{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,28]],"date-time":"2026-02-28T04:20:49Z","timestamp":1772252449403,"version":"3.50.1"},"reference-count":69,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2022,4,14]],"date-time":"2022-04-14T00:00:00Z","timestamp":1649894400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia","doi-asserted-by":"publisher","award":["LA\/P\/0045\/2020"],"award-info":[{"award-number":["LA\/P\/0045\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia","doi-asserted-by":"publisher","award":["UIDB\/50020\/2020"],"award-info":[{"award-number":["UIDB\/50020\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia","doi-asserted-by":"publisher","award":["UIDP\/50020\/2020"],"award-info":[{"award-number":["UIDP\/50020\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia","doi-asserted-by":"publisher","award":["POCI-01-0145-FEDER-031268"],"award-info":[{"award-number":["POCI-01-0145-FEDER-031268"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia","doi-asserted-by":"publisher","award":["UIDB\/50011\/2020"],"award-info":[{"award-number":["UIDB\/50011\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia","doi-asserted-by":"publisher","award":["UIDP\/50011\/2020"],"award-info":[{"award-number":["UIDP\/50011\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia","doi-asserted-by":"publisher","award":["CEECINST\/00102\/2018"],"award-info":[{"award-number":["CEECINST\/00102\/2018"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia","doi-asserted-by":"publisher","award":["UIDB\/50006\/2020"],"award-info":[{"award-number":["UIDB\/50006\/2020"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia","doi-asserted-by":"publisher","award":["CEECIND\/00383\/2017"],"award-info":[{"award-number":["CEECIND\/00383\/2017"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001871","name":"Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia","doi-asserted-by":"publisher","award":["CEECIND\/2020\/01867"],"award-info":[{"award-number":["CEECIND\/2020\/01867"]}],"id":[{"id":"10.13039\/501100001871","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001807","name":"S\u00e3o Paulo Research Foundation","doi-asserted-by":"publisher","award":["2018\/06908-8"],"award-info":[{"award-number":["2018\/06908-8"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["BioTech"],"abstract":"<jats:p>L-asparaginase (ASNase) is an aminohydrolase currently used in the pharmaceutical and food industries. Enzyme immobilization is an exciting option for both applications, allowing for a more straightforward recovery and increased stability. High surface area and customizable porosity make carbon xerogels (CXs) promising materials for ASNase immobilization. This work describes the influence of contact time, pH, and ASNase concentration on the immobilization yield (IY) and relative recovered activity (RRA) using the Central Composite Design methodology. The most promising results were obtained using CX with an average pore size of 4 nm (CX-4), reaching IY and RRA of 100%. At the optimal conditions (contact time 49 min, pH 6.73, and [ASNase] 0.26 mg\u00b7mL\u22121), the ASNase-CXs biocomposite was characterized and evaluated in terms of kinetic properties and operational, thermal, and pH stabilities. The immobilized ASNase onto CX-4 retained 71% of its original activity after six continuous reaction cycles, showed good thermal stability at 37 \u00b0C (RRA of 91% after 90 min), and was able to adapt to both acidic and alkaline environments. Finally, the results indicated a 3.9-fold increase in the immobilized ASNase affinity for the substrate, confirming the potential of CXs as a support for ASNase and as a cost-effective tool for subsequent use in the therapeutic and food sectors.<\/jats:p>","DOI":"10.3390\/biotech11020010","type":"journal-article","created":{"date-parts":[[2022,4,14]],"date-time":"2022-04-14T21:44:06Z","timestamp":1649972646000},"page":"10","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Immobilization and Characterization of L-Asparaginase over Carbon Xerogels"],"prefix":"10.3390","volume":"11","author":[{"given":"Rita A. M.","family":"Barros","sequence":"first","affiliation":[{"name":"LSRE-LCM\u2014Laboratory of Separation and Reaction Engineering-Laboratory of Catalysis and Materials, Faculty of Engineering, University of Porto, 4200-465 Porto, Portugal"},{"name":"ALiCE\u2014Associate Laboratory in Chemical Engineering, Faculty of Engineering, University of Porto, 4200-465 