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SENSORS AND ACTUATORS B-CHEMICAL · 2024 · IF 8

Cell membrane-inspired COF/AAO hybrid nanofluidic membrane with ion current rectification properties for ultrasensitive detection of E. coli

Mengyuan Chen,Chong Wang,Jing Tan,Jin Wang,Chen Wang

Nanofluidic membranes hold great prospects in sensitive and rapid detection of bioanalysts, while remaining lacking robust and general approaches for specificity. In this work, we introduce a cell membrane-inspired nanofluidic membrane with excellent ion current rectification properties as a simple and robust platform for label-free and ultrasensitive biosensing. The asymmetric nanofluidic membrane is prepared by coupling a two-dimensional covalent organic framework (COF) membrane to an anodic aluminum oxide (AAO) nanochannel via hydrogen bonding. The prepared COF/AAO membrane presents selective ion transport due to the asymmetric structure and anion selectivity of COF. With the large surface area and abundant functional modification sites of COF, further grafting of the recognition unit can be easily achieved to mimic the recognition process of receptors and ligands on cell membranes. As a proof of concept, the application in bacterial detection is demonstrated by covalently bonded Concanavalin A as recognition units. The highly amplified ionic current based on ion current rectification properties of the COF/AAO membrane allows sensitive and selective bioanalysis. Specifically, the proposed asymmetric nanofluidic membrane presents a linear detection range from 10 to 10 6 CFU/mL with an ultralow detection limit of 7 CFU/mL for E. coli . This work reveals the great potential of COF/AAO hybrid nanofluidic membrane as a sensor for rapid and precise bioanalysis.

研究领域: 生物传感器 · 材料科学 · 微生物学
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COLLOIDS AND SURFACES B-BIOINTERFACES · 2024 · IF 5.4

Evaluating the antibacterial and antibiofilm activities of chitosan derivatives containing six-membered heterocyclics against E. coli and S. aureus

Lin Wang,Yu Pang,Meihua Xin,Mingchun Li,Lulu Shi,Yangfan Mao

Chitosan exhibits good biocompatibility and some antibacterial activity , making it a popular choice in biomedicine, personal care products, and food packaging. Despite its advantages, the limited antibacterial effectiveness of chitosan hinders its widespread use. Introducing a six-membered heterocyclic structure through chemical modification can significantly enhance its antimicrobial properties and broaden its potential applications. In order to explore the effect of six-membered heterocyclic structure on the antibacterial and antibiofilm activities of chitosan. In this study, seven chitosan derivatives containing six-membered heterocyclics were prepared. They were characterized using Fourier transform infrared (FT-IR) spectroscopy, nuclear magnetic resonance (NMR) spectroscopy, and elemental analysis . Cell viability assay showed that they were non-toxic. The antibacterial and antibiofilm activities against Escherichia coli ( E. coli ) and Staphylococcus aureus ( S. aureus ) were evaluated. Our research findings demonstrate that increasing the hydrophobicity , alkalinity and charge density of the substitute groups improved the antibacterial and antibiofilm activities of chitosan. This study also offers valuable insights into the quantitative structure-activity relationships of chitosan derivatives in terms of antibacterial and antibiofilm activities.

研究领域: 生物医学 · 材料科学 · 微生物学
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JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY · 2024 · IF 5.7

Multiple Regulatory Mechanisms Synergistically Control the Soluble Expression of CsCE for Enhanced Enzymatic Productivity of Lactulose in E. coli

Chenlu Zhu,Renjiao Han,Bixuan Gu,Caiyun Wang,Han Liu,Xiaomei Lyu,Jian He,Ruijin Yang

The limited expression of cellobiose 2-epimerase poses a significant constraint on the industrial enzymatic production of lactulose. Extensive modifications to the expression cassette offer a means to enhance the yield of recombinant proteins. In this study, an integrated strategy, combining individual and collaborative approaches, is proposed to fine-tune each stage of the CsCE overexpression program. This strategy involves the multidimensional integration of standardized genetic elements at various levels, including transcription, translation, folding, and three-dimensional structure. The volumetric activity of the final recombinant strain was markedly increased by 12-fold compared to the wild-type strain, reaching 2260.62 U/L. The protein expression in the newly developed high-yield recombinant strain exhibited a significant enhancement, with a higher proportion of soluble protein compared to that of inclusion bodies. Our findings offer insights into the multifaceted synergistic regulation of protein expression processes, holding promising implications for the production of heterologous recombinant proteins.

