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Titel: Cascaded valorization of brown seaweed to produce l-lysine and value-added products using Corynebacterium glutamicum streamlined by systems metabolic engineering
VerfasserIn: Hoffmann, Sarah Lisa
Kohlstedt, Michael
Jungmann, Lukas
Hutter, Michael
Poblete-Castro, Ignacio
Becker, Judith
Wittmann, Christoph
Sprache: Englisch
Titel: Metabolic Engineering
Bandnummer: 67
Seiten: 293-307
Verlag/Plattform: Elsevier
Erscheinungsjahr: 2021
Freie Schlagwörter: Transhydrogenase
Fructokinase
Glyceraldehyde 3-phosphate dehydrogenase
Mannitol 2-dehydrogenase
Protein engineering
NADH
NADPH
Redox balancing
L-lysine
Oxidative pentose phosphate pathway
Fructose
Seaweed
Macro algae
DDC-Sachgruppe: 500 Naturwissenschaften
Dokumenttyp: Journalartikel / Zeitschriftenartikel
Abstract: Seaweeds emerge as promising third-generation renewable for sustainable bioproduction. In the present work, we valorized brown seaweed to produce l-lysine, the world's leading feed amino acid, using Corynebacterium glutamicum, which was streamlined by systems metabolic engineering. The mutant C. glutamicum SEA-1 served as a starting point for development because it produced small amounts of l-lysine from mannitol, a major seaweed sugar, because of the deletion of its arabitol repressor AtlR and its engineered l-lysine pathway. Starting from SEA-1, we systematically optimized the microbe to redirect excess NADH, formed on the sugar alcohol, towards NADPH, required for l-lysine synthesis. The mannitol dehydrogenase variant MtlD D75A, inspired by 3D protein homology modelling, partly generated NADPH during the oxidation of mannitol to fructose, leading to a 70% increased l-lysine yield in strain SEA-2C. Several rounds of strain engineering further increased NADPH supply and l-lysine production. The best strain, SEA-7, overexpressed the membrane-bound transhydrogenase pntAB together with codon-optimized gapN, encoding NADPH-dependent glyceraldehyde 3-phosphate dehydrogenase, and mak, encoding fructokinase. In a fed-batch process, SEA-7 produced 76 g L-1l-lysine from mannitol at a yield of 0.26 mol mol-1 and a maximum productivity of 2.1 g L-1 h-1. Finally, SEA-7 was integrated into seaweed valorization cascades. Aqua-cultured Laminaria digitata, a major seaweed for commercial alginate, was extracted and hydrolyzed enzymatically, followed by recovery and clean-up of pure alginate gum. The residual sugar-based mixture was converted to l-lysine at a yield of 0.27 C-mol C-mol-1 using SEA-7. Second, stems of the wild-harvested seaweed Durvillaea antarctica, obtained as waste during commercial processing of the blades for human consumption, were extracted using acid treatment. Fermentation of the hydrolysate using SEA-7 provided l-lysine at a yield of 0.40 C-mol C-mol-1. Our findings enable improvement of the efficiency of seaweed biorefineries using tailor-made C. glutamicum strains.
DOI der Erstveröffentlichung: 10.1016/j.ymben.2021.07.010
URL der Erstveröffentlichung: https://www.sciencedirect.com/science/article/pii/S1096717621001208
Link zu diesem Datensatz: urn:nbn:de:bsz:291--ds-380120
hdl:20.500.11880/34357
http://dx.doi.org/10.22028/D291-38012
ISSN: 1096-7176
Datum des Eintrags: 15-Nov-2022
Bezeichnung des in Beziehung stehenden Objekts: Appendix A. Supplementary data
In Beziehung stehendes Objekt: https://ars.els-cdn.com/content/image/1-s2.0-S1096717621001208-mmc1.pdf
https://ars.els-cdn.com/content/image/1-s2.0-S1096717621001208-mmc2.zip
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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