{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,20]],"date-time":"2026-01-20T05:33:15Z","timestamp":1768887195513,"version":"3.49.0"},"reference-count":56,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2022,8,17]],"date-time":"2022-08-17T00:00:00Z","timestamp":1660694400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100009708","name":"Novo Nordisk Foundation, Denmark","doi-asserted-by":"publisher","award":["NNF20SA0066233"],"award-info":[{"award-number":["NNF20SA0066233"]}],"id":[{"id":"10.13039\/501100009708","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100009708","name":"Novo Nordisk Foundation, Denmark","doi-asserted-by":"publisher","award":["2013\/08617-7"],"award-info":[{"award-number":["2013\/08617-7"]}],"id":[{"id":"10.13039\/501100009708","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100009708","name":"Novo Nordisk Foundation, Denmark","doi-asserted-by":"publisher","award":["2019\/03981-9"],"award-info":[{"award-number":["2019\/03981-9"]}],"id":[{"id":"10.13039\/501100009708","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100009708","name":"Novo Nordisk Foundation, Denmark","doi-asserted-by":"publisher","award":["298596"],"award-info":[{"award-number":["298596"]}],"id":[{"id":"10.13039\/501100009708","id-type":"DOI","asserted-by":"publisher"}]},{"name":"State of S\u00e3o Paulo Research Foundation, FAPESP, Brazil","award":["NNF20SA0066233"],"award-info":[{"award-number":["NNF20SA0066233"]}]},{"name":"State of S\u00e3o Paulo Research Foundation, FAPESP, Brazil","award":["2013\/08617-7"],"award-info":[{"award-number":["2013\/08617-7"]}]},{"name":"State of S\u00e3o Paulo Research Foundation, FAPESP, Brazil","award":["2019\/03981-9"],"award-info":[{"award-number":["2019\/03981-9"]}]},{"name":"State of S\u00e3o Paulo Research Foundation, FAPESP, Brazil","award":["298596"],"award-info":[{"award-number":["298596"]}]},{"DOI":"10.13039\/501100003141","name":"Mexican Council for Science and Technology, CONACYT, Mexico","doi-asserted-by":"publisher","award":["NNF20SA0066233"],"award-info":[{"award-number":["NNF20SA0066233"]}],"id":[{"id":"10.13039\/501100003141","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003141","name":"Mexican Council for Science and Technology, CONACYT, Mexico","doi-asserted-by":"publisher","award":["2013\/08617-7"],"award-info":[{"award-number":["2013\/08617-7"]}],"id":[{"id":"10.13039\/501100003141","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003141","name":"Mexican Council for Science and Technology, CONACYT, Mexico","doi-asserted-by":"publisher","award":["2019\/03981-9"],"award-info":[{"award-number":["2019\/03981-9"]}],"id":[{"id":"10.13039\/501100003141","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003141","name":"Mexican Council for Science and Technology, CONACYT, Mexico","doi-asserted-by":"publisher","award":["298596"],"award-info":[{"award-number":["298596"]}],"id":[{"id":"10.13039\/501100003141","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Fermentation"],"abstract":"<jats:p>This study reports the production of L-asparaginase (ASNase), an enzyme mainly used for the treatment of acute lymphoblastic leukemia, by Leucosporidiumscottii L115, a psychrotolerant yeast isolated from the Antarctic ecosystem. Focus was given to select the most appropriate medium components able to maximize the enzyme production by this yeast, as a first step for the development of a new process to produce ASNase. By combining knowledge in bioprocesses, statistical analysis and modeling, the medium composition that most favored enzyme production was established, which consisted of using a mixture of sucrose (28.34 g L\u22121) and glycerol (15.61 g L\u22121) as carbon sources, supplemented with proline (6.15 g L\u22121) and the following salts (g L\u22121): KCl, 0.52; MgSO4\u00b77H2O, 0.52; CuNO3\u00b73H2O, 0.001; ZnSO4\u00b77H2O, 0.001; and FeSO4\u00b77H2O, 0.001. By using this medium, enzyme production of 2850 U L\u22121 (productivity of 23.75 U L\u22121 h\u22121) was obtained, which represented a 28-fold increase in enzyme production per gram of cells (178 U gdcw\u22121) when compared to the control (non-optimized medium), and a 50-fold increase when compared to a reference medium used for ASNase production.