{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T04:06:35Z","timestamp":1790222795246,"version":"4.1.0"},"reference-count":59,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2023,9,6]],"date-time":"2023-09-06T00:00:00Z","timestamp":1693958400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"FuturEnzyme Project funded by the European Union\u2019s Horizon 2020 Research and Innovation Programme","award":["101000327"],"award-info":[{"award-number":["101000327"]}]},{"name":"FuturEnzyme Project funded by the European Union\u2019s Horizon 2020 Research and Innovation Programme","award":["PID2020-112758RB-I00"],"award-info":[{"award-number":["PID2020-112758RB-I00"]}]},{"name":"FuturEnzyme Project funded by the European Union\u2019s Horizon 2020 Research and Innovation Programme","award":["PDC2021-121534-I00"],"award-info":[{"award-number":["PDC2021-121534-I00"]}]},{"name":"FuturEnzyme Project funded by the European Union\u2019s Horizon 2020 Research and Innovation Programme","award":["TED2021-130544B-I00"],"award-info":[{"award-number":["TED2021-130544B-I00"]}]},{"name":"FuturEnzyme Project funded by the European Union\u2019s Horizon 2020 Research and Innovation Programme","award":["PID2019-106370RB-I00"],"award-info":[{"award-number":["PID2019-106370RB-I00"]}]},{"name":"FuturEnzyme Project funded by the European Union\u2019s Horizon 2020 Research and Innovation Programme","award":["FPU19\/00608"],"award-info":[{"award-number":["FPU19\/00608"]}]},{"name":"FuturEnzyme Project funded by the European Union\u2019s Horizon 2020 Research and Innovation Programme","award":["PRE2020-091825"],"award-info":[{"award-number":["PRE2020-091825"]}]},{"name":"Ministerio de Ciencia e Innovaci\u00f3n, Agencia Estatal de Investigaci\u00f3n (AEI) (Digital Object Identifier MCIN\/AEI\/10.13039\/501100011033)","award":["101000327"],"award-info":[{"award-number":["101000327"]}]},{"name":"Ministerio de Ciencia e Innovaci\u00f3n, Agencia Estatal de Investigaci\u00f3n (AEI) (Digital Object Identifier MCIN\/AEI\/10.13039\/501100011033)","award":["PID2020-112758RB-I00"],"award-info":[{"award-number":["PID2020-112758RB-I00"]}]},{"name":"Ministerio de Ciencia e Innovaci\u00f3n, Agencia Estatal de Investigaci\u00f3n (AEI) (Digital Object Identifier MCIN\/AEI\/10.13039\/501100011033)","award":["PDC2021-121534-I00"],"award-info":[{"award-number":["PDC2021-121534-I00"]}]},{"name":"Ministerio de Ciencia e Innovaci\u00f3n, Agencia Estatal de Investigaci\u00f3n (AEI) (Digital Object Identifier MCIN\/AEI\/10.13039\/501100011033)","award":["TED2021-130544B-I00"],"award-info":[{"award-number":["TED2021-130544B-I00"]}]},{"name":"Ministerio de Ciencia e Innovaci\u00f3n, Agencia Estatal de Investigaci\u00f3n (AEI) (Digital Object Identifier MCIN\/AEI\/10.13039\/501100011033)","award":["PID2019-106370RB-I00"],"award-info":[{"award-number":["PID2019-106370RB-I00"]}]},{"name":"Ministerio de Ciencia e Innovaci\u00f3n, Agencia Estatal de Investigaci\u00f3n (AEI) (Digital Object Identifier MCIN\/AEI\/10.13039\/501100011033)","award":["FPU19\/00608"],"award-info":[{"award-number":["FPU19\/00608"]}]},{"name":"Ministerio de Ciencia e Innovaci\u00f3n, Agencia Estatal de Investigaci\u00f3n (AEI) (Digital Object Identifier MCIN\/AEI\/10.13039\/501100011033)","award":["PRE2020-091825"],"award-info":[{"award-number":["PRE2020-091825"]}]},{"name":"Fondo Europeo de Desarrollo Regional (ERDF) A way of making Europe and the European Union NextGenerationEU\/PRTR","award":["101000327"],"award-info":[{"award-number":["101000327"]}]},{"name":"Fondo Europeo de Desarrollo Regional (ERDF) A way of making Europe and the European Union NextGenerationEU\/PRTR","award":["PID2020-112758RB-I00"],"award-info":[{"award-number":["PID2020-112758RB-I00"]}]},{"name":"Fondo Europeo de Desarrollo Regional (ERDF) A way of making Europe and the European Union NextGenerationEU\/PRTR","award":["PDC2021-121534-I00"],"award-info":[{"award-number":["PDC2021-121534-I00"]}]},{"name":"Fondo Europeo de Desarrollo Regional (ERDF) A way of making Europe and the European Union NextGenerationEU\/PRTR","award":["TED2021-130544B-I00"],"award-info":[{"award-number":["TED2021-130544B-I00"]}]},{"name":"Fondo Europeo de Desarrollo