{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,23]],"date-time":"2026-08-23T18:26:41Z","timestamp":1787509601586,"version":"build-2736575974"},"reference-count":50,"publisher":"Public Library of Science (PLoS)","issue":"2","license":[{"start":{"date-parts":[[2026,2,13]],"date-time":"2026-02-13T00:00:00Z","timestamp":1770940800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>\n                    In this study, L\u2012asparaginase production in several endophytic fungi was evaluated along with their L\u2012glutaminase and urease co-activities. The effect of L\u2012asparagine and different culture media on L-asparaginase production were also evaluated. Among the 62 investigated isolates, 49 isolates exhibited L\u2012asparaginase activity, and the maximum zone index (6.58) was observed in\n                    <jats:italic>Cladosporium perangustum<\/jats:italic>\n                    EL1. Evaluation of L\u2012glutaminase and urease co-activities in L\u2012asparaginase-positive isolates screened 19 isolates with no L\u2012glutaminase activity and four isolates with minimum urease production. L\u2012asparaginase activity was quantified in 12 selected isolates using the Nesslerization method.\n                    <jats:italic>Cladosporium cladosporioides<\/jats:italic>\n                    Kr5\u22122 exhibited the maximum L\u2012asparaginase activity (10.78 U mL\n                    <jats:sup>-1<\/jats:sup>\n                    ).\n                    <jats:italic>Alternaria brassicae<\/jats:italic>\n                    C showed high L\u2012asparaginase activity (7.07 U mL\n                    <jats:sup>-1<\/jats:sup>\n                    ) free of L\u2012glutaminase, and low urease co-activity (1.97 U mL\n                    <jats:sup>-1<\/jats:sup>\n                    ). Assessment of the effect of L\u2012asparagine on L\u2012asparaginase activity showed that the enzyme is inducible and substrate-regulated. Evaluation of ten different culture media showed that all isolates were able to produce L\u2012asparaginase on Mineral salts agar and Citrate agar culture media. Also, Cerelose ammonium nitrate agar, Kuehner basal culture medium, and Piefer, Humphrey, and Acree culture medium inhibited L\u2012asparaginase production in the majority of the isolates. This is the first report of L\u2012asparaginase production by endophytic fungi isolated from\n                    <jats:italic>Taxus baccata<\/jats:italic>\n                    ,\n                    <jats:italic>Pistacia vera<\/jats:italic>\n                    ,\n                    <jats:italic>Prunus avium<\/jats:italic>\n                    ,\n                    <jats:italic>Prunus cerasus<\/jats:italic>\n                    , and\n                    <jats:italic>Punica granatum<\/jats:italic>\n                    , as well as the investigation of their L\u2012glutaminase and urease co-activities. Among the evaluated culture media, Mineral salts agar and Citrate agar culture media are suggested here as alternate for MCD. Moreover,\n                    <jats:italic>Alternaria brassicae<\/jats:italic>\n                    C is recommended as a promising isolate for future commercial L\u2012asparaginase production.\n                  <\/jats:p>","DOI":"10.1371\/journal.pone.0339829","type":"journal-article","created":{"date-parts":[[2026,2,13]],"date-time":"2026-02-13T18:43:11Z","timestamp":1771008191000},"page":"e0339829","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":1,"title":["L\u2012asparaginase activity in some endophytic fungi: Glutaminase\u2012free and low urease co\u2012activities"],"prefix":"10.1371","volume":"21","author":[{"given":"Zahra","family":"Zaeimian","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4790-6171","authenticated-orcid":true,"given":"Khalil-Berdi","family":"Fotouhifar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohsen","family":"Farzaneh","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2026,2,13]]},"reference":[{"key":"pone.0339829.ref001","article-title":"Acute