{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,17]],"date-time":"2026-03-17T04:54:07Z","timestamp":1773723247290,"version":"3.50.1"},"reference-count":51,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2024,5,7]],"date-time":"2024-05-07T00:00:00Z","timestamp":1715040000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"\u201cExcellence Initiative\u2014Research University\u201d program, NCU in Toru\u0144","award":["9\/2022\/Debiuty3"],"award-info":[{"award-number":["9\/2022\/Debiuty3"]}]},{"name":"\u201cExcellence Initiative\u2014Research University\u201d program, NCU in Toru\u0144","award":["6\/2022\/Debiuty3"],"award-info":[{"award-number":["6\/2022\/Debiuty3"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The use of lipase immobilized on an octyl-agarose support to obtain the optically pure enantiomers of chiral drugs in reactions carried out in organic solvents is a great challenge for chemical and pharmaceutical sciences. Therefore, it is extremely important to develop optimal procedures to achieve a high enantioselectivity of the biocatalysts in the organic medium. Our paper describes a new approach to biocatalysis performed in an organic solvent with the use of CALB-octyl-agarose support including the application of a polypropylene reactor, an appropriate buffer for immobilization (Tris base\u2014pH 9, 100 mM), a drying step, and then the storage of immobilized lipases in a climatic chamber or a refrigerator. An immobilized lipase B from Candida antarctica (CALB) was used in the kinetic resolution of (R,S)-flurbiprofen by enantioselective esterification with methanol, reaching a high enantiomeric excess (eep = 89.6 \u00b1 2.0%). As part of the immobilization optimization, the influence of different buffers was investigated. The effect of the reactor material and the reaction medium on the lipase activity was also studied. Moreover, the stability of the immobilized lipases: lipase from Candida rugosa (CRL) and CALB during storage in various temperature and humidity conditions (climatic chamber and refrigerator) was tested. The application of the immobilized CALB in a polypropylene reactor allowed for receiving over 9-fold higher conversion values compared to the results achieved when conducting the reaction in a glass reactor, as well as approximately 30-fold higher conversion values in comparison with free lipase. The good stability of the CALB-octyl-agarose support was demonstrated. After 7 days of storage in a climatic chamber or refrigerator (with protection from humidity) approximately 60% higher conversion values were obtained compared to the results observed for the immobilized form that had not been stored. The new approach involving the application of the CALB-octyl-agarose support for reactions performed in organic solvents indicates a significant role of the polymer reactor material being used in achieving high catalytic activity.<\/jats:p>","DOI":"10.3390\/ijms25105084","type":"journal-article","created":{"date-parts":[[2024,5,7]],"date-time":"2024-05-07T09:03:38Z","timestamp":1715072618000},"page":"5084","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["A New Approach in Lipase-Octyl-Agarose Biocatalysis of 2-Arylpropionic Acid Derivatives"],"prefix":"10.3390","volume":"25","author":[{"given":"Joanna","family":"Si\u00f3dmiak","sequence":"first","affiliation":[{"name":"Department of Laboratory Medicine, Faculty of Pharmacy, Ludwik Rydygier Collegium Medicum, Nicolaus Copernicus University in Toru\u0144, 85-094 Bydgoszcz, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1322-3820","authenticated-orcid":false,"given":"Jacek","family":"Dul\u0119ba","sequence":"additional","affiliation":[{"name":"Department of Medicinal Chemistry, Faculty of Pharmacy, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Toru\u0144, 85-089 Bydgoszcz, Poland"},{"name":"Department of Pharmaceutical Technology, Faculty of Pharmacy, Medical Biotechnology and Laboratory Medicine, Pomeranian Medical University in Szczecin, 71-251 Szczecin, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9107-263X","authenticated-orcid":false,"given":"Natalia","family":"Kocot","sequence":"additional","affiliation":[{"name":"Department of Medicinal Chemistry, Faculty of