Engineered Vibrio natriegens with a Toxin–Antitoxin System for High-Productivity Biotransformation of l-Lysine to Cadaverine

Journal of Agricultural and Food Chemistry, Год журнала: 2025, Номер unknown

Опубликована: Фев. 22, 2025

Vibrio natriegens, a fast-growing bacterium, is an emerging chassis of next-generation industrial biotechnology capable thriving under open and continuous culture conditions. Cadaverine, valuable C5 platform chemical, has various chemical biological activities. This study found that V. natriegens exhibited superior tolerance to lysine, the substrate cadaverine production. For first time, synthesis pathway was introduced into for whole-cell catalysis from lysine. A high-efficiency cadaverine-producing strain harboring toxin–antitoxin system, (pSEVA341-pTac-ldcC-pHbpBC-hbpBC) with lysE (PN96_RS17440) inactivation, constructed. In 7 L bioreactors, titer increased 115 g/L in original 158 within 11 h biotransformation, exhibiting 37% increase Its productivity reached 14.4 g/L/h conversion rate as high 90%. These results confirm exceptional effective bioproduction.

Язык: Английский

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Low-biomass pyruvate production with engineered Vibrio natriegens is accompanied by parapyruvate formation

Microbial Cell Factories, Год журнала: 2025, Номер 24(1)

Опубликована: Март 28, 2025

Abstract Background Pyruvate is a precursor for various compounds in the chemical, drug, and food industries therefore an attractive target molecule microbial production processes. The fast-growing bacterium Vibrio natriegens excels with its specific substrate uptake rate as unconventional chassis industrial biotechnology. Here, we aim to exploit traits of V. pyruvate fermentations low biomass concentrations. Results We inactivated dehydrogenase complex Δ vnp12 , which harbors deletions prophage regions . resulting strain aceE was unable grow minimal medium glucose unless supplemented acetate. In shaking flasks, showed growth 1.16 ± 0.03 h − 1 produced 4.0 0.3 g Pyr L within 5 h. optimized parameters aerobic fermentation process applied constant maintenance feed 0.24 Ac resulted maximal concentration only 6.6 0.4 CDW yielded highly active resting cells (q S ) 3.5 0.2 Glc −1 41.0 1.8 volumetric productivity 4.1 Carbon balancing disclosed gap 30%, identified partly parapyruvate. Deletion ligK encoding HMG/CHA aldolase did not impact formation but plasmid-based overexpression negatively affected led 3-fold higher parapyruvate culture broth. Notably, also supernatants pyruvate-producing Corynebacterium glutamicum strain. Cell-free bioreactor experiments mimicking biological formation, pointing chemical reaction contributing synthesis. Conclusions engineered metabolically producing high at concentration. However, found that accompanied by well C. Parapyruvate seems be result conversion might supported biochemically reaction.

Язык: Английский

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β-nicotinamide mononucleotide production in Vibrio natriegens: a preliminary study

F1000Research, Год журнала: 2025, Номер 14, С. 408 - 408

Опубликована: Апрель 7, 2025

Background Nicotinamide mononucleotide (NMN), is a promising nutraceutical attracting much attention for its pharmacological and anti-aging efficacies. However, NMN-containing commercial products are very high-priced due to the lack of efficient facile methods industrial-scale production. To date, various metabolic engineering strategies have been successfully applied produce NMN in Escherichia coli. Recently, Vibrio natriegens has become host bioindustry thanks rapid growth capabilities broad substrate utilization. This study aims evaluate biosynthesis capability V. natriegens. Methods Firstly, mutant strain (Δdns::araC-T7RNAP-KanR ΔpncC::FtnadE-SmR ΔnadR) was generated via multiplex genome editing by natural transformation (MuGENT). Nampt genes encoding nicotinamide phosphoribosyltransferase from Chitinophaga pinensis, Sphingopyxis sp. C-1, Haemophilus ducreyi, Vibrio phage KVP40 were codon-optimized cloned into pACYCDuet™-1 under control T7 promoter. The recombinant plasmids electroporated strain. expression NAMPTs evaluated SDS-PAGE analysis intracellular concentrations quantified HPLC. Results After two rounds MuGENT, V54-33 generated. demonstrated that all strongly expressed HPLC revealed highest concentration obtained with NAMPT pinensis (44.5 μM), followed Vibrio (23.3 μM). Conclusion feasibility natriegens.

Язык: Английский

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Optimizing Monascus pigment production: Insights from mycelial morphology, gene expression, and transcriptomic analysis in simulated seawater fermentation

Food Microbiology, Год журнала: 2025, Номер unknown, С. 104797 - 104797

Опубликована: Апрель 1, 2025

Язык: Английский

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Small-scale fed-batch cultivations of Vibrio natriegens: overcoming challenges for early process development

Bioprocess and Biosystems Engineering, Год журнала: 2025, Номер unknown

Опубликована: Апрель 18, 2025

Abstract Vibrio natriegens is a fast-growing microbial workhorse with high potential for biotechnological applications. However, handling the bacterium in batch processes challenging due to its overflow metabolism and mixed acid formation under microaerobic conditions. For early process development, technologies enabling small-scale fed-batch cultivation of V. Vmax are needed. In this study, cultivations 96-well microtiter plates were successfully online-monitored first time µTOM device. Using oxygen transfer rate, scale up membrane-based shake flasks was performed. The efficiently minimized by choosing suitable feed rates, prevented. A glucose soft sensor using rate provided accurate estimates consumption throughout fermentation, eliminating need offline sampling. Analyzing impact inducer IPTG on recombinant production enzyme inulosucrase revealed concentration-dependent effects processes. contrast, operating mode resulted activity even without induction. Overall, an titer 80 U/mL achieved. conclusion, advantages supported have been demonstrated development Vmax.

