{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,17]],"date-time":"2026-02-17T10:12:17Z","timestamp":1771323137692,"version":"3.50.1"},"reference-count":77,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2026,2,17]],"date-time":"2026-02-17T00:00:00Z","timestamp":1771286400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100011033","name":"Ministerio de Ciencia e Innovaci\u00f3n and Agencia Estatal de Investigaci\u00f3n","doi-asserted-by":"publisher","award":["PID2022-136535OB-I00"],"award-info":[{"award-number":["PID2022-136535OB-I00"]}],"id":[{"id":"10.13039\/501100011033","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002850","name":"FONDECYT","doi-asserted-by":"publisher","award":["1231631(MU)"],"award-info":[{"award-number":["1231631(MU)"]}],"id":[{"id":"10.13039\/501100002850","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Beca Doctorado Nacional","award":["21212403"],"award-info":[{"award-number":["21212403"]}]},{"name":"Agencia Nacional de Investigaci\u00f3n y Desarrollo ANID"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>The lipase B from Candida antarctica was immobilized on octyl-agarose using low and high (one that saturated the support surface with enzyme) loadings. Then, both biocatalysts were aminated, and the aminated and non-aminated biocatalysts were used in further experiments. The enzyme activity was determined using substrates with different structures. The modification of the four biocatalysts with poly (ethylene-alt-maleic anhydride) revealed that only a marginal covalent reaction occurs. That way, the ion exchange of the polymer on the immobilized enzyme surface should be responsible for the enzyme functional changes. The modification of the biocatalysts with this polymer produced mixed results for enzyme activity (depending on the enzyme loading, use of aminated or non-aminated enzyme, polymer concentration and used substrate), in some instances more than doubling the activity, in others reducing it by 5\u20136 times the activity when compared to the unmodified biocatalyst. The effects on biocatalyst stability were also mixed, depending on the same factors; in some instances, great stabilization could be found (e.g., in inactivation of the highly loaded aminated biocatalyst at pH 7.0, the unmodified biocatalyst kept 5% of the initial activity, while the biocatalyst modified with 1% of the polymer maintained 80%), but in other instances, enzyme stability was reduced after modification. It was shown that one of the effects of the polymer modification was the prevention of the enzyme release during inactivation.<\/jats:p>","DOI":"10.3390\/molecules31040691","type":"journal-article","created":{"date-parts":[[2026,2,17]],"date-time":"2026-02-17T09:22:46Z","timestamp":1771320166000},"page":"691","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Poly (Ethylene-Alt-Maleic Anhydride) Ionic Modification of Lipase B from Candida antarctica Immobilized on Octyl Agarose Beads Alters Its Catalytic Properties"],"prefix":"10.3390","volume":"31","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-1409-2485","authenticated-orcid":false,"given":"Alex D.","family":"Gonzalez-Vasquez","sequence":"first","affiliation":[{"name":"Departamento de Biocatalisis, Instituto de Catalisis y Petrolooquimica-Consejo Superior de Investigaciones Cientificas, Campus de Excelencia Universidad Autonoma de Madrid-Consejo Superior de Investigaciones Cientificas, Cantoblanco, 28049 Madrid, Spain"},{"name":"Departamento de Qu\u00edmica, Facultad de Ciencias, Universidad de Chile, Las Palmeras 3425, Casilla 653, \u00d1u\u00f1oa, Santiago 7800003, Chile"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-8316-7013","authenticated-orcid":false,"given":"Pedro","family":"Abellanas-Perez","sequence":"additional","affiliation":[{"name":"Departamento de Biocatalisis, Instituto de Catalisis y Petrolooquimica-Consejo Superior de Investigaciones Cientificas, Campus de Excelencia Universidad Autonoma de Madrid-Consejo Superior de Investigaciones Cientificas, Cantoblanco, 28049 Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7681-5439","authenticated-orcid":false,"given":"Javier","family":"Rocha-Martin","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Molecular Biology, Faculty of Biology, Complutense University of Madrid, Jos\u00e9 Antonio Novais 12, 28040 Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2371-620X","authenticated-orcid":false,"given":"Marcela","family":"Urz\u00faa","sequence":"additional","affiliation":[{"name":"Departamento de Qu\u00edmica, Facultad de Ciencias, Universidad de Chile, Las Palmeras 3425, Casilla 653, \u00d1u\u00f1oa, Santiago 7800003, Chile"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4976-7096","authenticated-orcid":false,"given":"Roberto","family":"Fernandez-Lafuente","sequence":"additional","affiliation":[{"name":"Departamento de Biocatalisis, Instituto de Catalisis y Petrolooquimica-Consejo Superior de Investigaciones Cientificas, Campus de Excelencia Universidad Autonoma de Madrid-Consejo Superior de Investigaciones Cientificas, Cantoblanco, 28049 Madrid, Spain"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,2,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"3370","DOI":"10.1002\/bit.28245","article-title":"Fungal Lipases as Biocatalysts: A Promising Platform in Several Industrial Biotechnology Applications","volume":"119","author":"Mahfoudhi","year":"2022","journal-title":"Biotechnol. Bioeng."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"119219","DOI":"10.1016\/j.jenvman.2023.119219","article-title":"Agri-Residues and Agro-Industrial Waste Substrates Bioconversion by Fungal Cultures to Biocatalyst Lipase for Green Chemistry: A Review","volume":"348","author":"Singh","year":"2023","journal-title":"J. Environ. Manag."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"81","DOI":"10.3109\/07388551.2012.672319","article-title":"Lipase Applications in Oil Hydrolysis with a Case Study on Castor Oil: A Review","volume":"33","author":"Goswami","year":"2013","journal-title":"Crit. Rev. Biotechnol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"5696","DOI":"10.1039\/D1CY00696G","article-title":"Solvent-Free Esterifications Mediated by Immobilized Lipases: A Review from Thermodynamic and Kinetic Perspectives","volume":"11","author":"Sousa","year":"2021","journal-title":"Catal. Sci. Technol."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Cheng, W., and Nian, B. (2023). Computer-Aided Lipase Engineering for Improving Their Stability and Activity in the Food Industry: State of the Art. Molecules, 28.","DOI":"10.3390\/molecules28155848"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1016\/0141-0229(93)90123-J","article-title":"Lipase-Catalyzed Transesterification in Organic Solvents: Applications to the Preparation of Enantiomerically Pure Compounds","volume":"15","author":"Santaniello","year":"1993","journal-title":"Enzym. Microb. Technol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1016\/j.molcatb.2009.09.018","article-title":"Biodiesel Production through Lipase Catalyzed Transesterification: An Overview","volume":"62","author":"Bajaj","year":"2010","journal-title":"J. Mol. Catal. B Enzym."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1016\/j.procbio.2024.03.005","article-title":"Production of Structured Lipids via Batch and Continuous Acidolysis of Triglycerides Using the Hydration\u2013Aggregation-Pretreated Lipase Loaded onto Surface-Modified Glass Bead Supports","volume":"141","author":"Yoshioka","year":"2024","journal-title":"Process Biochem."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"142878","DOI":"10.1016\/j.ijbiomac.2025.142878","article-title":"A Comprehensive Review of Lipase-Catalyzed Acidolysis as a Method for Producing Structured Glycerides","volume":"309","author":"Tavano","year":"2025","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"305","DOI":"10.17113\/ftb.57.03.19.6025","article-title":"Lipase-Catalyzed Interesterification for the Synthesis of Medium-Long-Medium (MLM) Structured Lipids","volume":"57","author":"Utama","year":"2019","journal-title":"Food Technol. Biotechnol."