CAS / MDL: 4618-18-2
研究领域: 生物工程 · 分子生物学
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JOURNAL OF HAZARDOUS MATERIALS · 2025 · IF 11.3

Enhanced phenanthrene degradation by a synthetically engineered E. coli consortium utilizing the SCRaMbLE for the evolution of key genes

Yibin Yang,Rui Chen,Zhiqiang Zhang,Yuke Qin,Attiqa Parveen,He Bai,Xiaoqiang Jia

Phenanthrene, a pervasive polycyclic aromatic hydrocarbon (PAH), poses significant environmental and health risks due to its persistence and bioaccumulation. This study presents the development of a synthetic bacterial consortium comprising three engineered Escherichia coli BL21(DE3) strains, each harboring distinct catabolic modules encoded by pET-28(a) plasmids for the complete mineralization of phenanthrene. The consortium was engineered by integrating 17 genes critical for phenanthrene degradation, organized into three distinct modules that harness the collective metabolic potential of separate strains to enhance the degradation process. Each module was tailored to a specific phase of phenanthrene catabolism, ensuring the coordinated and efficient breakdown of the pollutant. Following the identification of rate-limiting enzymes within the up- and downstream degradation modules, the eight corresponding genes were subjected to laboratory evolution using the SCRaMbLE method to bolster the degradation capabilities of the strains. The consortium was then tested in a minimal salt medium (MSM) containing 100 mg/L phenanthrene. Following their evolution, the two SCRaMbLE strains E. coli SCRPHE033 and E. coli SCRKA065 exhibited enhanced degradation rates of 11.5 and 25.18 % for their respective substrates. Finally, the comprehensive phenanthrene degradation system, which includes the two SCRaMbLE strains and E. coli CRSA, was subjected to comprehensive optimization, after which it achieved a significant degradation efficiency of 84.33 % for 100 mg/L phenanthrene within a 120-hour timeframe. This research demonstrates the potential of synthetic biology in constructing microbial consortia for environmental bioremediation based on the SCRaMbLE method, offering a promising strategy for mitigating PAH pollution.

CAS / MDL: 129-00-0 85-01-8
研究领域: 合成生物学 · 环境微生物学 · 基因组工程 · 生物修复
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JOURNAL OF BIOTECHNOLOGY · 2016 · IF 2.667

Engineering of a novel carbonyl reductase with coenzyme regeneration in E. coli for efficient biosynthesis of enantiopure chiral alcohols

Ping Wei,Jia-Xin Gao,Gao-Wei Zheng,Hong Wu,Min-Hua Zong,Wen-Yong Lou

The novel anti-Prelog stereospecific carbonyl reductase from Acetobacter sp. CCTCC M209061 was successfully expressed in E. coli combined with glucose dehydrogenase (GDH) to construct an efficient whole-cell biocatalyst with coenzyme NADH regeneration. The enzymatic activity of GAcCR (AcCR with a GST tag) reached 304.9   U/g-dcw, even 9 folds higher than that of wild strain, and the activity of GDH for NADH regeneration recorded 46.0   U/mg-protein in the recombinant E. coli . As a whole-cell biocatalyst, the recombinant E. coli BL21(DE3)pLysS (pETDuet- gaccr - gdh ) possessed a broad substrate spectrum for kinds of carbonyl compounds with encouraging yield and stereoselectivity. Besides, the asymmetric reduction of ethyl 4-chloroacetoacetate (COBE) to optically pure ethyl 4-chloro-3-hydroxybutyrate (CHBE) catalyzed by the whole-cell biocatalyst was systematically investigated. Under the optimal reaction conditions, the optical purity of CHBE was over 99% e.e. for ( S )-enantiomer, and the initial rate and product yield reached 8.04   μmol/min and 99.4%, respectively. Moreover, the space-time yield was almost 20 folds higher than that catalyzed by the wild strain. Therefore, a new, high efficiency biocatalyst for asymmetric reductions was constructed successfully, and the enantioselective reduction of prochiral compounds using the biocatalyst was a promising approach for obtaining enantiopure chiral alcohols.

