Engineering de novo production of chlorinated polyketides in Pseudomonas putida

Project Information

DONNA

Grant agreement ID: 839839

DOI

10.3030/839839

Project closed

EC signature date 30 April 2019

Start date 1 February 2020

End date 31 January 2022

Funded under

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Total cost

€ 207 312,00

EU contribution

€ 207 312,00

Coordinated by

DANMARKS TEKNISKE UNIVERSITET
Denmark

Periodic Reporting for period 1 - DONNA (Engineering de novo production of chlorinated polyketides in Pseudomonas putida)

Reporting period: 2020-02-01 to 2022-01-31

Summary of the context and overall objectives of the project

Sustainable economic development demands new biocatalysts to carry out novel, selective synthesis reactions in an environmentally-friendly fashion. Halogenated organic compounds, key products due to their multiple biological activities and industrial applications, continue to be produced via traditional chemistry and their efficient and cost-effective biological synthesis has not been yet achieved. Currently, 30 % of drugs in clinical trials are halogenated, and 85 % of all pharmaceutical agents involve chlorine. The EU-funded DONNA project aims at the bio-based production of chlorinated metabolites via metabolic engineering of the model bacterium Pseudomonas putida. During the implementation of the project, the first steps of a roadmap towards a re-factored version of the microorganism capable of funneling chlorinated metabolites into new-to-nature polyketides by re-programmed polyketide synthases enzymes have been established. In addition, the portfolio of halogenated molecules that have been synthetized with the cell factories from this project (e.g. chlorinated sugars, nucleotides, and carboxylic acids) could contribute to the European knowledge-based bio-economy, enhancing the EU's competitiveness in White Biotechnology and benefiting the society as a whole.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

In this project, we have achieved several scientifically and industrially interesting results. For instance, we have built a collection of portable genetic modules (ChloroBricks) that has been applied during the construction of bacterial platforms for the production of high added-value chlorinated chemicals (including chlorinated sugars, nucleotides, and carboxylic acids). We have also made significant progress in the rational reprogramming of modular polyketide synthases to produce chlorinated polyketides (e.g. chlorinated triketide lactones) in both in vivo and in vitro assays. In a society that seeks more selective, ecological, and sustainable solutions for the manufacture of halogenated molecules, the biotechnological alternatives developed in this project could have some commercial relevance. Although no patent has been filed, for the time being, the receiving laboratory will continue to work on optimizing the results of the project and the subsequent exploitation of those results with exceptional commercial potential.

During the implementation of the project, we have carried out multiple activities aimed at communicating and disseminating the results of the research. One research article has been published and two additional articles are expected to be published in the coming months. All articles will be published in high-quality peer-reviewed journals under the Open Access framework and will include a reference to EU funding. The vision and results of the project have also been disseminated among scientific colleagues and collaborators at regular meetings and seminars, and among experts in the field at an international scientific conference. To bring research closer to non-specialized groups, we have collaborated in "Science is wonderful!", an outreach event organized by the European Commission aimed at European secondary school students

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

The potential for turning the final results of DONNA into commercial products can be expected to be high, considering the strong interest from chemical and pharmaceutical companies in sustainable alternative ways of producing added-value halogenated compounds. Halogenated compounds are currently synthesized by chemical processes that operate under harsh reaction conditions, sometimes with low product selectivity. Therefore, sustainable economic development requires novel catalysts for selective and environmentally friendly halogenation reactions. The results of this project might help to strengthen the economy and competitiveness of the European Union in the relevant White Biotechnology field. For instance, the triketide lactones (TKLs) (and their chlorinated analogs, Cl-TKLs) are very interesting molecules due to their multiple chiral centers, since they can be used as synthons in the chemical synthesis processes of more complex bioactive molecules. Although we have not managed to produce Cl-TKLs during the execution of the project due to time constraints, it is expected that the receiving laboratory will continue to investigate the synthesis of these compounds in the near future. If successful, the approach used here could be applied to any modular polyketide synthase to produce potentially any chlorinated polyketide à la carte. These types of approaches are essential to accelerate the development of new drug analogs (e.g. antibiotics). However, the portfolio of interesting molecules that have been produced in DONNA goes beyond the chlorinated polyketides. Halogenated building blocks have gained greater relevance during the last four decades due to the development of cross-coupling chemistry. Therefore, several chlorinated intermediates such as chlorinated sugars, nucleotides, and carboxylic acids could be used as building blocks for organic chemical synthesis in different industrial applications.

Overview of the project