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J. Mol. Sci.","IJMS"],"abstract":"<jats:p>Xylan, the principal component of hemicellulose, requires the concerted action of endo- and exo-acting xylanolytic enzymes for its complete depolymerization into fermentable sugars. This study reports the biochemical and thermodynamic characterization of a glycoside hydrolase family 10 endo-\u03b2-1,4-xylanase from the thermophilic bacterium Geobacillus stearothermophilus CECT43 (GsXynA), cloned and heterologously expressed in Escherichia coli BL21(DE3) and purified to homogeneity by immobilized metal-ion affinity chromatography. The 338-residue enzyme behaved as a monomer and showed maximal activity on xylan substrates at pH 6.5 and 75 \u00b0C, retaining over 90% residual activity between pH 5 and 9 and remaining stable up to 70 \u00b0C, after 60 min of incubation. Kinetic and thermodynamic analyses of thermal inactivation monitored by enzymatic activity measurements revealed an enthalpy-driven denaturation process, while circular dichroism indicated that this irreversible unfolding is kinetically, not thermodynamically, controlled. GsXynA hydrolyzed beechwood xylan mainly to xylobiose and xylotetraose, consistent with mixed endo and exo activity, and was uncompetitively inhibited by 4-hydroxybenzoic acid. Combined action with the \u03b2-xylosidase GsXynB2 markedly increased D-xylose release, reaching a 53.23% hydrolysis yield and a degree of synergy of 3.79 after 24 h. These results demonstrate that GsXynA is a thermostable, versatile xylanase with promising potential for lignocellulosic biomass bioconversion.<\/jats:p>","DOI":"10.3390\/ijms27167339","type":"journal-article","created":{"date-parts":[[2026,8,17]],"date-time":"2026-08-17T12:01:26Z","timestamp":1786968086000},"page":"7339","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Endo-\u03b2-1,4-Xylanase GH10 from Geobacillus stearothermophilus CECT43: Biochemical Characterization, Insights into Its Thermal Behavior, and Synergistic Effects with a \u03b2-Xylosidase"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-3271-2614","authenticated-orcid":false,"given":"Alba","family":"Carrillo-Moreno","sequence":"first","affiliation":[{"name":"Departamento de Qu\u00edmica y F\u00edsica, Universidad de Almer\u00eda, 04120 Almer\u00eda, Spain"},{"name":"CIAMBITAL, CeiA3, University of Almer\u00eda, 04120 Almer\u00eda, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-1526-7847","authenticated-orcid":false,"given":"Mar\u00eda Gabriela","family":"\u00c1lvarez-Rodr\u00edguez","sequence":"additional","affiliation":[{"name":"Instituto de Biotecnolog\u00eda Sanitaria de Elche (IDIBE), Universidad Miguel Hern\u00e1ndez, 03202 Elche, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6675-1550","authenticated-orcid":false,"given":"Sergio","family":"Mart\u00ednez-Rodr\u00edguez","sequence":"additional","affiliation":[{"name":"Departamento de Bioqu\u00edmica y Biolog\u00eda Molecular III e Inmunolog\u00eda, Universidad de Granada, 18071 Granada, Spain"},{"name":"Raw Materials, Human and Environmental Health, UGR, Associated Unit of the CSIC by the IACT-CSIC, 18100 Granada, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3696-1319","authenticated-orcid":false,"given":"Pablo","family":"Soriano-Maldonado","sequence":"additional","affiliation":[{"name":"Departamento de Qu\u00edmica y F\u00edsica, Universidad de Almer\u00eda, 04120 Almer\u00eda, Spain"},{"name":"CIAMBITAL, CeiA3, University of Almer\u00eda, 04120 Almer\u00eda, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9905-4185","authenticated-orcid":false,"given":"Josefa Mar\u00eda","family":"Clemente-Jim\u00e9nez","sequence":"additional","affiliation":[{"name":"Departamento de Qu\u00edmica y F\u00edsica, Universidad de Almer\u00eda, 04120 Almer\u00eda, Spain"},{"name":"CIAMBITAL, CeiA3, University of Almer\u00eda, 04120 Almer\u00eda, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5374-9088","authenticated-orcid":false,"given":"Francisco Javier Las","family":"Heras-V\u00e1zquez","sequence":"additional","affiliation":[{"name":"Departamento de Qu\u00edmica y F\u00edsica, Universidad de Almer\u00eda, 04120 Almer\u00eda, Spain"},{"name":"CIAMBITAL, CeiA3, University of Almer\u00eda, 04120 Almer\u00eda, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9424-9602","authenticated-orcid":false,"given":"Lellys M.","family":"Contreras","sequence":"additional","affiliation":[{"name":"Departamento de Qu\u00edmica y F\u00edsica, Universidad de Almer\u00eda, 04120 Almer\u00eda, Spain"},{"name":"CIAMBITAL, CeiA3, University of Almer\u00eda, 04120 Almer\u00eda, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1186\/s13068-020-01744-6","article-title":"Engineering microbial pathways for production of bio-based chemicals from lignocellulosic sugars: Current status and perspectives","volume":"13","author":"Francois","year":"2020","journal-title":"Biotechnol. 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