研究领域: 生物工程 · 合成生物学
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Frontiers of Materials Science · 2025 ·Vol. 19 ·期 3 ·1-11 · IF 2.3

Enhanced photodynamic therapy of porphyrin grafted Zn–Ag–In–S QDs/PMAO nanoparticles against melanoma cells and E. coli

Sheng Yang,Qing Danni,Zhang Peng,Sun Yixin,Zhang Rong

Tetra-aminophenyl porphyrin (TAPP)-grafted Zn–Ag–In–S quantum dots (ZAIS QDs)/poly(maleic anhydride- alt -1-octadecene) (PMAO) nanoparticles were synthesized and their photoluminescence properties as well as photodynamic properties were studied. ZAIS QDs showed the brightest photoluminescence and highest quantum yield at an optimized Zn feeding molar ratio of 20%. Those TAPP-grafted nanoparticles (i.e., ZAIS/PMAO- g -TAPP) were able to produce 1 O 2 in aqueous solution under light irradiation as indicated by the 1 O 2 indicator, 9,10-anthracenediyl-bis(methylene) dimalonic acid (ADMA). ZAIS/PMAO- g -TAPP nanoparticles also demonstrate good biocompatibility and low dark toxicity even at a concentration as high as 2.8 mg·mL −1 , whith can be applied as both a fluorescence probe and a photodynamic therapy (PDT) agent. The PDT treatment showed that the viability of melanoma A2058 cells was less than 10% after treatment with the 420 nm light irradiation for 15 min at a photosensitizer concentration of 1.7 mg·mL −1 . During the PDT treatment with Escherichia coli , the survival rate of the bacteria decreased by −95% after light irradiation at the same concentration. Such dual-functional ZAIS/PMAO- g -TAPP nanoparticles researched in this study demonstrate promising potential for fluorescence labeling as well as effective PDT treatment against cancer cells and bacteria.

研究领域: 纳米技术 · 光动力治疗 · 材料科学 · 生物医学工程 · 微生物学 · 肿瘤学
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DYES AND PIGMENTS · 2020 · IF 4.613

A novel AgI/BiOI/pg-C3N4 composite with enhanced photocatalytic activity for removing methylene orange, tetracycline and E. coli

Ding Yuan,Liying Huang,Yeping Li,Hao Wang,Xiuquan Xu,Chaobao Wang,Lei Yang

Making multiple heterojunction of photocatalysts is an effect approach to achieve high effective utilization of solar resource. A novel ternary AgI/BiOI/pg-C 3 N 4 nanocomposite was synthesized with a unique n-p-n heterojunction. In this work, an in-situ transformation method was used to fabricate AgI/BiOI/pg-C 3 N 4 heterostructure with different AgI/BiOI ratios. The photocatalytic activity of the AgI/BiOI/pg-C 3 N 4 was detected by the degradation of methylene orange (MO) and tetracycline (TC). The AgI/BiOI/pg-C 3 N 4 composites exhibited better photocatalytic activity than pg-C 3 N 4 and BiOI/pg-C 3 N 4 composite. The performance of photocatalytic antibacterial inactivation of E. coli indicated that the AgI/BiOI/pg-C 3 N 4 heterostructure is an ideal material for antibacterial applications. The ESR and trapping experiments confirmed that the degradation of organic pollution is completed through the co-operation of •O 2 − , •OH and h + . A possible mechanism of the degradation process is proposed.

