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Expanding the industrial use of Robust Oxidative Biocatalysts for the conversion and production of alcohols (ROBOX)

Deliverables

Completion of benchmarking protocol and flowsheets established for the case studies

Completion of benchmarking protocol and flowsheets established for the case studies

Stakeholder analysis

Stakeholder analysis

Identification of genes predicted to encode relevant robust BVMOs and oxidases

Identification of genes predicted to encode relevant robust BVMOs and oxidases

Public website and newsletter, first press releases on the project scopes

Public website and newsletter, first press releases on the project scopes

Overall analysis of environmental benefits and techno-economics of the processes and platform

Overall analysis of environmental benefits and techno-economics of the processes and platform

Structural elucidation of relevant of BVMOs e.g. at least 3 new enzyme structures

Structural elucidation of relevant of BVMOs e.g. at least 3 new enzyme structures

Project leaflet in several languages

Project leaflet in several languages

Communication and dissemination plan

Communication and dissemination plan

Quorum sensing technology and micro fluidics platform available

Quorum sensing technology and micro fluidics platform available

Optimized enzyme production processes for ROBOX enzymes in bioreactor including DSP

Optimized enzyme production processes for ROBOX enzymes in bioreactor including DSP

Set of P450 enzymes with improved performance for hydroxylation of aromatics, alkenes and APIs and a set of API-sensors to enable P450 improvement

Set of P450 enzymes with improved performance for hydroxylation of aromatics, alkenes and APIs and a set of API-sensors to enable P450 improvement

First generation production strains for robust BVMOs

First generation production strains for robust BVMOs

First generation production strains for microbial and human P450s

First generation production strains for microbial and human P450s

96-well capillary electrophoreses screening system and first flow cytometer based screening system available

96-well capillary electrophoreses screening system and first flow cytometer based screening system available

Generation of a first flow cytometer based screening system for quantification of one product in the BM3 demonstration process through fluorogenic sensors

Generation of a first flow cytometer based screening system for quantification of one product in the BM3 demonstration process through fluorogenic sensors

New vector and host systems for enzyme coexpression

New vector and host systems for enzyme coexpression

Biocatalytic and structural exploration of P450-AX as well as generation of improved variants

Biocatalytic and structural exploration of P450-AX as well as generation of improved variants

Installation of advisory boards

Installation of advisory boards

Publications

Production of diclofenac metabolites by applying cytochrome P450 technology

Author(s): Iwona Kaluzna, Jan Brummund, and Martin Schuermann
Published in: Chimica Oggi - Chemistry Today, Issue vol. 35(6) November/December 2017, 2017, Page(s) 55-58, ISSN 0392-839X

Same Substrate, Many Reactions: Oxygen Activation in Flavoenzymes

Author(s): Elvira Romero, J. Rubén Gómez Castellanos, Giovanni Gadda, Marco W. Fraaije, Andrea Mattevi
Published in: Chemical Reviews, Issue 118/4, 2018, Page(s) 1742-1769, ISSN 0009-2665
DOI: 10.1021/acs.chemrev.7b00650

Polycyclic Ketone Monooxygenase (PockeMO): A Robust Biocatalyst for the Synthesis of Optically Active Sulfoxides

Author(s): Gonzalo de Gonzalo, Maximilian Fürst, Marco Fraaije
Published in: Catalysts, Issue 7/12, 2017, Page(s) 288, ISSN 2073-4344
DOI: 10.3390/catal7100288

Coupled reactions by coupled enzymes: alcohol to lactone cascade with alcohol dehydrogenase–cyclohexanone monooxygenase fusions

Author(s): Friso S. Aalbers, Marco W. Fraaije
Published in: Applied Microbiology and Biotechnology, Issue 101/20, 2017, Page(s) 7557-7565, ISSN 0175-7598
DOI: 10.1007/s00253-017-8501-4

Overriding Traditional Electronic Effects in Biocatalytic Baeyer–Villiger Reactions by Directed Evolution

Author(s): Guangyue Li, Marc Garcia-Borràs, Maximilian J. L. J. Fürst, Adriana Ilie, Marco W. Fraaije, K. N. Houk, Manfred T. Reetz
Published in: Journal of the American Chemical Society, Issue 140/33, 2018, Page(s) 10464-10472, ISSN 0002-7863
DOI: 10.1021/jacs.8b04742

High performing immobilized Baeyer-Villiger monooxygenase and glucose dehydrogenase for the synthesis of Ɛ-caprolactone derivative

Author(s): Marie A.F. Delgove, Daniela Valencia, Jordi Solé, Katrien V. Bernaerts, Stefaan M.A. De Wildeman, Marina Guillén, Gregorio Álvaro
Published in: Applied Catalysis A: General, Issue 572, 2018, Page(s) 134-141, ISSN 0926-860X
DOI: 10.1016/j.apcata.2018.12.036

