SCIENCE
·
2025
·
IF 45.8
Zhifeng Zeng,Zeyu Hu,Ruiliang Zhao,Jikai Rao,Mario Rodríguez Mestre,Yanqiu Liu,Shunhang Liu,Hao Feng,Yu Chen,Huan He,Nuo Chen,Jinshui Zheng,Donghai Peng,Min Luo,Qunxin She,Rafael Pinilla-Redondo,Wenyuan Han
Signaling from pathogen sensing to effector activation is a fundamental principle of cellular immunity. While cyclic (oligo)nucleotides have emerged as key signaling molecules, the existence of other messengers remains largely unexplored. Here, we reveal a bacterial anti-phage system that mediates immune signaling through nucleobase modification. Immunity is triggered by phage nucleotide kinases, which, combined with the system-encoded adenosine deaminase, produce deoxyinosine 5′-triphosphate (dITP) as immune messengers. The dITP signal activates downstream effector to mediate cellular NAD+ depletion, resulting in population-level defense through the death of infected cells. To counteract immune signaling, phages deploy specialized enzymes that deplete cellular dAMP, the precursor of dITP messengers. Our findings uncover a nucleobase modification-based anti-phage signaling pathway, establishing noncanonical nucleotides as a new type of immune messengers in bacteria.
研究领域:
微生物学
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免疫学
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分子生物学
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DOI
PubMed
高影响因子
JOURNAL OF BIOTECHNOLOGY
·
2021
·
IF 3.307
Bo Gao,Ning Zhao,Jieying Deng,Yang Gu,Shiru Jia,Ying Hou,Xueqin Lv,Long Liu
Methanol is a promising green feedstock for producing fuels and chemicals because it is inexpensive, clean, environmentally friendly, and easily prepared. Thus, many studies have been devoted to engineering non-native methylotrophic platform microorganisms to utilize methanol. This study adopted a series of strategies to develop a synthetic methylotrophic Bacillus subtilis that can use methanol as the carbon source, including the heterologous expression of methanol dehydrogenase (Mdh), enhancement of the expressions of 3-hexulose-6-phosphate synthase (Hps) and 6-phospho-3-hexuloisomerase (Phi), regulation of the expressions of key enzymes at both the translational and transcriptional levels, stabilization of the key enzyme expression through a dual-system for expressing the target genes on both the plasmid and genome, and improvement of the catalytic activity of Mdh with a recycling strategy for NAD + . As a result, the methanol consumption of the synthetic methylotrophic B. subtilis reached 4.09 g/L, with the maximum OD 600 showing a 2.21-fold increase compared with the wild-type B. subtilis , which cannot use methanol. We further deleted the phosphoglucose isomerase (Pgi) and added co-substrates to increase the supply of ribulose-5-phosphate (Ru-5-P), and the specific methanol consumption rate increased by an additional 27.54%. Finally, we successfully constructed two strains that cannot grow in M9 medium with xylose or ribose unless methanol is utilized. The strategies used in this study are generally applicable to other studies on synthetic methylotrophy .
