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  1. Artikel ; Online: Engineering 5-hydroxymethylfurfural (HMF) oxidation in Pseudomonas boosts tolerance and accelerates 2,5-furandicarboxylic acid (FDCA) production.

    Lechtenberg, Thorsten / Wynands, Benedikt / Wierckx, Nick

    Metabolic engineering

    2023  Band 81, Seite(n) 262–272

    Abstract: Due to its tolerance properties, Pseudomonas has gained particular interest as host for oxidative upgrading of the toxic aldehyde 5-hydroxymethylfurfural (HMF) into 2,5-furandicarboxylic acid (FDCA), a promising biobased alternative to terephthalate in ... ...

    Abstract Due to its tolerance properties, Pseudomonas has gained particular interest as host for oxidative upgrading of the toxic aldehyde 5-hydroxymethylfurfural (HMF) into 2,5-furandicarboxylic acid (FDCA), a promising biobased alternative to terephthalate in polyesters. However, until now, the native enzymes responsible for aldehyde oxidation are unknown. Here, we report the identification of the primary HMF-converting enzymes of P. taiwanensis VLB120 and P. putida KT2440 by extended gene deletions. The key players in HMF oxidation are a molybdenum-dependent periplasmic oxidoreductase and a cytoplasmic dehydrogenase. Deletion of the corresponding genes almost completely abolished HMF oxidation, leading instead to aldehyde reduction. In this context, two HMF-reducing dehydrogenases were also revealed. These discoveries enabled enhancement of Pseudomonas' furanic aldehyde oxidation machinery by genomic overexpression of the respective genes. The resulting BOX strains (Boosted OXidation) represent superior hosts for biotechnological synthesis of FDCA from HMF. The increased oxidation rates provide greatly elevated HMF tolerance, thus tackling one of the major drawbacks of whole-cell catalysis with this aldehyde. Furthermore, the ROX (Reduced OXidation) and ROAR (Reduced Oxidation And Reduction) deletion mutants offer a solid foundation for future development of Pseudomonads as biotechnological chassis notably for scenarios where rapid HMF conversion is undesirable.
    Mesh-Begriff(e) Pseudomonas/genetics ; Furaldehyde ; Furans ; Dicarboxylic Acids
    Chemische Substanzen 5-hydroxymethylfurfural (70ETD81LF0) ; 2,5-furandicarboxylic acid (73C4JJ695C) ; Furaldehyde (DJ1HGI319P) ; Furans ; Dicarboxylic Acids
    Sprache Englisch
    Erscheinungsdatum 2023-12-26
    Erscheinungsland Belgium
    Dokumenttyp Journal Article
    ZDB-ID 1470383-x
    ISSN 1096-7184 ; 1096-7176
    ISSN (online) 1096-7184
    ISSN 1096-7176
    DOI 10.1016/j.ymben.2023.12.010
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  2. Buch ; Online ; Dissertation / Habilitation: Enabling mixed microbial upcycling of plastic monomers

    Ackermann, Yannic [Verfasser] / Wierckx, Nick [Gutachter] / Axmann, Ilka [Gutachter]

    2024  

    Verfasserangabe Yannic Sebastian Ackermann ; Gutachter: Nick Wierckx, Ilka Axmann
    Schlagwörter Technische Chemie ; Technical Chemistry
    Thema/Rubrik (Code) sg660
    Sprache Englisch
    Verlag Universitäts- und Landesbibliothek der Heinrich-Heine-Universität Düsseldorf
    Erscheinungsort Düsseldorf
    Dokumenttyp Buch ; Online ; Dissertation / Habilitation
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  3. Buch ; Online ; Dissertation / Habilitation: Strain- and process engineering for polyketides production with Pseudomonas taiwanensis VLB120 in two-phase cultivations

    Schwanemann, Tobias Philipp [Verfasser] / Wierckx, Nick [Gutachter] / Frunzke, Julia [Gutachter]

    2024  

    Verfasserangabe Tobias Philipp Schwanemann ; Gutachter: Nick Wierckx, Julia Frunzke
    Schlagwörter Technische Chemie ; Technical Chemistry
    Thema/Rubrik (Code) sg660
    Sprache Englisch
    Verlag Universitäts- und Landesbibliothek der Heinrich-Heine-Universität Düsseldorf
    Erscheinungsort Düsseldorf
    Dokumenttyp Buch ; Online ; Dissertation / Habilitation
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  4. Artikel: Editorial: Microbial Degradation of Plastics.

