{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T06:16:09Z","timestamp":1788416169372,"version":"build-2803163510"},"reference-count":28,"publisher":"Springer Science and Business Media LLC","issue":"4","license":[{"start":{"date-parts":[[2025,7,11]],"date-time":"2025-07-11T00:00:00Z","timestamp":1752192000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2025,7,11]],"date-time":"2025-07-11T00:00:00Z","timestamp":1752192000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100016367","name":"Vysok\u00e1 \u0160kola Chemicko-technologick\u00e1 v Praze","doi-asserted-by":"publisher","award":["A1_FPBT_2024_001"],"award-info":[{"award-number":["A1_FPBT_2024_001"]}],"id":[{"id":"10.13039\/501100016367","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100016367","name":"Vysok\u00e1 \u0160kola Chemicko-technologick\u00e1 v Praze","doi-asserted-by":"publisher","award":["A2_FPBT_2024_024"],"award-info":[{"award-number":["A2_FPBT_2024_024"]}],"id":[{"id":"10.13039\/501100016367","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002969","name":"Technologick\u00e1 Agentura \u010cesk\u00e9 Republiky","doi-asserted-by":"publisher","award":["TQ03000738"],"award-info":[{"award-number":["TQ03000738"]}],"id":[{"id":"10.13039\/501100002969","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001823","name":"Ministerstvo \u0160kolstv\u00ed, Ml\u00e1de\u017ee a T\u011blov\u00fdchovy","doi-asserted-by":"publisher","award":["LUC23140"],"award-info":[{"award-number":["LUC23140"]}],"id":[{"id":"10.13039\/501100001823","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001823","name":"Ministerstvo \u0160kolstv\u00ed, Ml\u00e1de\u017ee a T\u011blov\u00fdchovy","doi-asserted-by":"publisher","award":["LUC23140"],"award-info":[{"award-number":["LUC23140"]}],"id":[{"id":"10.13039\/501100001823","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001823","name":"Ministerstvo \u0160kolstv\u00ed, Ml\u00e1de\u017ee a T\u011blov\u00fdchovy","doi-asserted-by":"publisher","award":["LUC23140"],"award-info":[{"award-number":["LUC23140"]}],"id":[{"id":"10.13039\/501100001823","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100016367","name":"University of Chemistry and Technology, Prague","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100016367","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Folia Microbiol"],"published-print":{"date-parts":[[2026,8]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    <jats:sc>l<\/jats:sc>\n                    -Asparaginases are enzymes known for decades due to their use in medicine for the treatment of acute lymphoblastic leukemia. Recently, they have also found application in the food industry, and other possibilities are emerging in the treatment of infectious diseases or in the design of biosensors. For this reason, an ongoing effort has been made to find and characterize new enzymes with properties suitable for these specific applications. In this work,\n                    <jats:sc>l<\/jats:sc>\n                    -asparaginase from\n                    <jats:italic>Paenibacillus thiaminolyticus<\/jats:italic>\n                    (isoenzyme 1) belonging to the least explored group of\n                    <jats:sc>l<\/jats:sc>\n                    -asparaginases derived from\n                    <jats:sc>l<\/jats:sc>\n                    -asparaginase from\n                    <jats:italic>Rhizobium etli<\/jats:italic>\n                    was recombinantly produced with high yields (335\u00a0mg per L of culture medium) in\n                    <jats:italic>E. coli<\/jats:italic>\n                    cells and characterized:\n                    <jats:italic>K<\/jats:italic>\n                    <jats:sub>\n                      <jats:italic>M<\/jats:italic>\n                    <\/jats:sub>\n                    \u2009=\u2009(26\u2009\u00b1\u20098) mmol L\n                    <jats:sup>\u22121<\/jats:sup>\n                    , pH\n                    <jats:sub>opt<\/jats:sub>\n                    \u2009=\u200910.3, without\n                    <jats:sc>l<\/jats:sc>\n                    -glutaminase and urease activity. A probable oligomeric structure (homodimer) was derived by computer modeling and confirmed by gel chromatography experiments. The results of this work extend the current limited knowledge about the poorly described class of\n                    <jats:italic>R. etli<\/jats:italic>\n                    <jats:sc>l<\/jats:sc>\n                    -asparaginases. Moreover, this\n                    <jats:sc>l<\/jats:sc>\n                    -asparaginase exhibits suitable properties for use in biosensor construction because of the high yields during recombinant production,\n                    <jats:italic>K<\/jats:italic>\n                    <jats:sub>\n                      <jats:italic>M<\/jats:italic>\n                    <\/jats:sub>\n                    value, stability, and absence of\u00a0\n                    <jats:sc>l<\/jats:sc>\n                    -glutaminase activity.\n                  <\/jats:p>","DOI":"10.1007\/s12223-025-01296-y","type":"journal-article","created":{"date-parts":[[2025,7,11]],"date-time":"2025-07-11T03:47:20Z","timestamp":1752205640000},"page":"875-887","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["High-yield production and functional analysis of novel rhizobial-type glutaminase-free l-asparaginase from Paenibacillus thiaminolyticus"],"prefix":"10.1007","volume":"71","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9621-7516","authenticated-orcid":false,"given":"Tom\u00e1\u0161","family":"Podzimek","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-8824-9355","authenticated-orcid":false,"given":"Karol\u00edna","family":"Lou\u017eeck\u00e1","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-6468-993X","authenticated-orcid":false,"given":"Veronika","family":"Urb\u00e1nkov\u00e1","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5347-3966","authenticated-orcid":false,"given":"Jan","family":"Ber\u00e1nek","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2275-3479","authenticated-orcid":false,"given":"Petra","family":"Lipovov\u00e1","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3696-3517","authenticated-orcid":false,"given":"Eva","family":"Bene\u0161ov\u00e1","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,7,11]]},"reference":[{"key":"1296_CR1","doi-asserted-by":"publisher","first-page":"113177","DOI":"10.1016\/j.foodres.2023.113177","volume":"172","author":"E Abedi","year":"2023","unstructured":"Abedi E, Hashemi SMB, Ghiasi F (2023) Effective mitigation in the amount of acrylamide through enzymatic approaches. Food Res Int 172:113177. https:\/\/doi.org\/10.1016\/j.foodres.2023.113177","journal-title":"Food Res Int"},{"key":"1296_CR2","doi-asserted-by":"publisher","first-page":"493","DOI":"10.1038\/s41586-024-07487-w","volume":"630","author":"J Abramson","year":"2024","unstructured":"Abramson J, Adler J, Dunger J et al (2024) Accurate structure prediction of biomolecular interactions with AlphaFold 3. Nature 630:493\u2013500. https:\/\/doi.org\/10.1038\/s41586-024-07487-w","journal-title":"Nature"},{"key":"1296_CR3","doi-asserted-by":"publisher","first-page":"75","DOI":"10.1016\/s0928-0987(99)00047-0","volume":"9","author":"K Arii","year":"1999","unstructured":"Arii K, Kobayashi H, Kai T, Kokuba Y (1999) Degradation kinetics of l-glutamine in aqueous solution. Eur J Pharm Sci 9:75\u201378. https:\/\/doi.org\/10.1016\/s0928-0987(99)00047-0","journal-title":"Eur J Pharm Sci"},{"key":"1296_CR4","doi-asserted-by":"publisher","first-page":"375","DOI":"10.1134\/S0006297910030144","volume":"75","author":"S Bansal","year":"2010","unstructured":"Bansal S, Gnaneswari D, Mishra P et al (2010) Structural stability and functional analysis of l-asparaginase from Pyrococcus