研究领域:
微生物学
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代谢工程
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合成生物学
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DOI
PubMed
高影响因子
CHEMISTRY-A EUROPEAN JOURNAL
·
2021
·Vol. 28
·期 3
·e202103346
·
IF 5.236
Zhongxian Fan,Jiaen Xie,Tumpa Sadhukhan,Chao Liang,Can Huang,Wenqing Li,Tingxuan Li,Pingyu Zhang,Samya Banerjee,Krishnan Raghavachari,Huaiyi Huang
Graphical Four novel Ir(III) photo-catalysts ( Ir1 – Ir4 ) were synthesized and applied for photocatalytic cancer therapy. Upon blue and green light irradiation, these photocatalysts generated singlet oxygen and induced rapid photo-catalytic oxidation of the cellular coenzymes NAD(P)H. Ir2 triggered promising photo-induced anticancer activity via induction of intracellular redox imbalance and exhibited ideal in vivo photo-induced catalytic anticancer activity. Four photo-catalysts of the general formula [Ir(CO6/ppy) 2 (L)]Cl where CO6=coumarin 6 ( Ir1 – Ir3 ), ppy=2-phenylpyridine ( Ir4 ), L=4′-(3,5-di-tert-butylphenyl)-2,2′ : 6′,2′′-terpyridine ( Ir1 ), 4′-(3,5-bis(trifluoromethyl)phenyl)-2,2′ : 6′,2′′-terpyridine ( Ir2 and Ir4 ), and 4-([2,2′ : 6′,2′′-terpyridin]-4′-yl)-N,N-dimethylaniline ( Ir3 ) were synthesized and characterized. These photostable photo-catalysts ( Ir1 – Ir3 ) showed strong visible light absorption between 400–550 nm. Upon light irradiation (465 and 525 nm), Ir1 – Ir3 generated singlet oxygen and induced rapidly photo-catalytic oxidation of cellular coenzymes NAD(P)H. Ir1 – Ir3 showed time-dependent cellular uptake with excellent intracellular retention efficiency. Upon green light irradiation (525 nm), Ir2 provided a much higher photo-index (PI=793) than the clinically used photosensitizer, 5-aminolevulinicacid (5-ALA, PI>30) against HeLa cancer cells. The observed necro-apoptotic anticancer activity of Ir2 was due to the Ir2 triggered photo-induced intracellular redox imbalance (by NAD(P)H oxidation and ROS generation) and change in the mitochondrial membrane potential. Remarkably, Ir2 showed in vivo photo-induced catalytic anticancer activity in mouse models.
研究领域:
癌症研究
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光化学
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材料科学
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生物医学工程
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产品关联
DOI
PubMed
高影响因子
SCIENCE OF THE TOTAL ENVIRONMENT
·
2024
·
IF 8.2
Wenmin Li,Yikun Zan,Tong Wu,Sa Yang,Linlin Liu,Shanshan Li,Pingli Dai,Jing Gao
Chlorantraniliprole (CAP), a diamide insecticide, is extensively applied to combat pests in various crops. However, the widespread use of insecticides has raised concerns about their potential impact on pollinators. In the present study, we explored the toxic effects of CAP in two important honey bee species, Apis mellifera and Apis cerana . The 48 h LC 50 values of CAP for A. mellifera and A. cerana was 256.052 mg/L and 109.709 mg/L, implying that A. cerana is more sensitive to CAP. Prolonged exposure to 40 mg/L CAP significantly impaired sucrose responsiveness and climbing activity in both bee species. Both species showed a decrease in GR activity and GSH content with increasing CAP concentration. By contrast, the activities of GST, CAT, P450 and NAD-MDH were increased in both A. mellifera and A. cerana , but the differences between the 10 mg/L and 40 mg/L treatments were less pronounced in A. mellifera . Moreover, the immune related genes exhibited differential responses to CAP when comparing the two species. Low CAP concentrations led to down-regulation in expression of toll but up-regulation in expression of apideacin and hymeopatecin in A. mellifera , whereas A. cerana exhibited minimal changes in these genes. Additionally, CAP significantly inhibited the expression of ER stress response genes gp - 93 and P58 in A. mellifera , while 10 mg/L of CAP promoted P58 expression in A. cerana . Our results highlight species-specific effects with the possible, distinct detoxification mechanisms and immune responses between A. mellifera and A. cerana . These findings serve as a foundation for further evaluating the safety of CAP for honey bee species and offer insights into the scientific use of pesticides.
