{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,11]],"date-time":"2026-09-11T05:55:12Z","timestamp":1789106112971,"version":"build-2803163510"},"reference-count":73,"publisher":"MDPI AG","issue":"14","license":[{"start":{"date-parts":[[2025,7,10]],"date-time":"2025-07-10T00:00:00Z","timestamp":1752105600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Poltegor Institute","award":["order No. 337015\/N"],"award-info":[{"award-number":["order No. 337015\/N"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Materials"],"abstract":"<jats:p>Reprocessing lignocellulosic waste to obtain new products for industrial purposes is a vital part of circular economy. This paper reports the cellulase production by newly isolated Bacillus methylotrophicus cultured on lignocellulosic agro-industrial by-products, out of which brewer\u2019s spent grain (BSG) was selected as most beneficial. Plackett\u2013Burman design was used for screening medium components, while Box\u2013Behnken design was further applied to model the impact of the three most influential variables. The maximum approximated cellulase activity was 0.469 U\/mL (1 U = 1 \u00b5mol of reducing sugars\/1 min), at 48.6 g\/L substrate, 5.3 g\/L ammonium sulfate, pH 6.1. The partially purified cellulase was characterized, which demonstrated broad range of optimal pH (6.5\u20139.4), temperature (50\u201360 \u00b0C), and sensitivity to metals. Changes in lignin and pentosans content was demonstrated as a result of BSG hydrolysis with a cell-free cellulase preparation. The produced enzyme was used for hydrolysis of various chemically pretreated (NaOH and H2SO4) cellulosic substrates, where for reused alkali-pretreated BSG (after microbial enzyme production) the saccharification efficiency was at a level of 25%. The cellulolytic potential of the bacterial strain, along with its resistance to ethanol, present a beneficial combination, potentially applicable to aid saccharification of lignocellulosic by-products for biofuel production.<\/jats:p>","DOI":"10.3390\/ma18143256","type":"journal-article","created":{"date-parts":[[2025,7,11]],"date-time":"2025-07-11T08:15:47Z","timestamp":1752221747000},"page":"3256","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Cellulolytic Potential of Newly Isolated Alcohol-Tolerant Bacillus methylotrophicus"],"prefix":"10.3390","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2413-9923","authenticated-orcid":false,"given":"Anna","family":"Choi\u0144ska-Pulit","sequence":"first","affiliation":[{"name":"\u201cPoltegor-Institute\u201d, Opencast Mining Institute, Parkowa 25, 51-616 Wroc\u0142aw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3221-6979","authenticated-orcid":false,"given":"Justyna","family":"Sobolczyk-Bednarek","sequence":"additional","affiliation":[{"name":"\u201cPoltegor-Institute\u201d, Opencast Mining Institute, Parkowa 25, 51-616 Wroc\u0142aw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2068-3641","authenticated-orcid":false,"given":"Wojciech","family":"\u0141aba","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Food Microbiology, Faculty of Biotechnology and Food Science, Wroc\u0142aw University of Environmental and Life Sciences, Che\u0142mo\u0144skiego 37, 51-630 Wroc\u0142aw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,7,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.jcs.2005.06.001","article-title":"Brewers\u2019 spent grain: Generation, characteristics and potential applications","volume":"43","author":"Mussatto","year":"2006","journal-title":"J. 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