furiosus. Biochemistry (Mosc) 75:375\u2013381. https:\/\/doi.org\/10.1134\/S0006297910030144","journal-title":"Biochemistry (Mosc)"},{"key":"1296_CR5","doi-asserted-by":"publisher","first-page":"1268","DOI":"10.1124\/dmd.118.081539","volume":"46","author":"TE Bapiro","year":"2018","unstructured":"Bapiro TE, Sykes A, Martin S et al (2018) Complete substrate inhibition of cytochrome P450 2C8 by AZD9496, an oral selective estrogen receptor degrader. Drug Metab Dispos 46:1268\u20131276. https:\/\/doi.org\/10.1124\/dmd.118.081539","journal-title":"Drug Metab Dispos"},{"key":"1296_CR6","doi-asserted-by":"publisher","first-page":"14","DOI":"10.3390\/pharmaceutics13010014","volume":"13","author":"T Barros","year":"2021","unstructured":"Barros T, Brumano L, Freitas M et al (2021) Development of processes for recombinant l-asparaginase II production by Escherichia coli BL21 (DE3): from shaker to bioreactors. Pharmaceutics 13:14. https:\/\/doi.org\/10.3390\/pharmaceutics13010014","journal-title":"Pharmaceutics"},{"key":"1296_CR7","doi-asserted-by":"publisher","first-page":"97","DOI":"10.1016\/j.cell.2013.12.007","volume":"156","author":"M Baruch","year":"2014","unstructured":"Baruch M, Belotserkovsky I, Hertzog BB et al (2014) An extracellular bacterial pathogen modulates host metabolism to regulate its own sensing and proliferation. Cell 156:97\u2013108. https:\/\/doi.org\/10.1016\/j.cell.2013.12.007","journal-title":"Cell"},{"key":"1296_CR8","doi-asserted-by":"publisher","first-page":"900","DOI":"10.1007\/s12010-015-1917-3","volume":"178","author":"T Batool","year":"2016","unstructured":"Batool T, Makky EA, Jalal M et al (2016) A comprehensive review on l-asparaginase and its applications. Appl Biochem Biotechnol 178:900\u2013923. https:\/\/doi.org\/10.1007\/s12010-015-1917-3","journal-title":"Appl Biochem Biotechnol"},{"key":"1296_CR9","doi-asserted-by":"publisher","first-page":"99","DOI":"10.1084\/jem.118.1.121","volume":"118","author":"JD Broome","year":"1963","unstructured":"Broome JD (1963) Evidence that the l-asparaginase of guinea pig serum is responsible for its antilymphoma effects. I. Properties of the l-asparaginase of guinea pig serum in relation to those of the antilymphoma substance. J Exp Med 118:99\u2013120. https:\/\/doi.org\/10.1084\/jem.118.1.121","journal-title":"J Exp Med"},{"key":"1296_CR10","doi-asserted-by":"publisher","first-page":"4515","DOI":"10.1007\/s00253-021-11359-y","volume":"105","author":"D Castro","year":"2021","unstructured":"Castro D, Marques ASC, Almeida MR et al (2021) l-Asparaginase production review: bioprocess design and biochemical characteristics. Appl Microbiol Biotechnol 105:4515\u20134534. https:\/\/doi.org\/10.1007\/s00253-021-11359-y","journal-title":"Appl Microbiol Biotechnol"},{"key":"1296_CR11","doi-asserted-by":"publisher","first-page":"503","DOI":"10.1002\/bab.2127","volume":"69","author":"LS da Silva","year":"2022","unstructured":"da Silva LS, Doonan LB, Pessoa A Jr (2022) Structural and functional diversity of asparaginases: Overview and recommendations for a revised nomenclature. Biotechnol Appl Biochem 69:503\u2013513. https:\/\/doi.org\/10.1002\/bab.2127","journal-title":"Biotechnol Appl Biochem"},{"key":"1296_CR12","unstructured":"DSM company websites (2024)\u00a0https:\/\/www.dsm.com\/food-beverage\/en_US\/insights\/insights\/baking\/2018-07-10-dsm-introduces-preventase-xr-to-extend-use-of-enzymatic-acrylamide-reduction-solution-to-more-snacks-and-baked-goods.html. Accessed 29 Aug 2024"},{"key":"1296_CR13","doi-asserted-by":"publisher","first-page":"62","DOI":"10.4103\/0976-500X.184769","volume":"7","author":"RA Egler","year":"2016","unstructured":"Egler RA, Ahuja SP, Matloub Y (2016) l-Asparaginase in the treatment of patients with acute lymphoblastic leukemia. J Pharmacol Pharmacother 7:62\u201371. https:\/\/doi.org\/10.4103\/0976-500X.184769","journal-title":"J Pharmacol