Pharmacy, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Toru\u0144, 85-089 Bydgoszcz, Poland"},{"name":"Doctoral School of Medical and Health Sciences, Jagiellonian University, \u0141azarza 16, 31-530 Krak\u00f3w, Poland"},{"name":"Department of Pharmaceutical Biochemistry, Faculty of Pharmacy, Jagiellonian University Medical College, Medyczna 9, 30-688 Krak\u00f3w, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8111-8945","authenticated-orcid":false,"given":"Rafa\u0142","family":"Mastalerz","sequence":"additional","affiliation":[{"name":"Department of Medicinal Chemistry, Faculty of Pharmacy, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Toru\u0144, 85-089 Bydgoszcz, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5664-2745","authenticated-orcid":false,"given":"Gudmundur G.","family":"Haraldsson","sequence":"additional","affiliation":[{"name":"Science Institute, University of Iceland, 107 Reykjavik, Iceland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Micha\u0142 Piotr","family":"Marsza\u0142\u0142","sequence":"additional","affiliation":[{"name":"Department of Medicinal Chemistry, Faculty of Pharmacy, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Toru\u0144, 85-089 Bydgoszcz, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9232-5798","authenticated-orcid":false,"given":"Tomasz","family":"Si\u00f3dmiak","sequence":"additional","affiliation":[{"name":"Department of Medicinal Chemistry, Faculty of Pharmacy, Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Toru\u0144, 85-089 Bydgoszcz, Poland"},{"name":"Department of Pharmaceutical Technology, Faculty of Pharmacy, Medical Biotechnology and Laboratory Medicine, Pomeranian Medical University in Szczecin, 71-251 Szczecin, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,5,7]]},"reference":[{"key":"ref_1","first-page":"1550","article-title":"Enantioselective Release of Chiral Drugs","volume":"33","author":"Song","year":"2021","journal-title":"Prog. Chem."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"462914","DOI":"10.1016\/j.chroma.2022.462914","article-title":"Progress of molecular imprinting technique for enantioseparation of chiral drugs in recent ten years","volume":"1668","author":"Pu","year":"2022","journal-title":"J. Chromatogr. A"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1403","DOI":"10.1002\/chir.23498","article-title":"Biocatalysis: A smart and green tool for the preparation of chiral drugs","volume":"34","author":"Rossino","year":"2022","journal-title":"Chirality"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1002\/bit.26458","article-title":"Burkholderia cepacia lipase: A versatile catalyst in synthesis reactions","volume":"115","author":"Sanchez","year":"2018","journal-title":"Biotechnol. Bioeng."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2348","DOI":"10.1194\/jlr.M063388","article-title":"Open and closed states of Candida antarctica lipase B: Protonation and the mechanism of interfacial activation","volume":"56","author":"Stauch","year":"2015","journal-title":"J. Lipid Res."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"5969","DOI":"10.1021\/acs.biochem.5b00586","article-title":"Interfacial Activation of Candida antarctica Lipase B: Combined Evidence from Experiment and Simulation","volume":"54","author":"Zisis","year":"2015","journal-title":"Biochemistry"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2380","DOI":"10.1039\/C9CY00415G","article-title":"Novozym 435: The \u201cperfect\u201d lipase immobilized biocatalyst?","volume":"9","author":"Ortiz","year":"2019","journal-title":"Catal. Sci. Technol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1016\/j.enzmictec.2015.02.001","article-title":"Immobilization of lipases on hydrophobic supports involves the open form of the enzyme","volume":"71","author":"Manoel","year":"2015","journal-title":"Enzyme Microb. Technol."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Souza, P.M.P., Carballares, D., Goncalves, L.R.B., Fernandez-Lafuente, R., and Rodrigues, S. (2022). Immobilization of Lipase B from Candida antarctica in Octyl-Vinyl Sulfone Agarose: Effect of the Enzyme-Support Interactions on Enzyme Activity, Specificity, Structure and Inactivation Pathway. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms232214268"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1016\/j.jbiotec.2018.07.024","article-title":"Prediction of enantioselectivity of lipase catalyzed kinetic resolution using umbrella sampling","volume":"283","author":"Mathpati","year":"2018","journal-title":"J. Biotechnol."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Ferraccioli, R. (2021). Progress on the Stereoselective Synthesis of Chiral Molecules Based on Metal-Catalyzed Dynamic Kinetic Resolution of Alcohols with Lipases. Symmetry, 13.","DOI":"10.3390\/sym13091744"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"29682","DOI":"10.3390\/ijms161226191","article-title":"Recent Advances in Lipase-Mediated Preparation of Pharmaceuticals and Their Intermediates","volume":"16","author":"Carvalho","year":"2015","journal-title":"Int. J. Mol. Sci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"146","DOI":"10.1007\/s12010-020-03272-3","article-title":"Synthesizing Chiral Drug Intermediates by Biocatalysis","volume":"192","author":"Jiang","year":"2020","journal-title":"Appl. Biochem. Biotechnol."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Ceramella, J., Iacopetta, D., Franchini, A., De Luca, M., Saturnino, C., Andreu, I., Sinicropi, M.S., and Catalano, A. (2022). A Look at the Importance of Chirality in Drug Activity: Some Significative Examples. Appl. Sci., 12.","DOI":"10.3390\/app122110909"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"663","DOI":"10.1002\/chir.22362","article-title":"Kinetic Resolution of Profens by Enantioselective Esterification Catalyzed by Candida antarctica and Candida rugosa Lipases","volume":"26","author":"Sikora","year":"2014","journal-title":"Chirality"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"346","DOI":"10.1002\/bit.25361","article-title":"Simple Amino Acid Tags Improve Both Expression and Secretion of Candida antarctica Lipase B in Recombinant Escherichia coli","volume":"112","author":"Kim","year":"2015","journal-title":"Biotechnol. Bioeng."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Si\u00f3dmiak, T., Dul\u0119ba, J., Haraldsson, G.G., Si\u00f3dmiak, J., and Marsza\u0142\u0142, M.P. (2023). The Studies of Sepharose-Immobilized Lipases: Combining Techniques for the Enhancement of Activity and Thermal Stability. Catalysts, 13.","DOI":"10.3390\/catal13050887"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Si\u00f3dmiak, T., Haraldsson, G.G., Dul\u0119ba, J., Ziegler-Borowska, M., Si\u00f3dmiak, J., and Marsza\u0142\u0142, M.P. (2020). Evaluation of Designed Immobilized Catalytic Systems: Activity Enhancement of Lipase B from Candida antarctica. Catalysts, 10.","DOI":"10.3390\/catal10080876"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2769","DOI":"10.1007\/s12010-014-1455-4","article-title":"High Enantioselective Novozym 435-Catalyzed Esterification of (R,S)-Flurbiprofen Monitored with a Chiral Stationary Phase","volume":"175","author":"Mangelings","year":"2015","journal-title":"Appl. Biochem. Biotechnol."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Suster, C.R.L., Toledo, M.V., Matkovic, S.R., Morcelle, S.R., and Briand, L.E. (2023). Rational Design of a Biocatalyst Based on Immobilized CALB onto Nanostructured SiO2. Catalysts, 13.","DOI":"10.3390\/catal13030625"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Khiari, O., Bouzemi, N., S\u00e1nchez-Montero, J.M., and Alc\u00e1ntara, A.R. (2023). On the Use of Orthoformates as an Efficient Approach to Enhance the Enzymatic Enantioselective Synthesis of (S)-Ibuprofen. Catalysts, 13.","DOI":"10.3390\/catal13020251"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"e23628","DOI":"10.1002\/chir.23628","article-title":"Enantioselective adsorption of ibuprofen enantiomers using chiral-active carbon nanoparticles induced (S)-\u03b1-methylbenzylamine","volume":"36","author":"Zhao","year":"2023","journal-title":"Chirality"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Zdun, B., Cie\u015bla, P., Kutner, J., and Borowiecki, P. (2022). Expanding Access to Optically Active Non-Steroidal Anti-Inflammatory Drugs via Lipase-Catalyzed KR of Racemic Acids Using Trialkyl Orthoesters as Irreversible Alkoxy Group Donors. Catalysts, 12.","DOI":"10.3390\/catal12050546"},{"key":"ref_24","first-page":"53","article-title":"Stable and efficient biopolymeric nanocompozite of Candida antarctica Lipase B","volume":"68","author":"Spelmezan","year":"2023","journal-title":"Stud. Univ. Babes-Bolyai Chem."