<\/jats:p>","DOI":"10.3390\/fermentation8080398","type":"journal-article","created":{"date-parts":[[2022,8,17]],"date-time":"2022-08-17T21:23:56Z","timestamp":1660771436000},"page":"398","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Selection and Optimization of Medium Components for the Efficient Production of L-Asparaginase by Leucosporidium scottii L115\u2014A Psychrotolerant Yeast"],"prefix":"10.3390","volume":"8","author":[{"given":"Ignacio S.","family":"Moguel","sequence":"first","affiliation":[{"name":"Department of Biochemical-Pharmaceutical Technology, School of Pharmaceutical Sciences, University of S\u00e3o Paulo, Butant\u00e3, S\u00e3o Paulo 05508-000, Brazil"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2394-0364","authenticated-orcid":false,"given":"Celina K.","family":"Yamakawa","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Biomedicine, Technical University of Denmark, S\u00f8ltofts Plads, Building 223, 2800 Kongens Lyngby, Denmark"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1135-4827","authenticated-orcid":false,"given":"Larissa P.","family":"Brumano","sequence":"additional","affiliation":[{"name":"Department of Biochemical-Pharmaceutical Technology, School of Pharmaceutical Sciences, University of S\u00e3o Paulo, Butant\u00e3, S\u00e3o Paulo 05508-000, Brazil"},{"name":"Center for Natural and Human Sciences, Federal University of ABC, Av. dos Estados, 5001, Bang\u00fa, Santo Andr\u00e9 09210-580, Brazil"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5268-8690","authenticated-orcid":false,"suffix":"Jr.","given":"Adalberto","family":"Pessoa","sequence":"additional","affiliation":[{"name":"Department of Biochemical-Pharmaceutical Technology, School of Pharmaceutical Sciences, University of S\u00e3o Paulo, Butant\u00e3, S\u00e3o Paulo 05508-000, Brazil"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7182-6198","authenticated-orcid":false,"given":"Solange I.","family":"Mussatto","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Biomedicine, Technical University of Denmark, S\u00f8ltofts Plads, Building 223, 2800 Kongens Lyngby, Denmark"}]}],"member":"1968","published-online":{"date-parts":[[2022,8,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"341","DOI":"10.1046\/j.1469-8137.2001.00177.x","article-title":"Cold adaptation in Arctic and Antarctic fungi","volume":"151","author":"Robinson","year":"2001","journal-title":"New Phytol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1920","DOI":"10.3390\/md8061920","article-title":"Research and Application of Marine Microbial Enzymes: Status and Prospects","volume":"8","author":"Zhang","year":"2010","journal-title":"Mar. Drugs"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"813","DOI":"10.1111\/1567-1364.12158","article-title":"Marine yeast isolation and industrial application","volume":"14","author":"Zaky","year":"2014","journal-title":"FEMS Yeast Res."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1023","DOI":"10.1007\/s00792-013-0584-y","article-title":"Taxonomic assessment and enzymes production by yeasts isolated from marine and terrestrial Antarctic samples","volume":"17","author":"Duarte","year":"2013","journal-title":"Extremophiles"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1080\/10826068.2020.1815053","article-title":"Glutaminase-free L-asparaginase production by Leucosporidium muscorum isolated from Antarctic marine-sediment","volume":"51","author":"Freire","year":"2020","journal-title":"Prep. Biochem. Biotechnol."},{"key":"ref_6","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":"Ashok","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"967","DOI":"10.1007\/s00203-020-01814-1","article-title":"Microbial l-asparaginase: Purification, characterization and applications","volume":"202","author":"Muneer","year":"2020","journal-title":"Arch. Microbiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1016\/j.foodchem.2016.04.105","article-title":"The use of asparaginase to reduce acrylamide levels in cooked food","volume":"210","author":"Xu","year":"2016","journal-title":"Food Chem."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"841","DOI":"10.1007\/s00792-015-0763-0","article-title":"Reduction of acrylamide level through blanching with treatment by an extremely thermostable l-asparaginase during French fries processing","volume":"19","author":"Zuo","year":"2015","journal-title":"Extremophiles"},{"key":"ref_10","unstructured":"World Health Organization (2019). Model List of Essential Medicines, 21st List, World Health Organization."