JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2020
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IF 4.192
Jieyuan Wu,Wei Chen,Yutong Zhang,Xuanxuan Zhang,Jian-Ming Jin,Shuang-Yan Tang
The biosynthetic efficiency of curcumin, a highly bioactive compound from the plant Curcuma longa, needs to be improved. In this study, we performed host cell and biosynthetic pathway engineering to improve curcumin biosynthesis. Using in vivo-directed evolution, the expression level of curcuminoid synthase (CUS), the rate-limiting enzyme in the curcumin biosynthetic pathway, was significantly improved. Furthermore, as curcumin is a highly hydrophobic compound, two cell membrane engineering strategies were applied to optimize the biosynthetic efficiency. Curcumin storage was increased by overexpression of monoglucosyldiacylglycerol synthase from Acholeplasma laidlawii, which optimized the cell membrane morphology. Furthermore, unsaturated fatty acid supplementation was used to enhance membrane fluidity, which greatly ameliorated the damaging effect of curcumin on the cell membrane. These two strategies enhanced curcumin biosynthesis and demonstrated an additive effect.
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ACS Synthetic Biology
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2025
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IF 3.9
Maosen Chen,Yukun Wang,Shen Yu,Wenkang Yu,Bingmei Wang,Wenjin He,Chentao Lin,Zhibin Zhao,Jerry Wang,Feng Qi
Neoxanthin is a valuable carotenoid with antioxidant, chemopreventive, anticancer, and nephroprotective properties, showing potential for pharmaceutical, food, and cosmetic applications. Despite its valuable applications, neoxanthin’s industrial production is hindered by environmental concerns and very low extraction yields. In this study, de novo neoxanthin biosynthesis was engineered in E. coli. The key precursor β-carotene biosynthetic pathway was established and systematically optimized through modulation of multiple metabolic routes, including the endogenous MEP pathway and heterologous MVA and IU pathways. The enhanced IU pathway demonstrated significantly higher β-carotene biosynthetic efficiency compared to both the MVA and MEP pathways. Cofactor availability was enhanced by engineering central metabolism through overexpression of ATP/NADPH-generating genes and knockout of ATP/NADPH-consuming genes. Ultimately, a β-carotene-to-neoxanthin conversion pathway was successfully developed in engineered E. coli, achieving an enhanced neoxanthin production of 37.3 mg/L in a 5 L fermenter. This work provides an effective strategy for de novo microbial neoxanthin production.
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METABOLIC ENGINEERING
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2024
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IF 6.8
Qingchen Li,Chenxi Li,Jie Zhong,Yukun Wang,Qinghua Yang,Bingmei Wang,Wenjin He,Jianzhong Huang,Shengyuan Lin,Feng Qi
N-methylserotonin (NMS) is a valuable indole alkaloid with therapeutic potential for psychiatric and neurological disorders, and it is used in health foods, cosmetics, and weight loss supplements. However, environmental challenges and low reaction efficiencies significantly hinder cost-effective, large-scale production of NMS in plants or through chemical synthesis. Herein, we have successfully engineered Escherichia coli strains to enhance NMS production from L-tryptophan using whole-cell catalysis. We developed multiple biosynthesis pathways incorporating modules for serotonin (5-hydroxytryptamine, 5-HT), tetrahydromonapterin (MH₄), and S-adenosylmethionine (SAM) synthesis. To enhance MH₄ availability, we employed a high-activity Bacillus subtilis FolE and minimized carbon flux loss through targeted gene knockouts in competitive metabolic pathways, improving 5-HT production. Additionally, we constructed a comprehensive SAM biosynthesis module to facilitate transmethylation by a selected N-methyltransferase fused with ProS2. These engineered modules were coexpressed in two plasmids within the optimized strain NMS-19, producing 128.6 mg/L of NMS in a 5-L bioreactor using fed-batch cultivation—a 92-fold increase over the original strain. This study introduces a viable strategy for NMS production and provides insights into the biosynthesis of SAM-dependent methylated tryptamine derivatives.