A hydroquinone-specific screening system for directed P450 evolution

Author(s): Alexandra M. Weingartner, Daniel F. Sauer, Gaurao V. Dhoke, Mehdi D. Davari, Anna Joëlle Ruff, Ulrich Schwaneberg
Published in: Applied Microbiology and Biotechnology, Issue 102/22, 2018, Page(s) 9657-9667, ISSN 0175-7598
DOI: 10.1007/s00253-018-9328-3

Online Measurement of Oxygen-Dependent Enzyme Reaction Kinetics


Published in: ISSN 1439-4227
DOI: 10.1002/cbic.201700577

Scoping Biocatalyst Performance Using Reaction Trajectory Analysis


Published in: ISSN 1083-6160
DOI: 10.1021/acs.oprd.8b00119

Structure-Based Engineering of Phanerochaete chrysosporium Alcohol Oxidase for Enhanced Oxidative Power toward Glycerol

Author(s): Quoc-Thai Nguyen, Elvira Romero, Willem P. Dijkman, Suzan Pantaroto de Vasconcellos, Claudia Binda, Andrea Mattevi, Marco W. Fraaije
Published in: Biochemistry, Issue 57/43, 2018, Page(s) 6209-6218, ISSN 0006-2960
DOI: 10.1021/acs.biochem.8b00918

Enzyme Fusions in Biocatalysis: Coupling Reactions by Pairing Enzymes


Published in: ISSN 1439-4227
DOI: 10.1002/cbic.201800394

Beyond active site residues: overall structural dynamics control catalysis in flavin-containing and heme-containing monooxygenases

Author(s): Maximilian JLJ Fürst, Filippo Fiorentini, Marco W Fraaije
Published in: Current Opinion in Structural Biology, Issue 59, 2019, Page(s) 29-37, ISSN 0959-440X
DOI: 10.1016/j.sbi.2019.01.019

What to sacrifice? Fusions of cofactor regenerating enzymes with Baeyer-Villiger monooxygenases and alcohol dehydrogenases for self-sufficient redox biocatalysis

Author(s): Ángela Mourelle-Insua, Friso S. Aalbers, Iván Lavandera, Vicente Gotor-Fernández, Marco W. Fraaije
Published in: Tetrahedron, Issue 75/13, 2019, Page(s) 1832-1839, ISSN 0040-4020
DOI: 10.1016/j.tet.2019.02.015

Side-Chain Pruning Has Limited Impact on Substrate Preference in a Promiscuous Enzyme

Author(s): Maximilian J. L. J. Fürst, Elvira Romero, J. Rúben Gómez Castellanos, Marco W. Fraaije, Andrea Mattevi
Published in: ACS Catalysis, Issue 8/12, 2018, Page(s) 11648-11656, ISSN 2155-5435
DOI: 10.1021/acscatal.8b03793

Cloning, expression and characterisation of P450-Hal1 (CYP116B62) from Halomonas sp. NCIMB 172: A self-sufficient P450 with high expression and diverse substrate scope

Author(s): Joanne L. Porter, Selina Sabatini, Jack Manning, Michele Tavanti, James L. Galman, Nicholas J. Turner, Sabine L. Flitsch
Published in: Enzyme and Microbial Technology, Issue 113, 2018, Page(s) 1-8, ISSN 0141-0229
DOI: 10.1016/j.enzmictec.2018.02.005

A biocatalytic cascade for the amination of unfunctionalised cycloalkanes


Published in: ISSN 1477-0520
DOI: 10.1039/c7ob02569f

Characterization and Crystal Structure of a Robust Cyclohexanone Monooxygenase


Published in: ISSN 0044-8249
DOI: 10.1002/ange.201608951

Polycyclic Ketone Monooxygenase from the Thermophilic Fungus Thermothelomyces thermophila : A Structurally Distinct Biocatalyst for Bulky Substrates

Author(s): Maximilian J. L. J. Fürst, Simone Savino, Hanna M. Dudek, J. Rúben Gómez Castellanos, Cora Gutiérrez de Souza, Stefano Rovida, Marco W. Fraaije, Andrea Mattevi
Published in: Journal of the American Chemical Society, Issue 139/2, 2017, Page(s) 627-630, ISSN 0002-7863
DOI: 10.1021/jacs.6b12246

Increasing the solubility range of polyesters by tuning their microstructure with comonomers

Author(s): Marie A. F. Delgove, Juandré Luchies, Iris Wauters, Geert G. P. Deroover, Stefaan M. A. De Wildeman, Katrien V. Bernaerts
Published in: Polym. Chem., 2017, ISSN 1759-9954
DOI: 10.1039/C7PY00976C

Experimental Protocols for Generating Focused Mutant Libraries and Screening for Thermostable Proteins

Author(s): Max J.L.J. Fürst, Caterina Martin, Nikola Lončar, Marco W. Fraaije
Published in: Methods in Enzymology, Issue 2018 June 9, 2018, Page(s) 151-187
DOI: 10.1016/bs.mie.2018.04.007

Flavoenzymes at the crossroads of biocatalysis and structural biology

Author(s): Nguyen, Quoc-Thai
Published in: Issue 1, 2017