Metabolic reprogramming and machine learning-guided cofactor engineering to boost nicotinamide mononucleotide production in Escherichia coli

Bioresource Technology, Journal Year: 2025, Volume and Issue: unknown, P. 132350 - 132350

Published: March 1, 2025

Language: Английский

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Design of NAMPTs with Superior Activity by Dual-Channel Protein Engineering Strategy

Journal of Agricultural and Food Chemistry, Journal Year: 2024, Volume and Issue: 72(24), P. 13834 - 13845

Published: June 6, 2024

The nicotinamide phosphoribosyltransferase (NAMPT)-catalyzed substitution reaction plays a pivotal role in the biosynthesis of nucleotide compounds. However, industrial applications are hindered by low activity NAMPTs. In this study, novel dual-channel protein engineering strategy was developed to increase NAMPT enhancing substrate accessibility. best mutant (CpNAMPTY13G+Y15S+F76P) with remarkable 5-fold enzyme obtained. By utilizing CpNAMPTY13G+Y15S+F76P as biocatalyst, accumulation β-nicotinamide mononucleotide reached high 19.94 g L–1 within 3 h an impressive conversion rate 99.8%. Further analysis revealed that newly generated channel, formed through crack propagation, facilitated binding and enhanced byproduct tolerance. addition, three NAMPTs from different sources were designed based on strategy, corresponding mutants improved obtained, which proved effectiveness practicability approach.

Language: Английский

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Use of Nicotinamide Mononucleotide as Non-Natural Cofactor

Catalysts, Journal Year: 2025, Volume and Issue: 15(1), P. 37 - 37

Published: Jan. 3, 2025

Nicotinamide mononucleotide (NMN) has emerged as a promising non-natural cofactor with significant potential to transform biocatalysis, synthetic biology, and therapeutic applications. By modulating NAD⁺ metabolism, NMN offers unique advantages in enzymatic reactions, metabolic engineering, regenerative medicine. This review provides comprehensive analysis of NMN’s biochemical properties, mechanisms action, diverse Emphasis is placed on its role addressing challenges multi-enzyme cascades, biofuel production, the synthesis high-value chemicals. The paper also highlights critical research gaps, including need for scalable methods, improved integration into systems, toxicity studies use. Emerging technologies such AI-driven enzyme design CRISPR-based genome engineering are discussed transformative tools optimizing NMN-dependent pathways. Furthermore, synergistic biology innovations, cell-free systems dynamic regulatory networks, explored, paving way precise modular biotechnological solutions. Looking forward, versatility positions it pivotal tool advancing sustainable bioprocessing precision Addressing current limitations through interdisciplinary approaches will enable redefine boundaries innovation. serves roadmap leveraging across scientific industrial domains.

Language: Английский

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Improving Biosynthesis Efficiency of Nicotinamide Mononucleotide by ATP Recycling Engineering and Condition Optimization

Journal of Agricultural and Food Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: April 23, 2025

Nicotinamide mononucleotide (NMN) is a very important bioactive nucleotide that of great help to human health. However, its widespread application has been limited by high production costs, especially the cost core substrates, coenzyme, and enzymes. In this study, ADP/GDP-polyphosphate phosphotransferase RhPPK2 originating from Rhodobacter sphaeroides was successfully expressed in Escherichia coli with high-level solubility, enzyme activity lysate supernatant reached 21.9 ± 0.65 U/mL. And then, temperature profiles, pH kinetic parameters purified reRhPPK2 were systematically characterized, which demonstrate potential for enzymatic ATP regeneration systems. Furthermore, introduction significantly enhanced NMN efficiency, achieving 2.3-fold increase compared conventional supplementation method. Finally, efficiency further improved single-factor experiment L9(34) orthogonal design, yield up 14.6 0.51 g/L, about 5.4 times initial yield. This research substantially reduced costs established robust foundation industrial-scale production.

Language: Английский

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Efficient Escherichia coli Platform for Cannabinoid Precursor Olivetolic Acid Biosynthesis from Inexpensive Inputs

Journal of Agricultural and Food Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 2, 2025

Olivetolic acid (OLA), an initial precursor of cannabinoids, is catalyzed by type III polyketide synthase, which has a wide range pharmacological activities, such as antimicrobial and cytotoxic effects. Here, we applied systematic metabolic engineering to develop multienzyme cascade system produce OLA via two low-cost inputs. The synthase (OLS) cyclase enzymes (OAC), along with the best combination hexanoyl-CoA malonyl-CoA synthetases (AEE3 MatB), were first introduced into biocatalytic increase supply starting extender units. To drive catalysis smoothly, ATP regeneration CoA-sufficient incorporated biocatalysts provide enough cofactors. Furthermore, flux was redirected biosynthesis through delicate control fatty (FAB) pathway promoter engineering. Collectively, these strategies have led us at titer 102.1 mg/L productivity 25.5 mg/L/h using malonate hexanoate direct substrates. Our provides effective platform for production cannabinoid in Escherichia coli may be valuable reference development microbial cell factories that use important intermediates.

