Technologies of the Future for Low-Cost Enzymes for Environment-Friendly Products

Informations projet

FuturEnzyme

N° de convention de subvention: 101000327

DOI 10.3030/101000327

Date de signature de la CE 22 Avril 2021

Date de début 1 Juin 2021

Date de fin 31 Mai 2025

Financé au titre de

• SOCIETAL CHALLENGES - Food security, sustainable agriculture and forestry, marine, maritime and inland water research, and the bioeconomy

Coût total € 5 995 035,13

Contribution de l’UE € 5 995 035,13

Coordonné par

AGENCIA ESTATAL CONSEJO SUPERIOR DE INVESTIGACIONES CIENTIFICAS
Spain

Livrables

Websites, patent fillings, videos etc. (3)

Visual identity guidelines

This deliverable will consist in a series of visual elements to create distinction and a strong visual identity for the FuturEnzyme project as well as template for presentations posters and reports

FuturEnzyme website

This deliverable will consist in an open website to assure communication towards the outside world that will be maintained and regularly updated throughout the project The website will serve as the main interface for distributing project information and results as elaborated in all WPs

FuturEnzyme project leaflet and brochure

This deliverable will consist in a series of leaflet and brochure materials for disseminating and communicating Project objectives and outcomes for consumers policymakers This material will be available through the website

Publications

Peer reviewed articles (38)

Improving bioprocess conditions for the production of prodigiosin using a marine Serratia rubidaea strain

Auteurs: Ricardo F. S. Pereira, Carla C. C. R. de Carvalho
Publié dans: Marine Drugs, Numéro 22(4), 2024, Page(s) 142, ISSN 1660-3397
Éditeur: Multidisciplinary Digital Publishing Institute (MDPI)
DOI: 10.3390/md22040142

Metagenomic mining for esterases in the microbial community of Los Rueldos acid mine drainage formation

Auteurs: Paula Vidal, Mónica Martínez-Martínez, Laura Fernandez-Lopez, Sergi Roda, Celia Méndez-García, Olga V. Golyshina, Víctor Guallar, Ana I. Peláez, Manuel Ferrer
Publié dans: Frontiers in Microbiology, Numéro Volume 13, 2022, Page(s) 868839, ISSN 1664-302X
Éditeur: Frontiers Media
DOI: 10.3389/fmicb.2022.868839

An archaeal lid-containing feruloyl esterase degrades polyethylene terephthalate

Auteurs: Perez-Garcia, Pablo; Chow, Jennifer; Costanzi, Elisa; Gurschke, Marno; Dittrich, Jonas; Dierkes, Robert F.; Molitor, Rebecka; Applegate, Violetta; Feuerriegel, Golo; Tete, Prince; Danso, Dominik; Thies, Stephan; Schumacher, Julia; Pfleger, Christopher; Jaeger, Karl-Erich; Gohlke, Holger; Smits, Sander H. J.; Schmitz, Ruth A.; Streit, Wolfgang R.
Publié dans: Communications Chemistry, Numéro 6, 2023, Page(s) 193, ISSN 2399-3669
Éditeur: Nature Portfolio
DOI: 10.1038/s42004-023-00998-z

A Choice Experiment Model for Sustainable Consumer Goods: A Systematic Literature Review and Workflow Design

Auteurs: Saija, Maria Elena; Daniotti, Sara; Bosco, Diego; Re, Ilaria
Publié dans: A Choice Experiment Model for Sustainable Consumer Goods: A Systematic Literature Review and Workflow Design, Numéro 15, 2023, Page(s) 13183, ISSN 2071-1050
Éditeur: MDPI Open Access Publishing
DOI: 10.3390/su151713183

Process Development for Benzyl Alcohol Production by Whole-Cell Biocatalysis in Stirred and Packed Bed Reactors

Auteurs: Carlos Rodrigues; Carla De Carvalho
Publié dans: Microorganisms, Numéro Volume 10; Numéro 5, 2022, Page(s) 966, ISSN 2076-2607
Éditeur: MDPI
DOI: 10.3390/microorganisms10050966

A novel metagenome-derived viral RNA polymerase and its application in a cell-free expression system for metagenome screening