lymphoblastic leukemia","author":"JH-C Chang","year":"2021","journal-title":"Pediatr Blood Cancer"},{"issue":"9","key":"pone.0339829.ref002","doi-asserted-by":"crossref","first-page":"1926","DOI":"10.3390\/jcm10091926","article-title":"Advances in the diagnosis and treatment of pediatric acute lymphoblastic leukemia","volume":"10","author":"H Inaba","year":"2021","journal-title":"J Clin Med"},{"issue":"35","key":"pone.0339829.ref003","doi-asserted-by":"crossref","first-page":"3193","DOI":"10.1021\/acs.biochem.0c00354","article-title":"Improving the treatment of acute lymphoblastic leukemia","volume":"59","author":"A Radadiya","year":"2020","journal-title":"Biochemistry"},{"key":"pone.0339829.ref004","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.jbiotec.2023.10.001","article-title":"Screening of endophytic fungi from Antarctic mosses: Potential production for L-asparaginase free of glutaminase and urease activity","volume":"377","author":"G Afonso Kessler de Andrade","year":"2023","journal-title":"J Biotechnol"},{"issue":"3","key":"pone.0339829.ref005","article-title":"Selection of the optimal L-asparaginase II against acute lymphoblastic leukemia: an in silico approach","volume":"2","author":"A Baral","year":"2021","journal-title":"JMIRx Med"},{"issue":"6","key":"pone.0339829.ref006","doi-asserted-by":"crossref","first-page":"1549","DOI":"10.1158\/0008-5472.CAN-17-2106","article-title":"A novel l-asparaginase with low l-glutaminase coactivity is highly efficacious against both T- and B-cell acute lymphoblastic leukemias in vivo","volume":"78","author":"HA Nguyen","year":"2018","journal-title":"Cancer Res"},{"key":"pone.0339829.ref007","doi-asserted-by":"crossref","first-page":"111616","DOI":"10.1016\/j.biopha.2021.111616","article-title":"Circumventing the side effects of L-asparaginase","volume":"139","year":"2021","journal-title":"Biomed Pharmacother"},{"issue":"1","key":"pone.0339829.ref008","doi-asserted-by":"crossref","first-page":"17861","DOI":"10.1038\/s41598-021-97316-1","article-title":"Sequencing and characterization of an L-asparaginase gene from a new species of Penicillium section Citrina isolated from Cerrado","volume":"11","author":"KCR Andrade","year":"2021","journal-title":"Sci Rep"},{"issue":"4","key":"pone.0339829.ref009","doi-asserted-by":"crossref","first-page":"723","DOI":"10.3390\/cancers16040723","article-title":"Treatment of pediatric acute lymphoblastic leukemia: a historical perspective","volume":"16","author":"H Hayashi","year":"2024","journal-title":"Cancers (Basel)"},{"key":"pone.0339829.ref010","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1016\/j.enzmictec.2018.06.005","article-title":"In silico modelling and molecular dynamics simulation studies on L-Asparaginase isolated from bacterial endophyte of Ocimum tenuiflorum","volume":"117","author":"M Pola","year":"2018","journal-title":"Enzyme Microb Technol"},{"issue":"1","key":"pone.0339829.ref011","doi-asserted-by":"crossref","first-page":"29625","DOI":"10.1038\/s41598-024-77867-9","article-title":"Artificial intelligence-based optimization for extracellular L-glutaminase free L-asparaginase production by Streptomyces violaceoruber under solid state fermentation conditions","volume":"14","author":"NE-A El-Naggar","year":"2024","journal-title":"Sci Rep"},{"issue":"1","key":"pone.0339829.ref012","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1080\/10826068.2018.1536990","article-title":"Optimization of culture conditions and bench-scale production of anticancer enzyme L-asparaginase by submerged fermentation from Aspergillus terreus CCT 7693","volume":"49","author":"TA Costa-Silva","year":"2019","journal-title":"Prep Biochem Biotechnol"},{"issue":"1","key":"pone.0339829.ref013","doi-asserted-by":"crossref","first-page":"200","DOI":"10.3390\/catal13010200","article-title":"Anticancer