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This enzyme has significant biotechnological applications, functioning as an antineoplastic agent in the pharmaceutical industry and as a tool for reducing acrylamide formation in the food industry. This study aimed to identify a filamentous fungus strain capable of synthesizing a new\n                    <jats:sc>L<\/jats:sc>\n                    -asparaginase and to perform its biochemical characterisation. Among the fungal isolates screened using plate assays and submerged fermentation,\n                    <jats:italic>Cunninghamella echinulata<\/jats:italic>\n                    strain PA3S12MM was identified as a promising\n                    <jats:sc>L<\/jats:sc>\n                    -asparaginase producer. The highest enzymatic induction was achieved using 5% (w\/v) glucose as the carbon source and 0.4% (w\/v) chicken feather as an agro-industrial nitrogen source, under incubation for 120\u00a0h. The enzyme exhibited an apparent\n                    <jats:italic>K<\/jats:italic>\n                    <jats:sub>m app<\/jats:sub>\n                    of 0.011 mmol L\u207b\u00b9 and a\n                    <jats:italic>V<\/jats:italic>\n                    <jats:sub>max app<\/jats:sub>\n                    of 126.5 mmol L\u207b\u00b9 min\u207b\u00b9 for the hydrolysis of L-asparagine. Partial purification revealed that the enzyme displayed optimal activity at pH 7.0 and a temperature of 85\u00a0\u00b0C. In terms of substrate specificity, the enzyme demonstrated 100% relative activity toward L-asparagine and 32.9% toward L-glutamine. Enzymatic activity was enhanced by \u03b2-mercaptoethanol but inhibited by Mn\u00b2\u207a, L-cysteine, and dithiothreitol (DTT). These findings suggest that\n                    <jats:italic>C. echinulata<\/jats:italic>\n                    PA3S12MM is a promising candidate for large-scale biotechnological production of a new fungal\n                    <jats:sc>L<\/jats:sc>\n                    -asparaginase, with biochemical indicators of potential for applications in both pharmaceutical and food industries.\n                  <\/jats:p>","DOI":"10.1007\/s00203-026-04984-6","type":"journal-article","created":{"date-parts":[[2026,6,6]],"date-time":"2026-06-06T06:21:17Z","timestamp":1780726877000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Biochemical characterisation of a new thermotolerant L-asparaginase produced by Cunninghamella echinulata PA3S12MM isolated from the Atlantic Forest"],"prefix":"10.1007","volume":"208","author":[{"given":"Simone","family":"Von Groll","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leticia Claudia","family":"da Silva","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pedro Henrique Nascimento","family":"Souza","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"W\u00e1llison Justino","family":"da Silva","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tha\u00eds Duarte","family":"Bifano","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jos\u00e9 Luis da Concei\u00e7\u00e3o","family":"Silva","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rita de C\u00e1ssia Garcia","family":"Sim\u00e3o","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"family":"Sim\u00e3o","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marina Kimiko","family":"Kadowaki","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexandre","family":"Maller","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,6,6]]},"reference":[{"key":"4984_CR1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pep.2021.106006","volume":"190","author":"N Arumugam","year":"2022","unstructured":"Arumugam N, Thangavelu P (2022) Purification and anticancer activity of glutaminase and urease free intracellular L-asparaginase from Chaetomium sp. 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