Cascaded valorization of brown seaweed to produce L-lysine and value-added products using Corynebacterium glutamicum streamlined by systems metabolic engineering

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dc.contributor.authorHoffmann, Sarah Lisa
dc.contributor.authorKohlstedt, Michael
dc.contributor.authorJungmann, Lukas
dc.contributor.authorHutter, Michael
dc.contributor.authorPoblete-Castro, Ignacio
dc.contributor.authorBecker, Judith
dc.contributor.authorWittmann, Christoph
dc.date.accessioned2024-07-03T16:44:26Z
dc.date.available2024-07-03T16:44:26Z
dc.date.issued2021-09
dc.descriptionIndexación: Scopus.
dc.description.abstractSeaweeds 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−1 L-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.
dc.description.urihttps://www.sciencedirect.com/science/article/pii/S1096717621001208?via%3Dihub
dc.identifier.citationMetabolic Engineering Volume 67, Pages 293 - 307 September 2021
dc.identifier.doi10.1016/j.ymben.2021.07.010
dc.identifier.issn1096-7176
dc.identifier.urihttps://repositorio.unab.cl/handle/ria/58178
dc.language.isoen_US
dc.publisherAcademic Press Inc.
dc.rights.licenseATRIBUCIÓN-NOCOMERCIAL 4.0 INTERNACIONAL
dc.rights.urihttps://creativecommons.org/licenses/by-nc/4.0/deed.es
dc.subjectFructokinase
dc.subjectFructose
dc.subjectGlyceraldehyde 3-phosphate dehydrogenase
dc.subjectL-lysine
dc.subjectMacro algae
dc.subjectMannitol 2-dehydrogenase
dc.subjectNADH
dc.subjectNADPH
dc.subjectOxidative pentose phosphate pathway
dc.subjectProtein engineering
dc.subjectRedox balancing
dc.subjectSeaweed
dc.subjectTranshydrogenase
dc.titleCascaded valorization of brown seaweed to produce L-lysine and value-added products using Corynebacterium glutamicum streamlined by systems metabolic engineering
dc.typeArtículo
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