Enzyme Engineering

It was recently discovered that these tiny efficient machines - enzymes - can be modified to become robust enzymes that can revolutionize industrial processes. The ROBOX project specifically focuses on oxidative enzymes that can replace the chemical processes that the industry uses now. This is a complex challenge, but one that the ROBOX project has taken up - determined to make it a success.

Enzymes: What are they? What do they do?

Enzymes are biological catalysts: every form of life depends on many hundreds or thousands of different enzymes. They are molecular machines that facilitate or enable chemical reactions to occur - this is called catalysis. They can do this in astonishing specific and effective manner. Specific: they will only catalyze one type of reaction, converting one specific compound into something else, while not touching other compounds. Effective: they can easily speed up reactions by a million fold, converting billions of a specific compound molecules while just one enzyme molecule is needed. This means that they make things happen. 

What can we do with enzymes?

Enzymes are used in many, many products: washing powder, toothpaste, bread preparation, brewing beer. They are needed and used for: making medicines, sensors, plastics, biofuels, vitamins, improving food, producing antibiotics, developing healthcare products, and many more. Yet, the use of enzymes is limited, for a large part because enzymes from nature are not always suited for use in the final application. They have evolved to be active in their natural environment, and therefore often do not tolerate other conditions (e.g. high temperature, or acid conditions). As a consequence, enzymes often have to be tuned for their envisaged application. This can be done by enzyme engineering. 

What is enzyme engineering?

In the last few decades, techniques have been developed to study enzymes at molecular level. Recently, various methods have been developed that allow to engineer and redesign enzymes. We now know, for a fair number of enzymes, how they look, how they are able to perform chemical reactions and how they can be produced. With this it is possible to exchange each individual amino acid in an enzyme. This means that we can alter enzymes at atomic level at will. This is called enzyme engineering. 

ROBOX's results

ROBOX established a collection of stable enzymes, along with industrial conversion protocols. The project succeeded in: developing an enzyme capable of oxidizing glycerol, applying P450 enzymes to produce drug metabolites on a large scale, applying ADH and BVMO enzymes to produce novel fragrance molecules, as well as precursors for specialty and performance polymers.

In short: ROBOX has demonstrated the techno-economic viability of enzymatic bio-oxidation processes as a greener alternative to traditional chemical processes.

Work Packages

The ROBOX project has an integrated work-flow structure of 7 work packages. Find out more about the work packages below:  

Publications

Enzyme Fusions in Biocatalysis: Coupling Reactions by Pairing Enzymes

Aalbers, Friso S., and Marco W. Fraaije. 
ChemBioChem 20, no. 1 (2019): 20-28.
https://doi.org/10.1002/cbic.201800394

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

Fürst, Maximilian JLJ, Elvira Romero, J. Rúben Gómez Castellanos, Marco W. Fraaije, and Andrea Mattevi. 
ACS catalysis 8, no. 12 (2018): 11648-11656.
DOI: 10.1021/acscatal.8b03793

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

Li, Guangyue, Marc Garcia-Borràs, Maximilian JLJ Fürst, Adriana Ilie, Marco W. Fraaije, K. N. Houk, and Manfred T. Reetz. 
Journal of the American Chemical Society 140, no. 33 (2018): 10464-10472.
https://doi.org/10.1021/jacs.8b04742

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

Solé, Jordi, Gloria Caminal, Martin Schürmann, Gregorio Álvaro, and Marina Guillén. 
Journal of Chemical Technology & Biotechnology 94, no. 1 (2019): 244-255.
https://doi.org/10.1002/jctb.5770

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The crystal structure of P450-TT heme-domain provides the first structural insights into the versatile class VII P450s

Tavanti, Michele, Joanne L. Porter, Colin W. Levy, J. Rubén Gómez Castellanos, Sabine L. Flitsch, and Nicholas J. Turner.
Biochemical and biophysical research communications 501, no. 4 (2018): 846-850.
https://doi.org/10.1016/j.bbrc.2018.05.014

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Partners

A whopping 19 international partners (universities, companies and institutes) were involved in this groundbreaking research project. This just goes to show how huge this project’s scope was, with so many parties that benefited from the research. 

Team

Marco Fraaije
Coördinator

Marco Fraaije

Coördinator
University of Groningen
Margit Winkler
Team Member

Margit Winkler

Team Member
Austrian Centre of Industrial Biotechnology (acib)
Rolf Breinbauer
Team Member

Rolf Breinbauer

Team Member
Austrian Centre of Industrial Biotechnology (acib)
Geert Deroover
Team Member

Geert Deroover

Team Member
ChemStream 
Andreas Vogel
Team Member

Andreas Vogel

Team Member
c-LEcta
Sebastian Bartsch
Team Member

Sebastian Bartsch

Team Member
 c-LEcta
Sven Panke
Team Member

Sven Panke

Team Member
D-BSSE
Martin Held
Team Member

Martin Held

Team Member
D-BSSE
John M Woodley
Team Member

John M Woodley

Team Member
Biocatalysis group at Technical University of Denmark
Christian Leggewie
Team member

Christian Leggewie

Team member
evoXX
Robert Floor
Team Member

Robert Floor

Team Member
Genencor International B.V. 
Simon Ellwood
Team Member

Simon Ellwood

Team Member
Givaudan
Martin Schürmann
Team Member

Martin Schürmann

Team Member
InnoSyn B.V.
Arjan van Kampen
Team Member

Arjan van Kampen

Team Member
PNO
Ulrich Schwaneberg
Team Member

Ulrich Schwaneberg

Team Member
 Institute of Biotechnology at RWTH Aachen University
Anton Glieder
Team Member

Anton Glieder

Team Member
 Graz University of Technology
Gregorio Álvaro
Team Member

Gregorio Álvaro

Team Member
Universitat Autònoma de Barcelona
Andrea Mattevi
Team Member

Andrea Mattevi

Team Member
The University of Pavia
Stefaan De Wildeman
Team Member

Stefaan De Wildeman

Team Member
Maastricht University
John Whittall
Team Member

John Whittall

Team Member
The University of Manchester
Nick Turner
Team Member

Nick Turner

Team Member
The University of Manchester

Partners

Project Coordinator

This project is funded by
Grant no. 635734
EU

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