Auteurs: Yuchen Han, Birhanu M. Kinfu, Fabian Blombach, Gwenny Cackett, Hongli Zhang, Pablo Pérez‑García, Ines Krohn, Jesper Salomon, Volkan Besirlioglu, Tayebeh Mirzaeigarakani, Ulrich Schwaneberg, Jennifer Chow, Finn Werner, Wolfgang R. Streit
Publié dans: Scientific Reports, Numéro Volume 12, 2022, Page(s) 17882, ISSN 2045-2322
Éditeur: Nature Publishing Group
DOI: 10.1038/s41598-022-22383-x

Design and Characterization of In-One Protease-Esterase PluriZyme

Auteurs: Fernandez-Lopez, Laura; Roda, Sergi; Gonzalez-Alfonso, José L; Plou, Francisco J; Guallar, Víctor; Ferrer, Manuel
Publié dans: International Journal of Molecular Sciences, Numéro Volume 23, Numéro 21, 2022, Page(s) 13337, ISSN 1422-0067
Éditeur: Multidisciplinary Digital Publishing Institute (MDPI)
DOI: 10.3390/ijms232113337

Plasmonic photothermal activation of an organosilica shielded cold-adapted lipase coimmobilised with gold nanoparticles on silica particles

Auteurs: Giunta, Carolina I; Nazemi, Seyed Amirabbas; Olesinska, Magdalena; Shahgaldian, Patrick
Publié dans: Nanoscale Advances, Numéro 5, 2023, Page(s) 81-87, ISSN 2516-0230
Éditeur: Royal Society of Chemistry
DOI: 10.1039/d2na00605g

Enhancing the Hydrolytic Activity of a Lipase towards Larger Triglycerides through Lid Domain Engineering

Auteurs: Fernandez-Lopez, Laura; Roda, Sergi; Robles-Martín, A; Muñoz-Tafalla, Rubén; Almendral, David; Ferrer, Manuel; Guallar, Víctor
Publié dans: International Journal of Molecular Sciences, Numéro 24, 2023, Page(s) 18, ISSN 1422-0067
Éditeur: Multidisciplinary Digital Publishing Institute (MDPI)
DOI: 10.3390/ijms241813768

Sub-micro- and nano-sized polyethylene terephthalate deconstruction with engineered protein nanopores

Auteurs: Robles-Martín, Ana; Amigot-Sánchez, Rafael; Fernandez-Lopez, Laura; Gonzalez-Alfonso, Jose L; Roda, Sergi; Alcolea-Rodriguez, Víctor; Heras-Márquez, Diego; Almendral, David; Coscolín, Cristina; Plou, Francisco J; Portela, Raquel; Bañares, Miguel A; Martínez-del-Pozo, Álvaro; García-Linares, Sara; Ferrer, Manuel; Guallar, Víctor
Publié dans: Nature Catalysis, Numéro 6, 2023, Page(s) 174–1185, ISSN 2520-1158
Éditeur: Nature Portfolio
DOI: 10.1038/s41929-023-01048-6

Natranaeroarchaeum sulfidigenes gen. nov., sp. nov., carbohydrate-utilizing sulfur-respiring haloarchaeon from hypersaline soda lakes, a member of a new family Natronoarchaeaceae fam. nov. in the order Halobacteriales

Auteurs: Dimitry Y. Sorokin, Michail Yakimov, Enzo Messina, Alexander Y. Merkel, Michel Koenen, Nicole J. Bale, Jaap S. Sinninghe Damsté
Publié dans: ystematic and Applied Microbiology, Numéro Volume 45, 2022, Page(s) 126356, ISSN 0723-2020
Éditeur: Elsevier BV
DOI: 10.1016/j.syapm.2022.126356

Increased Thermostability of an Engineered Flavin-Containing Monooxygenase to Remediate Trimethylamine in Fish Protein Hydrolysates

Auteurs: Goris, Marianne; Cea-Rama, Isabel; Puntervoll, Pål; Ree, Rasmus; Almendral, David; Sanz-Aparicio, Julia; Ferrer, Manuel; Bjerga, Gro Elin Kjæreng
Publié dans: Applied and Environmental Microbiology, Numéro 2, 2023, ISSN 0099-2240
Éditeur: American Society for Microbiology
DOI: 10.1128/aem.00390-23