研究领域:
昆虫学
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毒理学
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环境科学
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DOI
PubMed
高影响因子
International Journal of Nanomedicine
·
2020
·
IF 5.115
Xiang Li,Qian Zou,Jing Zhang,Peng Zhang,Xiong Zhou,Satya Siva Kishan Yalamarty,Xinli Liang,Yali Liu,Qin Zheng,Jianqing Gao
Purpose Folic acid and cyclic arginylglycylaspartic acid peptides were introduced to the surface of negatively charged lipid-coated hybrid polydopamine-cysteine cores for the delivery of epirubicin (EPI) (E/PCF-NPs). The combined chemo-photothermal therapy using E/PCF-NPs for triple-negative breast cancer was evaluated. Materials and Methods The temperature elevation and thermal toxicity of nanoparticles were studied. The morphology and properties of E/PCF-NPs were characterized by transmission electron microscopy, scanning electron microscopy, and atomic force microscopy. Physicochemical properties, including particle size, zeta potential, drug loading, entrapment efficiency (EE%), stability and in vitro release, were determined. The cell viability, reactive oxygen species (ROS) levels, ratios of oxidized nicotinamide adenine dinucleotide to its reduced form (NAD + /NADH), apoptosis assays, and cellular uptake of E/PCF-NPs were determined on 4T1 cells. Pharmacokinetic studies and tissue distributions were performed and detected by an ultra-high performance liquid chromatography/mass spectrometry system. The antitumor effects of E/PCF-NPs under near-infrared (NIR) laser irradiation were also evaluated. Results The sphere-like morphology of E/PCF-NPs showed a high EE%, uniform size of 106.7 nm, remarkable stability, and highly improved cytotoxicity under NIR laser, when compared to that of photothermal treatment alone. In vitro release of EPI from E/PCF-NPs was pH sensitive, and a greater response was achieved under NIR laser irradiation. Compared to chemotherapy or photothermal treatment alone, the combined treatment in vitro significantly inhibited the survival rate of 4T1 cells to 17.7%, induced ROS generation, and reduced NAD + /NADH significantly. Treatment with E/PCF-NPs under irradiation induced 4T1 cell apoptosis in approximately 93.6% cells. In vitro cellular uptake of E/PCF-NPs was time-dependent. The long-circulating and higher tumor accumulation of E/PCF-NPs resulted in complete ablation of breast tumor tissue through the enhanced photothermal effect by NIR laser irradiation-mediated cell apoptosis. Conclusion E/PCF-NPs show enhanced anti-cancer effects due to synergistic effects of chemotherapy with photothermal therapy and may be potential therapeutic agents for cancer treatment.
研究领域:
肿瘤学
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纳米技术
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药学
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生物医学工程
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产品关联
DOI
PubMed
高影响因子
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
·
2021
·Vol. 105
·期 9
·3573-3586
·
IF 4.813
Liu Zhiyong,Zhang Rongzhen,Zhang Wenchi,Xu Yan
Abstract NAD(H)-dependent 7α-hydroxysteroid dehydrogenase catalyzes the oxidation of chenodeoxycholic acid to 7-oxolithocholic acid. Here, we designed mutations of Ile258 adjacent to the catalytic pocket of Brucella melitensis 7α-hydroxysteroid dehydrogenase. The I258M variant gave a 4.7-fold higher k cat , but 4.5-fold lower K M , compared with the wild type, resulting in a 21.8-fold higher k cat / K M value for chenodeoxycholic acid oxidation. It presented a 2.0-fold lower K M value with NAD + , suggesting stronger binding to the cofactor. I258M produced 7-oxolithocholic acid in the highest yield of 92.3% in 2 h, whereas the wild-type gave 88.4% in 12 h. The I258M mutation increased the half-life from 20.8 to 31.1 h at 30 °C. Molecular dynamics simulations indicated increased interactions and a modified tunnel improved the catalytic efficiency, and enhanced rigidity at three regions around the ligand-binding pocket increased the enzyme thermostability. This is the first report about significantly improved catalytic efficiency, cofactor affinity, and enzyme thermostability through single site-mutation of Brucella melitensis 7α-hydroxysteroid dehydrogenase. Key points • Sequence and structure analysis guided the site mutation design. • Thermostability, catalytic efficiency and 7 - oxo - LCA production were determined . • MD simulation was performed to indicate the improvement by I258M mutation .
研究领域:
生物化学
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酶学
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微生物学
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分子生物学
✓ 已验证
DOI
PubMed
高影响因子
Animal Cells and Systems
·
2023
·
IF 2.9
Weiming Yan,Yan Sun,Yutong Wang,Wangjiao Liang,Yuxin Xia,Weihua Yan,Meizhu Chen,Tao Chen,Dongliang Li
Upregulation of Sirtuin Type 1 (SIRT1), a nicotinamide adeninedinucleotide (NAD+)-dependent deacetylase, has been proved to protect against ample ocular diseases, while its effect on retinitis pigmentosa (RP) has not been illustrated. The study was aimed to explore the impacts of resveratrol (RSV), a SIRT1 activator, on the photoreceptor degeneration in a rat model of RP induced by N-methyl-N-nitrosourea (MNU), an alkylation. The rats were induced RP phenotypes via the intraperitoneal injection of MNU. The electroretinogram was conducted and revealed that RSV could not prevent the decline of retinal function in the RP rats. The optical coherence tomography (OCT) and the retinal histological examination were performed and showed that the reduced thickness of the outer nuclear layer (ONL) was not preserved by RSV intervention. The immunostaining technique was applied. Afther the MNU administration, the number of the apoptotic photoreceptors in the ONL throughout the retinasand the number of microglia cells present among the outer part throughout the retinas were not significantly reduced by RSV. Western blotting was also performed. The data showed that the level of SIRT1 protein was decreased after MNU administration, while RSV was not able to obviously alleviate the downregulation. Our data together indicated that RSV was not able to rescue the photoreceptor degeneration in the MNU-induced RP rats, which might be due to the MNU-induced consumption of the NAD+.
