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
ANALYST
·
2023
·Vol. 148
·期 10
·2308-2315
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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
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分子生物学
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高影响因子
WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
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2022
·Vol. 38
·期 5
·1-8
·
IF 4.253
Li Yudi,Chen Guiying,Ge Fanglan,Dang Ting,Ren Yao,Zeng Bei,Li Wei
γ-aminobutyric acid (GABA) has various physiological functions and is widely used in medicine, food, and other fields. Glutamate decarboxylase (GAD) is a key enzyme that catalyzes the decarboxylation of L-glutamate to synthesize GABA. However, the industrial application of microorganism-derived GAD is limited by its rapid loss of enzymatic activity with pH approaching neutrality. In this study, a novel glutamate decarboxylase, GAD MSM, from Mycobacterium smegmatis was overexpressed and purified. On the basis of homologous modeling and substrate molecular docking, several GAD MSM mutants were constructed, and their enzymatic properties were analyzed. The results showed that the optimal pH of wild-type GAD MSM is 5.4; at pH 6.2, 22.8% enzymatic activity was retained. The T211I replacement in GAD and C-terminal deletion mutant GAD MSMΔC showed relatively high catalytic activity in a pH range of 5.0–7.0. The V max and K m values of GAD MSMΔC were 14.69 and 5.70, respectively, at pH 5.5, and 9.87 and 6.17, respectively, at pH 7.0. Compared with the wild-type GAD, GAD MSMΔC maintained higher affinity and enzymatic activity of the substrate, maintaining 78.5% of the highest enzymatic activity even at pH 7.0, which is the highest reported activity retention for GAD under neutral pH condition. Therefore, GAD MSMΔC can be used for the transformation of high-yielding strains and industrial production of GABA.
研究领域:
微生物学
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酶工程
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生物化学
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高影响因子
BMC INFECTIOUS DISEASES
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2015
·Vol. 15
·期 1
·1-7
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IF 2.613
Zhang Zhe,Yan Jie,Xu Kaijin,Ji Zhongkang,Li Lanjuan
Background Tetrandrine is a natural chemical product purified from fourstamen stephania root which recently has been shown to act similarly as synthesized drug efflux pump inhibitor verapamil. The aim of the study is to examine whether tetrandrine could potentiate anti-tubercular drugs to which Mycobacterium tuberculosis (MTB) has turned resistant via efflux mechanisms.Methods We screened 200 MTB clinical isolates using drug sensitivity test to look for those who have turned resistant to the drugs most probably due to efflux mechanisms. We found 29 isoniazid (INH) and ethambutol (EMB) - dual resistant (IEDR) strains. Then we tested if treatment with tetrandrine or verapamil could reverse drug resistance to INH and/or EMB in IEDR isolates.Results There is a parallel resistance among EMB- and INH-resistant strains in the tested clinical isolates. Among INH-resistant strains, 65% was also EMB-resistant. This suggests an involvement of efflux mechanisms which can lead to dual drug resistance in IEDR clinical isolates. Similar to a synthesized efflux pump inhibitor verapamil, tetrandrine treatment together with INH or EMB brought down the MICs from the clinical level of drug resistance to the sensitive range of both drugs. The effective rate reached 82% among IEDR clinical isolates.Conclusion sCombinational application of tetrandrine with INH or EMB increased drug efficacy. Drugs like tetrandrine may help to reduce drug dosage thus alleviate side effects.
研究领域:
微生物学
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药理学
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传染病学
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高影响因子
MICROBIAL PATHOGENESIS
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2021
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IF 3.738
Wenmin Yang,Minqiang Liu,Xi Yu,Yu Huang,Jie Zeng,Yongdong Dai,Hongping Luo,Qinqin Huang,Lin Fan,Jianping Xie
Mycobacterium tuberculosis ( Mtb ) infection is the major cause of tuberculosis. Mtb regions of difference (RD) genes are vital for survival of the pathogen within hosts and for the attenuation of the bacillus Calmette–Guérin vaccine. However, the function of most RD proteins largely remains unexplored. In the present study, we focused on Rv1515c, an RD6 member from M. tuberculosis , and characterised it as a cell surface-associated protein that functions in disrupting the cytokine profile and promoting endoplasmic reticulum stress-mediated apoptosis . Rv1515c expression in M. smegmatis , a nonpathogenic species, resulted in enhanced resistance of the bacterium to various in vitro stressors (such as low pH, sodium dodecyl sulfate , oxidative pressure, and nitrogen intermediate) and its cellular survival within macrophages. Our study is the first to identify the role of Rv1515c in the physiology and pathogenesis of mycobacterium .
