Microbial Biotechnology
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2022
·Vol. 15
·期 5
·1610-1621
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IF 6.575
Xing Liu,Xiao Han,Yuan Peng,Chunlin Tan,Jing Wang,Hongsong Xue,Ping Xu,Fei Tao
Summary 3, 4-Dihydroxyphenyl- l -alanine ( l -DOPA) is a compound of high medical value and is considered effective as a treatment for Parkinson’s disease. Currently, bioproduction of l -DOPA is mainly carried out by whole-cell catalysis mediated by recombinant Escherichia coli carrying heterogeneous tyrosine phenol lyase. Vibrio natriegens is increasingly attracting attention owing to its superiority, including extremely rapid growth and high soluble protein expression capacity. In this study, we attempt to develop an efficient whole-cell catalyst for l -DOPA production using V. natriegens as the chassis. The maximum soluble protein expression by V. natriegens was accomplished in 4 h at 37°C, which was equivalent to that achieved by E. coli in 16 h at 16°C. Furthermore, the maximum productivity reached over 10.0 g l −1 h −1 in the early stage of biocatalysis, nearly two-fold higher than previously reported. Approximately 54.0 g l −1 l -DOPA was obtained with a catechol conversion rate greater than 95%. In conclusion, V. natriegens displays advantages, including rapid protein expression and catalytic rate in the catalysis process for l -DOPA production. These findings strongly suggest that V. natriegens has remarkable potential as a whole-cell catalysis chassis for the production of valuable chemicals.
研究领域:
生物工程
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医学
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微生物学
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PubMed
高影响因子
Synthetic and Systems Biotechnology
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2026
·
IF 4.8
Wangshuying Deng,Xinyu Zhang,Jiayin Liu,Wei Ye,Xianfeng Zhu,Weihong Jiang,Yang Gu
Formic acid (FA), a key one-carbon liquid compound derived directly from CO 2 , can serve as a dual-purpose substrate in microbial metabolism, supplying both carbon and energy. Its potential for green biomanufacturing is immense, yet its inherent toxicity and poor metabolizability to most microbes pose a major hurdle in developing efficient microbial cell factories for value-added chemical production. Building on our prior discovery of Vibrio natriegens as a naturally proficient formic acid utilizer, we demonstrate here that formate supplementation as an auxiliary substrate can dramatically boost pyruvate production of V. natriegens from sodium gluconate, achieving a 1.9-fold increase in titer. Transcriptomic analysis revealed that formate presence induces global changes in gene expression. By subsequently downregulating the pyruvate consumption pathway, we engineered a strain that, when co-fed with formate and sodium gluconate, achieved a 49.0% improvement in pyruvate synthesis. Isotopic tracer analysis confirmed substantial formate assimilation, with approximately 9.43% incorporated into biomass. In a fed-batch fermentation, the engineered V. natriegens strain consumed 82.8 g/L sodium gluconate and 37.4 g/L formate (HCOONa·2H 2 O) within 51 h, producing 56.4 g/L pyruvate at a rate of 1.1 g/L/h. This work elucidates the stimulatory role of formate in the pyruvate biosynthesis of V. natriegens and establishes a novel strategy for leveraging this feedstock in microbial production.
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BIORESOURCE TECHNOLOGY
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2023
·
IF 11.4
Yu Zhou,Bowen Shen,Shengping You,Qingdian Yin,Mengfan Wang,Nan Jiang,Rongxin Su,Wei Qi
Recently, clean PET biodegradation has gained widespread attention in tackling white pollution. Nonetheless, the development of industrial biotechnology is still impeded by its contamination susceptibility, high energy input, and consumption of substantial freshwater resources. Thus, a novel PET biodegradation process was developed based on host screening and by-product circulation to address the aforementioned issues. The fast-growth host halophilic Vibrio natriegens ( V. natriegens ) was used and exhibited an increased protein expression level of 87.3% compared to E. coli . Meanwhile, the new process utilized a seawater-based medium for fermentation under non-sterile conditions, leading to energy-saving (energy reduced by 2.48-fold) and cost-reduction (cost reduced by 47.9%). Moreover, the large amount of saline wastewater from terephthalic acid purification was ingeniously reused for the cultivation of V. natriegens , thereby avoiding resource wastage and secondary pollution. Therefore, an efficient, economical, energy-saving, and eco-friendly process was designed, potentially addressing the industrial bottleneck in PET bio-recycling.