研究领域:
眼科学
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神经科学
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药理学
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产品关联
DOI
PubMed
高影响因子
MOLECULAR CELL
·
2023
·
IF 16
Dongmei Tang,Yijun Chen,Hao Chen,Tingting Jia,Qiang Chen,Yamei Yu
In response to the persistent exposure to phage infection, bacteria have evolved diverse antiviral defense mechanisms. In this study, we report a bacterial two-component defense system consisting of a Sir2 NADase and a HerA helicase. Cryo-electron microscopy reveals that Sir2 and HerA assemble into a ∼1 MDa supramolecular octadecamer. Unexpectedly, this complex exhibits various enzymatic activities, including ATPase, NADase, helicase, and nuclease, which work together in a sophisticated manner to fulfill the antiphage function. Therefore, we name this defense system "Nezha" after a divine warrior in Chinese mythology who employs multiple weapons to defeat enemies. Our findings demonstrate that Nezha could sense phage infections, self-activate to arrest cell growth, eliminate phage genomes, and subsequently deactivate to allow for cell recovery. Collectively, Nezha represents a paradigm of sophisticated and multifaceted strategies bacteria use to defend against viral infections.
研究领域:
微生物学
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分子生物学
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结构生物学
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病毒学
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产品关联
DOI
PubMed
高影响因子
Synthetic and Systems Biotechnology
·
2024
·
IF 4.4
Zetian Qiu,Xiaohui Liu,Jie Yu,Yushuo Zhao,Guang-Rong Zhao,Shengying Li,Kun Liu,Lei Du,Li Ma
Benzyl and phenylpropanoid acids are widely used in organic synthesis of fine chemicals, such as pharmaceuticals and condiments. However, biocatalysis of these acids has received less attention than chemical synthesis. One of the main challenges for biological production is the limited availability of alcohol dehydrogenases and aldehyde dehydrogenases. Environmental microorganisms are potential sources of these enzymes. In this study, 129 alcohol dehydrogenases and 42 aldehyde dehydrogenases from Corynebacterium glutamicum , Pseudomonas aeruginosa , and Bacillus subtilis were identified and explored with various benzyl and phenylpropanoid alcohol and aldehyde substrates, among which four alcohol dehydrogenases and four aldehyde dehydrogenases with broad substrate specificity and high catalytic activity were obtained. Moreover, a cascade whole-cell catalytic system including ADH-90, ALDH-40, and the NAD(P)H oxidase LreNox was established, which showed high efficiency in converting cinnamyl alcohol and p -methylbenzyl alcohol into the respective carboxylic acids. Remarkably, this biocatalytic system can be easily scaled up to gram-level production, facilitating preparation purposes.
研究领域:
生物技术
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生物化学
·
有机化学
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DOI
PubMed
高影响因子
Journal of Materials Chemistry B
·
2024
·
IF 6.1
Mingye Li,Yuyu Dong,Zheng Wang,Yanjun Zhao,Yujie Dai,Baoxin Zhang
Glucose-6-phosphate dehydrogenase (G6PD) is a promising target in cancer therapy. However, poor cellular uptake and off-target toxicity impeded the clinical translation of a canonical G6PD inhibitor (6-aminonicotinamide/6AN). Here, we report a prodrug strategy to address this issue. The tailored 6AN prodrug contains an azo-bearing protection moiety. The hydrophobic prodrug showed increased cellular uptake than 6AN and is vulnerable to hypoxia, resulting in NAD(P)H quinone dehydrogenase 1 (NQO1)-triggered cleavage of azo bonds. Intriguingly, the prodrug shows configuration-dependent anti-cancer potency. Despite the lower thermodynamic stability, the cis isomer showed enhanced cellular uptake compared to the trans counterpart due to the increased aqueous solubility. Moreover, the boosted potency of cis isomer compared to the trans isomer resulted from the enhancement of NOQ1-catalyzed 6AN release under hypoxia, a hallmark of solid tumors. The discovery of hypoxia-responsive 6AN prodrugs in the current work opens new avenues of G6PD-targeting cancer medicines via redox manipulation.