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"212","DOI":"10.3389\/fbioe.2018.00212","article-title":"Development of L-Asparaginase Biobetters: Current Research Status and Review of the Desirable Quality Profiles","volume":"6","author":"Brumano","year":"2019","journal-title":"Front. Bioeng. Biotechnol."},{"key":"ref_12","unstructured":"Merck (2022, July 27). Elspar Asparaginase, Available online: https:\/\/www.accessdata.fda.gov\/drugsatfda_docs\/label\/2002\/aspamer080102LB.pdf."},{"key":"ref_13","unstructured":"European Medicines Agency (2022, August 08). Assessment Report Spectrila, Available online: https:\/\/www.ema.europa.eu\/en\/documents\/assessment-report\/spectrila-epar-public-assessment-report_en.pdf."},{"key":"ref_14","unstructured":"European Medicines Agency (2022, August 08). Assessment Report Oncaspar, Available online: https:\/\/www.ema.europa.eu\/en\/documents\/assessment-report\/oncaspar-epar-public-assessment-report_en.pdf."},{"key":"ref_15","unstructured":"Medicines Evaluation Board (2015). Public Assessmen Report Crisantaspase, European Medicines Agency. Available online: https:\/\/file.wuxuwang.com\/hma\/NL_H_3194_001_PARSummary.pdf."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"(2021). Asparaginase (Erwinia chrysanthemi) (Recombinant)-rywn. Am. J. Health Pharm., 78, 1919\u20131921.","DOI":"10.1093\/ajhp\/zxab309"},{"key":"ref_17","first-page":"627031","article-title":"Optimization of Culture Conditions for Production of the Anti-Leukemic Glutaminase Free L-Asparaginase by Newly Isolated Streptomyces olivaceus NEAE-119 Using Response Surface Methodology","volume":"2015","author":"Moawad","year":"2015","journal-title":"BioMed Res. Int."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"208","DOI":"10.1016\/j.critrevonc.2006.07.009","article-title":"Pharmacological and clinical evaluation of l-asparaginase in the treatment of leukemia","volume":"61","author":"Narta","year":"2007","journal-title":"Crit. Rev. Oncol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1007\/s13205-016-0544-1","article-title":"Isolation and screening of l-asparaginase free of glutaminase and urease from fungal sp","volume":"6","author":"Doriya","year":"2016","journal-title":"3 Biotech"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1016\/j.bjm.2016.10.004","article-title":"Current applications and different approaches for microbial l-asparaginase production","volume":"47","author":"Antunes","year":"2016","journal-title":"Braz. J. Microbiol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"119308","DOI":"10.1016\/j.jclepro.2019.119308","article-title":"Lipid and carotenoid production from wheat straw hydrolysates by different oleaginous yeasts","volume":"249","author":"Liu","year":"2020","journal-title":"J. Clean. Prod."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"508968","DOI":"10.1155\/2013\/508968","article-title":"Effect of C\/N Ratio and Media Optimization through Response Surface Methodology on Simultaneous Productions of Intra- and Extracellular Inulinase and Invertase from Aspergillus niger ATCC 20611","volume":"2013","author":"Dinarvand","year":"2013","journal-title":"BioMed Res. Int."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"540","DOI":"10.1016\/j.procbio.2008.01.013","article-title":"Establishment of the optimum initial xylose concentration and nutritional supplementation of brewer\u2019s spent grain hydrolysate for xylitol production by Candida guilliermondii","volume":"43","author":"Mussatto","year":"2008","journal-title":"Process. Biochem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1973","DOI":"10.1007\/s10529-007-9494-3","article-title":"Brewer\u2019s spent grain as raw material for lactic acid production by Lactobacillus delbrueckii","volume":"29","author":"Mussatto","year":"2007","journal-title":"Biotechnol. Lett."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"334","DOI":"10.1128\/MMBR.62.2.334-361.1998","article-title":"Yeast Carbon Catabolite Repression","volume":"62","author":"Gancedo","year":"1998","journal-title":"Microbiol. Mol. Biol. Rev."