{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,25]],"date-time":"2026-04-25T17:07:55Z","timestamp":1777136875079,"version":"3.51.4"},"reference-count":67,"publisher":"Springer Science and Business Media LLC","issue":"7","license":[{"start":{"date-parts":[[2025,3,31]],"date-time":"2025-03-31T00:00:00Z","timestamp":1743379200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2025,3,31]],"date-time":"2025-03-31T00:00:00Z","timestamp":1743379200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100002347","name":"Bundesministerium f\u00fcr Bildung und Forschung","doi-asserted-by":"publisher","award":["13FH027KX1"],"award-info":[{"award-number":["13FH027KX1"]}],"id":[{"id":"10.13039\/501100002347","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100008033","name":"Hochschule Niederrhein","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100008033","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Appl Biochem Biotechnol"],"published-print":{"date-parts":[[2025,7]]},"abstract":"<jats:title>Abstract<\/jats:title>\n          <jats:p>We investigated the mechanisms of polymer-lipase interactions that govern the catalytic activity of lipases in the presence of polymers. Using a combination of fluorescence correlation spectroscopy (FCS), activity analysis, fluorescence spectroscopy, and computational surface analysis, three model lipases\u2014<jats:italic>Thermomyces lanuginosus<\/jats:italic> lipase (TLL), <jats:italic>Candida antarctica<\/jats:italic> lipase B (CalB), and <jats:italic>Bacillus subtilis<\/jats:italic> lipase A (BSLA), with different degrees of hydrophobic active site exposure were studied. Low-molecular-weight poly(methyl methacrylate) (PMMA), synthesized via ARGET ATRP, was employed to study the effect of unstructured polymers in dispersed solution on lipase activity. PMMA significantly enhanced TLL and BSLA hydrolytic activity, while no CalB activation was observed. FCS analysis indicated that this activation was facilitated by polymer lipase binding, a phenomenon observed with TLL and BSLA but not with CalB. Computational analysis further revealed that the surface properties of the lipases were critical for the lipases\u2019 susceptibility to activation by PMMA. Although CalB exhibited the largest total hydrophobic surface area, its homogeneous distribution prevented activation, whereas strong, localized hydrophobic interactions allowed PMMA to bind and activate TLL and BSLA. Supported by the quantitative correlation between elevated 8-anilino-1-naphthalenesulfonic acid (ANS) fluorescence in the presence of PMMA and lipase activity, the activation was attributed to locally increased hydrophobicity of the lipases upon polymer binding. These findings provide critical insights into the role of polymer interactions in lipase activation and stabilization, highlighting the potential for designing tailored polymer carriers to optimize enzyme performance in industrial and biotechnological applications.<\/jats:p>","DOI":"10.1007\/s12010-025-05217-0","type":"journal-article","created":{"date-parts":[[2025,4,1]],"date-time":"2025-04-01T22:23:12Z","timestamp":1743546192000},"page":"4315-4331","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Lipase Activation by Poly(Methyl Methacrylate) in Dispersed Solution: Mechanistic Insights by Fluorescence Spectroscopy"],"prefix":"10.1007","volume":"197","author":[{"given":"Andr\u00e9","family":"Merz","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jonas","family":"Thelen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"J\u00fcrgen","family":"Linders","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christian","family":"Mayer","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7526-0953","authenticated-orcid":false,"given":"Kerstin","family":"Hoffmann-Jacobsen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2025,3,31]]},"reference":[{"key":"5217_CR1","doi-asserted-by":"publisher","first-page":"7399","DOI":"10.1007\/s00253-019-10027-6","volume":"103","author":"DG Filho","year":"2019","unstructured":"Filho, D. G., Silva, A. G., & Guidini, C. Z. (2019). Lipases: Sources, immobilization methods, and industrial applications. Applied Microbiology and Biotechnology, 103, 7399\u20137423. https:\/\/doi.org\/10.1007\/s00253-019-10027-6","journal-title":"Applied Microbiology and Biotechnology"},{"key":"5217_CR2","doi-asserted-by":"publisher","first-page":"88","DOI":"10.1002\/anie.202006648","volume":"60","author":"S