研究领域:
微生物学
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免疫学
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传染病学
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PubMed
高影响因子
ARCHIVES OF MICROBIOLOGY
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2021
·Vol. 204
·期 1
·1-16
·
IF 2.552
Haris Muhammad,Chen Changming,Wu Jing,Ramzan Muhammad Noman,Taj Ayaz,Sha Shanshan,Ullah Hayan,Ma Yufang
Tuberculosis (TB) causes millions of deaths each year across the globe. Multiple drug-resistant (MDR) and extensively drug-resistant (XDR) mycobacterial strains have made the treatment extremely difficult. To overcome this hurdle, the development of new drug targets and an effective treatment strategy are desperately needed. This can be achieved by deciphering the role of essential genes and enzymes which are involved in cell survival. One such enzyme is glyoxalase II. The glyoxalase system (glyoxalase I and glyoxalase II) has a pivotal role in cellular survival and detoxification by converting methylglyoxal (MG) into lactate. Otherwise, the increased concentration of MG then modifies DNA, proteins, and lipids, resulting in abnormalities and cell death. Interestingly, the function and physiological role of glyoxalase II have remained undetermined in mycobacteria. In this study, the functional activity of MSMEG_2975 (putative glyoxalase II) after heterologous cloning and expression was determined. And the knockdown strain Mycobacterium smegmatis KD for MSMEG_2975 was constructed with tetracycline-inducible vector pMIND. The inducible knockdown of MSMEG_2975 affected bacterial growth, biofilm formation, transcriptome, and enhanced the susceptibility to antibiotics. This work represents mycobacterial glyoxalase II as a potential drug target against mycobacterial pathogens and indicates the crucial regulatory role of glyoxalase II in mycobacteria.
研究领域:
微生物学
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分子生物学
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药理学
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传染病学
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高影响因子
Biotechnology Journal
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2021
·Vol. 17
·期 2
·2100441
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IF 4.677
Liuzhu Wang,Yangyang Sun,Shiqing Diao,Shuiqin Jiang,Hualei Wang,Dongzhi Wei
Background: Carboxylic acid reductases (CARs) represent useful tools for the production of aldehydes from ubiquitous organic carboxylic acids. However, the low catalytic efficiency of these enzymes hampers their application. Methods: Herein, a CAR originating from Mycobacterium smegmatis was redesigned through rational hinge engineering to enhance the catalytic efficiency. Results: Based on the unique domain architecture of CARs and their superfamily, a mutagenesis library of the hinge region was designed. The best mutant R505I/N506K showed a 6.57-fold improved catalytic efficiency. Molecular dynamics simulations showed the increased catalytic efficiency was due to the strong binding of the acyl-AMP complex with it. Meanwhile, the ε-nitrogen atom of Lys610 frequently interacted with the ribose-ring oxygen atom of the complex, the distance (d1) between them represents a great indicator for that. The d1 value was used as a nimble indicator to evaluate unexplored mutants of that region for enhanced activity by in silico mutational experiments. Overall, eight mutants were identified to show higher enhanced activity compared with wild-type enzyme and R505F/N506G showed the highest catalytic efficiency. Conclusion: Altogether, the two-step strategy used here provided useful references for the engineering of CARs and other similar multiple-domain enzymes. Graphical and Lay Summary Carboxylic acid reductases (CARs) provide useful tools for the production of aldehydes from organic carboxylic acids but the low catalytic efficiency hinders their application. In this study, the catalytic efficiency of CARs originating from Mycobacterium smegmatis was rationally enhanced through the two-step hinge region engineering. And the method used provides useful guidance for engineering other CARs and related enzymes.
研究领域:
生物化学
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酶工程
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计算生物学
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高影响因子
IONICS
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2024
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IF 2.4
Qiu Shengzhou,Wu Chenxi,Akter Sima,Hong Shaoli,Liu Huihong,Mahmud Sakil
This study presents the development of copper oxide–modified screen-printed carbon electrodes (CuO/SPCE) for the sensitive electrochemical detection of isoniazid (INZ). The electrodeposition of CuO on the surface of SPCE in a dilute CuSO 4 solution forms CuO amorphisms or clusters, which catalyze the oxidation of INZ. The sensor exhibited a linear response to INZ concentrations ranging from 4 to 200 μM with a correlation coefficient of 0.991 and a limit of detection (LoD) of 8.39 μM. Stability and reproducibility were confirmed through repetitive measurements with relative standard deviations of 0.27% and 1.20%, respectively. Interference studies demonstrated the sensor’s selectivity toward INZ, and a real sample analysis of pharmaceutical tablets yielded recovery rates ranging from 99.50 to 105.03%. Hence, regular monitoring of INZ dosage levels in human body fluids is critical, necessitating the development of sensitive, effective, and consistent methods to quantify INZ in clinical and pharmaceutical samples. Graphical Abstract
研究领域:
医学诊断
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分析化学
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生物传感技术
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高影响因子
ANALYTICAL CHEMISTRY
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2020
·
IF 6.785
Jiao Cao,Shanyu Zhang,Jinyue Zhang,Sha Wang,Wendong Jia,Shuanghong Yan,Yuqin Wang,Panke Zhang,Hong-Yuan Chen,Shuo Huang
Gold(I) compounds are known to bind sulfur-containing proteins, forming the basis in the design of gold(I)-based drugs. However, the intrinsic molecular mechanism of the chemical reaction is easily hidden when monitored in ensemble. We have previously demonstrated that Mycobacterium smegmatis porin A (MspA) can be engineered (MspA-M) to contain a specialized nanoreactor to probe chemical reactions involving tetrachloroaurate(III). Here, we provide further investigations of coordination interactions between dichloroaurate(I) and MspA-M. Gold compounds of different coordination geometry and valence states are as well probed and evaluated, demonstrating the generality of MspA-M. With single-molecule evidence, MspA-M demonstrates a preference for dichloroaurate(I) than tetrachloroaurate(III), an observation in a single molecule that has never been reported. By counting the maximum number of simultaneous ion bindings, the narrowly confined pore restriction also efficiently distinguishes dichloroaurate(I) and tetrachloroaurate(III) according to their differences in geometry or size. The above demonstration complemented a previous study by demonstrating other possible gold-based single-molecule chemical reactions observable by MspA. These observations bring insights in the understanding of gold-based coordination chemistry in a nanoscale.
