Frontiers in Microbiology
·
2019
·
IF 4.259
Han Xiuyan,Chen Changming,Yan Qiulong,Jia Liqiu,Taj Ayaz,Ma Yufang
Mycobacterium tuberculosis is one of most pathogenic microorganisms in the world. Previously, the bifunctional enzyme GlmU with glucosamine-1-phosphate acetyltransferase activity and N-acetylglucosamine-1-phosphate uridyltransferase activity has been suggested as a potential drug target; therefore, discovering compounds targeting GlmU acetyltransferase is necessary. The natural products were tested for inhibition of GlmU acetyltransferase activity. We found that dicumarol exhibited inhibitory effects on GlmU acetyltransferase, with a concentration achieving a 50% inhibition (IC50) value of 4.608 μg/ml (13.7 μM). The inhibition kinetics indicated that dicumarol uncompetitively inhibited acetyl CoA and showed mixed-type inhibition for glucosamine-1-phosphate (GlcN-1-P). The activity of dicumarol against M. tuberculosis H37Ra was evaluated with a minimum inhibitory concentration (MIC) value of 6.25 μg/ml (18.55 μM) in the Alamar blue assay. Dicumarol also exhibited inhibitory effects on several clinically sensitive M. tuberculosis strains and drug-resistant strains, with a range of MIC value of 6.25 to >100 μg/ml. Dicumarol increased the sensitivity of anti-tuberculosis drugs (isoniazid and rifampicin) when dicumarol was present at a low concentration. The transcriptome and proteome data of M. tuberculosis H37Ra treated by dicumarol showed that the affected genes were associated with cell wall synthesis, DNA damage and repair, metabolic processes, and signal transduction. These results provided the mechanism of dicumarol inhibition against GlmU acetyltransferase and M. tuberculosis and also suggested that dicumarol is a potential candidate for TB treatment.
研究领域:
微生物学
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药理学
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分子生物学
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结核病研究
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PubMed
高影响因子
ACS Sensors
·
2021
·
IF 7.711
Jinyue Zhang,Jiao Cao,Wendong Jia,Shanyu Zhang,Shuanghong Yan,Yuqin Wang,Panke Zhang,Hong-Yuan Chen,Wenfei Li,Shuo Huang
Protein nanopores can be engineered as nanoreactors to investigate single-molecule chemical reactions. Recent studies have demonstrated that Mycobacterium smegmatis porin A (MspA) nanopore is a superior engineering template acknowledging its geometrical advantages. However, reported engineering of MspA to form a nanoreactor has focused only on site 91 and mapping of other engineering sites have never been performed before. By taking tetrachloraurate(III) ([AuCl4]−) as a model reactant, potential engineering sites within the pore constriction of MspA have been thoroughly investigated. It is discovered that the produced event amplitude is inversely correlated to the cross-sectional diameter of the pore constriction size at the engineering site, providing evidence that site 91 is actually already the optimum place to introduce the chemical reactivity. Other unavailable engineering sites, which either significantly interfere with the pore assembly or produce reactive sites facing to the pore’s exterior instead of to the pore lumen, were also spotted and discussed. All results demonstrated above have provided a complete map of engineering sites within the constriction area of MspA and may be beneficial as a reference in future engineering of corresponding nanoreactors.
