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Titel: High-efficiency production of 5-hydroxyectoine using metabolically engineered Corynebacterium glutamicum
VerfasserIn: Jungmann, Lukas
Hoffmann, Sarah Lisa
Lang, Caroline
De Agazio, Raphaela
Becker, Judith
Kohlstedt, Michael
Wittmann, Christoph
Sprache: Englisch
Titel: Microbial Cell Factories
Bandnummer: 21
Heft: 1
Verlag/Plattform: BMC
Erscheinungsjahr: 2022
Freie Schlagwörter: Corynebacterium glutamicum
Extremolyte
Ectoine
5-Hydroxyectoine
Ectoine hydroxylase
Biotransformation
Intracellular metabolite
Proline
Trehalose
High-value product
DDC-Sachgruppe: 500 Naturwissenschaften
Dokumenttyp: Journalartikel / Zeitschriftenartikel
Abstract: Background: Extremolytes enable microbes to withstand even the most extreme conditions in nature. Due to their unique protective properties, the small organic molecules, more and more, become high-value active ingredients for the cosmetics and the pharmaceutical industries. While ectoine, the industrial extremolyte fagship, has been successfully commercialized before, an economically viable route to its highly interesting derivative 5-hydroxyectoine (hydroxyectoine) is not existing. Results: Here, we demonstrate high-level hydroxyectoine production, using metabolically engineered strains of C. glutamicum that express a codon-optimized, heterologous ectD gene, encoding for ectoine hydroxylase, to convert supplemented ectoine in the presence of sucrose as growth substrate into the desired derivative. Fourteen out of sixteen codon-optimized ectD variants from phylogenetically diverse bacterial and archaeal donors enabled hydroxyectoine production, showing the strategy to work almost regardless of the origin of the gene. The genes from Pseudomonas stutzeri (PST) and Mycobacterium smegmatis (MSM) worked best and enabled hydroxyectoine production up to 97% yield. Metabolic analyses revealed high enrichment of the ectoines inside the cells, which, inter alia, reduced the synthesis of other compatible solutes, including proline and trehalose. After further optimization, C. glutamicum Ptuf ectDPST achieved a titre of 74 g L−1 hydroxyectoine at 70% selectivity within 12 h, using a simple batch process. In a two-step procedure, hydroxyectoine production from ectoine, previously synthesized fermentatively with C. glutamicum ectABCopt, was successfully achieved without intermediate purifcation. Conclusions: C. glutamicum is a well-known and industrially proven host, allowing the synthesis of commercial products with granted GRAS status, a great beneft for a safe production of hydroxyectoine as active ingredient for cosmetic and pharmaceutical applications. Because ectoine is already available at commercial scale, its use as precursor appears straightforward. In the future, two-step processes might provide hydroxyectoine de novo from sugar.
DOI der Erstveröffentlichung: 10.1186/s12934-022-02003-z
URL der Erstveröffentlichung: https://doi.org/10.1186/s12934-022-02003-z
Link zu diesem Datensatz: urn:nbn:de:bsz:291--ds-388181
hdl:20.500.11880/35002
http://dx.doi.org/10.22028/D291-38818
ISSN: 1475-2859
Datum des Eintrags: 24-Jan-2023
Bezeichnung des in Beziehung stehenden Objekts: Supplementary Information
In Beziehung stehendes Objekt: https://static-content.springer.com/esm/art%3A10.1186%2Fs12934-022-02003-z/MediaObjects/12934_2022_2003_MOESM1_ESM.docx
Fakultät: NT - Naturwissenschaftlich- Technische Fakultät
Fachrichtung: NT - Biowissenschaften
Professur: NT - Prof. Dr. Christoph Wittmann
Sammlung:SciDok - Der Wissenschaftsserver der Universität des Saarlandes

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