Regional (ERDF) A way of making Europe and the European Union NextGenerationEU\/PRTR","award":["PID2019-106370RB-I00"],"award-info":[{"award-number":["PID2019-106370RB-I00"]}]},{"name":"Fondo Europeo de Desarrollo Regional (ERDF) A way of making Europe and the European Union NextGenerationEU\/PRTR","award":["FPU19\/00608"],"award-info":[{"award-number":["FPU19\/00608"]}]},{"name":"Fondo Europeo de Desarrollo Regional (ERDF) A way of making Europe and the European Union NextGenerationEU\/PRTR","award":["PRE2020-091825"],"award-info":[{"award-number":["PRE2020-091825"]}]},{"DOI":"10.13039\/501100004837","name":"Spanish Ministry of Science and Innovation for a PhD fellowship","doi-asserted-by":"publisher","award":["101000327"],"award-info":[{"award-number":["101000327"]}],"id":[{"id":"10.13039\/501100004837","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004837","name":"Spanish Ministry of Science and Innovation for a PhD fellowship","doi-asserted-by":"publisher","award":["PID2020-112758RB-I00"],"award-info":[{"award-number":["PID2020-112758RB-I00"]}],"id":[{"id":"10.13039\/501100004837","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004837","name":"Spanish Ministry of Science and Innovation for a PhD fellowship","doi-asserted-by":"publisher","award":["PDC2021-121534-I00"],"award-info":[{"award-number":["PDC2021-121534-I00"]}],"id":[{"id":"10.13039\/501100004837","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004837","name":"Spanish Ministry of Science and Innovation for a PhD fellowship","doi-asserted-by":"publisher","award":["TED2021-130544B-I00"],"award-info":[{"award-number":["TED2021-130544B-I00"]}],"id":[{"id":"10.13039\/501100004837","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004837","name":"Spanish Ministry of Science and Innovation for a PhD fellowship","doi-asserted-by":"publisher","award":["PID2019-106370RB-I00"],"award-info":[{"award-number":["PID2019-106370RB-I00"]}],"id":[{"id":"10.13039\/501100004837","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004837","name":"Spanish Ministry of Science and Innovation for a PhD fellowship","doi-asserted-by":"publisher","award":["FPU19\/00608"],"award-info":[{"award-number":["FPU19\/00608"]}],"id":[{"id":"10.13039\/501100004837","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004837","name":"Spanish Ministry of Science and Innovation for a PhD fellowship","doi-asserted-by":"publisher","award":["PRE2020-091825"],"award-info":[{"award-number":["PRE2020-091825"]}],"id":[{"id":"10.13039\/501100004837","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004837","name":"Spanish Ministry of Science and Innovation for a PhD fellowship","doi-asserted-by":"publisher","award":["101000327"],"award-info":[{"award-number":["101000327"]}],"id":[{"id":"10.13039\/501100004837","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004837","name":"Spanish Ministry of Science and Innovation for a PhD fellowship","doi-asserted-by":"publisher","award":["PID2020-112758RB-I00"],"award-info":[{"award-number":["PID2020-112758RB-I00"]}],"id":[{"id":"10.13039\/501100004837","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004837","name":"Spanish Ministry of Science and Innovation for a PhD fellowship","doi-asserted-by":"publisher","award":["PDC2021-121534-I00"],"award-info":[{"award-number":["PDC2021-121534-I00"]}],"id":[{"id":"10.13039\/501100004837","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004837","name":"Spanish Ministry of Science and Innovation for a PhD fellowship","doi-asserted-by":"publisher","award":["TED2021-130544B-I00"],"award-info":[{"award-number":["TED2021-130544B-I00"]}],"id":[{"id":"10.13039\/501100004837","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004837","name":"Spanish Ministry of Science and Innovation for a PhD fellowship","doi-asserted-by":"publisher","award":["PID2019-106370RB-I00"],"award-info":[{"award-number":["PID2019-106370RB-I00"]}],"id":[{"id":"10.13039\/501100004837","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004837","name":"Spanish Ministry of Science and Innovation for a PhD fellowship","doi-asserted-by":"publisher","award":["FPU19\/00608"],"award-info":[{"award-number":["FPU19\/00608"]}],"id":[{"id":"10.13039\/501100004837","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004837","name":"Spanish