Anaerobic carboxydotrophy in sulfur-respiring haloarchaea from hypersaline lakes

Auteurs: Dimitry Y. Sorokin, Alexander Y. Merkel, Enzo Messina, Claudia Tugui, Martin Pabst, Peter N. Golyshin & Michail M. Yakimov 
Publié dans: The ISME Journal, Numéro Volume 16, 2022, Page(s) 1534–1546, ISSN 1751-7362
Éditeur: Nature Publishing Group
DOI: 10.1038/s41396-022-01206-x

Exploiting cyclodextrins as artificial chaperones to enhance enzyme protection through supramolecular engineering

Auteurs: Foroutan Kalourazi, Ali; Amirabbas Nazemi, Seyed; Unniram Parambi, Ajmal Roshan; Muñoz-Tafalla, Ruben; Vidal, Paula; Shahangian, S. Shirin; Guallar, Víctor; Ferrer, Manuel; Shahgaldian, Patrick
Publié dans: Nanoscale, Numéro 16, 2024, Page(s) 5123-5129, ISSN 2040-3372
Éditeur: The Royal Society of Chemistry
DOI: 10.1039/D3NR06044F

Isolation and Characterization of a Serratia rubidaea from a Shallow Water Hydrothermal Vent

Auteurs: Pereira, Ricardo F. S.; Ferreira, Maria J.; Oliveira, M. Conceição; Serra, Maria C.; de Carvalho, Carla C. C. R.
Publié dans: Marine Drugs, Numéro 21, 2023, Page(s) 599, ISSN 1660-3397
Éditeur: Multidisciplinary Digital Publishing Institute (MDPI)
DOI: 10.3390/md21120599

Halopseudomonas species: Cultivation and molecular genetic tools

Auteurs: Kruse, Luzie; Loeschcke, Anita; de Witt, Jan; Wierckx, Nick; Jaeger, Karl-Erich; Thies, Stephan
Publié dans: Microbial Biotechnology, Numéro 17, 2024, Page(s) e14369, ISSN 1751-7915
Éditeur: Wiley
DOI: 10.1111/1751-7915.14369

Calm and Frenzy: marine obligate hydrocarbonoclastic bacteria sustain ocean wellness

Auteurs: Michail M. Yakimov; Rafael Bargiela; Peter N. Golyshin
Publié dans: Current Opinion in Biotechnology, Numéro Volume 73, 2022, Page(s) 337-345, ISSN 0958-1669
Éditeur: Elsevier BV
DOI: 10.1016/j.copbio.2021.09.015

Crystal structure of a family VIII β-lactamase fold hydrolase reveals the molecular mechanism for its broad substrate scope

Auteurs: Isabel Cea-Rama, Cristina Coscolín, Jose L. Gonzalez-Alfonso, Jog Raj, Marko Vasiljević, Francisco J. Plou, Manuel Ferrer, Julia Sanz-Aparicio
Publié dans: FEBS Journal, Numéro Volume 289, Numéro 21, 2022, Page(s) 6714-6730, ISSN 1742-464X
Éditeur: Blackwell Publishing Inc.
DOI: 10.1111/febs.16554

Transforming an esterase into an enantioselective catecholase through bioconjugation of a versatile metal-chelating inhibitor

Auteurs: Fernandez-Lopez, Laura; Cea-Rama, Isabel; Alvarez-Malmagro, Julia; Ressmann, Anna K; Gonzalez-Alfinso, Jose Luis; Coscolín, Cristina; Shahgaldian, Patrick; Plou, Francisco J; Modregger, Jan; Pita, Marcos; Sanz-Aparicio, Julia; Ferrer, Manuel
Publié dans: Chemical Communications, Numéro 1, 2023, ISSN 1359-7345
Éditeur: Royal Society of Chemistry
DOI: 10.1039/D3CC01946B

Thermophilic carboxylesterases from hydrothermal vents of the volcanic island of Ischia active on synthetic and biobased polymers and mycotoxins

Auteurs: Distaso, Marco; Chernikova, Tatyana; Bargiela, Rafael; Coscolín, Cristina; Stogios, Peter; Gonzalez-Alfonso, Jose L; Lemak, Sofia; Khusnutdinova, Anna; Plou, Francisco J; Evdokimova, Elena; Savchenko, Alexei; Lunev, Evgenii A; Yakimov, Michail M; Golyshina, Olga; Ferrer, Manuel; Yakunin, Alexander F; Golyshin, Peter N
Publié dans: Applied and Environmental Microbiology, 2023, ISSN 0099-2240
Éditeur: American Society for Microbiology
DOI: 10.1101/2022.09.17.508236