研究领域: 环境科学 · 材料科学
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FOOD CHEMISTRY · 2024 · IF 8.5

A HOF-101@AgNPs-based dual-signal mode aptasensor for electrochemiluminescence and fluorescence detection of E. coli O157:H7

Lanqing Yang,Xiaomin Wang,Xu Jing,Baoqing Bai,Tao Bo,Jinhua Zhang,Ligang Yu,Hailong Qian,Ying Gu,Yukun Yang

Escherichia coli O157:H7 ( E. coli O157:H7 ) emerges as a foodborne pathogen, emphasizing the imperative for creating precise detection tools. An ultrasensitive electrochemiluminescence/fluorescence (ECL/FL) dual-signal mode aptasensor was constructed for the detection of E. coli O157:H7 . Ultrafine silver nanoparticles were loaded on the surface of hydrogen-bonded organic skeleton materials by in-situ photoreduction method, and then combined with aptamers that can identify specific targets to prepare HOF-101@AgNPs@Apt, greatly simplifying the ECL/FL dual-signal mode probe fabrication process and improving sensing reliability. HOF-101@AgNPs@Apt had both strong ECL luminescence and fluorescence, resulting in a high detection sensitivity. The low limit of detection (LOD) for ECL and FL were 0.48 CFU mL −1 and 2.39 CFU mL −1 , respectively. Moreover, the proposed dual-signal mode aptasensor has been successfully applied to determine E. coli O157:H7 levels in tap water and milk with superior accuracy and high antiinterference capability, providing a promising method for food safety monitoring.

CAS / MDL: 1633-78-9 51805-45-9
研究领域: 食品科学 · 生物传感 · 分析化学
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BIOSENSORS & BIOELECTRONICS · 2020 · IF 10.257

Ultrasensitive magnetic DNAzyme-copper nanoclusters fluorescent biosensor with triple amplification for the visual detection of E. coli O157:H7

Ziqi Zhou,Yangzi Zhang,Mingzhang Guo,Kunlun Huang,Wentao Xu

A relatively-simple and high-efficient fluorescent magnetic biosensor based on DNAzyme was established for the detection of E. coli O157:H7. In order to solve the problem of weak signal and low sensitivity in the detection of foodborne pathogenic bacteria , we ingeniously designed a fluorescent sensor based on triple signal amplification of magnetic beads, DNAzyme and photoluminescence . In the detection process, the E. coli -specific RNA-cleaving DNAzyme can specifically identify the target protein in crude intracellular mixture (CIM), which caused its conformation changes and induced rolling circle amplification (RCA) to the generation and luminescence of copper nanoclusters (CuNCs). This cascade amplification design can capture weak signals in the sample. The biosensor also indicated a good linear range from 10  CFU  mL −1 to 1000 CFU mL −1 and obtained a limit of detection (LOD) of 1.57 CFU mL −1 , which showed a relatively high sensitivity compared with other studies. Furthermore, the biosensor displayed high-efficient detection capability in 1.5 h and good reproducibility (relative standard deviations < 2%). It has been proved that this sensor is feasible to the detection of E. coli O157:H7 in drinking water and apple juice. Moreover, we found that the final detection product can effectively wrap the magnetic beads and can be driven by the magnetic field. And this unexpected discovery will provide ideas for the development of biosensing robots.