Language: Английский

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Establishing a Novel Pathway for the Biosynthesis of Nicotinamide Mononucleotide

Enzyme and Microbial Technology, Journal Year: 2025, Volume and Issue: unknown, P. 110633 - 110633

Published: March 1, 2025

Language: Английский

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Systematic Engineering for High-level Production of β-nicotinamide mononucleotide from NAM and Ribose

Food Bioscience, Journal Year: 2024, Volume and Issue: unknown, P. 105725 - 105725

Published: Dec. 1, 2024

Language: Английский

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Β-Nicotinamide Mononucleotide Attenuates Oxidative Stress and Activates Steroidogenesis in Sheep Ovarian Granulosa Cells by Ampk/Ulk1/Mtor Pathway

Published: Jan. 1, 2024

Follicular granulosa cells (GCs) are responsive in the increase of ovulation number, thereby indirectly influencing litter size. Oxidative stress-induced GCs death represents a common reason for follicular atresia, although underlying metabolic molecules remains to be elucidated. Here, we provided first combined metabolomic fluid and cellular transcriptomic analysis sheep associated with high low prolificacy. Then explored effects β-Nicotinamide mononucleotide (NMN) on biological behaviors lipopolysaccharide (LPS)-induced potential mechanisms. Our results showed that NMN increased levels Nicotinamide Adenine Dinucleotide adenosine triphosphate (NAD+ ATP), suppressed LPS-induced cell apoptosis as indicated by enhancing expression BAX PARP1, well alleviated mitochondrial dysfunction through genes oxidative phosphorylation (Naufa1 Ndufs6). Moreover, attenuated stress inhibiting production reactive oxygen species (ROS), lipid peroxidation marker (MDA), promoted activity antioxidative system including CAT, GSH, T-AOC, SOD. Mechanically, found exposure LPS stimulated improved impaired estrogen (E2) progesterone (P4) along abundances steroidogenic enzyme mRNA transcripts, STAR, CYP19A1, 3β-HSD, then altered AMPK, mTOR, ULK1. Further, might play crucial role primary follicle formation ovary cortex cultures at tissue level. Findings indicate beneficial effect is mediated eliminating accumulated ROS suppress restoring function via AMPK/ULK1/mTOR signaling pathway.

Language: Английский

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β-Nicotinamide Mononucleotide Reduces Oxidative Stress and Improves Steroidogenesis in Granulosa Cells Associated with Sheep Prolificacy via Activating AMPK Pathway

Antioxidants, Journal Year: 2024, Volume and Issue: 14(1), P. 34 - 34

Published: Dec. 30, 2024

Oxidative stress is a significant factor in the death of granulosa cells (GCs), leading to follicular atresia and consequently limiting number dominant follicles that can mature ovulate within each wave. Follicular fluid contains diverse array metabolites play crucial roles regulating GCs’ proliferation oocyte maturation, which are essential for follicle development female fertility. However, mechanisms behind metabolite heterogeneity its effects on function remain poorly understood. Here, we identified elevated nicotinamide levels high-prolificacy sheep, correlated with oxidative GCs, by an integrated analysis. In vitro experiments demonstrated supplementation β-nicotinamide mononucleotide (NMN) significantly increased adenine dinucleotide (NAD+) adenosine triphosphate (ATP) GCs. NMN treatment effectively reduced Lipopolysaccharide (LPS)-induced apoptosis mitigated mitochondrial dysfunction, while also decreasing production reactive oxygen species (ROS), thereby enhancing activity antioxidant defense system. Importantly, improved impairments steroid hormone induced LPS. Mechanistically, protective against GCs were mediated via AMPK/mTOR pathway. Collectively, our findings elucidate metabolic characteristics associated sheep prolificacy demonstrate protects from LPS-induced dysfunction enhances ovarian responsiveness These position as potential novel biomarker function.

Language: Английский

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Harnessing lactic acid bacteria for nicotinamide mononucleotide biosynthesis: a review of strategies and future directions

Frontiers in Microbiology, Journal Year: 2024, Volume and Issue: 15

Published: Dec. 13, 2024

Nicotinamide mononucleotide (NMN), one of the crucial precursors nicotinamide adenine dinucleotide, has garnered considerable interest for its pharmacological and anti-aging effects, conferring potential health economic benefits humans. Lactic acid bacteria (LAB) are most important probiotics, which is commonly used in dairy industry. Due to probiotic properties, it presents an attractive platform food-grade NMN production. LAB have also been extensively utilized enhance functional properties pharmaceuticals cosmetics, making them promising candidates large-scale up synthesis NMN. This review provides in-depth analysis various metabolic engineering strategies, including enzyme optimization, pathway rewiring, fermentation process enhancements, increase yields LAB. It explores both CRISPR/Cas9 traditional methods manipulate key biosynthetic pathways. In particular, this study discussed future research directions, emphasizing application synthetic biology, systems AI-driven optimization further invaluable insights into developing scalable industrially relevant processes production meet growing market demand.

Language: Английский

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