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"283","DOI":"10.1111\/j.1751-7915.2007.00015.x","article-title":"Metabolic regulation and overproduction of primary metabolites","volume":"1","author":"Sanchez","year":"2008","journal-title":"Microb. Biotechnol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/S0378-1119(02)00558-9","article-title":"Nitrogen regulation in Saccharomyces cerevisiae","volume":"290","author":"Magasanik","year":"2002","journal-title":"Gene"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/0168-1656(94)90093-0","article-title":"Properties of cold-adapted microorganisms and their potential role in biotechnology","volume":"33","author":"Margesin","year":"1994","journal-title":"J. Biotechnol."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Segal-Kischinevzky, C., Romero-Aguilar, L., Alcaraz, L.D., L\u00f3pez-Ortiz, G., Mart\u00ednez-Castillo, B., Torres-Ram\u00edrez, N., Sandoval, G., and Gonz\u00e1lez, J. (2022). Yeasts Inhabiting Extreme Environments and Their Biotechnological Applications. Microorganisms, 10.","DOI":"10.3390\/microorganisms10040794"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"576511","DOI":"10.3389\/fbioe.2020.576511","article-title":"L-asparaginase Production by Leucosporidium scottii in a Bench-Scale Bioreactor with Co-production of Lipids","volume":"8","author":"Moguel","year":"2020","journal-title":"Front. Bioeng. Biotechnol."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Granato, D., and Calado, V.M.D.A. (2013). The use and importance of design of experiments (DOE) in process modelling in food science and technology. Mathematical and Statistical Methods in Food Science and Technology, John Wiley & Sons, Ltd.","DOI":"10.1002\/9781118434635.ch01"},{"key":"ref_32","first-page":"473","article-title":"Impact of carbon and nitrogen sources on L-asparaginase pro-duction by isolated Bacillus circulans (MTCC 8574): Application of saturated Plackett-Burman Design","volume":"24","author":"Hymavathi","year":"2010","journal-title":"Chem. Biochem. Eng. Q."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Analytical Methods Committee (2013). Experimental design and optimisation (4): Plackett\u2013Burman designs. Anal. Methods, 5, 1901\u20131903.","DOI":"10.1039\/C3AY90020G"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Freitas, M., Souza, P., Cardoso, S., Cruvinel, K., Abrunhosa, L.S., Filho, E.X.F., In\u00e1cio, J., Pinho, D.B., Pessoa, A., and Magalh\u00e3es, P.O. (2021). Filamentous Fungi Producing l-Asparaginase with Low Glutaminase Activity Isolated from Brazilian Savanna Soil. Pharmaceutics, 13.","DOI":"10.3390\/pharmaceutics13081268"},{"key":"ref_35","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":"Gulati","year":"1997","journal-title":"Lett. Appl. Microbiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1007\/s12088-008-0018-1","article-title":"L-asparaginase production by Streptomyces albidoflavus","volume":"48","author":"Narayana","year":"2008","journal-title":"Indian J. Microbiol."},{"key":"ref_37","first-page":"203","article-title":"Asparaginase II of Saccharomyces cerevisiae","volume":"Volume 63\u201365","author":"Davison","year":"1997","journal-title":"Biotechnology for Fuels and Chemicals"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"3065","DOI":"10.1128\/MCB.01084-06","article-title":"Effect of 21 Different Nitrogen Sources on Global Gene Expression in the Yeast Saccharomyces cerevisiae","volume":"27","author":"Godard","year":"2007","journal-title":"Mol. Cell. Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1007\/s00726-006-0407-x","article-title":"Free radical scavenging potential of L-proline: Evidence from in vitro assays","volume":"34","author":"Kaul","year":"2008","journal-title":"Amino Acids"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"919","DOI":"10.1016\/j.biortech.2008.07.062","article-title":"Medium optimization for the production of cyclic adenosine 3\u2032,5\u2032-monophosphate by Microbacterium sp. no. 205 using response surface methodology","volume":"100","author":"Chen","year":"2009","journal-title":"Bioresour. Technol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1007\/s00253-008-1698-5","article-title":"Proline as a stress protectant in yeast: Physiological functions, metabolic regulations, and biotechnological applications","volume":"81","author":"Takagi","year":"2008","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1186\/1475-2859-11-88","article-title":"Switching industrial production processes from complex to defined media: Method development and case study using the example of Penicillium chrysogenum","volume":"11","author":"Posch","year":"2012","journal-title":"Microb. Cell Fact."