    Wei, Ren / Wierckx, Nick

    Frontiers in microbiology

    2021  Band 12, Seite(n) 635621

    Sprache Englisch
    Erscheinungsdatum 2021-02-11
    Erscheinungsland Switzerland
    Dokumenttyp Editorial
    ZDB-ID 2587354-4
    ISSN 1664-302X
    ISSN 1664-302X
    DOI 10.3389/fmicb.2021.635621
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  5. Artikel ; Online: Improving 5-(hydroxymethyl)furfural (HMF) tolerance of

    Lechtenberg, Thorsten / Wynands, Benedikt / Müller, Moritz-Fabian / Polen, Tino / Noack, Stephan / Wierckx, Nick

    Metabolic engineering communications

    2024  Band 18, Seite(n) e00235

    Abstract: The aldehyde 5-(hydroxymethyl)furfural (HMF) is of great importance for a circular bioeconomy. It is a renewable platform chemical that can be converted into a range of useful compounds to replace petroleum-based products such as the green plastic ... ...

    Abstract The aldehyde 5-(hydroxymethyl)furfural (HMF) is of great importance for a circular bioeconomy. It is a renewable platform chemical that can be converted into a range of useful compounds to replace petroleum-based products such as the green plastic monomer 2,5-furandicarboxylic acid (FDCA). However, it also exhibits microbial toxicity for example hindering the efficient biotechnological valorization of lignocellulosic hydrolysates. Thus, there is an urgent need for tolerance-improved organisms applicable to whole-cell biocatalysis. Here, we engineer an oxidation-deficient derivative of the naturally robust and emerging biotechnological workhorse
    Sprache Englisch
    Erscheinungsdatum 2024-05-10
    Erscheinungsland Netherlands
    Dokumenttyp Journal Article
    ZDB-ID 2821894-2
    ISSN 2214-0301 ; 2214-0301
    ISSN (online) 2214-0301
    ISSN 2214-0301
    DOI 10.1016/j.mec.2024.e00235
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  6. Artikel ; Online: Microbial synthesis of the plant natural product precursor p-coumaric acid with Corynebacterium glutamicum.

    Mutz, Mario / Kösters, Dominic / Wynands, Benedikt / Wierckx, Nick / Marienhagen, Jan

    Microbial cell factories

    2023  Band 22, Heft 1, Seite(n) 209

    Abstract: Background: Phenylpropanoids such as p-coumaric acid represent important precursors for the synthesis of a broad range of plant secondary metabolites including stilbenoids, flavonoids, and lignans, which are of pharmacological interest due to their ... ...

    Abstract Background: Phenylpropanoids such as p-coumaric acid represent important precursors for the synthesis of a broad range of plant secondary metabolites including stilbenoids, flavonoids, and lignans, which are of pharmacological interest due to their health-promoting properties. Although extraction from plant material or chemical synthesis is possible, microbial synthesis of p-coumaric acid from glucose has the advantage of being less expensive and more resource efficient. In this study, Corynebacterium glutamicum was engineered for the production of the plant polyphenol precursor p-coumaric acid from glucose.
    Results: Heterologous expression of the tyrosine ammonia-lyase encoding gene from Flavobacterium johnsoniae enabled the conversion of endogenously provided tyrosine to p-coumaric acid. Product consumption was avoided by abolishing essential reactions of the phenylpropanoid degradation pathway. Accumulation of anthranilate as a major byproduct was eliminated by reducing the activity of anthranilate synthase through targeted mutagenesis to avoid tryptophan auxotrophy. Subsequently, the carbon flux into the shikimate pathway was increased, phenylalanine biosynthesis was reduced, and phosphoenolpyruvate availability was improved to boost p-coumaric acid accumulation. A maximum titer of 661 mg/L p-coumaric acid (4 mM) in defined mineral medium was reached. Finally, the production strain was utilized in co-cultivations with a C. glutamicum strain previously engineered for the conversion of p-coumaric acid into the polyphenol resveratrol. These co-cultivations enabled the synthesis of 31.2 mg/L (0.14 mM) resveratrol from glucose without any p-coumaric acid supplementation.
    Conclusions: The utilization of a heterologous tyrosine ammonia-lyase in combination with optimization of the shikimate pathway enabled the efficient production of p-coumaric acid with C. glutamicum. Reducing the carbon flux into the phenylalanine and tryptophan branches was the key to success along with the introduction of feedback-resistant enzyme variants.
    Mesh-Begriff(e) Resveratrol/metabolism ; Corynebacterium glutamicum/genetics ; Corynebacterium glutamicum/metabolism ; Tryptophan/metabolism ; Plants/genetics ; Glucose/metabolism ; Polyphenols ; Phenylalanine/metabolism ; Metabolic Engineering
    Chemische Substanzen Resveratrol (Q369O8926L) ; p-coumaric acid (IBS9D1EU3J) ; Tryptophan (8DUH1N11BX) ; shikimate ; Glucose (IY9XDZ35W2) ; Polyphenols ; Phenylalanine (47E5O17Y3R)
    Sprache Englisch
    Erscheinungsdatum 2023-10-13
    Erscheinungsland England
    Dokumenttyp Journal Article
    ZDB-ID 2091377-1
    ISSN 1475-2859 ; 1475-2859
    ISSN (online) 1475-2859
    ISSN 1475-2859
    DOI 10.1186/s12934-023-02222-y
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  7. Artikel ; Online: A Pseudomonas taiwanensis malonyl-CoA platform strain for polyketide synthesis.