研究领域:
纳米技术
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生物化学
·
分子生物学
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DOI
PubMed
高影响因子
JOURNAL OF BIOLOGICAL CHEMISTRY
·
2026
·
IF 4.1
Yunjie Liu,Yue Zhao,Weihan Sun,Nian Li,Yunjun Pan,Li Ma,Shengying Li
Steroids with anti-inflammatory, anti-allergic, endocrine-regulating, and other pharmaceutical activities represent the second most widely used class of drugs worldwide, following antibiotics. Their industrial production primarily relies on Mycobacteria -mediated biotransformation of sterols into key intermediates, followed by chemical or enzymatic modifications. While the sterol metabolic pathways in Mycobacteria have been intensively studied, the identification and functional characterization of key enzymes, particularly cytochrome P450 enzymes (CYPs or P450s) and their cognate redox partners, remain incomplete. Here, we heterologously expressed 24 P450s, 10 ferredoxin reductases (FdRs), and 12 ferredoxins (Fdxs) from Mycobacterium neoaurum ZC-1 in Escherichia coli . In vitro biochemical experiments identified five P450 enzymes (CYP125A76, CYP125A77, CYP125A78, CYP142A12, and CYP124A1) capable of catalyzing sterol side-chain terminal oxidation. Screening 120 redox partner combinations revealed FdR4662/Fdx4443 as the optimal cognate redox partners for all five P450 enzymes. With this redox partner pair, CYP142A12 achieved a conversion ratio of 89 % for 4-cholesten-3-one with NADH as the preferred cofactor. Structural analyses indicate that the electron-transfer efficiency is primarily governed by electrostatic complementarity around the Fe–S cluster, the redox-center distance between the Fe–S cluster and heme-iron, and the FAD-to-cluster distance within the FdR-Fdx complex. These findings highlight the critical role of redox partner selection in enhancing P450 catalytic efficiency and provide a solid foundation for engineering high-efficiency industrial strains to improve steroid biomanufacturing and reduce production costs.
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PubMed
高影响因子
BIOORGANIC CHEMISTRY
·
2025
·
IF 4.7
Xiuyan Han,Changming Chen,Renxiang Zhao,Jing Wu,Shujing Zhang,Yuzhuo Wu,Yufang Ma,Chao Wang,Xiaochi Ma
Euphorbia fischeriana has been traditionally used in Chinese medicine for tuberculosis (TB) treatment since ancient times. In this study, we first report the identification of an abietane-type diterpenoid, 17-hydroxy-jolkinolide B (HJKB), from E. fischeriana , which exhibits potent antimycobacterial activity against both Mycobacterium tuberculosis H37Ra strain and clinical isolates. The minimum inhibitory concentrations (MICs) of HJKB against diverse M. tuberculosis strains range from 1 to 12 μg/mL. Notably, HJKB demonstrates significant bactericidal activity against intracellular M. tuberculosis H37Ra in macrophage models, accompanied by anti-inflammatory effects at concentrations of 2–5 μg/mL. Using a combination of chemoproteomic analysis and pull-down assays, we explored the preliminary antimycobacterial mechanism of HJKB. Results indicate that HJKB interacts with the target proteins RpoB and RpoC in M. tuberculosis H37Ra, a finding further corroborated by molecular docking studies. RpoB and RpoC are essential subunits of the DNA-directed RNA polymerase holoenzyme, which is critical for bacterial ribosomal transcription regulation. In summary, HJKB represents a bioactive constituent of E. fischeriana with anti-TB efficacy, acting as a transcription inhibitor against M. tuberculosis . This study not only elucidates its antimycobacterial mechanism but also provides a preclinical foundation for the development of natural product-based TB therapeutics.
研究领域:
药理学
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微生物学
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天然产物研究
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药物化学
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生物化学
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DOI
PubMed
高影响因子
Food Science and Technology
·
2021
·
IF 1.718
SHI Min,YIN Pengyi,GUO Xiaobo,LI Qian,SUN Lin,CAO Xiaohua
Baicalein has strong anti-inflammatory activity, but its efficacy and mechanism in pneumonia of mice is still unclear. Herein, RAW264.7 cells were infected with M. tuberculosis H37Rv (MOI 1:10) after pre-incubation with 50 μM baicalein, and the result found that baicalein incubation ameliorated the increase of bacterial load, and reduced the levels of ROS, IL-12p40, TLR2 protein, phosphorylated JNK protein and M1 polarization markers (iNOS and CD11c), and increased the levels of IL-10, M2 polarization markers (Arg1, Mrc1 and CD206) and NEDD4L protein in H37Rv infected-RAW264.7 cells. And lentivirus-mediated NEDD4L overexpression vector (LV-NEDD4L) transfection had the same effect as baicalein, while small interfering RNA target NEDD4L (si-NEDD4L) transfection or JNK pathway inhibitor IQ 3 treatment reversed the effects of baicalein, and ubiquitin inhibitor MG132 treatment reversed the effects of LV-NEDD4L. Co-immunoprecipitation (Co-IP) and TLR2 ubiquitination assays found that the binding of NEDD4L and TLR2 promoted the ubiquitination of TLR2. In vivo, C57BL/6 mice were nasal infected with H37Rv (1000 ± 150 CFU/mouse) to establish a mouse pneumonia model, and then administered with baicalein. The result found that baicalein improved H37Rv-induced pneumonia in mice. In conclusion, baicalein increased NEDD4L-mediated ubiquitination of TLR2, promoting macrophage M2 polarization and alleviating H37Rv-induced pneumonia.