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Silva, F., Szemes, J., Mustashev, A., Tak\u00e1cs, O., Imarah, A.O., and Poppe, L. (2023). Immobilization of Lipase B from Candida antarctica on Magnetic Nanoparticles Enhances Its Selectivity in Kinetic Resolutions of Chiral Amines with Several Acylating Agents. Life, 13.","DOI":"10.3390\/life13071560"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"603","DOI":"10.1042\/bj20020404","article-title":"Multiple mutagenesis of non-universal serine codons of the Candida rugosa LIP2 gene and biochemical characterization of purified recombinant LIP2 lipase overexpressed in Pichia pastoris","volume":"366","author":"Lee","year":"2002","journal-title":"Biochem. Technol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"11510","DOI":"10.1021\/acs.langmuir.8b02255","article-title":"Effect of Surfactant Structure on the Superactivity of Candida rugosa Lipase","volume":"34","author":"Gabriele","year":"2018","journal-title":"Langmuir"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1016\/j.molcatb.2016.04.006","article-title":"Improving enantioselectivity of lipase from Candida rugosa by carrier-bound and carrier-free immobilization","volume":"130","year":"2016","journal-title":"J. Mol. Catal. B Enzym"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"382","DOI":"10.1002\/chir.22444","article-title":"Biocatalytic Approach for the Synthesis of Enantiopure Acebutolol as a \u03b21-Selective Blocker","volume":"27","author":"Banoth","year":"2015","journal-title":"Chirality"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"104613","DOI":"10.1016\/j.reactfunctpolym.2020.104613","article-title":"On the taught new tricks of enzymes immobilization: An all-inclusive overview","volume":"152","author":"Wahab","year":"2020","journal-title":"React. Funct. Polym."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"117579","DOI":"10.1016\/j.envres.2023.117579","article-title":"Enzyme-linked carbon nanotubes as biocatalytic tools to degrade and mitigate environmental pollutants","volume":"241","author":"Bilal","year":"2024","journal-title":"Environ. Res."},{"key":"ref_32","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_33","doi-asserted-by":"crossref","first-page":"927","DOI":"10.1021\/acs.biomac.0c01608","article-title":"Functionalization of Porous Cellulose with Glyoxyl Groups as a Carrier for Enzyme Immobilization and Stabilization","volume":"22","author":"Orrego","year":"2021","journal-title":"Biomacromolecules"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Alnoch, R.C., dos Santos, L.A., de Almeida, J.M., Krieger, N., and Mateo, C. (2020). Recent Trends in Biomaterials for Immobilization of Lipases for Application in Non-Conventional Media. Catalysts, 10.","DOI":"10.3390\/catal10060697"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Si\u00f3dmiak, T., Si\u00f3dmiak, J., Mastalerz, R., Kocot, N., Dul\u0119ba, J., Haraldsson, G.G., W\u0105tr\u00f3bska-\u015awietlikowska, D., and Marsza\u0142\u0142, M.P. (2023). Climatic Chamber Stability Tests of Lipase-Catalytic Octyl-Sepharose Systems. Catalysts, 13.","DOI":"10.3390\/catal13030501"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"108","DOI":"10.1016\/j.colsurfb.2015.04.041","article-title":"Conformation and activity of lipase B from Candida antarctica in bicontinuous microemulsions","volume":"131","author":"Subinya","year":"2015","journal-title":"Colloids Surf. B Biointerfaces"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1700266","DOI":"10.1002\/ejlt.201700266","article-title":"Immobilization of Candida antarctica Lipase B Onto ECR1030 Resin and its Application in the Synthesis of n-3 PUFA-Rich Triacylglycerols","volume":"119","author":"Li","year":"2017","journal-title":"Eur. J. Lipid Sci. Technol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"109461","DOI":"10.1016\/j.enzmictec.2019.109461","article-title":"Modulating the properties of the lipase from Thermomyces lanuginosus immobilized on octyl agarose beads by altering the immobilization conditions","volume":"133","author":"Lokha","year":"2020","journal-title":"Enzyme Microb. Technol."