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However, immunogenic side-effects restrict its application and frequently result in stopping treatment. There is a current need for the identification of novel L-asparaginase with improved properties and lower adverse effects compared to those available in the market. L-asparaginase gene from\n                    <jats:italic>Stenotrophomonas maltophilia<\/jats:italic>\n                    (\n                    <jats:italic>S. maltophilia<\/jats:italic>\n                    ), an isolated organism that was mentioned as a novel and excellent source for L- asparaginase, was cloned and expressed using\n                    <jats:italic>E. coli<\/jats:italic>\n                    DH5\u03b1 and\n                    <jats:italic>E. coli<\/jats:italic>\n                    BL21(DE3). Investigations of different conditions of expression of recombinant L-asparaginase in\n                    <jats:italic>E. coli<\/jats:italic>\n                    BL21(DE3) using Box\u2013Behnken design predicted maximum expression at 37\u00a0\u00b0C temperature, 250\u00a0rpm agitation, 0.83 mM isopropylthio-\u03b2-D-galactoside (IPTG) concentration after incubation for 17\u00a0h. The optimized expression conditions were validated using L-asparaginase activity assay. The obtained recombinant protein was purified using Ni-NTA spin column. SDS-PAGE demonstrated a single band of 17 KDa apparent molecular weight. The kinetic parameters were determined, and they exhibited a low Km value of 2.94\u2009\u00d7\u200910\n                    <jats:sup>\u2212\u20092<\/jats:sup>\n                    M and Vmax of 14.73 IU\/ml. Cytotoxicity on various cell lines was tested in relation to marketed\n                    <jats:italic>E. coli<\/jats:italic>\n                    L-asparaginase and exhibited low IC50 of 1.92 IU\/ml and 2.03 IU\/ml for HEP-G2 and K-562 cell lines, respectively. Additionally, mice treated with recombinant L-asparaginase displayed a significantly lower immunological response (IgG) compared to mice treated with marketed\n                    <jats:italic>E. coli<\/jats:italic>\n                    L-asparaginase (\n                    <jats:italic>p<\/jats:italic>\n                    -value\u2009&lt;\u20090.0001). These findings demonstrate the potentiality of recombinant L-asparaginase for its development as a chemotherapeutic drug.\n                  <\/jats:p>","DOI":"10.1186\/s12934-025-02856-0","type":"journal-article","created":{"date-parts":[[2025,12,19]],"date-time":"2025-12-19T06:05:54Z","timestamp":1766124354000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Recombinant L-asparaginase from Stenotrophomonas maltophilia: a promising low-immunogenic anticancer agent"],"prefix":"10.1186","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8082-2298","authenticated-orcid":false,"given":"Nada A.","family":"Abdelrazek","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4809-4451","authenticated-orcid":false,"given":"Sarra E.","family":"Saleh","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0629-9245","authenticated-orcid":false,"given":"Amal E.","family":"Ali","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0828-1420","authenticated-orcid":false,"given":"Mohammad M.","family":"Aboulwafa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5614-5127","authenticated-orcid":false,"given":"Marwa M.","family":"Raafat","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,12,19]]},"reference":[{"key":"2856_CR1","doi-asserted-by":"publisher","first-page":"68","DOI":"10.1016\/j.copbio.2020.12.002","volume":"69","author":"A Fasim","year":"2021","unstructured":"Fasim A, More VS, More SS. 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