Asparaginases: Perspectives in Using Filamentous Fungi as Cell Factories","volume":"13","author":"PHD Garcia","year":"2023","journal-title":"Catalysts"},{"key":"pone.0339829.ref014","doi-asserted-by":"crossref","first-page":"134547","DOI":"10.1016\/j.seppur.2025.134547","article-title":"Production, extraction, and purification of L-asparaginase from Penicillium sizovae: aqueous biphasic systems as an effective pre-concentration strategy","volume":"378","author":"MB Guimar\u00e3es","year":"2025","journal-title":"Separation and Purification Technology"},{"issue":"3","key":"pone.0339829.ref015","doi-asserted-by":"crossref","first-page":"100","DOI":"10.3390\/applmicrobiol5030100","article-title":"Characterization of L-asparaginase free of L-glutaminase and urease activity produced by the marine paraconiothyrium cyclothyrioides strain MABIK FU00000820","volume":"5","author":"W-J Yu","year":"2025","journal-title":"Applied Microbiology"},{"issue":"9","key":"pone.0339829.ref016","doi-asserted-by":"crossref","first-page":"1587","DOI":"10.1158\/1535-7163.MCT-18-1329","article-title":"Glutaminase activity of L-asparaginase contributes to durable preclinical activity against acute lymphoblastic leukemia","volume":"18","author":"W-K Chan","year":"2019","journal-title":"Mol Cancer Ther"},{"issue":"10","key":"pone.0339829.ref017","doi-asserted-by":"crossref","first-page":"152","DOI":"10.1007\/s11274-019-2731-9","article-title":"What makes a good new therapeutic L-asparaginase?","volume":"35","author":"A Beckett","year":"2019","journal-title":"World J Microbiol Biotechnol"},{"key":"pone.0339829.ref018","doi-asserted-by":"crossref","first-page":"130312","DOI":"10.1016\/j.ijbiomac.2024.130312","article-title":"Homology modeling and molecular docking studies to decrease glutamine affinity of Yarrowia lipolytica L-asparaginase","volume":"263","author":"F Darvishi","year":"2024","journal-title":"Int J Biol Macromol"},{"key":"pone.0339829.ref019","doi-asserted-by":"crossref","first-page":"102958","DOI":"10.1016\/j.bcab.2023.102958","article-title":"Antitumor activity of urease-free l-asparaginase with low glutaminase coactivity produced by marine-derived Aspergillus flavus","volume":"54","author":"Z Nur\u00e7e","year":"2023","journal-title":"Biocatalysis and Agricultural Biotechnology"},{"issue":"11","key":"pone.0339829.ref020","doi-asserted-by":"crossref","first-page":"4515","DOI":"10.1007\/s00253-021-11359-y","article-title":"L-asparaginase production review: bioprocess design and biochemical characteristics","volume":"105","author":"D Castro","year":"2021","journal-title":"Appl Microbiol Biotechnol"},{"key":"pone.0339829.ref021","doi-asserted-by":"crossref","first-page":"101398","DOI":"10.1016\/j.imu.2023.101398","article-title":"In-silico evaluation of fungal and bacterial L-asparaginases allergenicity","volume":"43","author":"T Sisay","year":"2023","journal-title":"Informatics in Medicine Unlocked"},{"issue":"1","key":"pone.0339829.ref022","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1186\/s12934-023-02118-x","article-title":"Bioactive compounds and biomedical applications of endophytic fungi: a recent review","volume":"22","author":"AH Hashem","year":"2023","journal-title":"Microb Cell Fact"},{"key":"pone.0339829.ref023","first-page":"1","article-title":"Secondary metabolites from endophytic fungi: Production, methods of analysis, and diverse pharmaceutical potential","author":"VK Singh","year":"2023","journal-title":"Symbiosis"},{"key":"pone.0339829.ref024","doi-asserted-by":"crossref","first-page":"102055","DOI":"10.1016\/j.fochx.2024.102055","article-title":"Recent advances in L-Asparaginase enzyme production and formulation development for acrylamide reduction during food processing","volume":"25","author":"A Jana","year":"2024","journal-title":"Food Chem