研究领域:
免疫学
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药理学
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微生物学
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分子生物学
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DOI
高影响因子
SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2026
·
IF 4.8
Xinyu Liu,Xinyi Zeng,Zhenyue Guo,Baoyin Yuan,Yinuo Yuan,Shanfeng Zhang
Tuberculosis (TB), a deadly infectious disease caused by Mycobacterium tuberculosis (Mtb), was the second leading cause of death from a single infectious agent after COVID-19 in 2022, killing nearly twice as many people as AIDS, and still affecting more than 10 million people every year. Traditional diagnostic methods have the problems of low sensitivity, poor specificity, long time consuming and high cost. MPT64 is a 24-kDa protein secreted by Mtb during its propagation. It only exists in the Mtb complex and is one of the important antigens for the serological diagnosis of tuberculosis. Based on the principle of one-step substrate capture and detection probe with aptamer specific recognition of MPT64, this study proposed an innovative and efficient strategy for Mtb detection through serum testing using an buffer system integrated with dual aptamer modified surface-enhanced Raman scattering (SERS) biosensor. By incubating NBA Raman reporter molecules on the surface of Gold-core silver-shell nanoparticles and coupling MPT64 aptamer A to form reporter probes; and modifying MPT64 aptamer-B onto the surface of magnetic beads to form capturing probes, which together constitute the detection system. This study constructed a curve showing the logarithmic relationship between the standard concentration of MPT64 and Raman intensity. Within the concentration range of 1 × 10 −3 to 1 × 10 4 ng/mL, a good linear relationship was obtained (R 2 = 0.9963), with a detection limit of 20 fg/mL. In 25 human serum samples (10 positive and 15 negative), the aptamer-SERS biosensor system achieved 100% sensitivity and specificity, and was capable of quantitatively detecting tuberculosis biomarkers, providing a reliable detection method for tuberculosis diagnosis.
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PubMed
高影响因子
MICROCHIMICA ACTA
·
2021
·Vol. 188
·期 11
·1-10
·
IF 5.833
Xie Jiaojiao,Mu Zhaode,Yan Bin,Wang Jie,Zhou Jing,Bai Lijuan
A label-free electrochemical aptasensor is reported for sensitive detection of the 6-kDa early secreted antigenic target (ESAT-6). For the first time, the bimetallic organic framework (b-MOF) of Zr-MOF-on-Ce-MOF was decorated with nitrogen-doped graphene (NG) and applied as the matrix for electroactive toluidine blue (Tb) to form the NG@Zr-MOF-on-Ce-MOF@Tb nanohybrid. The prepared nanohybrid with excellent hydrophilicity, dispersibility, and large specific surface exhibited significant electrochemical response. This nanohybrid could be directly used for anchoring ESAT-6 binding aptamers (EBA) through the interaction between the 5′-phosphate group (PO 4 3− ) of EBA and Zr 4+ of Zr-MOF. The signal response before and after incubating the ESAT-6 antigen has been evaluated by cyclic voltammetry at a scan rate of 100 mV s −1 from − 0.7 to 0.3 V (vs. SCE). Under optimal conditions, the proposed aptasensor displayed a wide linear range from 100 fg mL −1 to 10 ng mL −1 with a limit of detection (LOD) of 12 fg mL −1 . The developed method showed good reproducibility with a relative standard deviation (RSD) of 2.27%. The aptasensor showed favorable results in the analysis of the real samples. With these merits, the aptasensor has exceptional potential as a diagnostic tool for tuberculosis in clinical practice. Graphical abstract
研究领域:
分析化学
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生物传感器
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纳米技术
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医学诊断