Ministry of Science and Innovation for a PhD fellowship","doi-asserted-by":"publisher","award":["PRE2020-091825"],"award-info":[{"award-number":["PRE2020-091825"]}],"id":[{"id":"10.13039\/501100004837","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Lipases have valuable potential for industrial use, particularly those mostly active against water-insoluble substrates, such as triglycerides composed of long-carbon chain fatty acids. However, in most cases, engineered variants often need to be constructed to achieve optimal performance for such substrates. Protein engineering techniques have been reported as strategies for improving lipase characteristics by introducing specific mutations in the cap domain of esterases or in the lid domain of lipases or through lid domain swapping. Here, we improved the lipase activity of a lipase (WP_075743487.1, or LipMRD) retrieved from the Marine Metagenomics MarRef Database and assigned to the Actinoalloteichus genus. The improvement was achieved through site-directed mutagenesis and by substituting its lid domain (FRGTEITQIKDWLTDA) with that of Rhizopus delemar lipase (previously R. oryzae; UniProt accession number, I1BGQ3) (FRGTNSFRSAITDIVF). The results demonstrated that the redesigned mutants gain activity against bulkier triglycerides, such as glyceryl tridecanoate and tridodecanoate, olive oil, coconut oil, and palm oil. Residue W89 (LipMRD numbering) appears to be key to the increase in lipase activity, an increase that was also achieved with lid swapping. This study reinforces the importance of the lid domains and their amino acid compositions in determining the substrate specificity of lipases, but the generalization of the lid domain swapping between lipases or the introduction of specific mutations in the lid domain to improve lipase activity may require further investigation.<\/jats:p>","DOI":"10.3390\/ijms241813768","type":"journal-article","created":{"date-parts":[[2023,9,6]],"date-time":"2023-09-06T10:14:14Z","timestamp":1693995254000},"page":"13768","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":30,"title":["Enhancing the Hydrolytic Activity of a Lipase towards Larger Triglycerides through Lid Domain Engineering"],"prefix":"10.3390","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8861-3191","authenticated-orcid":false,"given":"Laura","family":"Fernandez-Lopez","sequence":"first","affiliation":[{"name":"Instituto de Catalisis y Petroleoquimica (ICP), CSIC, 28049 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0174-7435","authenticated-orcid":false,"given":"Sergi","family":"Roda","sequence":"additional","affiliation":[{"name":"Department of Life Sciences, Barcelona Supercomputing Center (BSC), 08034 Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6377-6338","authenticated-orcid":false,"given":"Ana","family":"Robles-Mart\u00edn","sequence":"additional","affiliation":[{"name":"Department of Life Sciences, Barcelona Supercomputing Center (BSC), 08034 Barcelona, Spain"},{"name":"PhD Programme, Faculty of Pharmacy and Food Science, Universitat de Barcelona (UB), 08007 Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4380-4888","authenticated-orcid":false,"given":"Rub\u00e9n","family":"Mu\u00f1oz-Tafalla","sequence":"additional","affiliation":[{"name":"Department of Life Sciences, Barcelona Supercomputing Center (BSC), 08034 Barcelona, Spain"},{"name":"PhD Programme, Faculty of Pharmacy and Food Science, Universitat de Barcelona (UB), 08007 Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1466-1067","authenticated-orcid":false,"given":"David","family":"Almendral","sequence":"additional","affiliation":[{"name":"Instituto de Catalisis y Petroleoquimica (ICP), CSIC, 28049 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4962-4714","authenticated-orcid":false,"given":"Manuel","family":"Ferrer","sequence":"additional","affiliation":[{"name":"Instituto de Catalisis y Petroleoquimica (ICP), CSIC, 28049 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4580-1114","authenticated-orcid":false,"given":"V\u00edctor","family":"Guallar","sequence":"additional","affiliation":[{"name":"Department