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Sun, M., Nie, K., Wang, F., and Deng, L. (2020). Optimization of the Lipase-Catalyzed Selective Amidation of Phenylglycinol. Front. Bioeng. Biotechnol., 7.","DOI":"10.3389\/fbioe.2019.00486"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"14419","DOI":"10.1021\/ja9052015","article-title":"Dynamic Asymmetric Multicomponent Resolution: Lipase-Mediated Amidation of a Double Dynamic Covalent System","volume":"131","author":"Vongvilai","year":"2009","journal-title":"J. Am. Chem. Soc."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2554","DOI":"10.1002\/cctc.201900397","article-title":"Biocatalytic Promiscuity of Lipases in Carbon-Phosphorus Bond Formation","volume":"11","author":"Koszelewski","year":"2019","journal-title":"ChemCatChem"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"555","DOI":"10.1016\/j.procbio.2012.01.011","article-title":"Lipase Promiscuity and Its Biochemical Applications","volume":"47","author":"Kapoor","year":"2012","journal-title":"Process Biochem."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Reyes-Reyes, A.L., Valero Barranco, F., and Sandoval, G. (2022). Recent Advances in Lipases and Their Applications in the Food and Nutraceutical Industry. Catalysts, 12.","DOI":"10.3390\/catal12090960"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"101509","DOI":"10.1016\/j.fbio.2021.101509","article-title":"Microbial Lipases: Propitious Biocatalysts for the Food Industry","volume":"45","author":"Salgado","year":"2022","journal-title":"Food Biosci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"119085","DOI":"10.1016\/j.renene.2023.119085","article-title":"Demystifying the Enzymatic Biodiesel: How Lipases Are Contributing to Its Technological Advances","volume":"216","author":"Wancura","year":"2023","journal-title":"Renew. Energy"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"108215","DOI":"10.1016\/j.biotechadv.2023.108215","article-title":"Biocatalytic Production of Biolubricants: Strategies, Problems and Future Trends","volume":"68","author":"Monteiro","year":"2023","journal-title":"Biotechnol. Adv."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1007\/s11274-022-03465-4","article-title":"Recent Advances and Future Prospects for Biolubricant Base Stocks Production Using Lipases as Environmentally Friendly Catalysts: A Mini-Review","volume":"39","author":"Mendes","year":"2023","journal-title":"World J. Microbiol. Biotechnol."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Verma, S., Choudhary, R.N., Kanadje, A.P., and Banerjee, U.C. (2021). Diversifying Arena of Drug Synthesis: In the Realm of Lipase Mediated Waves of Biocatalysis. Catalysts, 11.","DOI":"10.3390\/catal11111328"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Todea, A., Dreava, D.M., Benea, I.C., B\u00eetcan, I., Peter, F., and Boeriu, C.G. (2021). Achievements and Trends in Biocatalytic Synthesis of Specialty Polymers from Biomass-Derived Monomers Using Lipases. Processes, 9.","DOI":"10.3390\/pr9040646"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2484","DOI":"10.1007\/s10924-023-03118-z","article-title":"Use of Lipases as a Sustainable and Efficient Method for the Synthesis and Degradation of Polymers","volume":"32","year":"2024","journal-title":"J. Polym. Environ."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"012008","DOI":"10.1088\/1755-1315\/1163\/1\/012008","article-title":"Lipase-Producing Marine Actinomycetes Having Potential to Degrade PET Film","volume":"1163","author":"Agustriana","year":"2023","journal-title":"IOP Conf. Ser. Earth Environ. Sci."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"130871","DOI":"10.1016\/j.biortech.2024.130871","article-title":"Biodegradation of Oxidized Low Density Polyethylene by Pelosinus fermentans Lipase","volume":"403","author":"Kim","year":"2024","journal-title":"Bioresour. Technol."