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5165-4766","authenticated-orcid":false,"given":"Raquel O.","family":"Crist\u00f3v\u00e3o","sequence":"additional","affiliation":[{"name":"LSRE-LCM\u2014Laboratory of Separation and Reaction Engineering-Laboratory of Catalysis and Materials, Faculty of Engineering, University of Porto, 4200-465 Porto, Portugal"},{"name":"ALiCE\u2014Associate Laboratory in Chemical Engineering, Faculty of Engineering, University of Porto, 4200-465 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9913-4671","authenticated-orcid":false,"given":"S\u00f3nia A. C.","family":"Carabineiro","sequence":"additional","affiliation":[{"name":"LSRE-LCM\u2014Laboratory of Separation and Reaction Engineering-Laboratory of Catalysis and Materials, Faculty of Engineering, University of Porto, 4200-465 Porto, Portugal"},{"name":"LAQV-REQUIMTE, Department of Chemistry, NOVA School of Science and Technology, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5611-4080","authenticated-orcid":false,"given":"M\u00e1rcia C.","family":"Neves","sequence":"additional","affiliation":[{"name":"CICECO-Aveiro Institute of Materials, Department of Chemistry, University of Aveiro, 3810-193 Aveiro, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8895-0614","authenticated-orcid":false,"given":"Mara G.","family":"Freire","sequence":"additional","affiliation":[{"name":"CICECO-Aveiro Institute of Materials, Department of Chemistry, University of Aveiro, 3810-193 Aveiro, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6531-3978","authenticated-orcid":false,"given":"Joaquim L.","family":"Faria","sequence":"additional","affiliation":[{"name":"LSRE-LCM\u2014Laboratory of Separation and Reaction Engineering-Laboratory of Catalysis and Materials, Faculty of Engineering, University of Porto, 4200-465 Porto, Portugal"},{"name":"ALiCE\u2014Associate Laboratory in Chemical Engineering, Faculty of Engineering, University of Porto, 4200-465 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6666-6695","authenticated-orcid":false,"given":"Val\u00e9ria C.","family":"Santos-Ebinuma","sequence":"additional","affiliation":[{"name":"Department of Engineering Bioprocess and Biotechnology, School of Pharmaceutical Sciences, UNESP-University Estadual Paulista, Araraquara 14800-903, Brazil"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9128-6275","authenticated-orcid":false,"given":"Ana P. M.","family":"Tavares","sequence":"additional","affiliation":[{"name":"CICECO-Aveiro Institute of Materials, Department of Chemistry, University of Aveiro, 3810-193 Aveiro, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6469-4871","authenticated-orcid":false,"given":"Cl\u00e1udia G.","family":"Silva","sequence":"additional","affiliation":[{"name":"LSRE-LCM\u2014Laboratory of Separation and Reaction Engineering-Laboratory of Catalysis and Materials, Faculty of Engineering, University of Porto, 4200-465 Porto, Portugal"},{"name":"ALiCE\u2014Associate Laboratory in Chemical Engineering, Faculty of Engineering, University of Porto, 4200-465 Porto, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2022,4,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1598","DOI":"10.1021\/acs.bioconjchem.7b00217","article-title":"Immobilization of l-Asparaginase on Carrier Materials: A Comprehensive Review","volume":"28","author":"Ulu","year":"2017","journal-title":"Bioconjug. Chem."},{"key":"ref_2","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_3","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":"Extrem. Life Extrem. Cond."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1752","DOI":"10.1039\/C5FO00320B","article-title":"Acrylamide: Inhibition of formation in processed food and mitigation of toxicity in cells, animals, and humans","volume":"6","author":"Friedman","year":"2015","journal-title":"Food Funct."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1080\/02648725.2017.1357294","article-title":"Biotechnological production and practical application of L-asparaginase enzyme","volume":"33","author":"Vimal","year":"2017","journal-title":"Biotechnol. Genet. Eng. Rev."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"4782","DOI":"10.1021\/jf034180i","article-title":"Acrylamide formation mechanism in heated foods","volume":"51","author":"Zyzak","year":"2003","journal-title":"J. Agric. Food Chem."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"741","DOI":"10.1007\/s11947-013-1108-6","article-title":"Reduction of Acrylamide Formation in Sweet Bread with l-Asparaginase Treatment","volume":"7","author":"Shimray","year":"2014","journal-title":"Food Bioprocess Technol."