of Life Sciences, Barcelona Supercomputing Center (BSC), 08034 Barcelona, Spain"},{"name":"Institution for Research and Advanced Studies (ICREA), 08010 Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,9,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1007\/978-1-61779-600-5_2","article-title":"Lipases or esterases: Does it really matter? Toward a new bio-physico-chemical classification","volume":"Volume 861","author":"Ali","year":"2012","journal-title":"Lipases and Phospholipases"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1035","DOI":"10.1111\/febs.15071","article-title":"The modular structure of \u03b1\/\u03b2-hydrolases","volume":"287","author":"Bauer","year":"2020","journal-title":"FEBS J."},{"key":"ref_3","first-page":"221","article-title":"Adipose triglyceride lipase regulation: An overview","volume":"19","author":"Cerk","year":"2018","journal-title":"Curr. Protein Pept. Sci."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"513","DOI":"10.1016\/0006-3002(58)90097-0","article-title":"Action de la lipase pancr\u00e9atique sur les esters en \u00e9mulsion","volume":"30","author":"Sarda","year":"1958","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1016\/S0167-7799(96)10064-0","article-title":"Interfacial activation of lipases: Facts and artifacts","volume":"15","author":"Verger","year":"1997","journal-title":"Trends. Biotechnol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"767","DOI":"10.1038\/343767a0","article-title":"A serine protease triad forms the catalytic centre of a triacylglycerol lipase","volume":"343","author":"Brady","year":"1990","journal-title":"Nature"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"771","DOI":"10.1038\/343771a0","article-title":"Structure of human pancreatic lipase","volume":"343","author":"Winkler","year":"1990","journal-title":"Nature"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"491","DOI":"10.1038\/351491a0","article-title":"A model for interfacial activation in lipases from the structure of a fungal lipase-inhibitor complex","volume":"351","author":"Brzozowski","year":"1991","journal-title":"Nature"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"814","DOI":"10.1038\/362814a0","article-title":"Interfacial activation of the lipase-procolipase complex by mixed micelles revealed by X-ray crystallography","volume":"362","author":"Egloff","year":"1993","journal-title":"Nature"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1002\/prot.22462","article-title":"Insights into lid movements of Burkholderia cepacia lipase inferred from molecular dynamics simulations","volume":"77","author":"Barbe","year":"2009","journal-title":"Proteins"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1016\/j.cis.2008.06.001","article-title":"Lipases at interfaces: A review","volume":"147\u2013148","author":"Reis","year":"2009","journal-title":"Adv. Colloid Interface Sci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"615","DOI":"10.1038\/356615a0","article-title":"Fusarium solani cutinase is a lipolytic enzyme with a catalytic serine accessible to solvent","volume":"356","author":"Martinez","year":"1992","journal-title":"Nature"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"4702","DOI":"10.1021\/bi00069a003","article-title":"A structural domain (the lid) found in pancreatic lipases is absent in the guinea pig (phospho)lipase","volume":"32","author":"Hjorth","year":"1993","journal-title":"Biochemistry"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1016\/0014-5793(93)80501-K","article-title":"Topological characterization and modeling of the 3D structure of lipase from Pseudomonas aeruginosa","volume":"332","author":"Jaeger","year":"1993","journal-title":"FEBS Lett."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/0014-5793(93)80310-Q","article-title":"The Crystal structure of triacyl-glycerol lipase from Pseudomonas glumae reveals a partially redundant catalytic aspartate","volume":"331","author":"Noble","year":"1993","journal-title":"FEBS Lett."