研究领域:
环境科学
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生物技术
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PROCESS BIOCHEMISTRY
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2026
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IF 4.4
Lin Zhang,Yiwei Guo,Hailiang Liu,Aoqian Qiu,Jiaxing Zhang,Shengping You,Rongxin Su,Wei Qi
The increasing accumulation of waste poly(ethylene terephthalate) (PET) poses a significant threat to the ecological environment. Although current recycling technologies offer promising solutions, their widespread implementation is hindered by high costs, intensive energy consumption, and low efficiency. To address these challenges, we developed a catalytic system that integrated glycolysis electroconversion, and enzymatic hydrolysis, achieving a synergistic treatment of efficient PET conversion and the valorization of formate-containing wastewater. In this system, PET was efficiently depolymerized via KHCO 3 -catalyzed glycolysis, resulting in a total yield of 96.5% of bis(2-hydroxyethyl) terephthalate (BHET) and mono(2-hydroxyethyl) terephthalate (MHET). The ethylene glycol (EG) was further electrooxidized to formate using a Co, Cl-NiS catalyst, achieving high faradic efficiency (>90%) and formate yield (>80%). Then, this resulting formate served as an effective carbon source for cultivating Vibrio natriegens under non-sterile conditions to produce FAST-PETase and KL-MHETase. The crude enzyme extracts could efficiently catalyze the hydrolysis of 150 g/L crude BHET mixture. In summary, this study establishes an integrated catalytic platform that synergizes chemical, electrochemical, and biological processes. This presents a novel and sustainable pathway for closed-loop recycling of waste PET.
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Frontiers in Microbiology
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2021
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IF 5.64
Xu Jiaqi,Dong Feng,Wu Meixian,Tao Rongsheng,Yang Junjie,Wu Mianbin,Jiang Yu,Yang Sheng,Yang Lirong
Efficient and novel recombinant protein expression systems can further reduce the production cost of enzymes. Vibrio natriegens is the fastest growing free-living bacterium with a doubling time of less than 10 min, which makes it highly attractive as a protein expression host. Here, 196 pET plasmids with different genes of interest (GOIs) were electroporated into the V. natriegens strain VnDX, which carries an integrated T7 RNA polymerase expression cassette. As a result, 65 and 75% of the tested GOIs obtained soluble expression in V. natriegens and Escherichia coli, respectively, 20 GOIs of which showed better expression in the former. Furthermore, we have adapted a consensus “what to try first” protocol for V. natriegens based on Terrific Broth medium. Six sampled GOIs encoding biocatalysts enzymes thus achieved 50–128% higher catalytic efficiency under the optimized expression conditions. Our study demonstrated V. natriegens as a pET-compatible expression host with a spectrum of highly expressed GOIs distinct from E. coli and an easy-to-use consensus protocol, solving the problem that some GOIs cannot be expressed well in E. coli.
研究领域:
生物技术
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分子生物学
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蛋白质工程
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METABOLIC ENGINEERING
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2021
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IF 9.783
Ye Zhang,Zihua Li,Yu Liu,Xuecong Cen,Dehua Liu,Zhen Chen
The economic viability of current bio-production systems is often limited by its low productivity due to slow cell growth and low substrate uptake rate. The fastest-growing bacterium Vibrio natriegens is a highly promising next-generation workhorse of the biotechnology industry which can utilize various industrially relevant carbon sources with high substrate uptake rates. Here, we demonstrate the first systematic engineering example of V. natriegens for the heterologous production of 1,3-propanediol (1,3-PDO) from glycerol. Systems metabolic engineering strategies have been applied in this study to develop a superior 1,3-PDO producer, including: (1) heterologous pathway construction and optimization; (2) engineering cellular transcriptional regulators and global transcriptomic analysis; (3) enhancing intracellular reducing power by cofactor engineering; (4) reducing the accumulation of toxic intermediate by pathway engineering; (5) systematic engineering of glycerol oxidation pathway to eliminate byproduct formation. A final engineered strain can efficiently produce 1,3-PDO with a titer of 56.2 g/L, a yield of 0.61 mol/mol, and an average productivity of 2.36 g/L/h. The strategies described in this study would be useful for engineering V. natriegens as a potential chassis for the production of other useful chemicals and biofuels.