},{"key":"ref_25","first-page":"405","article-title":"Application of Lipases and Phospholipases in Bioremediation of Oil-Contaminated Environments\/Habitats","volume":"Volume 1","author":"Chakraborti","year":"2023","journal-title":"Phospholipases in Physiology and Pathology"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"15071","DOI":"10.1021\/bi0013905","article-title":"Structural Origins of the Interfacial Activation in Thermomyces (Humicola) lanuginosa Lipase","volume":"39","author":"Brzozowski","year":"2000","journal-title":"Biochemistry"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"6223","DOI":"10.1039\/C3CS60075K","article-title":"Enzyme Immobilisation in Biocatalysis: Why, What and How","volume":"42","author":"Sheldon","year":"2013","journal-title":"Chem. Soc. Rev."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"6437","DOI":"10.1039\/c3cs35506c","article-title":"Industrial Use of Immobilized Enzymes","volume":"42","author":"DiCosimo","year":"2013","journal-title":"Chem. Soc. Rev."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/S0065-2164(08)70352-6","article-title":"Stabilization of Enzymes against Thermal Inactivation","volume":"29","author":"Klibanov","year":"1983","journal-title":"Adv. Appl. Microbiol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"107821","DOI":"10.1016\/j.biotechadv.2021.107821","article-title":"Stabilization of Enzymes via Immobilization: Multipoint Covalent Attachment and Other Stabilization Strategies","volume":"52","author":"Rodrigues","year":"2021","journal-title":"Biotechnol. Adv."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"435","DOI":"10.1016\/j.biotechadv.2015.03.006","article-title":"Strategies for the One-Step Immobilization\u2013Purification of Enzymes as Industrial Biocatalysts","volume":"33","author":"Barbosa","year":"2015","journal-title":"Biotechnol. Adv."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"6290","DOI":"10.1039\/C2CS35231A","article-title":"Modifying Enzyme Activity and Selectivity by Immobilization","volume":"42","author":"Rodrigues","year":"2013","journal-title":"Chem. Soc. Rev."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2885","DOI":"10.1002\/adsc.201100163","article-title":"Potential of Different Enzyme Immobilization Strategies to Improve Enzyme Performance","volume":"353","author":"Rodrigues","year":"2011","journal-title":"Adv. Synth. Catal."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"66","DOI":"10.1016\/j.procbio.2019.11.026","article-title":"Parameters Necessary to Define an Immobilized Enzyme Preparation","volume":"90","author":"Boudrant","year":"2020","journal-title":"Process Biochem."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"6251","DOI":"10.1039\/D2CS00083K","article-title":"Is Enzyme Immobilization a Mature Discipline? Some Critical Considerations to Capitalize on the Benefits of Immobilization","volume":"51","author":"Bolivar","year":"2022","journal-title":"Chem. Soc. Rev."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"746","DOI":"10.1016\/j.biotechadv.2019.04.003","article-title":"Immobilization of Lipases on Hydrophobic Supports: Immobilization Mechanism, Advantages, Problems, and Solutions","volume":"37","author":"Rodrigues","year":"2019","journal-title":"Biotechnol. Adv."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"989","DOI":"10.1016\/j.ijbiomac.2019.03.163","article-title":"New Applications of Glyoxyl-Octyl Agarose in Lipases Co-Immobilization: Strategies to Reuse the Most Stable Lipase","volume":"131","author":"Rios","year":"2019","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"3259","DOI":"10.1002\/slct.201600274","article-title":"Advantages of Heterofunctional Octyl Supports: Production of 1,2-Dibutyrin by Specific and Selective Hydrolysis of Tributyrin Catalyzed by Immobilized Lipases","volume":"1","author":"Hirata","year":"2016","journal-title":"ChemistrySelect"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.molcatb.2016.03.002","article-title":"Reversible Immobilization of Lipases on Octyl-Glutamic Agarose Beads: A Mixed Adsorption That Reinforces Enzyme Immobilization","volume":"128","author":"Rueda","year":"2016","journal-title":"J. Mol. Catal. B Enzym."