-type":[{"value":"0021-8561","type":"print"},{"value":"1520-5118","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,27]]}}}