Wu","year":"2021","unstructured":"Wu, S., Snajdrova, R., Moore, J. C., et al. (2021). Biocatalysis: Enzymatic synthesis for industrial applications. Angewandte Chemie International Edition, 60, 88\u2013119.","journal-title":"Angewandte Chemie International Edition"},{"key":"5217_CR3","doi-asserted-by":"publisher","first-page":"155","DOI":"10.1385\/ABAB:118:1-3:155","volume":"118","author":"A Houde","year":"2004","unstructured":"Houde, A., Kademi, A., & Leblanc, D. (2004). Lipases and their industrial applications: An overview. Applied Biochemistry and Biotechnology, 118, 155\u2013170.","journal-title":"Applied Biochemistry and Biotechnology"},{"key":"5217_CR4","doi-asserted-by":"publisher","first-page":"169","DOI":"10.1186\/s12934-020-01428-8","volume":"19","author":"P Chandra","year":"2020","unstructured":"Chandra, P., Enespa, S. R., et al. (2020). Microbial lipases and their industrial applications: A comprehensive review. Microbial Cell Factories, 19, 169. https:\/\/doi.org\/10.1186\/s12934-020-01428-8","journal-title":"Microbial Cell Factories"},{"key":"5217_CR5","doi-asserted-by":"publisher","first-page":"29682","DOI":"10.3390\/ijms161226191","volume":"16","author":"ACLdM Carvalho","year":"2015","unstructured":"Carvalho, A. CLd. M., Fonseca, Td. S., de Mattos, M. C., et al. (2015). Recent advances in lipase-mediated preparation of pharmaceuticals and their intermediates. International Journal of Molecular Sciences, 16, 29682\u201329716.","journal-title":"International Journal of Molecular Sciences"},{"key":"5217_CR6","doi-asserted-by":"publisher","first-page":"2441","DOI":"10.1002\/slct.201702954","volume":"3","author":"BP Dwivedee","year":"2018","unstructured":"Dwivedee, B. P., Soni, S., Sharma, M., et al. (2018). Promiscuity of lipase-catalyzed reactions for organic synthesis: A recent update. ChemistrySelect, 3, 2441\u20132466.","journal-title":"ChemistrySelect"},{"key":"5217_CR7","doi-asserted-by":"publisher","first-page":"111","DOI":"10.1016\/j.molcatb.2006.02.010","volume":"40","author":"V Gotor-Fern\u00e1ndez","year":"2006","unstructured":"Gotor-Fern\u00e1ndez, V., Brieva, R., & Gotor, V. (2006). Lipases: Useful biocatalysts for the preparation of pharmaceuticals. Journal of Molecular Catalysis B: Enzymatic, 40, 111\u2013120.","journal-title":"Journal of Molecular Catalysis B: Enzymatic"},{"key":"5217_CR8","doi-asserted-by":"publisher","first-page":"1211","DOI":"10.3390\/catal11101211","volume":"11","author":"X Lyu","year":"2021","unstructured":"Lyu, X., Gonzalez, R., Horton, A., et al. (2021). Immobilization of enzymes by polymeric materials. Catalysts, 11, 1211. https:\/\/doi.org\/10.3390\/catal11101211","journal-title":"Catalysts"},{"key":"5217_CR9","doi-asserted-by":"publisher","first-page":"409","DOI":"10.1002\/9783527817290.ch12","volume-title":"Catalyst immobilization","author":"M Romero-Fern\u00e1ndez","year":"2020","unstructured":"Romero-Fern\u00e1ndez, M., & Paradisi, F. (2020). General overview on immobilization techniques of enzymes for biocatalysis. In M. Benaglia & A. Puglisi (Eds.), Catalyst immobilization (pp. 409\u2013435). Wiley."},{"key":"5217_CR10","doi-asserted-by":"publisher","first-page":"6223","DOI":"10.1039\/C3CS60075K","volume":"42","author":"RA Sheldon","year":"2013","unstructured":"Sheldon, R. A., & van Pelt, S. (2013). Enzyme immobilisation in biocatalysis: Why, what and how. Chemical Society Reviews, 42, 6223\u20136235.","journal-title":"Chemical Society Reviews"},{"key":"5217_CR11","doi-asserted-by":"publisher","first-page":"453","DOI":"10.1039\/b711564b","volume":"38","author":"U Hanefeld","year":"2009","unstructured":"Hanefeld, U., Gardossi, L., & Magner, E. (2009). Understanding enzyme immobilisation. Chemical Society Reviews, 38, 453\u2013468. https:\/\/doi.org\/10.1039\/b711564b","journal-title":"Chemical Society Reviews"},{"key":"5217_CR12","doi-asserted-by":"publisher","first-page":"104613","DOI":"10.1016\/j.reactfunctpolym.2020.104613","volume":"152","author":"RA Wahab","year":"2020","unstructured":"Wahab, R. A., Elias, N., Abdullah, F., et al. (2020). On the taught new tricks of enzymes immobilization: An all-inclusive overview. Reactive and Functional Polymers, 152, 104613. https:\/\/doi.org\/10.1016\/j.reactfunctpolym.2020.104613","journal-title":"Reactive and Functional