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

Deliverables

Documents, reports (8)

Completion of benchmarking protocol and flowsheets established for the case studies

Stakeholder analysis

Identification of genes predicted to encode relevant robust BVMOs and oxidases

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

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 eg at least 3 new enzyme structures

Project leaflet in several languages

Communication and dissemination plan

Demonstrators, pilots, prototypes (9)

Quorum sensing technology and micro fluidics platform available

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

First generation production strains for robust BVMOs

First generation production strains for microbial and human P450s

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

New vector and host systems for enzyme coexpression

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

Other (1)

Installation of advisory boards

Publications

Peer reviewed articles (30)

Exploring the Substrate Scope of Baeyer-Villiger Monooxygenases with Branched Lactones as Entry towards Polyesters

Author(s): Marie A. F. Delgove, Maximilian J. L. J. Fürst, Marco W. Fraaije, Katrien V. Bernaerts, Stefaan M. A. De Wildeman
Published in: ChemBioChem, 2017, ISSN 1439-4227
Publisher: John Wiley & Sons Ltd.
DOI: 10.1002/cbic.201700427

Production of diclofenac metabolites by applying cytochrome P450 technology

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

Manipulating the stereoselectivity of the thermostable Baeyer–Villiger monooxygenase TmCHMO by directed evolution

Author(s): Guangyue Li, Maximilian J. L. J. Fürst, Hamid Reza Mansouri, Anna K. Ressmann, Adriana Ilie, Florian Rudroff, Marko D. Mihovilovic, Marco W. Fraaije, Manfred T. Reetz
Published in: Organic & Biomolecular Chemistry, 15/46, 2017, Page(s) 9824-9829, ISSN 1477-0520
Publisher: Royal Society of Chemistry
DOI: 10.1039/C7OB02692G

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, 118/4, 2018, Page(s) 1742-1769, ISSN 0009-2665
Publisher: American Chemical Society
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, 7/12, 2017, Page(s) 288, ISSN 2073-4344
Publisher: Multidisciplinary Digital Publishing Institute (MDPI)
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, 101/20, 2017, Page(s) 7557-7565, ISSN 0175-7598
Publisher: Springer Verlag
DOI: 10.1007/s00253-017-8501-4

Application of a thermostable Baeyer-Villiger monooxygenase for the synthesis of branched polyester precursors

Author(s): Marie A. F. Delgove, Matthew T. Elford, Katrien V. Bernaerts, Stefaan M.A. De Wildeman
Published in: Journal of Chemical Technology & Biotechnology, 2018, ISSN 0268-2575
Publisher: John Wiley & Sons Inc.
DOI: 10.1002/jctb.5623

Biocatalytic Properties and Structural Analysis of Eugenol Oxidase from Rhodococcus jostii RHA1: A Versatile Oxidative Biocatalyst

Author(s): Quoc-Thai Nguyen, Gonzalo de Gonzalo, Claudia Binda, Ana Rioz-Martínez, Andrea Mattevi, Marco W. Fraaije
Published in: ChemBioChem, 17/14, 2016, Page(s) 1359-1366, ISSN 1439-4227
Publisher: John Wiley & Sons Ltd.
DOI: 10.1002/cbic.201600148

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, 140/33, 2018, Page(s) 10464-10472, ISSN 0002-7863
Publisher: American Chemical Society
DOI: 10.1021/jacs.8b04742

Toward Upscaled Biocatalytic Preparation of Lactone Building Blocks for Polymer Applications

Author(s): Marie A. F. Delgove, Matthew T. Elford, Katrien V. Bernaerts, Stefaan M. A. De Wildeman
Published in: Organic Process Research & Development, 22/7, 2018, Page(s) 803-812, ISSN 1083-6160
Publisher: American Chemical Society
DOI: 10.1021/acs.oprd.8b00079

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, 572, 2018, Page(s) 134-141, ISSN 0926-860X
Publisher: Elsevier BV
DOI: 10.1016/j.apcata.2018.12.036

A prospective life-cycle assessment (LCA) of monomer synthesis: Comparison of biocatalytic and oxidative chemistry

Author(s): Marie A. F. Delgove, Achille-B. Laurent, John M. Woodley, Stefaan M. A. De Wildeman, Katrien V. Bernaerts, Yvonne van der Meer
Published in: ChemSusChem, 2019, ISSN 1864-5631
Publisher: Wiley - V C H Verlag GmbbH & Co.
DOI: 10.1002/cssc.201900007

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, 102/22, 2018, Page(s) 9657-9667, ISSN 0175-7598
Publisher: Springer Verlag
DOI: 10.1007/s00253-018-9328-3

Online Measurement of Oxygen-Dependent Enzyme Reaction Kinetics

Author(s): Murray P. Meissner, Mathias Nordblad, John M. Woodley
Published in: ChemBioChem, 19/1, 2018, Page(s) 106-113, ISSN 1439-4227
Publisher: John Wiley & Sons Ltd.
DOI: 10.1002/cbic.201700577