Pharmacother"},{"key":"1296_CR14","doi-asserted-by":"publisher","first-page":"599","DOI":"10.1134\/S0003683823602937","volume":"60","author":"K Hosseini","year":"2024","unstructured":"Hosseini K, Zivari-Ghader T, Akbarzadehlaleh P et al (2024) A comprehensive review of l-asparaginase: production, applications and therapeutic potential in cancer treatment. Appl Biochem Microbiol 60:599\u2013613. https:\/\/doi.org\/10.1134\/S0003683823602937","journal-title":"Appl Biochem Microbiol"},{"key":"1296_CR15","doi-asserted-by":"publisher","first-page":"565","DOI":"10.1084\/jem.98.6.565","volume":"98","author":"JG Kidd","year":"1953","unstructured":"Kidd JG (1953) Regression of transplanted lymphomas induced in vivo by means of normal guinea pig serum. I. Course of transplanted cancers of various kinds in mice and rats given guinea pig serum, horse serum or rabbit serum. J Exp Med 98:565\u2013582. https:\/\/doi.org\/10.1084\/jem.98.6.565","journal-title":"J Exp Med"},{"key":"1296_CR16","doi-asserted-by":"publisher","first-page":"1208277","DOI":"10.3389\/fphar.2023.1208277","volume":"14","author":"N Lefin","year":"2023","unstructured":"Lefin N, Miranda J, Beltr\u00e1n JF et al (2023) Current state of molecular and metabolic strategies for the improvement of l-asparaginase expression in heterologous systems. Front Pharmacol 14:1208277. https:\/\/doi.org\/10.3389\/fphar.2023.1208277","journal-title":"Front Pharmacol"},{"key":"1296_CR17","doi-asserted-by":"publisher","first-page":"514","DOI":"10.1107\/S2052252521006011","volume":"8","author":"JI Loch","year":"2021","unstructured":"Loch JI, Jaskolski M (2021) Structural and biophysical aspects of L-asparaginases: a growing family with amazing diversity. IUCrJ 8:514\u2013531. https:\/\/doi.org\/10.1107\/S2052252521006011","journal-title":"IUCrJ"},{"key":"1296_CR18","doi-asserted-by":"publisher","first-page":"6717","DOI":"10.1038\/s41467-021-27105-x","volume":"12","author":"JI Loch","year":"2021","unstructured":"Loch JI, Imiolczyk B, Sliwiak J et al (2021) Crystal structures of the elusive Rhizobium etli l-asparaginase reveal a peculiar active site. Nat Commun 12:6717. https:\/\/doi.org\/10.1038\/s41467-021-27105-x","journal-title":"Nat Commun"},{"key":"1296_CR19","doi-asserted-by":"publisher","first-page":"902117","DOI":"10.3389\/fped.2022.902117","volume":"10","author":"L Maese","year":"2022","unstructured":"Maese L, Rau RE (2022) Current use of asparaginase in acute lymphoblastic leukemia\/lymphoblastic lymphoma. Front Pediatr 10:902117. https:\/\/doi.org\/10.3389\/fped.2022.902117","journal-title":"Front Pediatr"},{"key":"1296_CR20","doi-asserted-by":"publisher","first-page":"292","DOI":"10.4014\/jmb.1107.07047","volume":"22","author":"A Moreno-Enriquez","year":"2012","unstructured":"Moreno-Enriquez A, Evangelista-Martinez Z, Gonzalez-Mondragon EG et al (2012) Biochemical characterization of recombinant l-asparaginase (AnsA) from Rhizobium etli, a member of an increasing rhizobial-type family of l-asparaginases. J Microbiol Biotechnol 22:292\u2013300. https:\/\/doi.org\/10.4014\/jmb.1107.07047","journal-title":"J Microbiol Biotechnol"},{"key":"1296_CR21","doi-asserted-by":"publisher","first-page":"5827","DOI":"10.3390\/molecules25245827","volume":"25","author":"JCF Nunes","year":"2020","unstructured":"Nunes JCF, Crist\u00f3v\u00e3o RO, Freire MG et al (2020) Recent strategies and applications for l-asparaginase confinement. Molecules 25:5827. https:\/\/doi.org\/10.3390\/molecules25245827","journal-title":"Molecules"},{"key":"1296_CR22","doi-asserted-by":"publisher","first-page":"848","DOI":"10.3390\/encyclopedia1030065","volume":"1","author":"JCF Nunes","year":"2021","unstructured":"Nunes JCF, Crist\u00f3v\u00e3o RO, Santos-Ebinuma VC et al (2021) l-Asparaginase-based biosensors. Encyclopedia 1:848\u2013858. https:\/\/doi.org\/10.3390\/encyclopedia1030065","journal-title":"Encyclopedia"},{"key":"1296_CR23","first-page":"639","volume":"19","author":"A