    Schwanemann, Tobias / Otto, Maike / Wynands, Benedikt / Marienhagen, Jan / Wierckx, Nick

    Metabolic engineering

    2023  Band 77, Seite(n) 219–230

    Abstract: Malonyl-CoA is a central precursor for biosynthesis of a wide range of complex secondary metabolites. The development of platform strains with increased malonyl-CoA supply can contribute to the efficient production of secondary metabolites, especially if ...

    Abstract Malonyl-CoA is a central precursor for biosynthesis of a wide range of complex secondary metabolites. The development of platform strains with increased malonyl-CoA supply can contribute to the efficient production of secondary metabolites, especially if such strains exhibit high tolerance towards these chemicals. In this study, Pseudomonas taiwanensis VLB120 was engineered for increased malonyl-CoA availability to produce bacterial and plant-derived polyketides. A multi-target metabolic engineering strategy focusing on decreasing the malonyl-CoA drain and increasing malonyl-CoA precursor availability, led to an increased production of various malonyl-CoA-derived products, including pinosylvin, resveratrol and flaviolin. The production of flaviolin, a molecule deriving from five malonyl-CoA molecules, was doubled compared to the parental strain by this malonyl-CoA increasing strategy. Additionally, the engineered platform strain enabled production of up to 84 mg L
    Mesh-Begriff(e) Fatty Acids/metabolism ; Malonyl Coenzyme A/metabolism ; Polyketides/metabolism ; Pseudomonas/classification ; Pseudomonas/genetics ; Pseudomonas/metabolism ; Resveratrol/metabolism ; Secondary Metabolism ; Stilbenes/metabolism ; Coumaric Acids/metabolism ; Phenylalanine/metabolism ; Genome, Bacterial/genetics ; Sequence Deletion ; Acetyl Coenzyme A/metabolism ; Citrate (si)-Synthase/metabolism ; Pyruvic Acid/metabolism ; Phytoalexins/metabolism ; Naphthoquinones/metabolism
    Chemische Substanzen Fatty Acids ; flaviolin (479-05-0) ; Malonyl Coenzyme A (524-14-1) ; pinosylvin (881R434AIB) ; Polyketides ; Resveratrol (Q369O8926L) ; Stilbenes ; Coumaric Acids ; Phenylalanine (47E5O17Y3R) ; Acetyl Coenzyme A (72-89-9) ; Citrate (si)-Synthase (EC 2.3.3.1) ; Pyruvic Acid (8558G7RUTR) ; Phytoalexins ; Naphthoquinones
    Sprache Englisch
    Erscheinungsdatum 2023-04-08
    Erscheinungsland Belgium
    Dokumenttyp Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 1470383-x
    ISSN 1096-7184 ; 1096-7176
    ISSN (online) 1096-7184
    ISSN 1096-7176
    DOI 10.1016/j.ymben.2023.04.001
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  8. Buch ; Online ; Dissertation / Habilitation: Development of an itaconic acid production process on alternative feedstocks

    Niehoff, Paul-Joachim [Verfasser] / Büchs, Jochen [Akademischer Betreuer] / Wierckx, Nick [Akademischer Betreuer]