Polymers"},{"key":"5217_CR13","doi-asserted-by":"publisher","first-page":"46727","DOI":"10.1002\/app.46727","volume":"135","author":"EP Cipolatti","year":"2018","unstructured":"Cipolatti, E. P., Pinto, M. C. C., Robert, Jd. M., et al. (2018). Pilot-scale development of core-shell polymer supports for the immobilization of recombinant lipase B from Candida antarctica and their application in the production of ethyl esters from residual fatty acids. Journal of Applied Polymer Science, 135, 46727. https:\/\/doi.org\/10.1002\/app.46727","journal-title":"Journal of Applied Polymer Science"},{"key":"5217_CR14","doi-asserted-by":"publisher","first-page":"1239","DOI":"10.1002\/bit.28046","volume":"119","author":"DC Kabernick","year":"2022","unstructured":"Kabernick, D. C., Gostick, J. T., & Ward, V. C. A. (2022). Kinetic characterization and modeling of sequentially entrapped enzymes in 3D-printed PMMA microfluidic reactors for the synthesis of amorphadiene via the isopentenol utilization pathway. Biotechnology and Bioengineering, 119, 1239\u20131251. https:\/\/doi.org\/10.1002\/bit.28046","journal-title":"Biotechnology and Bioengineering"},{"key":"5217_CR15","doi-asserted-by":"publisher","first-page":"114204","DOI":"10.1016\/j.bios.2022.114204","volume":"207","author":"\u0395I Tzianni","year":"2022","unstructured":"Tzianni, \u0395I., Moutsios, I., Moschovas, D., et al. (2022). Smartphone paired SIM card-type integrated creatinine biosensor. Biosensors & Bioelectronics, 207, 114204. https:\/\/doi.org\/10.1016\/j.bios.2022.114204","journal-title":"Biosensors & Bioelectronics"},{"key":"5217_CR16","doi-asserted-by":"publisher","first-page":"3384","DOI":"10.1007\/s12010-022-03913-9","volume":"194","author":"DS Wunschik","year":"2022","unstructured":"Wunschik, D. S., Lorenz, A., Ingenbosch, K. N., et al. (2022). Activation and stabilization of lipase B from Candida antarctica by immobilization on polymer brushes with optimized surface structure. Applied Biochemistry and Biotechnology, 194, 3384\u20133399. https:\/\/doi.org\/10.1007\/s12010-022-03913-9","journal-title":"Applied Biochemistry and Biotechnology"},{"key":"5217_CR17","doi-asserted-by":"publisher","first-page":"4992","DOI":"10.1021\/acscatal.9b01176","volume":"9","author":"JS Weltz","year":"2019","unstructured":"Weltz, J. S., Kienle, D. F., Schwartz, D. K., et al. (2019). Dramatic increase in catalytic performance of immobilized lipases by their stabilization on polymer brush supports. ACS Catalysis, 9, 4992\u20135001. https:\/\/doi.org\/10.1021\/acscatal.9b01176","journal-title":"ACS Catalysis"},{"key":"5217_CR18","doi-asserted-by":"publisher","first-page":"4304","DOI":"10.1021\/acscatal.3c00264","volume":"13","author":"H S\u00e1nchez-Mor\u00e1n","year":"2023","unstructured":"S\u00e1nchez-Mor\u00e1n, H., Gon\u00e7alves, L. R. B., Schwartz, D. K., et al. (2023). Framework for optimizing polymeric supports for immobilized biocatalysts by computational analysis of enzyme surface hydrophobicity. ACS Catalysis, 13, 4304\u20134315. https:\/\/doi.org\/10.1021\/acscatal.3c00264","journal-title":"ACS Catalysis"},{"key":"5217_CR19","doi-asserted-by":"publisher","first-page":"26694","DOI":"10.1021\/acsami.1c02443","volume":"13","author":"H S\u00e1nchez-Mor\u00e1n","year":"2021","unstructured":"S\u00e1nchez-Mor\u00e1n, H., Weltz, J. S., Schwartz, D. K., et al. (2021). Understanding design rules for optimizing the interface between immobilized enzymes and random copolymer brushes. ACS Applied Materials & Interfaces, 13, 26694\u201326703. https:\/\/doi.org\/10.1021\/acsami.1c02443","journal-title":"ACS Applied Materials & Interfaces"},{"key":"5217_CR20","doi-asserted-by":"publisher","first-page":"491","DOI":"10.1038\/351491a0","volume":"351","author":"AM Brzozowski","year":"1991","unstructured":"Brzozowski, A. M., Derewenda, U., Derewenda, Z. S., et al. (1991). A model for interfacial activation in lipases from the structure of a fungal lipase-inhibitor complex. Nature, 351, 491\u2013494. https:\/\/doi.org\/10.1038\/351491a0","journal-title":"Nature"},{"key":"5217_CR21","doi-asserted-by":"publisher","first-page":"1532","DOI":"10.1021\/bi00120a034","volume":"31","author":"U Derewenda","year":"1992","unstructured":"Derewenda, U., Brzozowski, A. M., Lawson, D. M., et al. (1992). Catalysis at the interface: The anatomy of a conformational change in a triglyceride lipase. Biochemistry, 31, 