},{"key":"ref_16","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_17","doi-asserted-by":"crossref","first-page":"921","DOI":"10.1016\/j.jprot.2011.10.011","article-title":"Proteomics combines morphological, physiological and bio-chemical attributes to unravel the survival strategy of Anabaena sp. PCC7120 under arsenic stress","volume":"75","author":"Pandey","year":"2012","journal-title":"J. Proteom."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"235","DOI":"10.3109\/10242422.2016.1151416","article-title":"Biodiversity for biocatalysis: A review of the \u03b1\/\u03b2-hydrolase fold superfamily of esterases-lipases discovered in metagenomes","volume":"3","author":"Ferrer","year":"2015","journal-title":"Biocatal. Biotransform."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"247303","DOI":"10.3389\/fbioe.2017.00016","article-title":"The lid domain in lipases: Structural and functional determinant of enzymatic properties","volume":"5","author":"Khan","year":"2017","journal-title":"Front. Bioeng. Biotechnol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"25396","DOI":"10.1074\/jbc.270.43.25396","article-title":"Human hepatic and lipoprotein lipase. The loop covering the catalytic site mediates lipase substrate specificity","volume":"270","author":"Dugi","year":"1995","journal-title":"J. Biol. Chem."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1016\/S0009-3084(98)00030-9","article-title":"Anatomy of lipase binding sites: The scissile fatty acid binding site","volume":"93","author":"Pleiss","year":"1998","journal-title":"Chem. Phys. Lipids"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2312","DOI":"10.1110\/ps.0304003","article-title":"Sequence of the lid affects activity and specificity of Candida rugosa lipase isoenzymes","volume":"12","author":"Brocca","year":"2003","journal-title":"Protein Sci."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2383","DOI":"10.1016\/j.febslet.2005.03.037","article-title":"Mutations in the \u2018lid\u2019 region affect chain length specificity and thermostability of a Pseudomonas fragi lipase","volume":"579","author":"Santarossa","year":"2005","journal-title":"FEBS Lett."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"162","DOI":"10.1016\/j.molcatb.2009.06.006","article-title":"The effect of substitution of phe181 and phe182 with ala on activity, substrate specificity and stabilization of substrate at the active site of Bacillus thermocatenulatus lipase","volume":"61","author":"Karkhane","year":"2009","journal-title":"J. Mol. Catal. B Enzym."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1002\/ejlt.200900143","article-title":"Protein engineering and discovery of lipases","volume":"112","author":"Kourist","year":"2010","journal-title":"Eur. J. Lipid Sci. Technol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1330","DOI":"10.1039\/C4CC08477B","article-title":"Saturation mutagenesis in selected amino acids to shift Pseudomonas sp. acidic lipase lip i.3 substrate specificity and activity","volume":"51","author":"Panizza","year":"2014","journal-title":"Chem. Commun."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"520","DOI":"10.1002\/cbic.200800668","article-title":"Understanding the plasticity of the alpha\/beta hydrolase fold: Lid swapping on the Candida antarctica lipase B results in chimeras with interesting biocatalytic properties","volume":"10","author":"Chatterjee","year":"2009","journal-title":"Chembiochem"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1044","DOI":"10.1194\/jlr.M043950","article-title":"Conversion of a Rhizopus chinensis lipase into an esterase by lid swapping","volume":"55","author":"Yu","year":"2014","journal-title":"J. Lipid Res."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1193955","DOI":"10.3389\/fmicb.2023.1193955","article-title":"N-terminal lid swapping contributes to the substrate specificity and activity of thermophilic lipase TrLipE","volume":"14","author":"Fang","year":"2023","journal-title":"Front. Microbiol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1385\/ABAB:118:1-3:155","article-title":"Lipases and their industrial applications: An overview","volume":"118","author":"Houde","year":"2004","journal-title":"Appl. Biochem. Biotechnol."