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Porto, A.C.V., Freitas-Silva, O., Souza, E.F.d., and Gottschalk, L.M.F. (2019). Effect of Asparaginase Enzyme in the Reduction of Asparagine in Green Coffee. Beverages, 5.","DOI":"10.3390\/beverages5020032"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"4168","DOI":"10.1021\/jf900174q","article-title":"Evaluating the potential for enzymatic acrylamide mitigation in a range of food products using an asparaginase from Aspergillus oryzae","volume":"57","author":"Hendriksen","year":"2009","journal-title":"J. Agric. Food Chem."},{"key":"ref_10","unstructured":"FAO, and WHO (2021). Compendium of Food Additive Specifications. Joint FAO\/WHO Expert Committee on Food Additives (JECFA), WHO. FAO JECFA Monographs No. 26."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Jia, R., Wan, X., Geng, X., Xue, D., Xie, Z., and Chen, C. (2021). Microbial L-asparaginase for Application in Acrylamide Mitigation from Food: Current Research Status and Future Perspectives. Microorganisms, 9.","DOI":"10.3390\/microorganisms9081659"},{"key":"ref_12","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_13","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1016\/j.bcab.2018.11.018","article-title":"L-asparaginase\u2014A promising biocatalyst for industrial and clinical applications","volume":"17","author":"Shakambari","year":"2019","journal-title":"Biocatal. Agric. Biotechnol."},{"key":"ref_14","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":"Castro","year":"2021","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"309214","DOI":"10.1155\/2015\/309214","article-title":"Purification and Characterization of Asparaginase from Phaseolus vulgaris Seeds","volume":"2015","author":"Mohamed","year":"2015","journal-title":"Evid.-Based Complement. Altern. Med."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"442","DOI":"10.1111\/bjh.16254","article-title":"Acute lymphoblastic leukaemia patients treated with PEGasparaginase develop antibodies to PEG and the succinate linker","volume":"189","author":"Kloos","year":"2020","journal-title":"Br. J. Haematol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1016\/j.jconrel.2003.11.018","article-title":"Drug, enzyme and peptide delivery using erythrocytes as carriers","volume":"95","author":"Marinero","year":"2004","journal-title":"J. Control Release"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1007\/s12154-013-0102-9","article-title":"Enzyme immobilization: An update","volume":"6","author":"Homaei","year":"2013","journal-title":"J. Chem. Biol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1080\/13102818.2015.1008192","article-title":"An overview of technologies for immobilization of enzymes and surface analysis techniques for immobilized enzymes","volume":"29","author":"Mohamad","year":"2015","journal-title":"Biotechnol. Biotechnol. Equip."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Nunes, J.C.F., Crist\u00f3v\u00e3o, R.O., Freire, M.G., Santos-Ebinuma, V.C., Faria, J.L., Silva, C.G., and Tavares, A.P.M. (2020). Recent Strategies and Applications for l-Asparaginase Confinement. Molecules, 25.","DOI":"10.3390\/molecules25245827"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1570","DOI":"10.1002\/jctb.4698","article-title":"A strategy for improving peroxidase stability via immobilization on surface modified multi-walled carbon nanotubes","volume":"90","author":"Azevedo","year":"2015","journal-title":"J. Chem. Technol. Biotechnol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"31205","DOI":"10.1039\/D0RA05534D","article-title":"Development and characterization of a novel l-asparaginase\/MWCNT nanobioconjugate","volume":"10","author":"Almeida","year":"2020","journal-title":"RSC Adv."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"743","DOI":"10.1016\/j.carbon.2017.04.009","article-title":"Protein adsorption and activity on carbon xerogels with narrow pore size distributions covering a wide mesoporous range","volume":"118","author":"Concheso","year":"2017","journal-title":"Carbon"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"4747","DOI":"10.1039\/C4RA15171B","article-title":"Synthesis of carbon xerogel nanoparticles by inverse emulsion polymerization of resorcinol\u2013formaldehyde and their use as anode materials for lithium-ion battery","volume":"5","author":"Kakunuri","year":"2015","journal-title":"RSC Adv."