1532\u20131541. https:\/\/doi.org\/10.1021\/bi00120a034","journal-title":"Biochemistry"},{"key":"5217_CR22","doi-asserted-by":"publisher","first-page":"16","DOI":"10.3389\/fbioe.2017.00016","volume":"5","author":"FI Khan","year":"2017","unstructured":"Khan, F. I., Lan, D., Durrani, R., et al. (2017). The lid domain in lipases: Structural and functional determinant of enzymatic properties. Front Bioeng Biotechnol, 5, 16. https:\/\/doi.org\/10.3389\/fbioe.2017.00016","journal-title":"Front Bioeng Biotechnol"},{"key":"5217_CR23","doi-asserted-by":"publisher","first-page":"4814","DOI":"10.1016\/j.bpj.2009.03.040","volume":"96","author":"S Santini","year":"2009","unstructured":"Santini, S., Crowet, J. M., Thomas, A., et al. (2009). Study of Thermomyces lanuginosa lipase in the presence of tributyrylglycerol and water. Biophysical Journal, 96, 4814\u20134825. https:\/\/doi.org\/10.1016\/j.bpj.2009.03.040","journal-title":"Biophysical Journal"},{"key":"5217_CR24","doi-asserted-by":"publisher","first-page":"900","DOI":"10.1016\/j.ijbiomac.2016.07.097","volume":"92","author":"JJ Bassi","year":"2016","unstructured":"Bassi, J. J., Todero, L. M., Lage, F. A. P., et al. (2016). Interfacial activation of lipases on hydrophobic support and application in the synthesis of a lubricant ester. International Journal of Biological Macromolecules, 92, 900\u2013909. https:\/\/doi.org\/10.1016\/j.ijbiomac.2016.07.097","journal-title":"International Journal of Biological Macromolecules"},{"key":"5217_CR25","doi-asserted-by":"publisher","first-page":"6615","DOI":"10.1021\/bi972998p","volume":"37","author":"OG Berg","year":"1998","unstructured":"Berg, O. G., Cajal, Y., Butterfoss, G. L., et al. (1998). Interfacial activation of triglyceride lipase from Thermomyces (Humicola) lanuginosa: Kinetic parameters and a basis for control of the lid. Biochemistry, 37, 6615\u20136627. https:\/\/doi.org\/10.1021\/bi972998p","journal-title":"Biochemistry"},{"key":"5217_CR26","doi-asserted-by":"publisher","first-page":"1634","DOI":"10.1016\/S0006-3495(00)76715-4","volume":"78","author":"A Jutila","year":"2000","unstructured":"Jutila, A., Zhu, K., Patkar, S. A., et al. (2000). Detergent-induced conformational changes of Humicola lanuginosa lipase studied by fluorescence spectroscopy. Biophysical Journal, 78, 1634\u20131642. https:\/\/doi.org\/10.1016\/S0006-3495(00)76715-4","journal-title":"Biophysical Journal"},{"key":"5217_CR27","doi-asserted-by":"publisher","first-page":"272","DOI":"10.1016\/0005-2760(95)00131-u","volume":"1258","author":"M Martinelle","year":"1995","unstructured":"Martinelle, M., Holmquist, M., & Hult, K. (1995). On the interfacial activation of Candida antarctica lipase A and B as compared with Humicola lanuginosa lipase. Biochimica et Biophysica Acta, 1258, 272\u2013276. https:\/\/doi.org\/10.1016\/0005-2760(95)00131-u","journal-title":"Biochimica et Biophysica Acta"},{"key":"5217_CR28","doi-asserted-by":"publisher","first-page":"2410","DOI":"10.1016\/j.ijbiomac.2018.09.009","volume":"120","author":"M Mesa","year":"2018","unstructured":"Mesa, M., Perea\u00f1ez, J. A., Preciado, L. M., et al. (2018). How the Triton X-100 modulates the activity\/stability of the Thermomyces lanuginose lipase: Insights from experimental and molecular docking approaches. International Journal of Biological Macromolecules, 120, 2410\u20132417. https:\/\/doi.org\/10.1016\/j.ijbiomac.2018.09.009","journal-title":"International Journal of Biological Macromolecules"},{"key":"5217_CR29","doi-asserted-by":"publisher","first-page":"1719","DOI":"10.1021\/bi0479757","volume":"44","author":"JE Mogensen","year":"2005","unstructured":"Mogensen, J. E., Sehgal, P., & Otzen, D. E. (2005). Activation, inhibition, and destabilization of Thermomyces lanuginosus lipase by detergents. Biochemistry, 44, 1719\u20131730. https:\/\/doi.org\/10.1021\/bi0479757","journal-title":"Biochemistry"},{"key":"5217_CR30","doi-asserted-by":"publisher","first-page":"199","DOI":"10.1016\/j.molcatb.2015.08.015","volume":"122","author":"S Moreno-Perez","year":"2015","unstructured":"Moreno-Perez, S., Ghattas, N., Filice, M., et al. (2015). Dramatic hyperactivation of lipase of Thermomyces lanuginosa by a cationic surfactant: Fixation of the hyperactivated form by adsorption on sulfopropyl-sepharose. Journal of Molecular Catalysis B: Enzymatic, 122, 199\u2013203. https:\/\/doi.org\/10.1016\/j.molcatb.2015.08.015","journal-title":"Journal of Molecular Catalysis B: Enzymatic"},{"key":"5217_CR31","doi-asserted-by":"publisher","first-page":"15071","DOI":"10.1021\/bi0013905","volume":"39","author":"AM Brzozowski","year":"2000","unstructured":"Brzozowski, A. M., Savage, H., Verma, C. S., et al. (2000). Structural origins of the interfacial activation in Thermomyces (Humicola) lanuginosa lipase. Biochemistry, 39, 15071\u201315082. https:\/\/doi.org\/10.1021\/bi0013905","journal-title":"Biochemistry"},{"key":"5217_CR32","doi-asserted-by":"publisher","first-page":"20","DOI":"10.1016\/j.bbapap.2016.09.016","volume":"1865","author":"J Skjold-J\u00f8rgensen","year":"2017","unstructured":"Skjold-J\u00f8rgensen, J., Vind, J., Moroz, O. V., et al. (2017). Controlled lid-opening in Thermomyces lanuginosus lipase- An engineered switch for studying lipase function. Biochim Biophys Acta Proteins Proteom, 1865, 20\u201327. https:\/\/doi.org\/10.1016\/j.bbapap.2016.09.016","journal-title":"Biochim Biophys Acta Proteins Proteom"},{"key":"5217_CR33","doi-asserted-by":"publisher","first-page":"4152","DOI":"10.1021\/bi500233h","volume":"53","author":"J Skjold-J\u00f8rgensen","year":"2014","unstructured":"Skjold-J\u00f8rgensen, J., Vind, J., Svendsen, A., et al. (2014). Altering the activation mechanism in Thermomyces lanuginosus lipase. Biochemistry, 53, 4152\u20134160. https:\/\/doi.org\/10.1021\/bi500233h","journal-title":"Biochemistry"},{"key":"5217_CR34","doi-asserted-by":"publisher","first-page":"146","DOI":"10.1021\/acs.biochem.5b01114","volume":"55","author":"J Skjold-J\u00f8rgensen","year":"2016","unstructured":"Skjold-J\u00f8rgensen, J., Vind, J., Svendsen, A., et al. (2016). Lipases that activate at high solvent polarities. Biochemistry, 55, 146\u2013156. https:\/\/doi.org\/10.1021\/acs.biochem.5b01114","journal-title":"Biochemistry"},{"key":"5217_CR35","doi-asserted-by":"publisher","first-page":"543","DOI":"10.1093\/protein\/7.4.543","volume":"7","author":"DM Lawson","year":"1994","unstructured":"Lawson, D. M., Brzozowski, A. M., Rety, S., et al. (1994). Probing the nature of substrate binding in Humicola lanuginosa lipase through X-ray crystallography and intuitive modelling. Protein Engineering, 7, 543\u2013550. https:\/\/doi.org\/10.1093\/protein\/7.4.543","journal-title":"Protein Engineering"},{"key":"5217_CR36","doi-asserted-by":"publisher","first-page":"98","DOI":"10.1016\/S0006-3495(02)75152-7","volume":"83","author":"M\u00d8 Jensen","year":"2002","unstructured":"Jensen, M. \u00d8., Jensen, T. R., Kjaer, K., et al. (2002). Orientation and conformation of a lipase at an interface studied by molecular dynamics simulations. Biophysical Journal, 83, 98\u2013111. https:\/\/doi.org\/10.1016\/S0006-3495(02)75152-7","journal-title":"Biophysical Journal"},{"key":"5217_CR37","doi-asserted-by":"publisher","first-page":"137","DOI":"10.1093\/protein\/10.2.137","volume":"10","author":"GH Peters","year":"1997","unstructured":"Peters, G. H., Toxvaerd, S., Olsen, O. H., et al. (1997). Computational studies of the activation of lipases and the effect of a hydrophobic environment. Protein Engineering, 10, 137\u2013147. https:\/\/doi.org\/10.1093\/protein\/10.2.137","journal-title":"Protein Engineering"},{"key":"5217_CR38","doi-asserted-by":"publisher","first-page":"2122","DOI":"10.1002\/pro.493","volume":"19","author":"S Rehm","year":"2010","unstructured":"Rehm, S., Trodler, P., & Pleiss, J. (2010). Solvent-induced lid opening in lipases: A molecular dynamics study. Protein Science, 19, 2122\u20132130. https:\/\/doi.org\/10.1002\/pro.493","journal-title":"Protein Science"},{"key":"5217_CR39","doi-asserted-by":"publisher","first-page":"293","DOI":"10.1016\/S0969-2126(00)00031-9","volume":"2","author":"J Uppenberg","year":"1994","unstructured":"Uppenberg, J., Hansen, M. T., Patkar, S., et al. (1994). The sequence, crystal structure determination and refinement of two crystal forms of lipase B from Candida antarctica. Structure, 2, 293\u2013308. https:\/\/doi.org\/10.1016\/S0969-2126(00)00031-9","journal-title":"Structure"},{"key":"5217_CR40","doi-asserted-by":"publisher","first-page":"15709","DOI":"10.1039\/C7CP02198D","volume":"19","author":"B Luan","year":"2017","unstructured":"Luan, B., & Zhou, R. (2017). A novel self-activation mechanism of Candida antarctica lipase B. Physical Chemistry Chemical Physics: PCCP, 19, 15709\u201315714. https:\/\/doi.org\/10.1039\/C7CP02198D","journal-title":"Physical Chemistry Chemical Physics: PCCP"},{"key":"5217_CR41","doi-asserted-by":"publisher","first-page":"1168","DOI":"10.1107\/S090744490200714X","volume":"58","author":"K Kawasaki","year":"2002","unstructured":"Kawasaki, K., Kondo, H., Suzuki, M., et al. (2002). Alternate conformations observed in catalytic serine of Bacillus subtilis lipase determined at 1.3 A resolution. Acta Crystallographica. Section D, Biological Crystallography, 58, 1168\u20131174. https:\/\/doi.org\/10.1107\/S090744490200714X","journal-title":"Acta Crystallographica. Section D, Biological Crystallography"},{"key":"5217_CR42","doi-asserted-by":"publisher","first-page":"215","DOI":"10.1006\/jmbi.2001.4659","volume":"309","author":"G van Pouderoyen","year":"2001","unstructured":"van Pouderoyen, G., Eggert, T., Jaeger, K. E., et al. (2001). The crystal structure of Bacillus subtilis lipase: A minimal alpha\/beta hydrolase fold enzyme. Journal of Molecular Biology, 309, 215\u2013226. https:\/\/doi.org\/10.1006\/jmbi.2001.4659","journal-title":"Journal of Molecular Biology"},{"key":"5217_CR43","doi-asserted-by":"publisher","first-page":"5969","DOI":"10.1021\/acs.biochem.5b00586","volume":"54","author":"T Zisis","year":"2015","unstructured":"Zisis, T., Freddolino, P. L., Turunen, P., et al. (2015). Interfacial activation of Candida antarctica Lipase B: Combined evidence from experiment and simulation. Biochemistry, 54, 5969\u20135979. https:\/\/doi.org\/10.1021\/acs.biochem.5b00586","journal-title":"Biochemistry"},{"key":"5217_CR44","doi-asserted-by":"publisher","first-page":"3151","DOI":"10.1021\/acs.langmuir.6b04570","volume":"33","author":"SP Benson","year":"2017","unstructured":"Benson, S. P., & Pleiss, J. (2017). Self-assembly nanostructures of triglyceride-water interfaces determine functional conformations of Candida antarctica Lipase B. Langmuir, 33, 3151\u20133159. https:\/\/doi.org\/10.1021\/acs.langmuir.6b04570","journal-title":"Langmuir"},{"key":"5217_CR45","doi-asserted-by":"publisher","first-page":"48","DOI":"10.1016\/j.molcatb.2012.05.012","volume":"84","author":"CC Gruber","year":"2012","unstructured":"Gruber, C. C., & Pleiss, J. (2012). Lipase B from Candida antarctica binds to hydrophobic substrate\u2013water interfaces via hydrophobic anchors surrounding the active site entrance. Journal of Molecular Catalysis B: Enzymatic, 84, 48\u201354. https:\/\/doi.org\/10.1016\/j.molcatb.2012.05.012","journal-title":"Journal of Molecular Catalysis B: Enzymatic"},{"key":"5217_CR46","doi-asserted-by":"publisher","first-page":"1248","DOI":"10.1021\/acs.jpcb.6b11037","volume":"121","author":"D K\u00fcbler","year":"2017","unstructured":"K\u00fcbler, D., Bergmann, A., Weger, L., et al. (2017). Kinetics of detergent-induced activation and inhibition of a minimal lipase. The Journal of Physical Chemistry B, 121, 1248\u20131257. https:\/\/doi.org\/10.1021\/acs.jpcb.6b11037","journal-title":"The Journal of Physical Chemistry B"},{"key":"5217_CR47","doi-asserted-by":"publisher","first-page":"2977","DOI":"10.1021\/acs.jpclett.0c00470","volume":"11","author":"S Das","year":"2020","unstructured":"Das, S., Behera, S., & Balasubramanian, S. (2020). Orientational switch of the Lipase A enzyme at the oil-water interface: An order of magnitude increase in turnover rate with a single surfactant tag explained. Journal of Physical Chemistry Letters, 11, 2977\u20132982. https:\/\/doi.org\/10.1021\/acs.jpclett.0c00470","journal-title":"Journal of Physical Chemistry Letters"},{"key":"5217_CR48","doi-asserted-by":"publisher","first-page":"10876","DOI":"10.1021\/acscatal.8b02935","volume":"8","author":"JM Thiele","year":"2018","unstructured":"Thiele, J. M., Davari, M. D., K\u00f6nig, M., et al. (2018). Enzyme\u2013polyelectrolyte complexes boost the catalytic performance of enzymes. ACS Catal, 8, 10876\u201310887. https:\/\/doi.org\/10.1021\/acscatal.8b02935","journal-title":"ACS Catal"},{"key":"5217_CR49","doi-asserted-by":"publisher","first-page":"361","DOI":"10.1002\/bies.201100111","volume":"34","author":"J Ries","year":"2012","unstructured":"Ries, J., & Schwille, P. (2012). Fluorescence correlation spectroscopy. BioEssays, 34, 361\u2013368. https:\/\/doi.org\/10.1002\/bies.201100111","journal-title":"BioEssays"},{"key":"5217_CR50","unstructured":"Schwille, P., & Haustein, E. (2002). Fluorescence correlation spectroscopy: An introduction to its concepts and applications. https:\/\/citeseerx.ist.psu.edu\/document?repid=rep1&type=pdf&doi=11a9cb045cc94ce2a9407de3269cb71d747a7ed7. Accessed 22 Jul 2024."