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BhGH18 displayed maximal activity at pH 7.5 and 40 \u00b0C, and activity was enhanced by Mg2+\/Zn2+ but inhibited by Cu2+\/Fe2+\/Ni2+. After further purification, the enzyme exhibited markedly improved kinetics toward pNP-(GlcNAc)2 (Km = 0.103 mM, Vmax = 0.217 \u03bcmol\/min). Importantly, on the natural substrate colloidal chitin, BhGH18 gave an apparent Km of 3.51 mg\/mL and Vmax of 1.69 \u03bcmol\/min (for colloidal chitin). The enzyme displayed an endohydrolytic preference, converting ultrasonically pretreated chitin predominantly into chitobiose (DP2). Ultrasonication at 500 W increased the hydrolysis yield nearly 6-fold (23.86% conversion). Molecular docking identified Trp157, Glu197, Tyr198 and Arg333 as key substrate-binding residues. This work demonstrates an integrated ultrasound-enzyme strategy using BhGH18 for sustainable production of DP2-enriched chitin oligosaccharides.<\/jats:p>","DOI":"10.1021\/acs.jafc.6c07475","type":"journal-article","created":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T12:02:38Z","timestamp":1786708958000},"page":"26610-26624","source":"Crossref","is-referenced-by-count":2,"title":["Purification and\nCharacterization of a Newly Identified\nGH18 Chitinase BhGH18 from\n                    <i>Bacillus haynesii<\/i>\n                    and Its Ultrasound-Assisted Hydrolysis of Chitin"],"prefix":"10.1021","volume":"74","author":[{"ORCID":"https:\/\/orcid.org\/0009-0006-3188-6440","authenticated-orcid":true,"given":"Bicheng","family":"Liang","sequence":"first","affiliation":[{"name":"Chinese Academy of Sciences , , ,","place":["Qingdao, China, 266071"]},{"name":"University of Chinese Academy of Sciences , ,","place":["Beijing, China, 100000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chen","family":"Zhang","sequence":"additional","affiliation":[{"name":"Chinese Academy of Sciences , , ,","place":["Qingdao, China, 266071"]},{"name":"University of Chinese Academy of Sciences , ,","place":["Beijing, China, 100000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guantian","family":"Li","sequence":"additional","affiliation":[{"name":"Chinese Academy of Sciences , , ,","place":["Qingdao, China, 266071"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haoyue","family":"Yang","sequence":"additional","affiliation":[{"name":"Chinese Academy of Sciences , , ,","place":["Qingdao, China, 266071"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8862-9943","authenticated-orcid":true,"given":"Kecheng","family":"Li","sequence":"additional","affiliation":[{"name":"Chinese Academy of Sciences , , ,","place":["Qingdao, China, 266071"]},{"name":"Qingdao Marine Science and Technology Center , , ,","place":["Qingdao, China, 266237"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9271-833X","authenticated-orcid":true,"given":"Rongfeng","family":"Li","sequence":"additional","affiliation":[{"name":"Chinese Academy of Sciences , , ,","place":["Qingdao, China, 266071"]},{"name":"Qingdao Marine Science and Technology Center , , ,","place":["Qingdao, China, 266237"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Song","family":"Liu","sequence":"additional","affiliation":[{"name":"Chinese Academy of Sciences , , ,","place":["Qingdao, China, 266071"]},{"name":"Qingdao Marine Science and Technology Center , , ,","place":["Qingdao, China, 266237"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huahua","family":"Yu","sequence":"additional","affiliation":[{"name":"Chinese Academy of Sciences , , ,","place":["Qingdao, China, 266071"]},{"name":"Qingdao Marine Science and Technology Center , , ,","place":["Qingdao, China, 266237"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9745-8808","authenticated-orcid":true,"given":"Ronge","family":"Xing","sequence":"additional","affiliation":[{"name":"Chinese Academy of Sciences , , ,","place":["Qingdao, China, 266071"]},{"name":"Qingdao Marine Science and Technology Center , , ,","place":["Qingdao, China, 266237"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,14]]},"reference":[{"key":"2026090104480308700_cit1","doi-asserted-by":"publisher","first-page":"3642","DOI":"10.1080\/10408398.2019.1703170","article-title":"The potential of\ninsects as food sources\u2013a review","volume":"60","author":"de Carvalho","year":"2020","journal-title":"Crit.\nRev. 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