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Bornscheuer, U.T., and Kazlauskas, R.J. (2006). Hydrolases in Organic Synthesis: Regio- and Stereoselective Biotransformations, Wiley-VCH Verlag GmbH & Co. KGaA. [2nd ed.].","DOI":"10.1002\/3527607544"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1016\/j.enzmictec.2005.10.016","article-title":"Industrial applications of microbial lipases","volume":"39","author":"Hasan","year":"2006","journal-title":"Enzyme Microb. Technol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"D692","DOI":"10.1093\/nar\/gkx1036","article-title":"The MAR databases: Development and implementation of databases specific for marine metagenomics","volume":"46","author":"Klemetsen","year":"2018","journal-title":"Nucleic Acids Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1093\/oxfordjournals.jbchem.a021442","article-title":"The crystal structure of lipase II from Rhizopus niveus at 2.2 A resolution","volume":"120","author":"Kohno","year":"1996","journal-title":"J. Biochem."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1016\/S0167-4781(98)00104-3","article-title":"Cloning, expression, characterization and role of the leader sequence of a lipase from Rhizopus oryzae","volume":"1399","author":"Beer","year":"1998","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1705","DOI":"10.1007\/s10482-017-0920-9","article-title":"Acti-noalloteichus fjordicus sp. nov. isolated from marine sponges: Phenotypic, chemotaxonomic and genomic characterisation","volume":"110","author":"Nouioui","year":"2017","journal-title":"Antonie Leeuwenhoek"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2317","DOI":"10.1002\/(SICI)1521-3765(19981102)4:11<2324::AID-CHEM2324>3.0.CO;2-I","article-title":"Rapid quantitative screening of hydrolases using pH indicators. Finding enantioselective hydrolases","volume":"4","author":"Janes","year":"1998","journal-title":"Chem. Eur. J."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1016\/S0167-7012(98)00008-6","article-title":"A colorimetric assay for detecting haloalkane dehalogenase activity","volume":"32","author":"Holloway","year":"1998","journal-title":"J. Microbiol. Methods"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"e202207344","DOI":"10.1002\/anie.202207344","article-title":"A Plurizyme with transaminase and hydrolase activity catalyzes cascade reactions","volume":"61","author":"Roda","year":"2022","journal-title":"Angew. Chem. Int. Ed. Engl."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"W320","DOI":"10.1093\/nar\/gkx246","article-title":"GalaxyHomomer: A web server for protein homo-oligomer structure prediction from a monomer sequence or structure","volume":"45","author":"Baek","year":"2017","journal-title":"Nucleic Acids Res."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1007\/s11745-997-0016-1","article-title":"Altered acyl chain length specificity of Rhizopus delemar lipase through mutagenesis and molecular modeling","volume":"32","author":"Klein","year":"1997","journal-title":"Lipids"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1304","DOI":"10.1021\/ct0501811","article-title":"PELE: Protein Energy Landscape Exploration. A novel Monte Carlo based technique","volume":"1","author":"Borrelli","year":"2005","journal-title":"J. Chem. Theory Comput."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"7655","DOI":"10.1021\/acs.jctc.0c00925","article-title":"AquaPELE: A Monte Carlo-based Algorithm to sample the effects of buried water molecules in proteins","volume":"16","author":"Municoy","year":"2020","journal-title":"J. Chem. Theory Comput."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41598-017-08445-5","article-title":"Adaptive simulations, towards interactive protein-ligand modeling","volume":"7","author":"Lecina","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1739","DOI":"10.1021\/jm0306430","article-title":"Glide: A new approach for rapid, accurate docking and scoring. 1. Method and assessment of docking accuracy","volume":"47","author":"Friesner","year":"2004","journal-title":"J. Med. Chem."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1109\/MCSE.2007.55","article-title":"Matplotlib: A 2D graphics environment","volume":"9","author":"Hunter","year":"2007","journal-title":"Comput. Sci. Eng."