},{"key":"5217_CR51","doi-asserted-by":"publisher","unstructured":"Yu, L., Lei, Y., Ma, Y., et al. (2021). A Comprehensive review of fluorescence correlation spectroscopy. Front Phys 9. https:\/\/doi.org\/10.3389\/fphy.2021.644450","DOI":"10.3389\/fphy.2021.644450"},{"key":"5217_CR52","doi-asserted-by":"publisher","first-page":"1669","DOI":"10.1021\/acs.joc.1c01136","volume":"87","author":"KN Ingenbosch","year":"2021","unstructured":"Ingenbosch, K. N., Vieyto-Nu\u00f1ez, J. C., Ruiz-Blanco, Y. B., et al. (2021). Effect of organic solvents on the structure and activity of a minimal lipase. The Journal of Organic Chemistry, 87, 1669\u20131678.","journal-title":"The Journal of Organic Chemistry"},{"key":"5217_CR53","doi-asserted-by":"publisher","first-page":"610","DOI":"10.1107\/S0907444903029640","volume":"60","author":"J Newman","year":"2004","unstructured":"Newman, J. (2004). Novel buffer systems for macromolecular crystallization. Acta Crystallographica. Section D, Biological Crystallography, 60, 610\u2013612. https:\/\/doi.org\/10.1107\/S0907444903029640","journal-title":"Acta Crystallographica. Section D, Biological Crystallography"},{"key":"5217_CR54","doi-asserted-by":"publisher","first-page":"1098","DOI":"10.1021\/ed100461v","volume":"88","author":"JU Izunobi","year":"2011","unstructured":"Izunobi, J. U., & Higginbotham, C. L. (2011). Polymer molecular weight analysis by 1 H NMR spectroscopy. Journal of Chemical Education, 88, 1098\u20131104. https:\/\/doi.org\/10.1021\/ed100461v","journal-title":"Journal of Chemical Education"},{"key":"5217_CR55","doi-asserted-by":"publisher","first-page":"655","DOI":"10.1007\/s00249-015-1061-6","volume":"44","author":"D K\u00fcbler","year":"2015","unstructured":"K\u00fcbler, D., Ingenbosch, K. N., Bergmann, A., et al. (2015). Fluorescence spectroscopic analysis of the structure and dynamics of Bacillus subtilis lipase A governing its activity profile under alkaline conditions. European Biophysics Journal, 44, 655\u2013665. https:\/\/doi.org\/10.1007\/s00249-015-1061-6","journal-title":"European Biophysics Journal"},{"key":"5217_CR56","doi-asserted-by":"publisher","first-page":"93102","DOI":"10.1063\/1.4986235","volume":"88","author":"L Weger","year":"2017","unstructured":"Weger, L., & Hoffmann-Jacobsen, K. (2017). A total internal reflection-fluorescence correlation spectroscopy setup with pulsed diode laser excitation. Review of Scientific Instruments, 88, 93102. https:\/\/doi.org\/10.1063\/1.4986235","journal-title":"Review of Scientific Instruments"},{"key":"5217_CR57","doi-asserted-by":"publisher","first-page":"507","DOI":"10.1038\/nmeth.1208","volume":"5","author":"R Roy","year":"2008","unstructured":"Roy, R., Hohng, S., & Ha, T. (2008). A practical guide to single-molecule FRET. Nature Methods, 5, 507\u2013516. https:\/\/doi.org\/10.1038\/nmeth.1208","journal-title":"Nature Methods"},{"key":"5217_CR58","doi-asserted-by":"publisher","first-page":"7021","DOI":"10.1021\/acs.langmuir.8b00660","volume":"34","author":"L Weger","year":"2018","unstructured":"Weger, L., Weidmann, M., Ali, W., et al. (2018). Polymer diffusion in the interphase between surface and solution. Langmuir, 34, 7021\u20137027. https:\/\/doi.org\/10.1021\/acs.langmuir.8b00660","journal-title":"Langmuir"},{"key":"5217_CR59","doi-asserted-by":"publisher","first-page":"963","DOI":"10.1038\/NMETH1104","volume":"4","author":"SA Kim","year":"2007","unstructured":"Kim, S. A., Heinze, K. G., & Schwille, P. (2007). Fluorescence correlation spectroscopy in living cells. Nature Methods, 4, 963\u2013973. https:\/\/doi.org\/10.1038\/NMETH1104","journal-title":"Nature Methods"},{"key":"5217_CR60","unstructured":"Krieger, J. W., & Langowski, J. (2012). QuickFit 3.0: A data evaluation application for biophysics. https:\/\/github.com\/jkriege2\/QuickFit3"},{"key":"5217_CR61","doi-asserted-by":"publisher","first-page":"6964","DOI":"10.1021\/acs.jcim.3c01490","volume":"63","author":"VJ Hoerschinger","year":"2023","unstructured":"Hoerschinger, V. J., Waibl, F., Pomarici, N. D., et al. (2023). PEP-patch: Electrostatics in protein-protein recognition, specificity, and antibody developability. Journal of Chemical Information and Modeling, 63, 6964\u20136971. https:\/\/doi.org\/10.1021\/acs.jcim.3c01490","journal-title":"Journal of Chemical Information and