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

Total Cost €


EC-Contrib. €






 ROBOX project word cloud

Explore the words cloud of the ROBOX project. It provides you a very rough idea of what is the project "ROBOX" about.

temperature    energy    catalyzed    nutrition    segment    pharma    commercial    benign    materials    lower    industrial    price    mainly    50    expression    molecular    reductions    groups    markets    adh    1000    raw    sme    ktons    enzymes    p450    catalytic    kg    technologies    widening    lab    plug    transformations    emissions    justifies    villiger    oxidation    robox    oxidative    sustainability    efficiency    16    baeyer    functional    ghg    enzyme    oxygen    dehydrogenase    p450s    replication    types    broad    computational    enzymatic    optimal    air    players    11    screening    chemical    pressure    routes    gt    bulk    highest    60    economics    limitations    introduction    bio    ambient    mild    specialty    viability    reactions    prepare    efficient    substantial    segments    industry    alcohol    economic    ph    platform    joint    added    oxidase    bug    monooxygenase    monooxygenases    volume    bvmos    aox    validated    plants    chemicals    innovative    functionalities    fine    larger    techno   

Project "ROBOX" data sheet

The following table provides information about the project.


Organization address
address: Broerstraat 5
postcode: 9712CP

contact info
title: n.a.
name: n.a.
surname: n.a.
function: n.a.
email: n.a.
telephone: n.a.
fax: n.a.

 Coordinator Country Netherlands [NL]
 Project website
 Total cost 11˙159˙493 €
 EC max contribution 8˙181˙676 € (73%)
 Programme 1. H2020-EU.2.1.4. (INDUSTRIAL LEADERSHIP - Leadership in enabling and industrial technologies – Biotechnology)
 Code Call H2020-LEIT-BIO-2014-1
 Funding Scheme IA
 Starting year 2015
 Duration (year-month-day) from 2015-04-01   to  2019-03-31


Take a look of project's partnership.

# participants  country  role  EC contrib. [€] 
2    DSM CHEMICAL TECHNOLOGY R & D BV NL (SITTARD) participant 762˙832.00
5    TECHNISCHE UNIVERSITAET GRAZ AT (GRAZ) participant 675˙415.00
7    INNOSYN BV NL (GELEEN) participant 478˙362.00
10    UNIVERSITA DEGLI STUDI DI PAVIA IT (PAVIA) participant 447˙205.00
11    EVOXX TECHNOLOGIES GMBH DE (MONHEIM) participant 312˙025.00
12    CHEMSTREAM BVBA BE (EDEGEM) participant 298˙812.00
13    ACIB GMBH AT (GRAZ) participant 282˙278.00
14    C-LECTA GMBH DE (LEIPZIG) participant 274˙952.00
15    GENENCOR INTERNATIONAL BV NL (LEIDEN) participant 265˙759.00
17    PNO CONSULTANTS BV NL (RIJSWIJK) participant 97˙370.00
18    DYADIC NEDERLAND BV NL (Wageningen) participant 89˙577.00
19    LentiKat's a.s. CZ (STRAZ POD RALSKEM) participant 55˙810.00
21    GIVAUDAN SUISSE SA CH (VERNIER) participant 0.00


 Project objective

Oxygen functionalities are key functional groups in many of today’s chemicals and materials. The efficient introduction of oxygen-functionalities into raw materials are key chemical transformations in bulk and fine chemicals. Innovative bio-catalytic oxidation routes using molecular oxygen (from air) under benign and mild (pH) conditions such as ambient temperature and pressure can greatly improve the sustainability and economics of processes, but were so far mainly been applied in the pharma segments. In this segment, the enzyme-catalyzed step often represents the highest added value and the high price of the end-product (> €1000/kg) justifies less than optimal enzyme production and limitations in its catalytic efficiency.

In order to achieve the widening of industrial application of enzymatic bio-oxidation processes to also larger volume but lower price chemical markets, ROBOX will demonstrate the techno-economic viability of bio-transformations of four types of robust oxidative enzymes: P450 monooxygenases (P450s), Baeyer-Villiger MonoOxygenase (BVMOs), Alcohol DeHydrogenase (ADH) and Alcohol OXidase (AOX) for which target reactions have already been validated on lab-scale in pharma, nutrition, fine & specialty chemicals and materials applications. ROBOX will demonstrate 11 target reactions on large scale for these markets in order to prepare them for scale up to commercial-scale plants.