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Arana-Pena, S., Lokha, Y., and Fernandez-Lafuente, R. (2019). Immobilization on octyl-agarose beads and some catalytic features of commercial preparations of lipase A from Candida antarctica (Novocor ADL): Comparison with immobilized lipase B from Candida antarctica. Biotechnol. Prog., 35.","DOI":"10.1002\/btpr.2735"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1016\/j.procbio.2023.08.003","article-title":"The nature of the buffer alters the effects of the chemical modification on the stability of immobilized lipases","volume":"133","author":"Carballares","year":"2023","journal-title":"Process Biochem."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"2843","DOI":"10.1007\/s12010-021-03570-4","article-title":"Effect of Tris Buffer in the Intensity of the Multipoint Covalent Immobilization of Enzymes in Glyoxyl-Agarose Beads","volume":"193","author":"Braham","year":"2021","journal-title":"Appl. Biochem. Biotechnol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"919","DOI":"10.1002\/2211-5463.12097","article-title":"Effect of Tris, MOPS, and phosphate buffers on the hydrolysis of polyethylene terephthalate films by polyester hydrolases","volume":"6","author":"Schmidt","year":"2016","journal-title":"FEBS Open Bio."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Abellanas-Perez, P., Carballares, D., Rocha-Martin, J., and Fernandez-Lafuente, R. (2024). The Effects of Buffer Nature on Immobilized Lipase Stability Depend on Enzyme Support Loading. Catalysts, 14.","DOI":"10.3390\/catal14020105"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"11499","DOI":"10.1021\/acs.langmuir.1c01726","article-title":"Molecular Mechanistic Insights into the Ionic-Strength-Controlled Interfacial Behavior of Proteins on a TiO2 Surface","volume":"37","author":"Dong","year":"2021","journal-title":"Langmuir"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"436","DOI":"10.1002\/jctb.6876","article-title":"Polypropylene as a selective support for the immobilization of lipolytic enzymes: Hyper-activation, purification and biotechnological applications","volume":"97","author":"Nadda","year":"2022","journal-title":"J. Chem. Technol. Biotechnol."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Godoy, C.A. (2017). New Strategy for the Immobilization of Lipases on Glyoxyl-Agarose Supports: Production of Robust Biocatalysts for Natural Oil Transformation. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18102130"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1016\/j.molcatb.2014.03.005","article-title":"Enantioselective resolution of racemic ibuprofen esters using different lipases immobilized on octyl sepharose","volume":"104","author":"Yousefi","year":"2014","journal-title":"J. Mol. Catal. B Enzym."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1016\/j.molcatb.2016.02.016","article-title":"Synthesis of butyl butyrate in batch and continuous enzymatic reactors using Thermomyces lanuginosus lipase immobilized in Immobead 150","volume":"127","author":"Matte","year":"2016","journal-title":"J. Mol. Catal. B Enzym."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"2507","DOI":"10.1007\/s12010-013-0702-4","article-title":"Immobilization of Thermomyces lanuginosus Lipase by Different Techniques on Immobead 150 Support: Characterization and Applications","volume":"172","author":"Matte","year":"2014","journal-title":"Appl. Biochem. Biotechnol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"6350","DOI":"10.1021\/acscatal.6b02073","article-title":"A Correlation between the Activity of Candida antarctica Lipase B and Differences in Binding Free Energies of Organic Solvent and Substrate","volume":"6","author":"Banik","year":"2016","journal-title":"ACS Catal."},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Si\u00f3dmiak, T., Dul\u0119ba, J., Kocot, N., W\u0105trobska-\u015awietlikowska, D., and Marsza\u0142\u0142, M.P. (2022). The High \u2018Lipolytic Jump\u2019 of Immobilized Amano A Lipase from Aspergillus niger in Developed \u2018ESS Catalytic Triangles\u2019 Containing Natural Origin Substrates. 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