ChemPhotoChem
·
2022
·Vol. 6
·期 8
·e202200067
·
IF 3.679
Yaoqiang Wang,Yu Jin,Zishuai Wang,Gang Xiao,Haijia Su
Graphical Cascading : A two-pot, light-dark cascade NADH regeneration procedure was developed. First, g-C 3 N 4 nanosheets photocatalyzed the conversion of triethanolamine (TEOA) to glycolaldehyde, which reduced NAD+ to NADH in the dark reaction step, thus avoiding the photo-deactivation of NADH. Photocatalytic regeneration of NAD(P)H is a green and promising method which utilizes clean solar energy. However, we found in this study that light exposure of NADH will cause the deactivation of the regenerated NADH. Therefore, we used ultrathin g-C 3 N 4 nanosheets as photocatalyst and developed a two-pot light-dark cascade procedure for NADH regeneration to avoid the light exposure and the deactivation of NADH. This two-pot procedure included a first light reaction step where triethanolamine was converted to glycolaldehyde through visible light photocatalysis which could reduce NAD + to NADH through a non-catalytic process in the followed dark reaction step. Using this procedure, the NADH productivity reached 1.625 mmol ⋅ h −1 ⋅ g −1 and was about 29 times higher that obtained in the one-pot method. The regenerated NADH using the two-pot procedure could act as cofactor of alcohol dehydrogenase to catalyze the reduction of acetaldehyde, illustrating the application potential of this procedure to aid enzyme catalyzed redox reactions.
研究领域:
化学
·
材料科学
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生物技术
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环境科学
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DOI
高影响因子
ANALYTICAL CHEMISTRY
·
2021
·
IF 6.986
Qiong Hu,Luofeng Su,Zhuohua Chen,Yanyu Huang,Dongdong Qin,Li Niu
Trypsin is a key proteolytic enzyme in the digestive system and its abnormal levels are indicative of some pancreatic diseases. Taking advantage of the coenzyme-mediated electrografting of ferrocenyl polymers as a novel strategy for signal amplification, herein, a signal-on cleavage-based electrochemical biosensor is reported for the highly selective interrogation of trypsin activity at ultralow levels. The construction of the trypsin biosensor involves (i) the immobilization of peptide substrates (without free carboxyl groups) via the N-terminus, (ii) the tryptic cleavage of peptide substrates, (iii) the site-specific labeling of the reversible addition–fragmentation chain transfer (RAFT) agents, and (iv) the grafting of ferrocenyl polymers through the electro-RAFT (eRAFT) polymerization, which is mediated by potentiostatic reduction of nicotinamide adenine dinucleotide (NAD+) coenzymes. Through the NAD+-mediated eRAFT (NAD+-eRAFT) polymerization of ferrocenylmethyl methacrylate (FcMMA), the presence of a few tryptic cleavage events can eventually result in the recruitment of a considerable amount of ferrocene redox tags. Obviously, the NAD+-eRAFT polymerization is low-cost and easy to operate as a highly efficient strategy for signal amplification. As expected, the as-constructed biosensor is highly selective and sensitive toward the signal-on interrogation of trypsin activity. Under optimal conditions, the detection limit can be as low as 18.2 μU/mL (∼72.8 pg/mL). The results also demonstrate that the as-constructed electrochemical trypsin biosensor is applicable to inhibitor screening and the interrogation of enzyme activity in the presence of complex sample matrices. Moreover, it is low-cost, less susceptible to false-positive results, and relatively easy to fabricate, thus holding great potential in diagnostic and therapeutic applications.