X"},{"issue":"10","key":"pone.0339829.ref025","volume":"21","year":"2023","journal-title":"EFSA J"},{"issue":"4","key":"pone.0339829.ref026","article-title":"Safety evaluation of the food enzyme asparaginase from the genetically modified Aspergillus oryzae strain NZYM-OA","volume":"21","year":"2023","journal-title":"EFSA J"},{"issue":"7","key":"pone.0339829.ref027","article-title":"Safety evaluation of the food enzyme asparaginase from the genetically modified Aspergillus niger strain ASP","volume":"22","year":"2024","journal-title":"EFSA J"},{"issue":"2","key":"pone.0339829.ref028","doi-asserted-by":"crossref","first-page":"360","DOI":"10.3390\/microorganisms10020360","article-title":"Endophytic fungi: key insights, emerging prospects, and challenges in natural product drug discovery","volume":"10","author":"P Tiwari","year":"2022","journal-title":"Microorganisms"},{"key":"pone.0339829.ref029","doi-asserted-by":"crossref","first-page":"100255","DOI":"10.1016\/j.scenv.2025.100255","article-title":"Production and characterization of L-asparaginase by Aspergillus niger: A sustainable use of pitaya (Hylocereus spp.) by-products","volume":"10","author":"JNB Honfoga","year":"2025","journal-title":"Sustainable Chemistry for the Environment"},{"key":"pone.0339829.ref030","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.ejbt.2024.05.002","article-title":"Isolation and characterization of fungi producing L-asparaginase with reduced L-glutaminase activity from soil samples","volume":"71","author":"T Sisay","year":"2024","journal-title":"Electronic Journal of Biotechnology"},{"key":"pone.0339829.ref031","author":"SJ Ashkezari","year":"2013"},{"key":"pone.0339829.ref032","author":"HK Dolatabad","year":"2017","journal-title":"Evaluation of antifungal of endophytic fungi isolated from some plants; improvement of efficiency of selected strains by protoplast fusion method. Dissertation, University of Tehran"},{"key":"pone.0339829.ref033","author":"MR Tarhani","year":"2017"},{"key":"pone.0339829.ref034","author":"SA Aghdam","year":"2016"},{"issue":"1","key":"pone.0339829.ref035","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1046\/j.1472-765X.1997.00331.x","article-title":"A rapid plate assay for screening L-asparaginase producing micro-organisms","volume":"24","author":"R Gulati","year":"1997","journal-title":"Lett Appl Microbiol"},{"issue":"1","key":"pone.0339829.ref036","doi-asserted-by":"crossref","first-page":"1423","DOI":"10.1038\/s41598-018-38094-1","article-title":"Microbes Producing L-Asparaginase free of glutaminase and urease isolated from extreme locations of antarctic soil and moss","volume":"9","author":"A Ashok","year":"2019","journal-title":"Sci Rep"},{"key":"pone.0339829.ref037","doi-asserted-by":"crossref","DOI":"10.7717\/peerj.8309","article-title":"Isolation and identification of L-asparaginase-producing endophytic fungi from the Asteraceae family plant species of Iran","volume":"8","author":"S Hatamzadeh","year":"2020","journal-title":"PeerJ"},{"issue":"1","key":"pone.0339829.ref038","doi-asserted-by":"crossref","first-page":"6192","DOI":"10.1038\/s41598-021-84949-5","article-title":"Bioprospecting of the agaricomycete Ganoderma australe GPC191 as novel source for L-asparaginase production","volume":"11","author":"M Chakraborty","year":"2021","journal-title":"Sci Rep"},{"issue":"3","key":"pone.0339829.ref039","article-title":"Antioxidant and L-asparaginase activities of culturable endophytic fungi from ornamental Dendrobium orchids","volume":"77","author":"RW Chua","year":"2024","journal-title":"Lett Appl Microbiol"},{"issue":"1","key":"pone.0339829.ref040","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1099\/00221287-76-1-85","article-title":"Asparaginase and glutaminase activities of micro-organisms","volume":"76","author":"A Imada","year":"1973","journal-title":"J Gen Microbiol"},{"key":"pone.0339829.ref041","article-title":"Molecular phylogenetic