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ANTIMICROBIAL AGENTS AND CHEMOTHERAPY
·
2021
·Vol. 65
·期 4
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IF 5.191
Lewis F. Neville,Itamar Shalit,Peter A. Warn,Marc H. Scheetz,Jiuzhi Sun,Madeline B. Chosy,Paul A. Wender,Lynette Cegelski,Jacob T. Rendell
The ability of vancomycin-arginine (V-r) to extend the spectrum of activity of glycopeptides to Gram-negative bacteria was investigated. Its MIC towards Escherichia coli, including β-lactamase expressing Ambler classes A, B, and D, was 8 to 16 μg/ml. Addition of 8 times the MIC of V-r to E. coli was acutely bactericidal and associated with a low frequency of resistance (<2.32 × 10−10). In vivo, V-r markedly reduced E. coli burden by >7 log10 CFU/g in a thigh muscle model. These data warrant further development of V-r in combatting E. coli, including resistant forms.
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JACS Au
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2025
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IF 8.7
Han Zu,Fang Pan,Heng-Li Fan,Zhi-Gang Qian,Xiao-Xia Xia
Hierarchical structures of biomolecular condensates in living cells are essential for cellular functions, particularly in orchestrating complex reactions through synergistic cooperation. However, our understanding of these structures remains limited, because controlling the hierarchical structures of condensates in living cells can be difficult. Here we create an artificial condensate system in model prokaryotic Escherichia coli cells via coexpressing two distinct intrinsically disordered proteins. By modulating the homotypic and heterotypic interactions between the two proteins, condensates with differential structural organization (cophase and multiphase separation) were successfully constructed and dynamically controlled via physical or chemical cues from the environments. In vitro reconstitutions further supported the underlying physicochemical principles of dynamic condensate organization. This study not only deepens our understanding of the structural organization of biomolecular condensates but also provides a useful strategy for dynamic regulation of synthetic condensates in living cells.
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LWT-FOOD SCIENCE AND TECHNOLOGY
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2025
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IF 6.6
Shudong Peng,Jiajun Li,Yating Long,Ying Zheng,Dongming Lan,Yonghua Wang
Foodborne Escherichia coli is one of the health-threatening pathogens existing on the surface of vegetables and fruits. Although antibiotics are the first line of defence against bacteria, increasing concerns are awake about the multidrug resistance caused by overuse and misuse of antibiotics. Therefore, alternative antibacterials are urgently necessary. In this context, we identified, cloned and expressed a novel endolysin LysD6E derived from marine phage D6E for its potential antibacterial action against E. coli . LysD6E consists of an Amidase_3 domain belonging to the N-acetylmuramoyl-L-alanine amidase family. LysD6E was stabile between 4 and 70 °C, and pH between 6 and 10. It had strong lysogenic activity against E. coli and Salmonella typhi . Furthermore, when applying 0.5 μM of LysD6E together with 5 mM EDTA on lettuce for 2 h, viable counts of E. coli ATCC 25922 reduced by 2.05 Log CFU/mL. Our findings suggest that phage-derived LysD6E has the potential against foodborne E. coli in raw vegetables.
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INTERNATIONAL JOURNAL OF MEDICAL MICROBIOLOGY
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2026
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IF 3.6
Hui Yu,Nan Wang,Changjiang Sun,Bingfeng Leng,Liping Sun,Xuming Deng,Lei Xu,Zihao Teng,Jianfeng Wang,Lin Wang
Enterotoxigenic Escherichia coli (ETEC), a predominant pathogen responsible for neonatal animal diarrhea, poses a significant threat to global health and results in considerable economic losses in animal husbandry. The pathogenicity of ETEC is primarily mediated by its adherence to intestinal epithelial cells via fimbriae adhesins and the subsequent secretion of enterotoxins. We developed a high-throughput screening platform for chorionic inhibitors. By employing semi-solid motility assays, hemagglutination inhibition assays, and cell adhesion assays, we screened a panel of natural compounds and identified Isogliquiritin (ISL) as the active compound. Subsequent validation using transmission electron microscopy and reverse transcription-polymerase chain reaction (qRT-PCR) revealed that ISL significantly inhibits the synthesis and assembly of ETEC fimbriae, thereby reducing bacterial adhesion to host cells. After assessing its safety, we demonstrated through in vivo experiments that ISL attenuates ETEC-induced diarrhea. In the Galleria mellonella larval model, ISL increased the survival rate, meanwhile in a mouse diarrhea model, ISL reduced inflammation caused by ETEC C83902 and upregulated the expression of tight junction protein (ZO-1, claudin-1, and occluding) genes. Collectively, our findings highlight that ISL is a promising natural compound for the prevention and treatment of ETEC-induced diarrhea. By targeting bacterial virulence factors rather than bacterial growth, ISL provides a novel strategy against antimicrobial resistance and offers a safer alternative for managing ETEC infections in livestock.
