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Work package 1: Enzyme engineering and identification of oxidative enzymes

 H2020 ROBOX RuG

Aim: We will exploit the current knowledge of structural and mechanistic properties of monooxygenases and oxidases to develop enzyme redesign to deliver robust enzymes using state-of-the art methodologies for predicting beneficial mutations, creating mutant libraries and developing effective screening methods. As well as tuning existing monooxygenases and oxidases towards robust biocatalysts some novel oxidative biocatalysts (alcohol oxidases and dehydrogenases) will be identified using sequence and structural information.

The research in Work Package 1 will result in the generation of a set of robust, productive and selective oxidative biocatalysts for which target reactions of interest for industrial applications have been identified for development in ROBOX. The reactions involve the generation and/or oxidation of alcohols through selective oxidations. The ROBOX biocatalysts generated in this Work Package will be delivered to the partners in the subsequent Work Packages where they will be evaluated and scaled-up for use in oxidation reactions.

Marco FraaijePartners: DSM Chemical Technology R&D BV, Dyadic Nederland BV, c-LEcta GmbH, Evocatal GmbH, Technical University Graz, University of Groningen, University of Pavia, Rheinisch-Westfälische Technische Hochschule Aachen, University of Manchester, Fraunhofer Gesellschaft zur Förderung der Angewandten Forschung EV, Eidgenössische Technische Hochschule Zurich, InnoSyn R&D.

Lead contact: Prof.Dr.Ir. Marco W. Fraaije, This email address is being protected from spambots. You need JavaScript enabled to view it. University of Groningen

 

 

 

 

“The research for this work has received funding from the European Union (EU) project ROBOX (grant agreement n° 635734) under EU’s Horizon 2020 Programme Research and Innovation actions H2020-LEIT BIO-2014-1”

Work package 2: Enzyme Production


Objectives of  WP2


A.GliederBy innovative expression system development and benchmarking of existing technologies WP2 will identify, advance and apply the most competitive expression systems for all individual enzyme classes of ROBOX to enable competitive production processes for robust redox enzymes by robust production strains. The efficiency will be demonstrated by bioreactor cultivations on lab scale at the same time providing the necessary amounts of enzymes in g-scale for process development.  Prof. Dr. Anton Glieder coordinates WP2 activities of 4 industrial and 6 academic partners. He is an experienced scientist of TU Graz and also cofounder of ACIB, acting primarily on the interface of academia and industry.

Contact: This email address is being protected from spambots. You need JavaScript enabled to view it.

 

 

 

 

 

“The research for this work has received funding from the European Union (EU) project ROBOX (grant agreement n° 635734) under EU’s Horizon 2020 Programme Research and Innovation actions H2020-LEIT BIO-2014-1”

Work Package 3: Process design and validation

Objectives:

The main objective of the WP3 is the overall design and validation at small pilot scale of the biocatalytic reactions which are central to the ROBOX project. Within this WP, for each of the biocatalytic reactions to be demonstrated, a reliable, efficient and scalable process will need to be developed. The work package builds on the robust enzymes developed in WP1 for which also efficient productions have been validated in WP2. The design of the biocatalytic reactions is a key step in preparing the demonstrations as it defines the conditions under which the developed biocatalysts can be deployed most effectively, but at the same time most efficiently.

Aiming to achieve this main objective, WP3 has several specific objectives:

  • To test the enzyme stability under different immobilization conditions. Moreover, the most appropriate method for enzyme/whole cell immobilization technique will be chosen and the characterization of the obtained biocatalytic derivatives will be carried out.
  • To define and optimize a suitable reaction for cofactor regeneration system.
  • To select a compatible medium for each biotransformation and cofactor recycling system.
  • To optimize reaction conditions (temperature pH, substrate and enzyme concentration, etc.).
  • To develop downstream processes for isolation/purification of target products.
  • To develop reactor design by engineering solutions. The air/O2 supply flow rate and substrate feeding.

WP leader: Universitat Autònoma de Barcelona - This email address is being protected from spambots. You need JavaScript enabled to view it.

WP participants: DSM, Givaudan, ChemStream, LentiKat’s, ACIB, The University of Manchester, Danmarks Tekniske Universitet, Universitat Autònoma de Barcelona and Universiteit Maastricht

“The research for this work has received funding from the European Union (EU) project ROBOX (grant agreement n° 635734) under EU’s Horizon 2020 Programme Research and Innovation actions H2020-LEIT BIO-2014-1”


 

Work Package 4: Demonstration of Robust Biocatalytic Oxidations and Biooxidation Catalysts

 

Martin SchurmannThe demonstration of the biooxidation enzymes and biocatalytic oxidation reactions developed in the work packages 1, 2 and 3 on pilot scale is the objective of this work package. The information, data and conclusions collected in WP4 will feed WP5 (Process benchmarking and evaluation) and deliver important feedback for the WPs 1-3. WP leader and main contact for the Demonstration WP4 is This email address is being protected from spambots. You need JavaScript enabled to view it. who is currently Senior Scientist at InnoSyn R&D – Innovative Synthesis and has almost 15 years’ experience in the development and scale-up of biocatalysts and biocatalytic processes in chemical industry. 