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Rueda, N., Albuquerque, T., Bartolome-Cabrero, R., Fernandez-Lopez, L., Torres, R., Ortiz, C., dos Santos, J., Barbosa, O., and Fernandez-Lafuente, R. (2016). Reversible Immobilization of Lipases on Heterofunctional Octyl-Amino Agarose Beads Prevents Enzyme Desorption. Molecules, 21.","DOI":"10.3390\/molecules21050646"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"11212","DOI":"10.1039\/C4RA13338B","article-title":"Improved Performance of Lipases Immobilized on Heterofunctional Octyl-Glyoxyl Agarose Beads","volume":"5","author":"Rueda","year":"2015","journal-title":"RSC Adv."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"2055","DOI":"10.1016\/j.procbio.2016.09.016","article-title":"Improved Catalytic Properties of Candida antarctica Lipase B Multi-Attached on Tailor-Made Hydrophobic Silica Containing Octyl and Multifunctional Amino-Glutaraldehyde Spacer Arms","volume":"51","author":"Vescovi","year":"2016","journal-title":"Process Biochem."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"865","DOI":"10.1016\/j.procbio.2016.04.002","article-title":"Easy Stabilization of Interfacially Activated Lipases Using Heterofunctional Divinyl Sulfone Activated-Octyl Agarose Beads. Modulation of the Immobilized Enzymes by Altering Their Nanoenvironment","volume":"51","author":"Albuquerque","year":"2016","journal-title":"Process Biochem."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1016\/j.enzmictec.2017.07.001","article-title":"Improved Stability of Immobilized Lipases via Modification with Polyethylenimine and Glutaraldehyde","volume":"106","author":"Zaak","year":"2017","journal-title":"Enzym. Microb. Technol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1016\/j.molcatb.2012.03.012","article-title":"Influence of Different Immobilization Techniques for Candida cylindracea Lipase on Its Stability and Fish Oil Hydrolysis","volume":"78","author":"Pizarro","year":"2012","journal-title":"J. Mol. Catal. B Enzym."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1016\/j.procbio.2016.12.018","article-title":"Physical Crosslinking of Lipase from Rhizomucor miehei Immobilized on Octyl Agarose via Coating with Ionic Polymers: Avoiding Enzyme Release from the Support","volume":"54","author":"Pedrero","year":"2017","journal-title":"Process Biochem."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"125853","DOI":"10.1016\/j.ijbiomac.2023.125853","article-title":"Glutaraldehyde Modification of Lipases Immobilized on Octyl Agarose Beads: Roles of the Support Enzyme Loading and Chemical Amination of the Enzyme on the Final Enzyme Features","volume":"248","author":"Carballares","year":"2023","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1511","DOI":"10.1016\/j.procbio.2014.05.009","article-title":"Stabilizing Hyperactivated Lecitase Structures through Physical Treatment with Ionic Polymers","volume":"49","author":"Rodrigues","year":"2014","journal-title":"Process Biochem."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1016\/j.molcatb.2005.12.007","article-title":"Stabilisation of Oxygen-Labile Nitrilases via Co-Aggregation with Poly(Ethyleneimine)","volume":"38","author":"Mateo","year":"2006","journal-title":"J. Mol. Catal. B Enzym."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1016\/j.molcatb.2007.02.005","article-title":"Hydrophilization of Immobilized Model Enzymes Suggests a Widely Applicable Method for Enhancing Protein Stability in Polar Organic Co-Solvents","volume":"46","author":"Irazoqui","year":"2007","journal-title":"J. Mol. Catal. B Enzym."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"136102","DOI":"10.1016\/j.ijbiomac.2024.136102","article-title":"Designing Mixed Cationic\/Anionic Supports to Covalently Immobilize\/Stabilize Enzymes with High Isoelectric Point by Enzyme Adsorption and Support-Enzyme Glutaraldehyde Crosslinking","volume":"280","author":"Hocine","year":"2024","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1002\/(SICI)1097-0290(20000405)68:1<98::AID-BIT12>3.0.CO;2-T","article-title":"Reversible Enzyme Immobilization via a Very Strong and Nondistorting Ionic Adsorption on Support-Polyethylenimine Composites","volume":"68","author":"Mateo","year":"2000","journal-title":"Biotechnol. Bioeng."