ROBOX is industry-driven with 2 major industrial players and 6 SME’s. It will assess the potential of technologies applied to become platform technologies technologies (multi-parameter screening systems, computational methodologies, “plug bug” expression systems) for broad replication throughout the chemical industry. The markets addressed within ROBOX represent a joint volume of over 6.000 ktons/year. The introduction of bio-oxidation processes is expected to bring substantial reductions in cost (up to -50%), energy use (-60%), chemicals (-16%) and GHG-emissions (-50%).


List of deliverables.
Optimized enzyme production processes for ROBOX enzymes in bioreactor including DSP Demonstrators, pilots, prototypes 2020-04-22 10:11:11
Overall analysis of environmental benefits and techno-economics of the processes and platform Documents, reports 2020-04-22 10:11:11
96-well capillary electrophoreses screening system and first flow cytometer based screening system available Demonstrators, pilots, prototypes 2020-04-22 10:11:11
Biocatalytic and structural exploration of P450-AX as well as generation of improved variants Demonstrators, pilots, prototypes 2020-04-22 10:11:11
Quorum sensing technology and micro fluidics platform available Demonstrators, pilots, prototypes 2020-04-22 10:11:10
Set of P450 enzymes with improved performance for hydroxylation of aromatics, alkenes and APIs and a set of API-sensors to enable P450 improvement Demonstrators, pilots, prototypes 2020-04-22 10:11:10
Generation of a first flow cytometer based screening system for quantification of one product in the BM3 demonstration process through fluorogenic sensors Demonstrators, pilots, prototypes 2020-04-22 10:11:10
Public website and newsletter, first press releases on the project scopes Documents, reports 2020-04-22 10:11:09
Project leaflet in several languages Documents, reports 2020-04-22 10:11:09
Completion of benchmarking protocol and flowsheets established for the case studies Documents, reports 2020-04-22 10:11:09
New vector and host systems for enzyme coexpression Demonstrators, pilots, prototypes 2020-04-22 10:11:10
First generation production strains for robust BVMOs Demonstrators, pilots, prototypes 2020-04-22 10:11:09
Identification of genes predicted to encode relevant robust BVMOs and oxidases Documents, reports 2020-04-22 10:11:09
Structural elucidation of relevant of BVMOs e.g. at least 3 new enzyme structures Documents, reports 2020-04-22 10:11:10
Communication and dissemination plan Documents, reports 2020-04-22 10:11:09
Installation of advisory boards Other 2020-04-22 10:11:09
First generation production strains for microbial and human P450s Demonstrators, pilots, prototypes 2020-04-22 10:11:09
Stakeholder analysis Documents, reports 2020-04-22 10:11:10

Take a look to the deliverables list in detail:  detailed list of ROBOX deliverables.