Язык: Английский

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Rapid, high-titer biosynthesis of melanin using the marine bacterium Vibrio natriegens

Frontiers in Bioengineering and Biotechnology, Год журнала: 2023, Номер 11

Опубликована: Сен. 13, 2023

Melanin is one of the most abundant natural biomolecules on Earth. These macromolecular biopolymers display several unique physical and chemical properties have garnered interest as biomaterials for various commercial industrial applications. To this end, extensive research has gone into refining methods synthesis extraction melanin from recombinant sources. In study, we developed refined a procedure using microbial system biosynthesis tyrosinase enzyme Tyr1 tyrosine substrate. Using emergent chassis organisms Vibrio natriegens , achieved maximal yields 7.57 g/L, highest reported volumetric productivities 473 mg L −1 h with 100% conversion rates in an optimized, minimally defined medium. Additionally, identified investigated use native copper responsive promoter V. stringent regulation heterologous protein expression cost effective alternative to traditional IPTG-based induction. This represents promising advancement towards green, rapid, economical biomanufacture melanin.

Язык: Английский

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Investigation of exopolysaccharide formation and its impact on anaerobic succinate production with Vibrio natriegens

Microbial Biotechnology, Год журнала: 2023, Номер 17(1)

Опубликована: Май 31, 2023

Abstract Vibrio natriegens is an emerging host for biotechnology due to its high growth and substrate consumption rates. In industrial processes typically fed‐batch are applied obtain space‐time yields. this study, we established aerobic glucose‐limited fermentation with the wild type (wt) of V. which yielded biomass concentrations up 28.4 g X L −1 . However, observed that viscosity culture broth increased by a factor 800 at end cultivation formation 157 ± 20 mg exopolysaccharides (EPS) Analysis genomic repertoire revealed several genes gene clusters associated EPS formation. Deletion transcriptional regulator cpsR in wt did not reduce formation, however, it resulted constantly low altered carbohydrate content EPS. A mutant lacking cps cluster secreted two‐fold less compared accompanied overall changed composition. When cultivated succinate producer Δ lldh dldh pfl ald dns::pyc Cg (Succ1) under anaerobic conditions on glucose, also cultivation. Succ1 reduced five‐ six‐fold remained same level start achieved final 51 46 volumetric productivity 8.5 7.7 Suc h , respectively. Both strains showed product yield about 1.4 mol Glc 27% higher corresponds 81% theoretical maximum.

Язык: Английский

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Strain and process engineering toward continuous industrial fermentation

Frontiers of Chemical Science and Engineering, Год журнала: 2023, Номер 17(10), С. 1336 - 1353

Опубликована: Апрель 20, 2023

Abstract Most current biotechnology industries are based on batch or fed-batch fermentation processes, which often show low productivity and high production costs compared to chemical processes. To increase the economic competitiveness of biological continuous technologies being developed that offer significant advantages in comparison with batch/fed-batch including: (1) removal potential substrates product inhibition, (2) prolonging microbial exponential growth phase enhancing productivity, (3) avoiding repeated preparation lowering operation installation costs. However, several key challenges should be addressed for industrial application including contamination system, degeneration strains, relatively titer. In this study, we reviewed discussed metabolic engineering synthetic biology strategies address these issues.

Язык: Английский

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Carotenoids synthesis affects the salt tolerance mechanism of Rhodopseudomonas palustris

Frontiers in Microbiology, Год журнала: 2023, Номер 14

Опубликована: Ноя. 22, 2023

Rhodopseudomonas palustris CGA009 is a Gram-negative, purple non-sulfur, metabolically diverse bacterium with wide-ranging habitats. The extraordinary ability of R. to decompose variety raw materials and convert them into high-value products makes it an attractive host for biotechnology industrial applications. However, being freshwater has limited application in highly-saline environments. Therefore, great significance obtain the salt-tolerant strain understand its tolerance mechanism. In this study, was successfully evolved eight strains using adaptive laboratory evolution technique. RPAS-11 (R. anti-salt 11) selected as best used further studies explore salt-tolerance expression most genes associated carotenoid synthesis increased significantly under high concentration salt stress, suggesting that one reasons RPAS-11. Gene overexpression knockout experiments were performed get clear about role carotenoids stress tolerance. RPAS-11-IDI, mutant IDI (Isopentenyl diphosphate isomerase) exhibited enhanced tolerance, whereas CGA009-∆crtI showed decline addition, results indicated rhodopin, compound, key pigment responsible palustris. Furthermore, production lycopene, widely-used carotenoid, also increased. Taken together, our research helps deepen understanding mechanism widens

Язык: Английский

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High-Performance Production of N-Acetyl-d-Neuraminic Acid with Whole Cells of Fast-Growing Vibrio natriegens via a Thermal Strategy

Journal of Agricultural and Food Chemistry, Год журнала: 2023, Номер 71(50), С. 20198 - 20209

Опубликована: Дек. 5, 2023

High performance is the core objective that biotechnologists pursue, of which low efficiency, titer, and side products are chief obstacles. Here, a thermal strategy proposed for simultaneously addressing obstacles whole-cell catalysis widely applied in food industry. The strategy, by combining fast-growing Vibrio natriegens, thermophilic enzymes, high-temperature catalysis, was successfully high-performance production N-acetyl-d-neuraminic acid (Neu5Ac) plays essential roles fields (infant formulas), healthcare, medicine. By using this we realized highest Neu5Ac titer productivity 126.1 g/L up to 71.6 g/(L h), respectively, 7.2-fold higher than Escherichia coli. major byproduct acetic also eliminated via quenching complex metabolic reactions enabled temperature elevation. This study offers broadly applicable producing chemicals relevant industry, providing insights its future development.

Язык: Английский

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