Modeling"},{"key":"5217_CR62","doi-asserted-by":"publisher","first-page":"2981","DOI":"10.1093\/bioinformatics\/btu426","volume":"30","author":"E Krieger","year":"2014","unstructured":"Krieger, E., & Vriend, G. (2014). YASARA View - molecular graphics for all devices - from smartphones to workstations. Bioinformatics, 30, 2981\u20132982. https:\/\/doi.org\/10.1093\/bioinformatics\/btu426","journal-title":"Bioinformatics"},{"key":"5217_CR63","doi-asserted-by":"publisher","first-page":"4904","DOI":"10.1021\/acscatal.7b00838","volume":"7","author":"J Kari","year":"2017","unstructured":"Kari, J., Andersen, M., Borch, K., et al. (2017). An inverse Michaelis-Menten approach for interfacial enzyme kinetics. ACS Catalysis, 7, 4904\u20134914. https:\/\/doi.org\/10.1021\/acscatal.7b00838","journal-title":"ACS Catalysis"},{"key":"5217_CR64","doi-asserted-by":"publisher","first-page":"1644","DOI":"10.1002\/ejlt.201600059","volume":"118","author":"J Skjold-J\u00f8rgensen","year":"2016","unstructured":"Skjold-J\u00f8rgensen, J., Vind, J., Svendsen, A., et al. (2016). Understanding the activation mechanism of Thermomyces lanuginosus lipase using rational design and tryptophan-induced fluorescence quenching. European Journal of Lipid Science and Technology, 118, 1644\u20131660. https:\/\/doi.org\/10.1002\/ejlt.201600059","journal-title":"European Journal of Lipid Science and Technology"},{"key":"5217_CR65","doi-asserted-by":"publisher","first-page":"4256","DOI":"10.1021\/acs.biochem.7b00420","volume":"56","author":"JK Madsen","year":"2017","unstructured":"Madsen, J. K., Kaspersen, J. D., Andersen, C. B., et al. (2017). Glycolipid biosurfactants activate, dimerize, and stabilize Thermomyces lanuginosus Lipase in a pH-dependent fashion. Biochemistry, 56, 4256\u20134268. https:\/\/doi.org\/10.1021\/acs.biochem.7b00420","journal-title":"Biochemistry"},{"key":"5217_CR66","doi-asserted-by":"publisher","first-page":"88","DOI":"10.1016\/j.bpc.2013.12.001","volume":"185","author":"KM Gon\u00e7alves","year":"2014","unstructured":"Gon\u00e7alves, K. M., Barbosa, L. R. S., Lima, L. M. T. R., et al. (2014). Conformational dissection of Thermomyces lanuginosus lipase in solution. Biophysical Chemistry, 185, 88\u201397. https:\/\/doi.org\/10.1016\/j.bpc.2013.12.001","journal-title":"Biophysical Chemistry"},{"key":"5217_CR67","doi-asserted-by":"publisher","first-page":"7545","DOI":"10.1021\/acs.jcim.3c01681","volume":"63","author":"S Behera","year":"2023","unstructured":"Behera, S., & Balasubramanian, S. (2023). Lipase A from Bacillus subtilis: Substrate binding, conformational dynamics, and signatures of a lid. Journal of Chemical Information and Modeling, 63, 7545\u20137556. https:\/\/doi.org\/10.1021\/acs.jcim.3c01681","journal-title":"Journal of Chemical Information and Modeling"}],"container-title":["Applied Biochemistry and Biotechnology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s12010-025-05217-0.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1007\/s12010-025-05217-0\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s12010-025-05217-0.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,9,6]],"date-time":"2025-09-06T08:29:26Z","timestamp":1757147366000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1007\/s12010-025-05217-0"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3,31]]},"references-count":67,"journal-issue":{"issue":"7","published-print":{"date-parts":[[2025,7]]}},"alternative-id":["5217"],"URL":"https:\/\/doi.org\/10.1007\/s12010-025-05217-0","relation":{},"ISSN":["0273-2289","1559-0291"],"issn-type":[{"value":"0273-2289","type":"print"},{"value":"1559-0291","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,3,31]]},"assertion":[{"value":"13 March 2025","order":1,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"31 March 2025","order":2,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"Not applicable.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethical Approval"}},{"value":"Not applicable.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent to Participate"}},{"value":"All authors accept that the copyright of the manuscript will be transferred to the journal upon acceptance for publication.","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for Publication"}},{"value":"The authors declare no competing interests.","order":5,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing Interests"}}]}}