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2887","DOI":"10.1002\/adma.201100283","article-title":"Advances in Tailoring Resorcinol-Formaldehyde Organic and Carbon Gels","volume":"23","author":"ElKhatat","year":"2011","journal-title":"Adv. Mater."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"10231","DOI":"10.1021\/la502394u","article-title":"Gelation Mechanism of Resorcinol-Formaldehyde Gels Investigated by Dynamic Light Scattering","volume":"30","author":"Taylor","year":"2014","journal-title":"Langmuir"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"4583","DOI":"10.1016\/j.watres.2011.06.008","article-title":"Adsorption of ciprofloxacin on surface-modified carbon materials","volume":"45","author":"Carabineiro","year":"2011","journal-title":"Water Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"449","DOI":"10.1016\/j.colsurfb.2019.01.032","article-title":"Repression of melanoma tumor in vitro and in vivo by photothermal effect of carbon xerogel nanoparticles","volume":"176","author":"Gorgizadeh","year":"2019","journal-title":"Colloids Surf. B Biointerfaces"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1021\/ja01269a023","article-title":"Adsorption of Gases in Multimolecular Layers","volume":"60","author":"Brunauer","year":"1938","journal-title":"J. Am. Chem. Soc."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"373","DOI":"10.1021\/ja01145a126","article-title":"The Determination of Pore Volume and Area Distributions in Porous Substances. I. Computations from Nitrogen Isotherms","volume":"73","author":"Barrett","year":"1951","journal-title":"J. Am. Chem. Soc."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1209","DOI":"10.1002\/jctb.506","article-title":"Activated carbon surface modifications by adsorption of bacteria and their effect on aqueous lead adsorption","volume":"76","year":"2001","journal-title":"J. Chem. Technol. Biotechnol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"6985","DOI":"10.1007\/s00216-018-1326-x","article-title":"A critical analysis of L-asparaginase activity quantification methods-colorimetric methods versus high-performance liquid chromatography","volume":"410","author":"Magri","year":"2018","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"134","DOI":"10.12793\/tcp.2018.26.3.134","article-title":"Development of a UPLC-MS\/MS method for the therapeutic monitoring of L-asparaginase","volume":"26","author":"Jeong","year":"2018","journal-title":"Transl. Clin. Pharm."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"773","DOI":"10.1007\/s13213-010-0195-0","article-title":"Optimization of xylanase production by Streptomyces sp. P12-137 using response surface methodology and central composite design","volume":"61","author":"Coman","year":"2011","journal-title":"Ann. Microbiol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"94","DOI":"10.14302\/issn.2576-6694.jbbs-18-2280","article-title":"Graphical Approach to Compare Concentration Constants of Hill and Michaelis-Menten Equations","volume":"1","author":"Elena","year":"2018","journal-title":"J. Biotechnol. Biomed. Sci."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"5607","DOI":"10.1021\/jm020467f","article-title":"Enzymes. A Practical Introduction to Structure, Mechanism, and Data Analysis. Second Edition By Robert A. Copeland. Wiley-VCH, New York. 2000. xvi + 397 pp. 16 \u00d7 24.5 cm. ISBN 0-471-35929-7. $99.95","volume":"45","author":"Walters","year":"2002","journal-title":"J. Med. Chem."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"619","DOI":"10.1016\/j.carbon.2003.12.072","article-title":"Porous carbon xerogels with texture tailored by pH control during sol\u2013gel process","volume":"42","author":"Job","year":"2004","journal-title":"Carbon"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1231","DOI":"10.1016\/j.carbon.2004.12.016","article-title":"The effects of different synthetic conditions on the porous properties of carbon cryogel microspheres","volume":"43","author":"Yamamoto","year":"2005","journal-title":"Carbon"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1007\/s10450-010-9272-8","article-title":"Adsorption of dyes on carbon xerogels and templated carbons: Influence of surface chemistry","volume":"17","author":"Figueiredo","year":"2011","journal-title":"Adsorption"},{"key":"ref_40","first-page":"2277","article-title":"Carbon xerogel\/Carbon Nanotubes Nanohybrid Doped with Ti for Removal of Methylene Blue Dye","volume":"62","author":"Fathy","year":"2019","journal-title":"Egypt. J. Chem."