Reiff","year":"2001","unstructured":"Reiff A, Zastrow M, Sun BC et al (2001) Treatment of collagen induced arthritis in DBA\/1 mice with l-asparaginase. Clin Exp Rheumatol 19:639\u2013646","journal-title":"Clin Exp Rheumatol"},{"key":"1296_CR24","doi-asserted-by":"publisher","first-page":"876","DOI":"10.1093\/jnci\/94.12.876","volume":"94","author":"T Reynolds","year":"2002","unstructured":"Reynolds T (2002) Acrylamide and cancer: tunnel leak in Sweden prompted studies. J Natl Cancer Inst 94:876\u2013878. https:\/\/doi.org\/10.1093\/jnci\/94.12.876","journal-title":"J Natl Cancer Inst"},{"key":"1296_CR25","doi-asserted-by":"publisher","first-page":"213","DOI":"10.1016\/j.bcab.2018.11.018","volume":"17","author":"G Shakambari","year":"2019","unstructured":"Shakambari G, Ashokkumar B, Varalakshmi P (2019) l-Asparaginase \u2013 a promising biocatalyst for industrial and clinical applications. Biocatal Agric Biotechnol 17:213\u2013224. https:\/\/doi.org\/10.1016\/j.bcab.2018.11.018","journal-title":"Biocatal Agric Biotechnol"},{"key":"1296_CR26","doi-asserted-by":"publisher","first-page":"93","DOI":"10.1016\/j.ijbiomac.2016.11.115","volume":"96","author":"R Shi","year":"2017","unstructured":"Shi R, Liu Y, Mu Q et al (2017) Biochemical characterization of a novel l-asparaginase from Paenibacillus barengoltzii being suitable for acrylamide reduction in potato chips and mooncakes. Int J Biol Macromol 96:93\u201399. https:\/\/doi.org\/10.1016\/j.ijbiomac.2016.11.115","journal-title":"Int J Biol Macromol"},{"key":"1296_CR27","doi-asserted-by":"publisher","first-page":"1373312","DOI":"10.3389\/fchem.2024.1373312","volume":"12","author":"J Sliwiak","year":"2024","unstructured":"Sliwiak J, Worsztynowicz P, Pokrywka K et al (2024) Biochemical characterization of l-asparaginase isoforms from Rhizobium etli-the boosting effect of zinc. Front Chem 12:1373312. https:\/\/doi.org\/10.3389\/fchem.2024.1373312","journal-title":"Front Chem"},{"key":"1296_CR28","doi-asserted-by":"publisher","first-page":"278","DOI":"10.1007\/s13205-018-1282-3","volume":"8","author":"A Vimal","year":"2018","unstructured":"Vimal A, Kumar A (2018) l-Asparaginase: a feasible therapeutic molecule for multiple diseases. 3 Biotech 8:278. https:\/\/doi.org\/10.1007\/s13205-018-1282-3","journal-title":"3 Biotech"}],"container-title":["Folia Microbiologica"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s12223-025-01296-y.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1007\/s12223-025-01296-y","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s12223-025-01296-y.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T05:45:59Z","timestamp":1788414359000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1007\/s12223-025-01296-y"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7,11]]},"references-count":28,"journal-issue":{"issue":"4","published-print":{"date-parts":[[2026,8]]}},"alternative-id":["1296"],"URL":"https:\/\/doi.org\/10.1007\/s12223-025-01296-y","relation":{},"ISSN":["0015-5632","1874-9356"],"issn-type":[{"value":"0015-5632","type":"print"},{"value":"1874-9356","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,7,11]]},"assertion":[{"value":"4 April 2025","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"26 June 2025","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"11 July 2025","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"No humans or animals were involved in the research described here.","order":1,"name":"Ethics","label":"Human\/animal ethics statement","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The character of the work did not require informed consent.","order":2,"name":"Ethics","label":"Informed consent","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The authors declare no competing interests.","order":3,"name":"Ethics","label":"Competing interests","group":{"name":"EthicsHeading","label":"Declarations"}}]}}