Efficient Natural Plasmid Transformation ofVibrio natriegensEnables Zero-capital Molecular Biology DOI Open Access
David A. Specht, Timothy J. Sheppard, Finn Kennedy

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: Aug. 13, 2023

Abstract The fast-growing microbe Vibrio natriegens is capable of natural transformation where it draws DNA in from media via an active process under physiological conditions. Using engineered strain with a genomic copy the master competence regulator tfoX cholera combination new minimal (MCM) that uses acetate as energy source, we demonstrate naturally competent cells which are created, transformed, and recovered entirely same media, without exchange or addition media. Cells to plasmids, recombination linear DNA, co-transformation both select for scarless markerless edits. entire simple inexpensive, requiring no capital equipment room temperature (Zero Capital protocol, 10 4 cfu/ µ g), just incubator (High Efficiency 5–6 g). These retain their state when frozen transformable immediately upon thawing like typical chemical electrochemical cell. Since optimized protocol requires only 50 minutes hands-on time, V. grows quickly even on plates, started at 9 AM yields abundant culturable single colonies by 5 PM. Further, because all stages occur can be arbitrarily scaled volume, this could ideal automated directed evolution applications. As result, compete E. coli excellent chassis low-cost highly scalable synthetic biology.

Language: Английский

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

Wenqian Zhao,

Tao Li,

Pang Mei

et al.

Food Microbiology, Journal Year: 2025, Volume and Issue: unknown, P. 104797 - 104797

Published: April 1, 2025

Language: Английский

Citations

0
Small-scale fed-batch cultivations of Vibrio natriegens: overcoming challenges for early process development DOI Creative Commons
Clara Lüchtrath, Eva Forsten,

Romeos Polis

et al.

Bioprocess and Biosystems Engineering, Journal Year: 2025, Volume and Issue: unknown

Published: April 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.

Language: Английский

Citations

0
Development of a novel “4E” polyethylene terephthalate bio-recycling process with the potential for industrial application: Efficient, economical, energy-saving, and eco-friendly DOI
Yu Zhou, Bowen Shen, Shengping You

et al.

Bioresource Technology, Journal Year: 2023, Volume and Issue: 391, P. 129913 - 129913

Published: Oct. 30, 2023

Language: Английский

Citations

9
Unraveling the impact of pH, sodium concentration, and medium osmolality on Vibrio natriegens in batch processes DOI Creative Commons
Eva Forsten, Steffen Gerdes,

René Petri

et al.

BMC Biotechnology, Journal Year: 2024, Volume and Issue: 24(1)

Published: Sept. 23, 2024

Vibrio natriegens, a halophilic marine γ-proteobacterium, holds immense biotechnological potential due to its remarkably short generation time of under ten minutes. However, the highest growth rates have been primarily observed on complex media, which often suffer from batch-to-batch variability affecting process stability and performance. Consistent bioprocesses necessitate use chemically defined are usually optimized for fermenters with pH dissolved oxygen tension (DOT) regulation, both not applied during early-stage cultivations in shake flasks or microtiter plates. Existing studies V. natriegens' mineral media report partially conflicting results, comprehensive study examining combined effects buffering, sodium concentration, medium osmolality is lacking.

Language: Английский

Citations

3
The new chassis in the flask: Advances in Vibrio natriegens biotechnology research DOI Creative Commons

Matthew Lima,

Charandatta Muddana,

Zhengyang Xiao

et al.

Biotechnology Advances, Journal Year: 2024, Volume and Issue: 77, P. 108464 - 108464

Published: Oct. 9, 2024

Biotechnology has been built on the foundation of a small handful well characterized and well-engineered organisms. Recent years have seen breakout performer gain attention as new entrant into bioengineering toolbox: Vibrio natriegens. This review covers recent research efforts making V. natriegens biotechnology platform, using large language model (LLM) knowledge graph to expedite literature survey process. Scientists made advancements in pertaining fundamental metabolic characteristics natriegens, development characterization synthetic biology tools, systems analysis modeling, bioproduction engineering, microbial ecology. Each these subcategories relevance future for applications. In this review, we cover offer context impact they may field, highlighting benefits drawbacks organism. From examining research, it appears that is precipice becoming platform bacterium biotechnology.

Language: Английский

Citations

3
Rapid, high-titer biosynthesis of melanin using the marine bacterium Vibrio natriegens DOI Creative Commons
Aaron D. Smith, Tanya Tschirhart, Jaimee R. Compton

et al.

Frontiers in Bioengineering and Biotechnology, Journal Year: 2023, Volume and Issue: 11

Published: Sept. 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.

Language: Английский

Citations

8
Investigation of exopolysaccharide formation and its impact on anaerobic succinate production with Vibrio natriegens DOI Creative Commons
Clarissa Schulze, Maurice Hädrich,

Jennifer Borger

et al.

Microbial Biotechnology, Journal Year: 2023, Volume and Issue: 17(1)

Published: May 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.

Language: Английский

Citations

7
Strain and process engineering toward continuous industrial fermentation DOI Creative Commons
Yufei Dong, Ye Zhang, Dehua Liu

et al.

Frontiers of Chemical Science and Engineering, Journal Year: 2023, Volume and Issue: 17(10), P. 1336 - 1353

Published: April 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.

Language: Английский

Citations

6
Carotenoids synthesis affects the salt tolerance mechanism of Rhodopseudomonas palustris DOI Creative Commons
Meijie Li, Tongtong Zhu,

Rumeng Yang

et al.

Frontiers in Microbiology, Journal Year: 2023, Volume and Issue: 14

Published: Nov. 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

Language: Английский

Citations

6
High-Performance Production of N-Acetyl-d-Neuraminic Acid with Whole Cells of Fast-Growing Vibrio natriegens via a Thermal Strategy DOI
Peng Yuan, Lina Ma, Ping Xu

et al.

Journal of Agricultural and Food Chemistry, Journal Year: 2023, Volume and Issue: 71(50), P. 20198 - 20209

Published: Dec. 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.

Language: Английский

Citations

6