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"18013","DOI":"10.1038\/s41598-018-36161-1","article-title":"Cloning and expression of L-asparaginase from Bacillus tequilensis PV9W and therapeutic efficacy of Solid Lipid Particle formulations against cancer","volume":"8","author":"Shakambari","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"357","DOI":"10.1146\/annurev-matsci-070115-032140","article-title":"Raman Studies of Carbon Nanostructures","volume":"46","author":"Jorio","year":"2016","journal-title":"Annu. Rev. Mater. Res."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"14095","DOI":"10.1103\/PhysRevB.61.14095","article-title":"Interpretation of Raman spectra of disordered and amorphous carbon","volume":"61","author":"Ferrari","year":"2000","journal-title":"Phys. Rev. B"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"2847","DOI":"10.1007\/s10008-014-2548-2","article-title":"Effect of the resorcinol\/catalyst ratio in the capacitive performance of carbon xerogels with potential use in sodium chloride removal from saline water","volume":"18","author":"Zafra","year":"2014","journal-title":"J. Solid State Electrochem."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1016\/S0927-796X(02)00005-0","article-title":"Diamond-like amorphous carbon","volume":"37","author":"Robertson","year":"2002","journal-title":"Mater. Sci. Eng. R Rep."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"413","DOI":"10.1080\/00018732.2011.582251","article-title":"Raman spectroscopy of graphene and carbon nanotubes","volume":"60","author":"Saito","year":"2011","journal-title":"Adv. Phys."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"974","DOI":"10.1016\/j.cej.2018.08.178","article-title":"Enhanced biocatalytic sustainability of laccase by immobilization on functionalized carbon nanotubes\/polysulfone membranes","volume":"355","author":"Costa","year":"2019","journal-title":"Chem. Eng. J."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"114204","DOI":"10.1016\/j.ab.2021.114204","article-title":"Covalent conjugation of single-walled carbon nanotube with CYP101 mutant for direct electrocatalysis","volume":"626","author":"Ray","year":"2021","journal-title":"Anal. Biochem."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1095","DOI":"10.1016\/j.procbio.2012.03.021","article-title":"Immobilization of commercial laccase on spent grain","volume":"47","author":"Tavares","year":"2012","journal-title":"Process Biochem."},{"key":"ref_50","first-page":"5001","article-title":"Evaluation of the absorption capacity of the natural clay from Bikougou (Gabon) to remove Mn(II) from aqueous solution","volume":"2","author":"Eba","year":"2010","journal-title":"Int. J. Eng. Sci. Technol."},{"key":"ref_51","first-page":"5","article-title":"Isolation and Purification of High Efficiency L-asparaginase by Quantitative Preparative Continuous-elution SDS PAGE Electrophoresis","volume":"3","author":"Senthil","year":"2011","journal-title":"J. Microb. Biochem. Technol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"1432","DOI":"10.1007\/s12010-020-03276-z","article-title":"Immobilization of L-Asparaginase on Magnetic Nanoparticles for Cancer Treatment","volume":"191","author":"Orhan","year":"2020","journal-title":"Appl. Biochem. Biotechnol."},{"key":"ref_53","first-page":"956","article-title":"Formulation of PEG-ylated L-asparaginase loaded poly (lactide-co-glycolide) nanoparticles: Influence of Pegylation on enzyme loading, activity and in vitro release","volume":"66","author":"Vasudev","year":"2011","journal-title":"Die Pharm."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"3751","DOI":"10.1016\/j.biomaterials.2003.10.019","article-title":"Immobilization of l-asparaginase on the microparticles of the natural silk sericin protein and its characters","volume":"25","author":"Zhang","year":"2004","journal-title":"Biomaterials"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"1679","DOI":"10.1007\/s10562-019-03069-y","article-title":"The Carboxylated Multi-walled Carbon Nanotubes\/l-Asparaginase Doped Calcium-Alginate Beads: Structural and Biocatalytic Characterization","volume":"150","author":"Ulu","year":"2020","journal-title":"Catal. Lett."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"443","DOI":"10.1016\/j.ijbiomac.2019.09.116","article-title":"Maltose functionalized magnetic core\/shell Fe3O4@Au nanoparticles for an efficient l-asparaginase immobilization","volume":"142","author":"Tarhan","year":"2020","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"1153","DOI":"10.1016\/j.cej.2018.08.058","article-title":"Immobilization of l-asparaginase on aspartic acid functionalized graphene oxide nanosheet: Enzyme kinetics and stability studies","volume":"354","author":"Monajati","year":"2018","journal-title":"Chem. Eng. J."