Questions, Feedback and Comments on any aspect of the ROBOX project are welcomed This email address is being protected from spambots. You need JavaScript enabled to view it..

 

 

“The research for this work has received funding from the European Union (EU) project ROBOX (grant agreement n° 635734) under EU’s Horizon 2020 Programme Research and Innovation actions H2020-LEIT BIO-2014-1”

Work package 5: Process benchmarking & evaluation

Partners involved in work package 5:
Technical University of Denmark (DTU, WP leader This email address is being protected from spambots. You need JavaScript enabled to view it.), DSM Chemical (DSM), Givaudan Suisse S.A. (GIV), ChemStream BVBA (CS), c-LEcta GmbH (CLE), Technische Universitaet Graz (TUG), Rijksuniversiteit Groningen (RUG).

j woodley2Description of work
Benchmarking and evaluation of new process for the production of chemicals is an essential stage in the development of a new chemical product and/or production route. For established chemical technology using conventional homogeneous or heterogeneous catalysts, procedures are already established to achieve such benchmarking. A primary driver in many of these processes is the improvement of selectivity. In contrast, for biocatalytic processes the very high selectivity is normally without question, but instead targets such as improvement of productivity (e.g. space-time yield) are critical. This means that the development targets for biocatalytic processes follow a different path. It is not surprising, given that biocatalysts used in industry will be under conditions very far from those in nature. For example non-natural reactants will be used at concentrations far in excess of anything ever seen by the enzyme before. Moreover an extra degree of freedom is possible in biocatalytic processes since we have the possibility not only to develop the process itself (e.g. via alternative reactors or technologies) but also via improvement of the biocatalyst properties (e.g. via protein engineering). The combination is very powerful but also requires an effective procedure. Hence in this WP the first task will be to develop a suitable protocol, followed by exemplifying this on the case studies.

Objectives
This work package addresses the benchmarking and evaluation of the demonstrated biocatalytic processes, but also individual enzyme performance and production system performance in order evaluate the technology platform. The benchmarking and evaluation are required at 2 stages of development – at the start to establish targets for processes and biocatalyst improvement – at the end to establish process feasibility. The process is iterative in that developments at each stage can be used to re-evaluate the process.

Specific objectives

  1. Establish a protocol for benchmarking and evaluation of the case studies in biooxidation in this proposal.
  2. Use the protocol to establish early stage targets for process development.
  3. Evaluate processes at the completion of development to assess feasibility for scale-up.
  4. Evaluating the economic feasibility of the processes and the platform compared to conventional processes.
  5. Assessing the environmental benefits of the processes.

 “The research for this work has received funding from the European Union (EU) project ROBOX (grant agreement n° 635734) under EU’s Horizon 2020 Programme Research and Innovation actions H2020-LEIT BIO-2014-1”

Work Package 6: Project Exploitation, Intellectual Property Management, Dissemination and Public Engagement.

Dr J WhittallObjectives of WP6

This  WP  defines a structured plan for  the foreground exploitation, IP management  and  technology  transfer  of  project  results  along with communication to  all the stakeholders (including policy makers, industry and society) to maximize impacts and deliver maximum value for the investment. Main contact for dissemination and leading WP6 will be This email address is being protected from spambots. You need JavaScript enabled to view it. who is Research Exploitation Manager at CoEBio3 and has 40 years’ experience working in the research environment at the academic-industry interface. He is Dissemination and Exploitation Officer (DEO) of ROBOX, with the role of communication, dissemination and exploitation of project results.

Questions, Feedback and Comments on any aspect of the ROBOX project are welcomed.  

Contact: This email address is being protected from spambots. You need JavaScript enabled to view it.

 

 “The research for this work has received funding from the European Union (EU) project ROBOX (grant agreement n° 635734) under EU’s Horizon 2020 Programme Research and Innovation actions H2020-LEIT BIO-2014-1”

 


 

 

Work Package 7: Project Management

Objectives of WP7:

This Work package supports the integral management of the ROBOX project, and enables and stimulates efficient exchange of information and collaboration
Specific objectives include:

-    Ensuring proper implementation of Work Plan and achievement of ROBOX objectives
-    Efficient organisation and day-to-day management of the project
-    Preparing and managing the Consortium Agreement
-    Monitoring and ensuring on-time delivery of scientific and financial reports
-    Establish effective communication with the Commission
-    Ensuring meaningful interactions between collaborating Partners to enhance project outputs.
-    Organisation of project meetings
-    Stimulate / facilitate dissemination of results

Rob Meier

Partners DSM, TU Graz, University of Groningen, University of Manchester, Danish Technical University, and Universitat Autonoma de Barcelona , representing the Coordinator and all 6 work package leaders.

 

Questions, Feedback and Comments on any aspect of the ROBOX project are welcomed

 

Contact: This email address is being protected from spambots. You need JavaScript enabled to view it.

 

“The research for this work has received funding from the European Union (EU) project ROBOX (grant agreement n° 635734) under EU’s Horizon 2020 Programme Research and Innovation actions H2020-LEIT BIO-2014-1”