characterization of L-asparaginase-producing endophytic fungi inhabiting Prunus africana and Periploca linearifolia: effect of incubation time and pH on enzyme production","volume":"26","author":"DK Cheruiyot","year":"2024","journal-title":"Sci Afr"},{"key":"pone.0339829.ref042","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1016\/j.procbio.2019.11.012","article-title":"A novel multiple reactor system for the long-term production of L-asparaginase by Penicillium sp. LAMAI 505","volume":"90","author":"WF Vieira","year":"2020","journal-title":"Process Biochemistry"},{"issue":"3","key":"pone.0339829.ref043","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1080\/10826068.2021.1931881","article-title":"L-asparaginase from Aspergillus oryzae spp.: effects of production process and biochemical parameters","volume":"52","author":"MC da Cunha","year":"2022","journal-title":"Prep Biochem Biotechnol"},{"key":"pone.0339829.ref044","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1016\/S0580-9517(09)70007-8","article-title":"Chapter II fungal culture media","author":"C Booth","year":"1971","journal-title":"Methods in microbiology. Academic Press"},{"issue":"3","key":"pone.0339829.ref045","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1093\/infdis\/39.3.209","article-title":"A culture medium for differentiating organisms of typhoid-colon aerogenes groups and for isolation of certain fungi: with colored plate","volume":"39","author":"JS Simmons","year":"1926","journal-title":"Journal of Infectious Diseases"},{"key":"pone.0339829.ref046","doi-asserted-by":"crossref","first-page":"106737","DOI":"10.1016\/j.psep.2024.12.118","article-title":"Bioprospecting of microbial L-asparaginase: Sustainable production from waste and its application for acrylamide reduction in food industry","volume":"195","author":"S Mudaliar","year":"2025","journal-title":"Process Safety and Environmental Protection"},{"issue":"5","key":"pone.0339829.ref047","doi-asserted-by":"crossref","first-page":"2161","DOI":"10.1002\/bab.2276","article-title":"Purification and anticancer activity of glutaminase and urease-free l-asparaginase from novel endophyte Chaetomium sp","volume":"69","author":"N Arumugam","year":"2022","journal-title":"Biotechnol Appl Biochem"},{"issue":"3","key":"pone.0339829.ref048","article-title":"L-asparaginase from the mangrove endophyte Penicillium citrinum MEF 455: a focus on cancer surveillance gene expression in tumor cell lines HL-60 and NCI-H 460","volume":"14","author":"N Job","year":"2025","journal-title":"Toxicol Res (Camb)"},{"issue":"3","key":"pone.0339829.ref049","doi-asserted-by":"crossref","first-page":"254","DOI":"10.1080\/10242422.2025.2470140","article-title":"Purification and characterization of the L-asparaginase enzyme derived from Aspergillus flavus HK03 along with its anti-proliferative activity","volume":"43","author":"K Luhana","year":"2025","journal-title":"Biocatalysis and Biotransformation"},{"issue":"1","key":"pone.0339829.ref050","first-page":"62","article-title":"The nutritional and cultural conditions in shake flask culture for improved production of L-Asparaginase from endophytic fungus Fusarium sp. LCJ324: A sequential statistical method","volume":"52","author":"E Udayan","year":"2023","journal-title":"Ital J Mycol"}],"container-title":["PLOS One"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0339829","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,13]],"date-time":"2026-02-13T18:43:20Z","timestamp":1771008200000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0339829"}},"subtitle":[],"editor":[{"given":"Shashi Kant","family":"Bhatia","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,2,13]]},"references-count":50,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2026,2,13]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0339829","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,2,13]]}}}