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"354","DOI":"10.1016\/j.ijbiomac.2018.02.147","article-title":"Improvement of stability and reusability of \u03b1-amylase immobilized on naringin functionalized magnetic nanoparticles: A robust nanobiocatalyst","volume":"113","author":"Defaei","year":"2018","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"4440","DOI":"10.1039\/D0NJ00127A","article-title":"Comparative study of ASNase immobilization on tannic acid-modified magnetic Fe3O4\/SBA-15 nanoparticles to enhance stability and reusability","volume":"44","author":"Noma","year":"2020","journal-title":"New J. Chem."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"106980","DOI":"10.1016\/j.polymertesting.2020.106980","article-title":"l-asparaginase immobilized p(HEMA-GMA) cryogels: A recent study for biochemical, thermodynamic and kinetic parameters","volume":"93","author":"Acet","year":"2021","journal-title":"Polym. Test."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"70","DOI":"10.2174\/0929867324666170823143634","article-title":"Enhancement in Biological Activity of L-Asparginase by its Conjugation on Silica Nanoparticles","volume":"12","author":"Golestaneh","year":"2018","journal-title":"Recent Pat. Nanotechnol."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"43","DOI":"10.5604\/17331331.1197274","article-title":"Comparison of Free and Immobilized L-asparaginase Synthesized by Gamma-Irradiated Penicillium cyclopium","volume":"65","author":"Shafei","year":"2016","journal-title":"Pol. J. Microbiol."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"151","DOI":"10.3109\/14756368609020113","article-title":"Interaction between L-aspartic acid and L-asparaginase from Escherichia coli: Binding and inhibition studies","volume":"1","author":"Jayaram","year":"1986","journal-title":"J. Enzym. Inhib."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"12672","DOI":"10.1038\/s41598-019-49206-w","article-title":"Immobilisation of \u03b1-amylase on activated amidrazone acrylic fabric: A new approach for the enhancement of enzyme stability and reusability","volume":"9","author":"Almulaiky","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"426","DOI":"10.1016\/j.cej.2013.11.059","article-title":"Enhanced stability and catalytic activity of immobilized \u03b1-amylase on modified Fe3O4 nanoparticles","volume":"240","author":"Sohrabi","year":"2014","journal-title":"Chem. Eng. J."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1016\/j.enzmictec.2014.05.006","article-title":"New insights into the effectiveness of alpha-amylase enzyme presentation on the Bacillus subtilis spore surface by adsorption and covalent immobilization","volume":"64\u201365","author":"Gashtasbi","year":"2014","journal-title":"Enzym. Microb. Technol."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"217","DOI":"10.1016\/j.cbpa.2005.02.014","article-title":"Immobilised enzymes: Science or art?","volume":"9","author":"Cao","year":"2005","journal-title":"Curr. Opin. Chem. Biol."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"1891","DOI":"10.1007\/s00289-015-1583-1","article-title":"Synthesis and characterization of biodegradable pHEMA-starch composites for immobilization of L-asparaginase","volume":"73","author":"Ulu","year":"2016","journal-title":"Polym. Bull."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"794","DOI":"10.1016\/j.jmb.2007.03.061","article-title":"Crystal structure and allosteric regulation of the cytoplasmic Escherichia coli L-asparaginase I","volume":"369","author":"Yun","year":"2007","journal-title":"J. Mol. Biol."}],"container-title":["BioTech"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2673-6284\/11\/2\/10\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:53:51Z","timestamp":1760136831000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2673-6284\/11\/2\/10"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,4,14]]},"references-count":69,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2022,6]]}},"alternative-id":["biotech11020010"],"URL":"https:\/\/doi.org\/10.3390\/biotech11020010","relation":{"has-preprint":[{"id-type":"doi","id":"10.20944\/preprints202204.0049.v1","asserted-by":"object"}]},"ISSN":["2673-6284"],"issn-type":[{"value":"2673-6284","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,4,14]]}}}