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"524","DOI":"10.1016\/S0022-2275(20)41203-9","article-title":"Conformational lability of lipases observed in the absence of an oil-water interface: Crystallographic studies of enzymes from the fungi Humicola lanuginosa and Rhizopus delemar","volume":"35","author":"Derewenda","year":"1994","journal-title":"J. Lipid Res."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"e0180722","DOI":"10.1128\/aem.01807-22","article-title":"The mobility of the cap domain is essential for the substrate promiscuity of a family iv esterase from Sorghum rhizosphere microbiome","volume":"89","author":"Distaso","year":"2023","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"11607","DOI":"10.1002\/anie.202003635","article-title":"Sequence-based prediction of promiscuous acyltransferase activity in hydrolases","volume":"59","author":"Becker","year":"2020","journal-title":"Angew. Chem. Int. Ed. Engl."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"6815","DOI":"10.1074\/jbc.M307738200","article-title":"A substrate-induced switch in the reaction mechanism of a thermophilic esterase: Kinetic evidences and structural basis","volume":"279","author":"Mandrich","year":"2004","journal-title":"J. Biol. Chem."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"761","DOI":"10.1006\/jmbi.2000.4195","article-title":"A snapshot of a transition state analogue of a novel thermophilic esterase belonging to the subfamily of mammalian hormone-sensitive lipase","volume":"303","author":"Galdiero","year":"2000","journal-title":"J. Mol. Biol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"1721","DOI":"10.1002\/prot.21877","article-title":"Functional and structural features of the oxyanion hole in a thermophilic esterase from Alicyclobacillus acidocaldarius","volume":"71","author":"Mandrich","year":"2008","journal-title":"Proteins"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"3570","DOI":"10.1111\/febs.15680","article-title":"Crystal structures of a novel family IV esterase in free and substrate-bound form","volume":"288","author":"Bollinger","year":"2021","journal-title":"FEBS J."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"2383","DOI":"10.1021\/acs.jcim.1c00152","article-title":"Promiscuous esterases counterintuitively are less flexible than specific ones","volume":"61","author":"Nutschel","year":"2021","journal-title":"J. Chem. Inf. Model."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"583","DOI":"10.1038\/s41586-021-03819-2","article-title":"Highly accurate protein structure prediction with AlphaFold","volume":"596","author":"Jumper","year":"2021","journal-title":"Nature"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"1752","DOI":"10.1002\/jcc.20292","article-title":"Integrated modeling program, Applied Chemical Theory (IMPACT)","volume":"26","author":"Banks","year":"2005","journal-title":"J. Comput. Chem."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"2110","DOI":"10.1002\/qua.24400","article-title":"Quantum mechanics approaches to drug research in the era of structural chemogenomics","volume":"113","author":"Ilatovskiy","year":"2013","journal-title":"Int. J. Quantum Chem."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1016\/S0006-3495(01)76033-X","article-title":"Anisotropy of fluctuation dynamics of proteins with an elastic network model","volume":"80","author":"Atilgan","year":"2001","journal-title":"Biophys. J."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1146\/annurev.physchem.51.1.129","article-title":"Generalized born models of macromolecular solvation effects","volume":"51","author":"Bashford","year":"2000","journal-title":"Annu. Rev. Phys. Chem."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/18\/13768\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T20:46:23Z","timestamp":1760129183000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/18\/13768"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,9,6]]},"references-count":59,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2023,9]]}},"alternative-id":["ijms241813768"],"URL":"https:\/\/doi.org\/10.3390\/ijms241813768","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,9,6]]}}}