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Journal of Biological Research-Thessaloniki
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2020
·Vol. 27
·期 1
·1-10
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IF 2.121
Cao Yujin,Mu Hui,Guo Jing,Liu Hui,Zhang Rubing,Liu Wei,Xian Mo,Liu Huizhou
Background The fuel ethanol industry has made tremendous progress in the last decades. Ethanol can be obtained by fermentation using a variety of biomass materials as the feedstocks. However, few studies have been conducted on ethanol utilization by microorganisms. The price of petroleum-derived ethanol, easily made by the hydrolysis of ethylene, is even lower than that of bioethanol. If ethanol can be metabolized by microorganisms to produce value-added chemicals, it will open a new door for the utilization of inexpensive ethanol resources.Results We constructed an engineered Escherichia coli strain which could utilize ethanol as the sole carbon source. The alcohol dehydrogenase and aldehyde dehydrogenase from Aspergillus nidulans was introduced into E. coli and the recombinant strain acquired the ability to grow on ethanol. Cell growth continued when ethanol was supplied after glucose starvation and 2.24 g L−1 of ethanol was further consumed during the shake-flasks fermentation process. Then ethanol was further used for the production of mevalonic acid by heterologously expressing its biosynthetic pathway. Deuterium-labeled ethanol-D6 as the feedstock confirmed that mevalonic acid was synthesized from ethanol.Conclusion sThis study demonstrated the possibility of using ethanol as the carbon source by engineered E. coli strains. It can serve as the basis for the construction of more robust strains in the future though the catabolic capacity of ethanol should be further improved.
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Microbial Cell Factories
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2016
·Vol. 15
·期 1
·1-9
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IF 3.744
Yang Jianming,Nie Qingjuan
Background Under aerobic conditions, acetic acid is the major byproduct produced by E. coli during the fermentation. And acetic acid is detrimental to cell growth as it destroys transmembrane pH gradients. Hence, how to reduce the production of acetic acid and how to utilize it as a feedstock are of intriguing interest. In this study, we provided an evidence to produce β-caryophyllene by the engineered E. coli using acetic acid as the only carbon source. Results Firstly, to construct the robust acetate-utilizing strain, acetyl-CoA synthases from three different sources were introduced and screened in the E. coli . Secondly, to establish the engineered strains converting acetic acid to β-caryophyllene, acetyl-CoA synthase (ACS), β-caryophyllene synthase (QHS1) and geranyl diphosphate synthase (GPPS2) were co-expressed in the E. coli cells. Thirdly, to further enhance β-caryophyllene production from acetic acid, the heterologous MVA pathway was introduced into the cells. What’s more, acetoacetyl-CoA synthase (AACS) was also expressed in the cells to increase the precursor acetoacetyl-CoA and accordingly resulted in the increase of β-caryophyllene. The final genetically modified strain, YJM67, could accumulate the production of biomass and β-caryophyllene up to 12.6 and 1.05 g/L during 72 h, respectively, with a specific productivity of 1.15 mg h −1 g −1 dry cells, and the conversion efficiency of acetic acid to β-caryophyllene (gram to gram) reached 2.1 %. The yield of β-caryophyllene on acetic acid of this strain also reached approximately 5.6 % of the theoretical yield. Conclusions In the present study, a novel biosynthetic pathway for β-caryophyllene has been investigated by means of conversion of acetic acid to β-caryophyllene using an engineered Escherichia coli . This was the first successful attempt in β-caryophyllene production by E. coli using acetic acid as the only carbon source. Therefore, we have provided a new metabolic engineering tool for β-caryophyllene synthesis.