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1134","DOI":"10.1021\/bp0499449","article-title":"Reversible and Strong Immobilization of Proteins by Ionic Exchange on Supports Coated with Sulfate-Dextran","volume":"20","author":"Fuentes","year":"2004","journal-title":"Biotechnol. Prog."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"111269","DOI":"10.1016\/j.eurpolymj.2022.111269","article-title":"Blends Based on Amino Acid Functionalized Poly (Ethylene-Alt-Maleic Anhydride) Polyelectrolytes and PEO for Nanofiber Elaboration: Biocompatible and Angiogenic Polyelectrolytes","volume":"173","author":"Leal","year":"2022","journal-title":"Eur. Polym. J."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1016\/j.eurpolymj.2018.03.014","article-title":"Multifunctional Crosslinkable Itaconic Acid Copolymers for Enzyme Immobilization","volume":"102","author":"Torger","year":"2018","journal-title":"Eur. Polym. J."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"5333","DOI":"10.1021\/acsabm.3c00603","article-title":"Conformational Changes of Poly(Maleic Anhydride-Alt-Styrene) Modified with Amino Acids in an Aqueous Medium and Their Effect on Cytocompatibility and Hemolytic Response","volume":"6","author":"Maine","year":"2023","journal-title":"ACS Appl. Bio Mater."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"181","DOI":"10.3109\/10242429809003198","article-title":"One Biocatalyst\u2014Many Applications: The Use of Candida antarctica B-Lipase in Organic Synthesis","volume":"16","author":"Anderson","year":"1998","journal-title":"Biocatal. Biotransform."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1016\/S0969-2126(00)00031-9","article-title":"The Sequence, Crystal Structure Determination and Refinement of Two Crystal Forms of Lipase B from Candida antarctica","volume":"2","author":"Uppenberg","year":"1994","journal-title":"Structure"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"147310","DOI":"10.1016\/j.ijbiomac.2025.147310","article-title":"Preparation and Characterization of Different Immobilized and Chemically Modified Preparations of Lipase B from Candida antarctica: Is It the Activation Energy a Good Indicator of the Biocatalyst Expressed Activity?","volume":"323","author":"Hackenhaar","year":"2025","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1016\/j.enzmictec.2016.12.002","article-title":"Effect of Protein Load on Stability of Immobilized Enzymes","volume":"98","author":"Pedrero","year":"2017","journal-title":"Enzym. Microb. Technol."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"616","DOI":"10.1016\/S0076-6879(72)25060-1","article-title":"Carbodiimide Modification of Proteins","volume":"25","author":"Carraway","year":"1972","journal-title":"Methods Enzymol."},{"key":"ref_62","doi-asserted-by":"crossref","unstructured":"Plata, E., Ruiz, M., Ruiz, J., Ortiz, C., Castillo, J.J., and Fern\u00e1ndez-Lafuente, R. (2020). Chemoenzymatic Synthesis of the New 3-((2,3-Diacetoxypropanoyl)Oxy)Propane-1,2-Diyl Diacetate Using Immobilized Lipase B from Candida antarctica and Pyridinium Chlorochromate as an Oxidizing Agent. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21186501"},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"3893","DOI":"10.1007\/s00253-018-8894-8","article-title":"Biocatalytic Production of Mandelic Acid and Analogues: A Review and Comparison with Chemical Processes","volume":"102","year":"2018","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1016\/0022-5193(73)90153-7","article-title":"Effect of Internal Diffusion in Heterogeneous Enzyme Systems: Evaluation of True Kinetic Parameters and Substrate Diffusivity","volume":"42","author":"Engasser","year":"1973","journal-title":"J. Theor. Biol."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"146777","DOI":"10.1016\/j.ijbiomac.2025.146777","article-title":"Multiple Co-Interactions of Different Parameters on the Functional Properties of Immobilized Lipases","volume":"322","author":"Abellanas","year":"2025","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"e3394","DOI":"10.1002\/btpr.3394","article-title":"The Effects of the Chemical Modification on Immobilized Lipase Features Are Affected by the Enzyme Crowding in the Support","volume":"40","author":"Abellanas","year":"2024","journal-title":"Biotechnol. Prog."