研究领域:
药学
·
癌症研究
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药物递送系统
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氧化还原生物学
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产品关联
DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2024
·
IF 13.3
Wenting Li,Jihui Zhou,Yangyang Zhang,Shikai Liu,Rumin Li,Shili Gai,He Ding,Lei Zhong,Piaoping Yang
Photothermal-enhanced chemodynamic therapy (CDT) based on the Fenton reaction has been widely studied as a safe, tumor-specific, and highly effective nanomedicine therapy. Herein, a collective hyperthermia-facilitated glutathione (GSH) inhibition strategy for boosted oxidative stress treatment was presented using mesoporous Prussian blue (MPB NPs) as a multifunctional carrier and loading β-lapachone (Lap) as an oxidative mediator. A phase transfer catalyst (lauric acid, La) was co-packed for preventing drug leakage and achieving temperature-controlled drug release. After biocompatible decoration by external encapsulation with polyethylene glycol (PEG), the MPB-Lap/La@PEG nanocatalysts (Mlalp NCs) were rationally constructed. Upon 808 nm light irradiation, the Mlalp NCs showed a good photothermal effect with temperature increases for directly damaging tumor cells and accelerating the release of Lap. Significantly, the expression of NAD(P)H: quinone oxidoreductase-1 (NQO1) in cells could increase and generate more hydrogen peroxide (H 2 O 2 ) with Lap and reduce the level of GSH concurrently, forming a hyperthermia-facilitated glutathione inhibition strategy. Most importantly, the hyperthermia also accelerated the Fenton reaction rate, resulting in an enhanced CDT effect. In addition, the Mlalp NCs also equipped with excellent T 1 -weighted magnetic resonance imaging properties. To sum up, this work displays a collective hyperthermia-facilitated glutathione inhibition strategy for boosted oxidative stress treatment.
研究领域:
纳米医学
·
癌症治疗
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材料科学
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生物医学工程
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产品关联
DOI
高影响因子
XENOBIOTICA
·
2018
·
IF 1.857
Yue Li,Yujuan Fan,Huizong Su,Qian Wang,Guo-Fu Li,Yiyang Hu,Jian Jiang,Bo Tan,Furong Qiu
Tanshinone I (TSI) is a lipophilic diterpene in Salvia miltiorrhiza with versatile pharmacological activities. However, metabolic pathway of TSI in human is unknown.In this study, we determined major metabolites of TSI using a preparation of human liver microsomes (HLMs) by HPLC-UV and Q-Trap mass spectrometer. A total of 6 metabolites were detected, which indicated the presence of hydroxylation, reduction as well as glucuronidation.Selective chemical inhibition and purified cytochrome P450 (CYP450) isoform screening experiments revealed that CYP2A6 was primarily responsible for TSI Phase I metabolism. Part of generated hydroxylated TSI was glucuronidated via several glucuronosyltransferase (UGT) isoforms including UGT1A1, UGT1A3, UGT1A7, UGT1A9, as well as extrahepatic expressed isoforms UGT1A8 and UGT1A10. TSI could be reduced to a relatively unstable hydroquinone intermediate by NAD(P)H: quinone oxidoreductase 1 (NQO1), and then immediately conjugated with glucuronic acid by a panel of UGTs, especially UGT1A9, UGT1A1 and UGT1A8. Additionally, NQO1 could also reduce hydroxylated TSI to a hydroquinone intermediate, which was immediately glucuronidated by UGT1A1.The study demonstrated that hydroxylation, reduction as well as glucuronidation were the major pathways for TSI biotransformation, and six metabolites generated by CYPs, NQO1 and UGTs were found in HLMs and S9 subcellular fractions.
研究领域:
药理学
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代谢
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药物代谢
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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）