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DOI
PubMed
高影响因子
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
·
2023
·Vol. 107
·期 7
·2321-2333
·
IF 5
Chi Huibing,Zhu Xiaoyu,Shen Juan,Lu Zhaoxin,Lu Fengxia,Lyu Yunbin,Zhu Ping
Abstract Acrylamide alleviation in food has represented as a critical issue due to its neurotoxic effect on human health. L-Asparaginase (ASNase, EC 3.5.1.1) is considered a potential additive for acrylamide alleviation in food. However, low thermal stability hinders the application of ASNase in thermal food processing. To obtain highly thermal stable ASNase for its industrial application, a consensus-guided approach combined with site-directed saturation mutation (SSM) was firstly reported to engineer the thermostability of Mycobacterium gordonae L-asparaginase (GmASNase). The key residues Gly97, Asn159, and Glu249 were identified for improving thermostability. The combinatorial triple mutant G97T/N159Y/E249Q (TYQ) displayed significantly superior thermostability with half-life values of 61.65 ± 8.69 min at 50 °C and 5.12 ± 1.66 min at 55 °C, whereas the wild-type was completely inactive at these conditions. Moreover, its T m value increased by 8.59 °C from parent wild-type. Interestingly, TYQ still maintained excellent catalytic efficiency and specific activity. Further molecular dynamics and structure analysis revealed that the additional hydrogen bonds, increased hydrophobic interactions, and favorable electrostatic potential were essential for TYQ being in a more rigid state for thermostability enhancement. These results suggested that our strategy was an efficient engineering approach for improving fundamental properties of GmASNase and offering GmASNase as a potential agent for efficient acrylamide mitigation in food industry. Key points
• The thermostability of GmASNase was firstly improved by consensus-guided engineering.
• The half-life and T
m
value of triple mutant TYQ were significantly increased.
• Insight on improved thermostability of TYQ was revealed by MD and structure analysis.
研究领域:
生物化学
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食品科学
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蛋白质工程
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DOI
PubMed
高影响因子
ANALYST
·
2023
·Vol. 148
·期 10
·2308-2315
·
IF 4.2
Ming Zhou,Xin Li,Herui Wen,Bin Huang,Jiali Ren,Jialin Zhang
The simple and efficient detection of nucleic acids is important in the diagnosis of tuberculosis (TB) caused by Mycobacterium tuberculosis (M. tuberculosis). However, base mismatch will lead to false positive and false negative nucleic acid test, which seriously interferes with the accuracy of the final results. Herein, we demonstrated a CRISPR/Cas-9-mediated fluorescent strategy utilizing fluorescence resonance energy transfer (FRET) for the detection of bacteria. High-variable region of M. tuberculosis 16S rDNA fragment was used as the target, and CRISPR/Cas9 was used as the recognition element. The binding of the P1 probe of upconversion nanoparticles (UCNPs) @SiO2-P1 and the P2 probe of Fe3O4@Au-P2 caused the fluorescence quenching of UCNPs. In the presence of the target, the P2 probe hybridized with the target to form double-stranded DNA (dsDNA), which was recognized and cleaved by CRISPR/Cas9, resulting in the breaking of the P1–P2 duplex linkage. UCNPs moved away from Fe3O4@Au under a magnetic field, and the fluorescence signal was restored; bacteria were detected under the excitation of a 980 nm laser source. Using the CRISPR/Cas-9-mediated system, the sensor could distinguish single-base mismatches in 10 bases from the protospacer adjacent motif (PAM) region. The limit of detection (LOD) was 20 CFU mL−1 and the detection time was 2 h. It developed a new way of accurate nucleic acid detection for disease diagnosis.