AsiteDesign: a Semirational Algorithm for an Automated EnzymeDesign

Auteurs: Roda, Sergi; Terholsen, Henrik; Heike Meyer, Jule Ruth; Cañellas-Solé, Albert; Guallar, Víctor; Bornscheuer, Uwe; Kazemi, Masoud
Publié dans: The Journal of Physical Chemistry B, Numéro 127, 2023, Page(s) 2661−2670, ISSN 1520-6106
Éditeur: American Chemical Society
DOI: 10.1021/acs.jpcb.2c07091

The metagenome-derived esterase PET40 is highlypromiscuous and hydrolyses polyethyleneterephthalate (PET)

Auteurs: Zhang, Hongli; Dierkes, Robert F.; Perez-Garcia, Pablo; Costanzi, Elisa; Dittrich, Jonas; Cea, Pablo A.; Gurschke, Marno; Applegate, Violetta; Partus, Kristina; Schmeisser, Christel; Pfleger, Christopher; Gohlke, Holger; Smits, Sander H. J.; Jennifer Chow; Wolfgang R. Streit
Publié dans: The FEBS Journal, 2023, ISSN 1742-464X
Éditeur: Blackwell Publishing Inc.
DOI: 10.1111/febs.16924

EP-Pred: A Machine Learning Tool for Bioprospecting Promiscuous Ester Hydrolases

Auteurs: Xiang, R; Fernandez-Lopez, Laura; Robles-Martín, Ana; Ferrer, Manuel; Guallar, Víctor
Publié dans: Biomolecules, Numéro Volume 12, issue 10, 2022, Page(s) 1529, ISSN 2218-273X
Éditeur: Multidisciplinary Digital Publishing Institute (MDPI)
DOI: 10.3390/biom12101529

Interplay of intracellular and trans-cellular DNA methylation in natural archaeal consortia

Auteurs: Reva, Oleg N.; La Cono, Violetta; Crisafi, Francesca; Smedile, Francesco; Mudaliyar, Manasi; Ghosal, Debnath; Giuliano, Laura; Krupovic, Mart; Yakimov, Michail M.
Publié dans: Environmental Microbiology Reports, Numéro 16, 2024, Page(s) e13258, ISSN 1758-2229
Éditeur: Wiley-Blackwell
DOI: 10.1111/1758-2229.13258

A Plurizyme with Transaminase and Hydrolase Activity Catalyzes Cascade Reactions

Auteurs: Sergi Roda, Laura Fernandez-Lopez, Marius Benedens, Alexander Bollinger, Stephan Thies, Julia Schumacher, Cristina Coscolín, Masoud Kazemi, Gerard Santiago, Christoph G. W. Gertzen, Jose L. Gonzalez-Alfonso, Francisco J. Plou, Karl-Erich Jaeger, Sander H. J. Smits, Manuel Ferrer, Víctor Guallar
Publié dans: Angewandte Chemie International Edition, Numéro Volume 61, Numéro 37, 2022, Page(s) e202207344, ISSN 1433-7851
Éditeur: John Wiley & Sons Ltd.
DOI: 10.1002/anie.202207344

Cultivation of a vampire: ‘Candidatus Absconditicoccus praedator’

Auteurs: Michail M. Yakimov; Alexander Y. Merkel; Vasil A. Gaisin; Martin Pilhofer; Enzo Messina; John E. Hallsworth; Alexandra A. Klyukina; Ekaterina N. Tikhonova; Vladimir M. Gorlenko
Publié dans: Environmental Microbiology, Numéro Volume 24, Numéro 1, 2021, Page(s) 30-49, ISSN 1462-2920
Éditeur: Wiley-Blackwell
DOI: 10.1111/1462-2920.15823