year authors and title journal last update
List of publications.
2017 Gonzalo de Gonzalo, Maximilian Fürst, Marco Fraaije
Polycyclic Ketone Monooxygenase (PockeMO): A Robust Biocatalyst for the Synthesis of Optically Active Sulfoxides
published pages: 288, ISSN: 2073-4344, DOI: 10.3390/catal7100288
Catalysts 7/12 2020-04-22
2017 Friso S. Aalbers, Marco W. Fraaije
Coupled reactions by coupled enzymes: alcohol to lactone cascade with alcohol dehydrogenase–cyclohexanone monooxygenase fusions
published pages: 7557-7565, ISSN: 0175-7598, DOI: 10.1007/s00253-017-8501-4
Applied Microbiology and Biotechnology 101/20 2020-04-22
2017 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
Manipulating the stereoselectivity of the thermostable Baeyer–Villiger monooxygenase TmCHMO by directed evolution
published pages: 9824-9829, ISSN: 1477-0520, DOI: 10.1039/C7OB02692G
Organic & Biomolecular Chemistry 15/46 2020-04-22
2017 Iwona Kaluzna, Jan Brummund, and Martin Schuermann
Production of diclofenac metabolites by applying cytochrome P450 technology
published pages: 55-58, ISSN: 0392-839X, DOI:
Chimica Oggi - Chemistry Today vol. 35(6) November/December 20 2020-04-22
2018 Marie A. F. Delgove, Matthew T. Elford, Katrien V. Bernaerts, Stefaan M.A. De Wildeman
Application of a thermostable Baeyer-Villiger monooxygenase for the synthesis of branched polyester precursors
published pages: , ISSN: 0268-2575, DOI: 10.1002/jctb.5623
Journal of Chemical Technology & Biotechnology 2020-04-22
2017 Marie A. F. Delgove, Maximilian J. L. J. Fürst, Marco W. Fraaije, Katrien V. Bernaerts, Stefaan M. A. De Wildeman
Exploring the Substrate Scope of Baeyer-Villiger Monooxygenases with Branched Lactones as Entry towards Polyesters
published pages: , ISSN: 1439-4227, DOI: 10.1002/cbic.201700427
ChemBioChem 2020-04-22
2018 Astrid Weninger, Jasmin E. Fischer, Hana Raschmanová, Claudia Kniely, Thomas Vogl, Anton Glieder
Expanding the CRISPR/Cas9 toolkit for Pichia pastoris with efficient donor integration and alternative resistance markers
published pages: 3183-3198, ISSN: 0730-2312, DOI: 10.1002/jcb.26474
Journal of Cellular Biochemistry 119/4 2020-04-22
2019 Jordi Solé, Gloria Caminal, Martin Schürmann, Gregorio Álvaro, Marina Guillén
Co-immobilization of P450 BM3 and glucose dehydrogenase on different supports for application as a self-sufficient oxidative biocatalyst
published pages: 244-255, ISSN: 0268-2575, DOI: 10.1002/jctb.5770
Journal of Chemical Technology & Biotechnology 94/1 2020-04-22
2017 Nguyen, Quoc-Thai
Flavoenzymes at the crossroads of biocatalysis and structural biology
published pages: , ISSN: , DOI:
1 2020-04-22
2018 Joanne L. Porter, Selina Sabatini, Jack Manning, Michele Tavanti, James L. Galman, Nicholas J. Turner, Sabine L. Flitsch
Cloning, expression and characterisation of P450-Hal1 (CYP116B62) from Halomonas sp. NCIMB 172: A self-sufficient P450 with high expression and diverse substrate scope
published pages: 1-8, ISSN: 0141-0229, DOI: 10.1016/j.enzmictec.2018.02.005
Enzyme and Microbial Technology 113 2020-04-22
2017 Michele Tavanti, Juan Mangas-Sanchez, Sarah L. Montgomery, Matthew P. Thompson, Nicholas J. Turner
A biocatalytic cascade for the amination of unfunctionalised cycloalkanes
published pages: 9790-9793, ISSN: 1477-0520, DOI: 10.1039/c7ob02569f
Organic & Biomolecular Chemistry 15/46 2020-04-22
2018 Joanne L. Porter, Jack Manning, Selina Sabatini, Michele Tavanti, Nicholas J. Turner, Sabine L. Flitsch
Characterisation of CYP102A25 from Bacillus marmarensis and CYP102A26 from Pontibacillus halophilus : P450 Homologues of BM3 with Preference towards Hydroxylation of Medium-Chain Fatty Acids
published pages: 513-520, ISSN: 1439-4227, DOI: 10.1002/cbic.201700598
ChemBioChem 19/5 2020-04-22
2018 Alexandra M. Weingartner, Daniel F. Sauer, Gaurao V. Dhoke, Mehdi D. Davari, Anna Joëlle Ruff, Ulrich Schwaneberg
A hydroquinone-specific screening system for directed P450 evolution
published pages: 9657-9667, ISSN: 0175-7598, DOI: 10.1007/s00253-018-9328-3
Applied Microbiology and Biotechnology 102/22 2020-04-22
2018 Murray P. Meissner, Mathias Nordblad, John M. Woodley
Online Measurement of Oxygen-Dependent Enzyme Reaction Kinetics
published pages: 106-113, ISSN: 1439-4227, DOI: 10.1002/cbic.201700577
ChemBioChem 19/1 2020-04-22
2018 Quoc-Thai Nguyen, Elvira Romero, Willem P. Dijkman, Suzan Pantaroto de Vasconcellos, Claudia Binda, Andrea Mattevi, Marco W. Fraaije