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CHEMICAL ENGINEERING JOURNAL
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2018
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IF 6.735
Shaofeng Zhou,Shaobin Huang,Xiaohu Li,Irini Angelidaki,Yifeng Zhang
Water quality deterioration caused by a wide variety of recalcitrant organics and pathogenic microorganisms has become a serious concern worldwide. Bio-electro-Fenton systems have been considered as cost-effective and highly efficient water treatment platform technology. While it has been extensively studied for recalcitrant organics removal, its application potential towards water disinfection (e.g., inactivation of pathogens) is still unknown. This study investigated the inactivation of Escherichia coli in a microbial electrolysis cell based bio-electro-Fenton system (renamed as microbial electrolytic-Fenton cell) with the aim to broad the application of microbial electrochemistry. Results showed that a 4-log reduction of Escherichia coli (10 7 to hundreds CFU/mL) was achieved with an external applied voltage of 0.2 V, 0.3 mM Fe 2+ and cathodic pH of 3.0. However, non-notable inactivation was observed in the control experiments without external voltage or Fe 2+ dose. The disinfection effect was enhanced when cathode air flow rate increased from 7 to 41 mL/min and was also in proportion to the increase of Fe 2+ concentration from 0.15 to 0.45 mmol/mL. Fatal cell membrane destruction by OH was identified as one potential mechanism for disinfection. This study successfully demonstrated the feasibility of bio-electro-Fenton process for pathogens inactivation, which offers insight for the future development of sustainable, efficient, and cost-effective biological water treatment technology.
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ACS Synthetic Biology
·
2025
·
IF 3.9
Baodong Hu,Jingwen Zhou,Jianghua Li,Jian Chen,Guocheng Du,Fang Zhong,Yucheng Zhao,Xinrui Zhao
Marmesin, a plant dihydrofuranocoumarin, is an important intermediate in the synthesis of linear furanocoumarins and exhibits a variety of pharmacological activities. However, due to the lack of efficient prenyltransferases, the incompatibility of redox partners for P450 enzymes, and the insufficient supply of precursor (DMAPP), the microbial synthesis of marmesin remained at an extremely low level. Here, we report the efficient biosynthesis of marmesin in Escherichia coli by screening the robust 6-prenyltransferase PpPT1 and marmesin synthase PpDCΔ2–29 from Peucedanum praeruptorum. Next, the activities of PpPT1 and PpDCΔ2–29 were enhanced using fusion protein tags and redox partner engineering, respectively. In addition, the synthesis of marmesin was further improved by strengthening the methylerythritol phosphate (MEP) pathway to increase the availability of DMAPP and by optimizing the modular pathway in the engineered strain. Finally, the titer of marmesin reached 203.69 mg L–1 in the fed-batch fermentation with a molar conversion rate of umbelliferone of 81.4%, which is the highest titer for marmesin production using engineered microorganisms. The applied strategy and marmesin-producing strain constructed in this study lay the foundation for the green production of valuable complex furanocoumarins.
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METABOLIC ENGINEERING
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2025
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IF 6.8
Dan He,Yan Chen,Junfeng Shen,Han Yu,Jay D. Keasling,Xiaozhou Luo
10-hydroxy-2-decenoic acid (10-HDA), a unique unsaturated fatty acid present in royal jelly, has attracted considerable interest due to its potential medical applications. However, its low concentration in royal jelly and complex conformational structure present challenges for large-scale production. In this study, we designed and constructed a de novo biosynthetic pathway for 10-HDA in Escherichia coli . Initially, we introduced the heterologous thioesterase UaFatB1 to hydrolyze trans-2-decenoyl ACP to produce trans-2-decenoic acid, a key precursor for 10-HDA. Subsequently, we employed the bacterial cytochrome P450 enzyme CYP153AMaq to catalyze the terminal hydroxylation of trans-2-decenoic acid. Furthermore, through redox partner engineering and directed evolution, we identified the optimal combination for 10-HDA production: CYP153AMaq Q129R/V141L mutant with redox partner FdR0978/Fdx0338. Finally, we optimized the fermentation conditions and achieved a 10-HDA titer of 18.8 mg/L using glucose as primary carbon source. Our work establishes a platform for producing α , β -unsaturated fatty acids and their derivatives, facilitating further studies on these compounds.
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