Enzymes for consumer products to achieve climate neutrality

Auteurs: Molina-Espeja, Patricia; Sanz-Aparicio, Julia; Golyshin, Peter N; Robles-Martín, Ana; Guallar, Victor; Beltrametti, Fabrizio; Müller, Markus; Yakimov, Mikhail; Modregger, Jan; van Logchem, Moniec; Corvini, Philippe; Shahgaldian, Patrick; Degering, Christian; Wieland, Susanne; Timm, Anne; de Carvalho, Carla CCR; Ilaria, Re; Daniotti, Sara; Thies, Stephan; Jaeger, Karl-Erich; Chow, Jennifer; Strei
Publié dans: Oxford Open Climate Change, Numéro 3, 2023, Page(s) kgad003, ISSN 2634-4068
Éditeur: Oxford University Press
DOI: 10.1093/oxfclm/kgad003

An archaeal lid-containing feruloyl-esterase degrades polyethylene terephthalate (PET)

Auteurs: Perez-Garcia, Pablo; Chow, Jennifer; Costanzi, Elisa; Gurschke, Marno; Dittrich, Jonas; Dierkes, Robert F.; Molitor, Rebecka; Applegate, Violetta; Feuerriegel, Golo; Tete, Prince; Danso, Dominik; Thies, Stephan; Schumacher, Julia; Pfleger, Christopher; Jaeger, Karl-Erich; Gohlke, Holger; Smits, Sander H. J.; Schmitz, Ruth A.; Streit, Wolfgang R.
Publié dans: Communications chemistry, Numéro 6, 2023, Page(s) 193, ISSN 2399-3669
Éditeur: Nature Portfolio
DOI: 10.21203/rs.3.rs-3196380/v1

The mobility of the cap domain is essential for the substrate promiscuity of a family IV esterase from sorghum rhizosphere microbiome

Auteurs: Distaso, Marco; Cea-Rama, Isabel; Coscolín, Cristina; Chernikova, Tatyana; Tran, Hai; Ferrer, Manuel; Sanz-Aparicio, Julia; Golyshin, Peter
Publié dans: Applied and Environmental Microbiology, 2023, ISSN 0099-2240
Éditeur: American Society for Microbiology
DOI: 10.1128/aem.01807-22

Nanohaloarchaea as beneficiaries of xylan degradation by haloarchaea

Auteurs: La Cono, Violetta; Messina, Enzo; Reva, Olga; Smedile, Francesco; La Spada, Gina; Crisafi, Francesca; Marturano, Laura; Miguez, Noa; Ferrer, Manuel; Selivanova, Elena A; Golyshina, Olga V; Golyshin, Peter N; Rohde, Manfred; Krupovic, Mart; Merkel, Alexander Y; Sorokin, Dimitry Y; Hallsworth, John E; Yakimov, Michail M
Publié dans: Microbial Biotechnology, 2023, ISSN 1751-7915
Éditeur: Wiley-Blackwell
DOI: 10.1111/1751-7915.14272

Halapricum desulfuricans sp. nov., carbohydrate-utilizing, sulfur-respiring haloarchaea from hypersaline lakes

Auteurs: Dimitry Y. Sorokin, Michail M. Yakimov, Enzo Messina, Alexander Y. Merkel, Michel Koenen, Nicole J. Bale, Jaap S. Sinninghe D
Publié dans: Systematic and Applied Microbiology, Numéro 44, 2021, Page(s) 126249, ISSN 0723-2020
Éditeur: Elsevier BV
DOI: 10.1016/j.syapm.2021.126249

Marine Bioprospecting, Biocatalysis and Process Development

Auteurs: Rodrigues, Carlos J.C.; de Carvalho, Carla C.C.R.
Publié dans: Microorganisms, Numéro Volume 10, issue 10, 2022, Page(s) 1965, ISSN 2076-2607
Éditeur: Multidisciplinary Digital Publishing Institute (MDPI)
DOI: 10.3390/microorganisms10101965

Functional diversity of nanohaloarchaea within xylan-degrading consortia

Auteurs: Oleg, Reva; Enzo, Messina; Violetta, La Cono; Francesca, Crisafi; Francesco, Smedile; Gina, La Spada; Laura, Marturano; Elena A, Selivanova; Manfred, Rohde; Mart, Krupovic; Michail M, Yakimov
Publié dans: Frontiers in Microbiology, Numéro 14, 2023, Page(s) 1182464, ISSN 1664-302X
Éditeur: Frontiers Media
DOI: 10.3389/fmicb.2023.1182464

Nanobiocatalysts with inbuilt cofactor recycling for oxidoreductase catalysis in organic solvents