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1016\/j.biotechadv.2008.07.006","article-title":"Glycerol: A promising and abundant carbon source for industrial microbiology","volume":"27","author":"Mack","year":"2009","journal-title":"Biotechnol. Adv."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1297","DOI":"10.1016\/S0021-9258(17)38144-9","article-title":"Characterization of two forms of asparaginase in Saccharomyces cerevisiae","volume":"253","author":"Dunlop","year":"1978","journal-title":"J. Biol. Chem."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"798","DOI":"10.2478\/s11696-011-0072-8","article-title":"Statistical and evolutionary optimisation of operating conditions for enhanced production of fungal l-asparaginase","volume":"65","author":"Baskar","year":"2011","journal-title":"Chem. Pap."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"295","DOI":"10.1016\/j.ejar.2015.10.002","article-title":"Optimization of production of anti-tumor l-asparaginase by free and immobilized marine Aspergillus terreus","volume":"41","author":"Farag","year":"2015","journal-title":"Egypt. J. Aquat. Res."},{"key":"ref_47","first-page":"218","article-title":"L-Asparaginase and glutaminase activities in the culture filtrates of Aspergillus nidulans","volume":"50","author":"Saxena","year":"1981","journal-title":"Curr. Sci."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"194","DOI":"10.1016\/j.critrevonc.2017.11.006","article-title":"Optimization and purification of l-asparaginase from fungi: A systematic review","volume":"120","author":"Souza","year":"2017","journal-title":"Crit. Rev. Oncol. Hematol."},{"key":"ref_49","unstructured":"Moguel, I.S. (2018). Production of L-Asparaginase of Pharmaceutical Nterest from Yeasts Isolated from the Antarctic continent. [Ph.D. Thesis, University of S\u00e3o Paulo]."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"356","DOI":"10.1080\/00021369.1972.10860270","article-title":"Production of Extracellularl-Asparaginases by Microorganisms","volume":"36","author":"Arima","year":"1972","journal-title":"Agric. Biol. Chem."},{"key":"ref_51","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":"Imada","year":"1973","journal-title":"J. Gen. Microbiol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"749","DOI":"10.1271\/bbb.59.749","article-title":"Extraction of Extracellular L-Asparaginase from Candida utilis","volume":"59","author":"Kil","year":"1995","journal-title":"Biosci. Biotechnol. Biochem."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"316","DOI":"10.1139\/m96-047","article-title":"Characterization of an extracellular asparaginase of Rhodosporidium toruloides CBS14 exhibiting unique physicochemical properties","volume":"42","author":"Ramakrishnan","year":"1996","journal-title":"Can. J. Microbiol."},{"key":"ref_54","first-page":"286","article-title":"Cloning, functional expression and characterization of L-asparaginase II from E. coli MTCC 739","volume":"49","author":"Vidya","year":"2011","journal-title":"Food Technol. Biotechnol."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1016\/j.pep.2015.12.012","article-title":"Saccharomyces cerevisiae asparaginase II, a potential antileukemic drug: Purification and characterization of the enzyme expressed in Pichia pastoris","volume":"120","author":"Sobral","year":"2016","journal-title":"Protein Expr. Purif."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"10049","DOI":"10.1021\/bi801177k","article-title":"Cold Adaptation of Enzyme Reaction Rates","volume":"47","author":"Bjelic","year":"2008","journal-title":"Biochemistry"}],"container-title":["Fermentation"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2311-5637\/8\/8\/398\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T00:11:04Z","timestamp":1760141464000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2311-5637\/8\/8\/398"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,8,17]]},"references-count":56,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2022,8]]}},"alternative-id":["fermentation8080398"],"URL":"https:\/\/doi.org\/10.3390\/fermentation8080398","relation":{},"ISSN":["2311-5637"],"issn-type":[{"value":"2311-5637","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,8,17]]}}}