研究领域:
生物技术
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医学诊断
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纳米技术
·
分子生物学
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DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2026
·
IF 13.2
Zhuojun He,Deliang Liu,Yang Zhou,Yao Zhao,Guiqin Dai,Zhiqiang Lin,Siwei Zhang,Xiafei Dai,Yuying Chen,Jinzhi Pan,Huiling Liu,Hong Yu,Pengfei Zhao,Mingbin Zheng,Hongzhou Lu
Cellulose, as a critical structural component of bacterial biofilms, forms a natural barrier that hinders drug penetration and immune cell invasion, and contributes to persistent tuberculosis (TB) infections and antibiotic resistance. Although Generally Recognized as Safe (GRAS)-affirmed cellulase can hydrolyze cellulose matrix, its clinical application is limited by poor biological availability and anti-biofilm activity. Herein, we developed Nano-GlycOTAC technology, constructing a nano-glycolytic targeting chimera (lactoferrin-cellulase nanoparticles, LCNP) composed of biofilm-binding lactoferrin and cellulase to eradicate biofilm-encased Mycobacterium tuberculosis ( Mtb ). LCNP effectively adhered to biofilms via lactoferrin, released cellulase in response to glutathione (GSH) to degrade cellulose, and subsequently disrupted biofilm and exposed the pathogen. Concurrently, the iron-binding capacity of LCNP blocked Mtb iron uptake and promoted macrophage M1 polarization, thereby enhancing host immunity for bacterial clearance. Consequently, LCNP treatment completely eliminated pulmonary hemorrhage and inflammation in bacteria-infected mice, robustly enhancing the therapeutic efficacy and biosafety of cellulase. In addition, LCNP significantly enhanced biofilm degradation and achieved significant reduction of bacterial load in ex vivo patient samples, including sputum, bronchoalveolar lavage fluid (BALF) and pus. The Nano-GlycOTAC approach combined biofilm disruption, metabolism rectification and immunity activation to normalize Mtb metabolism and remodel host immunity. It emerges as a powerful platform for regulating bacteria fate and activity, and combating biofilm-associated and refractory bacterial infections.
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高影响因子
Microbiology Spectrum
·
2026
·
IF 4.1
Yuwei Qiu,Xu Dong,Dan Cao,Xiuzhi Jiang,Zhongkang Ji,Pusheng Xu,Yi Li,Kaijing Xu,Ying Zhang
Mycobacterium avium-intracellulare complex (MAC) infections are the most common nontuberculous mycobacterial infections, and ethambutol (EMB) is one of the main therapeutic agents used to treat MAC infections. However, the EMB resistance profile and its resistance mechanisms in M. avium remain poorly understood. This study determined the minimum inhibitory concentration (MIC) of 40 M. avium clinical strains, revealing that 97.5% (39/40) of the strains were intermediate (MIC = 4 µg/mL) or resistant (MIC ≥ 8 µg/mL) to EMB. One susceptible clinical isolate strain 245 (MIC = 2 µg/mL) was picked for EMB-resistant mutant isolation, and a total of 121 resistant mutants were isolated and subjected to whole-genome sequencing or Sanger sequencing. Integrated analysis revealed that 94.21% (114/121) of the mutants carried mutations in the ubiA gene, which encodes decaprenylphosphoryl-β-D-5-phosphoribose (DPPR) synthase—an enzyme involved in cell wall biosynthesis that has been associated with high-level EMB resistance in Mycobacterium tuberculosis. Complementation with the wild-type ubiA gene restored EMB susceptibility in EMB-resistant mutants and clinical strain 322 (EMB MIC = 64 µg/mL) with ubiA mutation, reducing the MICs from 32 to 4 μg/mL and from 64 to 8 μg/mL, respectively. This study indicates that the ubiA mutation is the major mechanism of EMB resistance in M. avium, which should facilitate the development of molecular tests for rapid detection of resistance in this organism.
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PubMed
高影响因子
mSphere
·
2025
·
IF 3.1
Xinda Li,Xiaoyi Luo,Bin Wang,Lei Fu,Xi Chen,Yu Lu
Drug-resistant tuberculosis (DR-TB) urgently requires safer, more accessible alternatives to bedaquiline (BDQ), which faces critical flaws like cardiotoxicity, high costs, and emerging resistance. WX-081, a promising BDQ alternative, has demonstrated superior anti-TB activity and improved safety in clinical studies. However, its mechanism of action remains unexplored, underscoring the need for further research to optimize its potential in advancing global TB elimination efforts. This study reveals WX-081’s dual mechanisms: targeting atpE to disrupt ATP synthase and proton motive force via resistance screening, gene sequencing, and functional assays while enhancing host immunity through macrophage transcriptomics. Molecular docking confirmed atpE binding sites, and immune activation pathways (NF-κB/MAPK) were identified, positioning WX-081 as a potent, safe anti-DR-TB candidate despite unresolved mechanistic details.
研究领域:
微生物学
·
药物研发
·
免疫学
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）