Auteurs: Sahlin, Jenny; Wu, Congyu; Buscemi, Andrea; Schärer, Claude; Nazemi, Seyed Amirabbas; Rejaul, S. K.; Herrera-Reinoza, Nataly; Jung, Thomas A.; Shahgaldian, Patrick
Publié dans: Nanoscale Advances, Numéro 5, 2023, Page(s) 5036-5044, ISSN 2516-0230
Éditeur: The Royal Society of Chemistry
DOI: 10.1039/D3NA00413A

Enzyme adaptation to habitat thermal legacy shapes the thermal plasticity of marine microbiomes

Auteurs: Marasco, Ramona; Fusi, Marco; Coscolín, Cristina; Barozzi, Alan; Almendral, David; Bargiela, Rafael; Gohlke neé Nutschel, Christina; Pfleger, Christopher; Dittrich, Jonas; Gohlke, Holger; Matesaz, Ruth; Sanchez-Carrillo, Sergio; Mapelli, Francesca; Chernikova, Tatynana N; Golyshin, Peter N; Ferrer, Manuel; Daffonchio, Daniele
Publié dans: Nature Communications, Numéro 14, 2023, Page(s) 1045, ISSN 2041-1723
Éditeur: Nature Publishing Group
DOI: 10.1038/s41467-023-36610-0

Behind the Scenes of PluriZyme Designs

Auteurs: Robles-Martín, Ana; Roda, Sergi; Muñoz-Tafalla, Rubén; Guallar, Víctor
Publié dans: Eng, Numéro 5(1), 2024, Page(s) 91-103, ISSN 2673-4117
Éditeur: MDPI
DOI: 10.3390/eng5010006

Harnessing extremophilic carboxylesterases for applications in polyester depolymerisation and plastic waste recycling

Auteurs: Williams, Gwion B; Ma, Hairong; Khusnutdinova, Anna N.; Yakunin, Alexander F.; Golyshin, Peter N
Publié dans: Essays in Biochemistry, Numéro 67, 2023, Page(s) 715-729, ISSN 0071-1365
Éditeur: Portland Press, Ltd.
DOI: 10.1042/EBC20220255

Cultivating marine bacteria under laboratory conditions: Overcoming the “unculturable” dogma

Auteurs: Carlos J. C. Rodrigues, Carla C. C. R. de Carvalho
Publié dans: Frontiers in Bioengineering and Biotechnology, Numéro Volume 10, 2022, Page(s) 964589, ISSN 2296-4185
Éditeur: Frontiers Media S.A.
DOI: 10.3389/fbioe.2022.964589

Other (2)

Fantastic enzymes: where and how to find them

Auteurs: Ferrer, Manuel; Molina-Espeja, Patricia; Daniotti, Sara; Müller, Markus
Publié dans: The FEBS Network, 2022
Éditeur: Wiley-Blackwell Publishing Ltd
DOI: 10.5281/zenodo.7260738

Se buscan enzimas, razón: el medio ambiente

Auteurs: Manuel Ferrer, Cristina Coscolín Galán, Patricia Molina Espeja
Publié dans: The Conversation España, 2021, ISSN 2201-5639
Éditeur: The Conversation
DOI: 10.5281/zenodo.5566131

Non-peer reviewed articles (2)

The first archaeal PET-degrading enzyme belongs to the feruloyl-esterase family

Auteurs: Perez-Garcia, Pablo; Chow, Jennifer; Costanzi, Elisa; Gurschke, Marno F; Dittrich, Jonas; Dierkes, Robert F; Applegate, Violetta; Feuerriegel, Golo; Tete, Prince; Danso, Dominik; Schumacher, Julia; Pfleger, Christopher; Gohlke, Holger; Smits, Sander HJ; Schmitz, Ruth A; Streit, Wolfgang
Publié dans: bioRvix, 2022, ISSN 2692-8205
Éditeur: Cold Spring Harbor Laboratory
DOI: 10.1101/2022.10.14.512230

El uso de enzimas para conseguir productos de consumo diario más sostenibles

Auteurs: Ferrer, Manuel; Molina, Patricia; Peñalver, Carolina
Publié dans: Revista Ambienta, Numéro 128 (Innovation), 2021, ISSN 2605-4787
Éditeur: Gobierno de España, Ministerio para la Transición Ecológica y el Reto Demográfico
DOI: 10.5281/zenodo.5384608