研究领域:
生物技术
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代谢工程
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微生物生物技术
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PubMed
高影响因子
ACS Synthetic Biology
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2023
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IF 4.7
Hui-Hui Li,Jie Wu,Jia-Qi Liu,Qi-Zhong Wu,Ru−Li He,Zhou-Hua Cheng,Jun-Lu Lv,Wei-Qiang Lin,Jing Wu,Dong-Feng Liu,Wen-Wei Li
Polyhydroxybutyrate (PHB) is an attractive biodegradable polymer that can be produced through the microbial fermentation of organic wastes or wastewater. However, its mass production has been restricted by the poor utilization of organic wastes due to the presence of inhibitory substances, slow microbial growth, and high energy input required for feedstock sterilization. Here, Vibrio natriegens, a fast-growing bacterium with a broad substrate spectrum and high tolerance to salt and toxic substances, was genetically engineered to enable efficient PHB production from nonsterilized fermentation of organic wastes. The key genes encoding the PHB biosynthesis pathway of V. natriegens were identified through base editing and overexpressed. The metabolically engineered strain showed 166-fold higher PHB content (34.95 wt %) than the wide type when using glycerol as a substrate. Enhanced PHB production was also achieved when other sugars were used as feedstock. Importantly, it outperformed the engineered Escherichia coli MG1655 in PHB productivity (0.053 g/L/h) and tolerance to toxic substances in crude glycerol, without obvious activity decline under nonsterilized fermentation conditions. Our work demonstrates the great potential of engineered V. natriegens for low-cost PHB bioproduction and lays a foundation for exploiting this strain as a next-generation model chassis microorganism in synthetic biology.
研究领域:
生物技术
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微生物学
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合成生物学
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PubMed
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ACS Synthetic Biology
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2026
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IF 4.5
Ruiying Zhu,Zehua Bao
Genetic diversification serves as the foundation for in vivo molecular evolution. The fusion of a cytidine deaminase with T7 RNA polymerase (MutaT7) enables efficient diversification of DNA sequences downstream of the T7 promoter in model microbial hosts E. coli and S. cerevisiae. Vibrio natriegens, currently the fastest-growing nonpathogenic bacterium with a doubling time half that of E. coli, emerged as a promising alternative chassis for synthetic biology. Here, we report the establishment of MutaT7 in V. natriegens (dubbed VnMutaT7), achieving rapid evolution of the model protein TEM-1 to confer high-level ceftazidime resistance in just 36 h. This work establishes the potential of V. natriegens as an accelerated platform for protein engineering.
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高影响因子
Nature Communications
·
2023
·Vol. 14
·期 1
·1-12
·
IF 16.6
Tian Jinzhong,Deng Wangshuying,Zhang Ziwen,Xu Jiaqi,Yang Guiling,Zhao Guoping,Yang Sheng,Jiang Weihong,Gu Yang
Formic acid (FA) has emerged as a promising one-carbon feedstock for biorefinery. However, developing efficient microbial hosts for economically competitive FA utilization remains a grand challenge. Here, we discover that the bacterium Vibrio natriegens has exceptional FA tolerance and metabolic capacity natively. This bacterium is remodeled by rewiring the serine cycle and the TCA cycle, resulting in a non-native closed loop (S-TCA) which as a powerful metabolic sink, in combination with laboratory evolution, enables rapid emergence of synthetic strains with significantly improved FA-utilizing ability. Further introduction of a foreign indigoidine-forming pathway into the synthetic V. natriegens strain leads to the production of 29.0 g · L −1 indigoidine and consumption of 165.3 g · L −1 formate within 72 h, achieving a formate consumption rate of 2.3 g · L −1 · h −1 . This work provides an important microbial chassis as well as design rules to develop industrially viable microorganisms for FA biorefinery.
研究领域:
生物技术
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代谢工程
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微生物学
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PubMed
高影响因子
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