    2024  

    Verfasserangabe Paul-Joachim Niehoff ; Jochen Büchs, Nick Wierckx
    Schlagwörter Technische Chemie ; Technical Chemistry
    Thema/Rubrik (Code) sg660
    Sprache Englisch
    Verlag Universitätsbibliothek der RWTH Aachen
    Erscheinungsort Aachen
    Dokumenttyp Buch ; Online ; Dissertation / Habilitation
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  9. Buch ; Online ; Dissertation / Habilitation: Electrochemical pH-swing extraction for carboxylic acid purification

    Gausmann, Marcel [Verfasser] / Jupke, Andreas [Akademischer Betreuer] / Wierckx, Nick [Akademischer Betreuer]

    2024  

    Verfasserangabe Marcel Gausmann ; Andreas Jupke, Nick Wierckx
    Schlagwörter Technische Chemie ; Technical Chemistry
    Thema/Rubrik (Code) sg660
    Sprache Englisch
    Verlag Universitätsbibliothek der RWTH Aachen
    Erscheinungsort Aachen
    Dokumenttyp Buch ; Online ; Dissertation / Habilitation
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  10. Artikel ; Online: Engineering a Pseudomonas taiwanensis 4-coumarate platform for production of para-hydroxy aromatics with high yield and specificity

    Wynands, Benedikt / Kofler, Franziska / Sieberichs, Anka / da Silva, Nadine / Wierckx, Nick

    Metabolic Engineering. 2023 July, v. 78 p.115-127

    2023  

    Abstract: Aromatics are valuable bulk or fine chemicals with a myriad of important applications. Currently, their vast majority is produced from petroleum associated with many negative aspects. The bio-based synthesis of aromatics contributes to the much-required ... ...

    Abstract Aromatics are valuable bulk or fine chemicals with a myriad of important applications. Currently, their vast majority is produced from petroleum associated with many negative aspects. The bio-based synthesis of aromatics contributes to the much-required shift towards a sustainable economy. To this end, microbial whole-cell catalysis is a promising strategy allowing the valorization of abundant feedstocks derived from biomass to yield de novo-synthesized aromatics. Here, we engineered tyrosine-overproducing derivatives of the streamlined chassis strain Pseudomonas taiwanensis GRC3 for efficient and specific production of 4-coumarate and derived aromatics. This required pathway optimization to avoid the accumulation of tyrosine or trans-cinnamate as byproducts. Although application of tyrosine-specific ammonia-lyases prevented the formation of trans-cinnamate, they did not completely convert tyrosine to 4-coumarate, thereby displaying a significant bottleneck. The use of a fast but unspecific phenylalanine/tyrosine ammonia-lyase from Rhodosporidium toruloides (RtPAL) alleviated this bottleneck, but caused phenylalanine conversion to trans-cinnamate. This byproduct formation was greatly reduced through the reverse engineering of a point mutation in prephenate dehydratase domain-encoding pheA. This upstream pathway engineering enabled efficient 4-coumarate production with a specificity of >95% despite using an unspecific ammonia-lyase, without creating an auxotrophy. In shake flask batch cultivations, 4-coumarate yields of up to 21.5% (Cmol/Cmol) from glucose and 32.4% (Cmol/Cmol) from glycerol were achieved. Additionally, the product spectrum was diversified by extending the 4-coumarate biosynthetic pathway to enable the production of 4-vinylphenol, 4-hydroxyphenylacetate, and 4-hydroxybenzoate with yields of 32.0, 23.0, and 34.8% (Cmol/Cmol) from glycerol, respectively.
    Schlagwörter 4-hydroxybenzoic acid ; Pseudomonas ; Rhodosporidium toruloides ; biochemical pathways ; biomass ; byproducts ; catalytic activity ; culture flasks ; feedstocks ; glucose ; glycerol ; petroleum ; phenylalanine ; point mutation ; tyrosine ; tyrosine ammonia-lyase ; Aromatics ; Biocatalysis ; 4-Coumarate ; Metabolic engineering ; Phenylalanine/tyrosine ammonia-lyase
    Sprache Englisch
    Erscheinungsverlauf 2023-07
    Umfang p. 115-127.
    Erscheinungsort Elsevier Inc.
    Dokumenttyp Artikel ; Online
    ZDB-ID 1470383-x
    ISSN 1096-7184 ; 1096-7176
    ISSN (online) 1096-7184
    ISSN 1096-7176
    DOI 10.1016/j.ymben.2023.05.004
    Datenquelle NAL Katalog (AGRICOLA)

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