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- Title
Bio-upcycling of even and uneven medium-chain-length diols and dicarboxylates to polyhydroxyalkanoates using engineered Pseudomonas putida.
- Authors
Ackermann, Yannic S.; de Witt, Jan; Mezzina, Mariela P.; Schroth, Christoph; Polen, Tino; Nikel, Pablo I.; Wynands, Benedikt; Wierckx, Nick
- Abstract
Bio-upcycling of plastics is an emerging alternative process that focuses on extracting value from a wide range of plastic waste streams. Such streams are typically too contaminated to be effectively processed using traditional recycling technologies. Medium-chain-length (mcl) diols and dicarboxylates (DCA) are major products of chemically or enzymatically depolymerized plastics, such as polyesters or polyethers. In this study, we enabled the efficient metabolism of mcl-diols and -DCA in engineered Pseudomonas putida as a prerequisite for subsequent bio-upcycling. We identified the transcriptional regulator GcdR as target for enabling metabolism of uneven mcl-DCA such as pimelate, and uncovered amino acid substitutions that lead to an increased coupling between the heterologous β-oxidation of mcl-DCA and the native degradation of short-chain-length DCA. Adaptive laboratory evolution and subsequent reverse engineering unravelled two distinct pathways for mcl-diol metabolism in P. putida, namely via the hydroxy acid and subsequent native β-oxidation or via full oxidation to the dicarboxylic acid that is further metabolized by heterologous β-oxidation. Furthermore, we demonstrated the production of polyhydroxyalkanoates from mcl-diols and -DCA by a single strain combining all required metabolic features. Overall, this study provides a powerful platform strain for the bio-upcycling of complex plastic hydrolysates to polyhydroxyalkanoates and leads the path for future yield optimizations.
- Subjects
PSEUDOMONAS putida; POLYHYDROXYALKANOATES; BIOLOGICAL evolution; HYDROXY acids; GLYCOLS; DICARBOXYLIC acids; POLYESTERS
- Publication
Microbial Cell Factories, 2024, Vol 23, Issue 1, p1
- ISSN
1475-2859
- Publication type
Article
- DOI
10.1186/s12934-024-02310-7