Structure-Based Engineering of Phanerochaete chrysosporium Alcohol Oxidase for Enhanced Oxidative Power toward Glycerol
published pages: 6209-6218, ISSN: 0006-2960, DOI: 10.1021/acs.biochem.8b00918
Biochemistry 57/43 2020-04-22
2019 Ángela Mourelle-Insua, Friso S. Aalbers, Iván Lavandera, Vicente Gotor-Fernández, Marco W. Fraaije
What to sacrifice? Fusions of cofactor regenerating enzymes with Baeyer-Villiger monooxygenases and alcohol dehydrogenases for self-sufficient redox biocatalysis
published pages: 1832-1839, ISSN: 0040-4020, DOI: 10.1016/j.tet.2019.02.015
Tetrahedron 75/13 2020-04-22
2018 Marie A. F. Delgove, Matthew T. Elford, Katrien V. Bernaerts, Stefaan M. A. De Wildeman
Toward Upscaled Biocatalytic Preparation of Lactone Building Blocks for Polymer Applications
published pages: 803-812, ISSN: 1083-6160, DOI: 10.1021/acs.oprd.8b00079
Organic Process Research & Development 22/7 2020-04-22
2019 Friso S. Aalbers, Marco W. Fraaije
Design of Artificial Alcohol Oxidases: Alcohol Dehydrogenase-NADPH Oxidase Fusions for Continuous Oxidations
published pages: 51-56, ISSN: 1439-4227, DOI: 10.1002/cbic.201800421
ChemBioChem 20/1 2020-04-22
2019 Friso S. Aalbers, Marco W. Fraaije
Enzyme Fusions in Biocatalysis: Coupling Reactions by Pairing Enzymes
published pages: 20-28, ISSN: 1439-4227, DOI: 10.1002/cbic.201800394
ChemBioChem 20/1 2020-04-22
2018 Mathias Nordblad, Mafalda Dias Gomes, Murray P. Meissner, Hemalata Ramesh, John M. Woodley
Scoping Biocatalyst Performance Using Reaction Trajectory Analysis
published pages: 1101-1114, ISSN: 1083-6160, DOI: 10.1021/acs.oprd.8b00119
Organic Process Research & Development 22/9 2020-04-22
2018 Guangyue Li, Marc Garcia-Borràs, Maximilian J. L. J. Fürst, Adriana Ilie, Marco W. Fraaije, K. N. Houk, Manfred T. Reetz
Overriding Traditional Electronic Effects in Biocatalytic Baeyer–Villiger Reactions by Directed Evolution
published pages: 10464-10472, ISSN: 0002-7863, DOI: 10.1021/jacs.8b04742
Journal of the American Chemical Society 140/33 2020-04-22
2018 Marie A.F. Delgove, Daniela Valencia, Jordi Solé, Katrien V. Bernaerts, Stefaan M.A. De Wildeman, Marina Guillén, Gregorio Álvaro
High performing immobilized Baeyer-Villiger monooxygenase and glucose dehydrogenase for the synthesis of Ɛ-caprolactone derivative
published pages: 134-141, ISSN: 0926-860X, DOI: 10.1016/j.apcata.2018.12.036
Applied Catalysis A: General 572 2020-04-22
2019 Marie A. F. Delgove, Achille-B. Laurent, John M. Woodley, Stefaan M. A. De Wildeman, Katrien V. Bernaerts, Yvonne van der Meer
A prospective life-cycle assessment (LCA) of monomer synthesis: Comparison of biocatalytic and oxidative chemistry
published pages: , ISSN: 1864-5631, DOI: 10.1002/cssc.201900007
ChemSusChem 2020-04-22
2017 Michele Tavanti, Fabio Parmeggiani, Jose Rubén Gómez Castellanos, Andrea Mattevi, Nicholas John Turner
One-pot Biocatalytic Double Oxidation of α-Isophorone for the Synthesis of Ketoisophorone
published pages: , ISSN: 1867-3880, DOI: 10.1002/cctc.201700620
ChemCatChem 2020-04-22
2017 Marie A. F. Delgove, Juandré Luchies, Iris Wauters, Geert G. P. Deroover, Stefaan M. A. De Wildeman, Katrien V. Bernaerts
Increasing the solubility range of polyesters by tuning their microstructure with comonomers
published pages: , ISSN: 1759-9954, DOI: 10.1039/C7PY00976C
Polym. Chem. 2020-04-22
2017 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
Polycyclic Ketone Monooxygenase from the Thermophilic Fungus Thermothelomyces thermophila : A Structurally Distinct Biocatalyst for Bulky Substrates
published pages: 627-630, ISSN: 0002-7863, DOI: 10.1021/jacs.6b12246
Journal of the American Chemical Society 139/2 2020-04-22
2016 Quoc-Thai Nguyen, Gonzalo de Gonzalo, Claudia Binda, Ana Rioz-Martínez, Andrea Mattevi, Marco W. Fraaije
Biocatalytic Properties and Structural Analysis of Eugenol Oxidase from Rhodococcus jostii RHA1: A Versatile Oxidative Biocatalyst
published pages: 1359-1366, ISSN: 1439-4227, DOI: 10.1002/cbic.201600148
ChemBioChem 17/14 2020-04-22
2016 Elvira Romero, J. Rubén Gómez Castellanos, Andrea Mattevi, Marco W. Fraaije
Characterization and Crystal Structure of a Robust Cyclohexanone Monooxygenase
published pages: 16084-16087, ISSN: 0044-8249, DOI: 10.1002/ange.201608951
Angewandte Chemie 128/51 2020-04-22

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