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"852","DOI":"10.1021\/bm0343895","article-title":"Co-Aggregation of Penicillin G Acylase and Polyionic Polymers: An Easy Methodology To Prepare Enzyme Biocatalysts Stable in Organic Media","volume":"5","author":"Wilson","year":"2004","journal-title":"Biomacromolecules"},{"key":"ref_68","doi-asserted-by":"crossref","unstructured":"Guastaferro, M., Baldino, L., Reverchon, E., and Cardea, S. (2021). Production of Porous Agarose-Based Structures: Freeze-Drying vs. Supercritical CO2 Drying. Gels, 7.","DOI":"10.3390\/gels7040198"},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"1455","DOI":"10.1002\/(SICI)1522-2683(19990601)20:7<1455::AID-ELPS1455>3.0.CO;2-L","article-title":"Effect of Gelation Conditions on the Gel Structure and Resolving Power of Agarose-Based DNA Sequencing Gels","volume":"20","author":"Kusukawa","year":"1999","journal-title":"Electrophoresis"},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1016\/0003-2697(76)90527-3","article-title":"A Rapid and Sensitive Method for the Quantitation of Microgram Quantities of Protein Utilizing the Principle of Protein-Dye Binding","volume":"72","author":"Bradford","year":"1976","journal-title":"Anal. Biochem."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1016\/0005-2744(81)90328-4","article-title":"Effect of Alcohols on the Hydrolysis Catalyzed by Human Pancreatic Carboxylic-Ester Hydrolase","volume":"657","author":"Lombardo","year":"1981","journal-title":"Biochim. Biophys. Acta (BBA)-Enzymol."},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"688","DOI":"10.1016\/j.ijbiomac.2022.03.151","article-title":"Chemical Amination of Immobilized Enzymes for Enzyme Coimmobilization: Reuse of the Most Stable Immobilized and Modified Enzyme","volume":"208","author":"Carballares","year":"2022","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"680","DOI":"10.1038\/227680a0","article-title":"Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4","volume":"227","author":"Laemmli","year":"1970","journal-title":"Nature"},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"510","DOI":"10.1016\/j.enzmictec.2011.02.005","article-title":"Hydrolysis of Triacetin Catalyzed by Immobilized Lipases: Effect of the Immobilization Protocol and Experimental Conditions on Diacetin Yield","volume":"48","author":"Hernandez","year":"2011","journal-title":"Enzym. Microb. Technol."},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1080\/10242422.2016.1278212","article-title":"Optimization of the coating of octyl-CALB with ionic polymers to improve stability and decrease enzyme leakage","volume":"36","author":"Pedrero","year":"2018","journal-title":"Biocatal. Biotransf."},{"key":"ref_76","doi-asserted-by":"crossref","unstructured":"Virgen-Ort\u00edz, J.J., Pedrero, S.G., Fernandez-Lopez, L., L\u00f3pez-Carrobles, N., Gorines, B.C., Otero, C., and Fern\u00e1ndez-Lafuente, R. (2017). Desorption of lipases immobilized on octyl-agarose beads and coated with ionic polymers after thermal inactivation. Stronger adsorption of polymers\/unfolded protein composites. Molecules, 22.","DOI":"10.3390\/molecules22010091"},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"226","DOI":"10.1016\/j.procbio.2008.10.005","article-title":"Enhancement of Novozym-435 catalytic properties by physical or chemical modification","volume":"44","author":"Cabrera","year":"2009","journal-title":"Process Biochem."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/31\/4\/691\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,17]],"date-time":"2026-02-17T09:23:17Z","timestamp":1771320197000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/31\/4\/691"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,2,17]]},"references-count":77,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2026,2]]}},"alternative-id":["molecules31040691"],"URL":"https:\/\/doi.org\/10.3390\/molecules31040691","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,2,17]]}}}