CAS / MDL: 134-03-2
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Analytical Methods · 2024 ·Vol. 16 ·期 28 ·4865-4872 · IF 2.7

An efficient ratiometric fluorescence and colorimetric dual-mode probe for convenient determination of nitrite in real samples and E. coli

Yujie Wen,Cong Tang,Qing Shen,Shuqing Dong,Yaya Wang,Yunchun Li,Shijun Shao

Developing an effective and convenient nitrite detection method is indispensable in food safety, environmental monitoring, clinical diagnosis of diseases, and many other areas. Herein, a dicyanoisophorone derivative, TMN-NH2 with large Stokes shift and near-infrared (NIR) emission, was proposed as a ratiometric fluorescence and colorimetric dual-mode probe for the rapid determination of NO2− in acidic media, showing excellent selectivity and high sensitivity. The sensing mechanism is based on the diazotization of TMN-NH2 with NO2− and subsequent diazonium salt hydrolysis to form a hydroxyl-substituted product (TMN-OH). Under the optimized conditions of reaction and detection, a new quantitative analysis method based on TMN-NH2 was established for NO2− detection, exhibiting good linear relationships to NO2− in the range of 0.5 to 15 μM with practical detection limits of 26.6 nM and 17.6 nM for the colorimetric and fluorescent readout, respectively. The quantitative detection of NO2− in real samples demonstrated satisfactory recoveries and repeatability. Moreover, TMN-NH2 was successfully applied for monitoring NO2− in Escherichia coli by confocal fluorescence imaging.

研究领域: 分析化学 · 食品安全 · 环境科学 · 医学诊断
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JACS Au · 2026 · IF 8.5

Engineering Plant-Derived P450 Enables the Efficient Production of Berberine through a Concise and Modular Enzymatic Cascade in E. coli

Yu Qin,Huiling Liu,Yan Zhang,Fei Li,Zhiwei Deng,Zhengshan Luo,Yuhao Zhuang,Kai Fu,Zhenbo Yuan,Yijian Rao

Sustainable and scalable production of plant natural products remains a major challenge at the interface of synthetic biology, metabolic engineering, and drug discovery, often constrained by plant-membrane-bound cytochrome P450s. To overcome the bottleneck in P450-catalyzed methylenedioxy-bridge (MDB) formation, we designed a concise, modular Escherichia coli pathway to the alkaloid berberine that bypasses multiple native steps and enables high-level accumulation of the P450-dependent intermediate. We then implemented integrated engineering of CjCYP719A1, including expression optimization, structure-guided design, consensus mutagenesis, and machine learning, to obtain a variant with 8.9-fold higher activity and improved thermostability. This enabled a berberine titer of 353 mg/L in scaled-up preparations, the highest reported to date, demonstrating industrial potential. Furthermore, mechanistic studies reveal their improved catalytic activity and thermostability. Together, this work provides a practical framework for functional expression and systematic optimization of plant-derived P450s to enable efficient production of plant natural products in E. coli.

CAS / MDL: 2086-83-1
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CHEMISTRY & BIOLOGY · 2015 ·Vol. 22 ·期 11 ·1461-1469 · IF 6.645

Development of a Clickable Probe for Profiling of Protein Glutathionylation in the Central Cellular Metabolism of E. coli and Drosophila

Shan Feng,Yuling Chen,Fan Yang,Lei Zhang,Yiyi Gong,Gulishana Adilijiang,Yan Gao,Haiteng Deng

Protein glutathionylation is an important post-translational modification that regulates many cellular processes, including energy metabolism, signal transduction, and protein homeostasis. Global profiling of glutathionylated proteins (denoted as glutathionylome) is crucial for understanding redox-regulated signal transduction. Here, we developed a novel method based on click reaction and proteomics to enrich and identify the glutathionylated peptides in Escherichia coli and Drosophila lysates, in which 937 and 1,930 potential glutathionylated peptides were identified, respectively. Bioinformatics analysis showed that the cysteine residue next to negatively charged amino acid residues has a higher frequency of glutathionylation. Importantly, we found that most proteins associated with metabolic pathways were glutathionylated and that the glutathionylation sites of metabolic enzymes were highly conserved among different species. Our results indicate that the glutathione analog is a useful tool to characterize protein glutathionylation, and glutathionylation of metabolic enzymes, which play important roles in regulating cellular metabolism, is conserved.

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