Scoping Biocatalyst Performance Using Reaction Trajectory Analysis

Author(s): Mathias Nordblad, Mafalda Dias Gomes, Murray P. Meissner, Hemalata Ramesh, John M. Woodley
Published in: Organic Process Research & Development, 22/9, 2018, Page(s) 1101-1114, ISSN 1083-6160
Publisher: American Chemical Society
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, 57/43, 2018, Page(s) 6209-6218, ISSN 0006-2960
Publisher: American Chemical Society
DOI: 10.1021/acs.biochem.8b00918

Enzyme Fusions in Biocatalysis: Coupling Reactions by Pairing Enzymes

Author(s): Friso S. Aalbers, Marco W. Fraaije
Published in: ChemBioChem, 20/1, 2019, Page(s) 20-28, ISSN 1439-4227
Publisher: John Wiley & Sons Ltd.
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, 59, 2019, Page(s) 29-37, ISSN 0959-440X
Publisher: Elsevier BV
DOI: 10.1016/j.sbi.2019.01.019

Design of Artificial Alcohol Oxidases: Alcohol Dehydrogenase-NADPH Oxidase Fusions for Continuous Oxidations

Author(s): Friso S. Aalbers, Marco W. Fraaije
Published in: ChemBioChem, 20/1, 2019, Page(s) 51-56, ISSN 1439-4227
Publisher: John Wiley & Sons Ltd.
DOI: 10.1002/cbic.201800421

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, 75/13, 2019, Page(s) 1832-1839, ISSN 0040-4020
Publisher: Pergamon Press Ltd.
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, 8/12, 2018, Page(s) 11648-11656, ISSN 2155-5435
Publisher: American Chemical Society
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, 113, 2018, Page(s) 1-8, ISSN 0141-0229
Publisher: Elsevier BV
DOI: 10.1016/j.enzmictec.2018.02.005

Characterisation of CYP102A25 from Bacillus marmarensis and CYP102A26 from Pontibacillus halophilus : P450 Homologues of BM3 with Preference towards Hydroxylation of Medium-Chain Fatty Acids

Author(s): Joanne L. Porter, Jack Manning, Selina Sabatini, Michele Tavanti, Nicholas J. Turner, Sabine L. Flitsch
Published in: ChemBioChem, 19/5, 2018, Page(s) 513-520, ISSN 1439-4227
Publisher: John Wiley & Sons Ltd.
DOI: 10.1002/cbic.201700598

A biocatalytic cascade for the amination of unfunctionalised cycloalkanes

Author(s): Michele Tavanti, Juan Mangas-Sanchez, Sarah L. Montgomery, Matthew P. Thompson, Nicholas J. Turner
Published in: Organic & Biomolecular Chemistry, 15/46, 2017, Page(s) 9790-9793, ISSN 1477-0520
Publisher: Royal Society of Chemistry
DOI: 10.1039/c7ob02569f

Expanding the CRISPR/Cas9 toolkit for Pichia pastoris with efficient donor integration and alternative resistance markers

Author(s): Astrid Weninger, Jasmin E. Fischer, Hana Raschmanová, Claudia Kniely, Thomas Vogl, Anton Glieder
Published in: Journal of Cellular Biochemistry, 119/4, 2018, Page(s) 3183-3198, ISSN 0730-2312
Publisher: John Wiley & Sons Inc.
DOI: 10.1002/jcb.26474

Co-immobilization of P450 BM3 and glucose dehydrogenase on different supports for application as a self-sufficient oxidative biocatalyst

Author(s): Jordi Solé, Gloria Caminal, Martin Schürmann, Gregorio Álvaro, Marina Guillén
Published in: Journal of Chemical Technology & Biotechnology, 94/1, 2019, Page(s) 244-255, ISSN 0268-2575
Publisher: John Wiley & Sons Inc.
DOI: 10.1002/jctb.5770

One-pot Biocatalytic Double Oxidation of α-Isophorone for the Synthesis of Ketoisophorone

Author(s): Michele Tavanti, Fabio Parmeggiani, Jose Rubén Gómez Castellanos, Andrea Mattevi, Nicholas John Turner
Published in: ChemCatChem, 2017, ISSN 1867-3880
Publisher: Wiley - VCH Verlag GmbH & CO. KGaA
DOI: 10.1002/cctc.201700620

Characterization and Crystal Structure of a Robust Cyclohexanone Monooxygenase

Author(s): Elvira Romero, J. Rubén Gómez Castellanos, Andrea Mattevi, Marco W. Fraaije
Published in: Angewandte Chemie, 128/51, 2016, Page(s) 16084-16087, ISSN 0044-8249
Publisher: John Wiley & Sons Ltd.
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, 139/2, 2017, Page(s) 627-630, ISSN 0002-7863
Publisher: American Chemical Society
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
Publisher: Royal Society of Chemistry
DOI: 10.1039/C7PY00976C

Book chapters (1)

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, 2018 June 9, 2018, Page(s) 151-187, ISBN 978-0-12-815148-8
Publisher: Elsevier
DOI: 10.1016/bs.mie.2018.04.007

Thesis dissertations (1)

Flavoenzymes at the crossroads of biocatalysis and structural biology

Author(s): Nguyen, Quoc-Thai
Published in: 1, 2017
Publisher: Universityof Groningen