研究领域:
化学
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纳米技术
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生物物理学
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材料科学
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高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
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2024
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IF 3.8
Qiao Yinuo,Wang Xiaoyan,Fan Zhenlin,Song Yuzhu,Zhang Jinyang,Han Qinqin
Tuberculosis is a highly infectious disease caused by the bacterium Mycobacterium tuberculosis , and the spread of this agent has caused serious health problems worldwide. The rapid and accurate detection of M. tuberculosis is essential for controlling the spread of infection and for preventing the emergence of multidrug-resistant strains. In this study, the powerful trans-cleavage ability of CRISPR-Cas12a for ssDNA was combined with a surface-enhanced Raman spectroscopy (SERS)–based strategy to establish a CRISPR-SERS sensor for the hypersensitive detection of M. tuberculosis DNA. We observed a linear relationship between the concentration of M. tuberculosis DNA and the output signal over the range of 5 to 100 pM. The equation describing the standard curve was y = 24.10 x + 1594, with R 2 = 0.9914. The limit of detection was as low as 4.42 pM for genomic DNA, and a plasmid containing an M. tuberculosis –specific sequence was detected at 5 copy/μL. A detection accuracy of 100% was achieved in the analysis of DNA isolated from the sputum of hospitalized patients with tuberculosis. The entire detection process is simple to deploy and only takes 50 min and results in the sensitive and specific detection of M. tuberculosis DNA. This study provides a new method for the detection of tuberculosis. The tool is stable and can be utilized on-site, and it thus broadens the diagnostic application of CRISPR-Cas12a-based sensor technology.
研究领域:
医学诊断
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分子生物学
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PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2025
·
IF 8.5
Cuiying Lin,Yang Liu,Weiqi Wan,Junbo Gao,Lianli Sun
As the number of tuberculosis cases worldwide has increased, the enzyme hypoxanthine-guanine phosphoribosyltransferase from Mycobacterium tuberculosis ( Mt HGPRT) has emerged as a promising target for drug development because of its role in purine salvage and nucleotide homeostasis. However, the bidirectional catalytic mechanism remains poorly understood, posing challenges for rational drug design. Here, residue-specific functional mapping via systematic mutagenesis coupled with molecular dynamics reveals key residues governing the forward and reverse catalysis. D123 stabilizes α-D-5-phosphoribosyl-1-pyrophosphate binding in the forward reaction, whereas V124 and K154 enhance purine ring stability in the reverse reaction. In addition, D126, V176, L181, and D182 regulate substrate coordination and active site conformation. Cross-species validation confirmed the conserved roles of D123 and V124. These findings provide a structural blueprint for enhancing our understanding of the catalytic mechanism of Mt HGPRT and offer insights into drug design targeting HGPRT-related diseases and anti-tuberculosis therapies.
研究领域:
结构生物学
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生物化学
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分子生物学
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药物研发
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微生物学
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计算生物学
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PubMed
高影响因子
INFECTION GENETICS AND EVOLUTION
·
2021
·
IF 3.342
M.A. Abo-Kadoum,Mohammed Assad,Moure UAE,Stech A.E. Nzaou,Zhen Gong,Asmaa Moaaz,Samson Teweldebrhan,Adel Eltoukhy,Ai Xuefeng,Yu Chen,Jianping Xie
Mycobacterium tuberculosis (Mtb) survival and virulence largely reside on its ability to manipulate the host immune response . We have previously shown that M. tuberculosis Raf kinase inhibitor protein (RKIP) Rv2140c regulates diverse phosphorylation events in M. smegmatis . However, its role during infection is unknown. In this report, we show that Rv2140c can mimic the mammalian RKIP function. Rv2140c inhibit the activation of extracellular signal-regulated kinase (ERK) and nuclear factor κB (NF-κB) via decreasing the phosphorylation capacity of upstream mediators MEK1, ERK1/2, and IKKα/β, thus leading to a reduction in pro-inflammatory cytokines IL-1β, IL-6, and TNF-α. This effect can be reversed by RKIP inhibitor locostatin. Furthermore Rv2140c mediates apoptosis associated with activation of caspases cascades. This modulation enhances the intracellular survival of M. smegmatis within macrophage. We propose that Rv2140c is a multifunctional virulence factor and a promising novel anti-Tuberculosis drug target.
研究领域:
微生物学
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免疫学
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传染病学
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PubMed
高影响因子
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