研究领域:
分析化学
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生物化学
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生物技术
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电化学
·
酶学
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产品关联
DOI
PubMed
高影响因子
Chemical Biology & Drug Design
·
2020
·Vol. 96
·期 6
·1433-1446
·
IF 2.548
Yibing Guo,Liancheng Xu,Changchun Ling,Tao Yang,Wenjie Zheng,Jin Lv,Qingsong Guo,Bohua Chen
Eighteen new β-carboline-based indole-4,7-quinone derivatives ( 12a–i and 13a–i ) were designed and synthesized, and their in vitro and in vivo antiproliferative activities were studied. Most of target compounds showed strong inhibition on three human tumor cells' proliferation. In particular, the most active compound 13g not only displayed more prominent antiproliferative activities than β-lapachone, a clinical antitumor candidate, but also exerted significant NAD(P)H: quinone-oxidoreductase-1 (NQO1) inhibitory activity and NQO1-dependent cytotoxicity in HT29 cells. Furthermore, 13g dose-dependently induced high ROS levels in HT29 cells, and selectively inhibited cancer cell but not non-tumor colon cell proliferation in vitro. Importantly, 13g promoted HT29 cell apoptosis and DNA damage by regulating relative apoptotic proteins and H2AX expression. Finally, 13g displayed significant growth inhibition of HT29 human colorectal adenocarcinoma xenograft in mice without overt toxicity.
研究领域:
药理学
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肿瘤学
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药物化学
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DOI
PubMed
高影响因子
RSC Advances
·
2013
·Vol. 3
·期 44
·21848-21855
·
IF 2.562
Wan-Qian Guo,Shan-Shan Yang,Ji-Wei Pang,Jie Ding,Xian-Jiao Zhou,Xiao-Chi Feng,He-Shan Zheng,Nan-Qi Ren
In order to increase hydrogen yield and shorten the lag time, low frequency ultrasonic (LFU) pretreatment was applied to stimulate the bio-activity of the seed bacteria, using waste activated sludge as the inoculum. A pretreated, sterilized sludge sample was seeded as the substrate for bio-hydrogen production. A Unisense microsensor multimeter with a hydrogen microelectrode was employed to examine the hydrogenase activities at different ultrasonic densities and exposure times. The optimum conditions were achieved at an ultrasonic density of 125 W L−1 and a reaction time of 10 s. A maximum hydrogen yield of 13.03 mL H2/g TS was obtained by LFU pretreatment under these optimal conditions, which was 18% higher than that obtained when seeding the raw sludge sample. To gain a better understanding of the end product distribution, three-dimensional excitation–emission matrix (3D-EEM) fluorescence spectroscopy was performed to characterize the dissolved organic matter (DOM) and extracellular polymeric substances (EPS) in the sludge samples during the anaerobic fermentation process. Hydrogen production from NADH/NAD+ conversion was also proved by EEM.
研究领域:
环境工程
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生物技术
·
微生物学
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DOI
高影响因子
GREEN CHEMISTRY
·
2024
·
IF 9.3
Lin Fan,Shangshang Sun,Zhidan Zhang,Yanmei Qin,Peter Ruhdal Jensen,Chun You
In conjunction with the pressing global issue of climate change and the associated concern over global warming, increasing interest has emerged in the exploration of carbon dioxide (CO2) as a resource for the generation of a diverse array of products intended to serve societal needs. This study presents the development of an ATP-free and NAD-balanced in vitro synthetic enzymatic biosystem (ivSEB), which comprises only five cascade thermophilic enzymes, designed for the synthesis of L-malate through CO2 fixation powered by the utilization of D-glucose as a substrate. This designed ivSEB yields two moles of L-malate from one mole of D-glucose and two moles of CO2. Through meticulous refinement of reaction conditions and enzyme loading amounts, this ivSEB has demonstrated its capability to produce 6.85 mM of L-malate via CO2 fixation from an initial 5 mM of D-glucose with a molar product yield of 68.5%, and 2.45 mM of L-lactate as a byproduct. In the pursuit of assessing the industrial feasibility of this ivSEB, the study further subjected the system to the utilization of a high concentration (45.70 mM) of D-glucose. Although this endeavor necessitates additional optimization for enhanced efficiency, the present findings herald the emergence of an alternative avenue for the sustainable production of L-malate through CO2 fixation, thus bearing substantial promise for addressing ecological and industrial imperatives.
研究领域:
生物技术
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环境科学
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化学工程
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可持续化学
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酶学
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DOI
高影响因子
TOXICOLOGY AND APPLIED PHARMACOLOGY
·
2025
·
IF 3.4
Ya-Xin Yao,Xue Lu,Zhao Li,Hao-Yu Liao,Zhi-Bing Liu,Hui Zhao,Hua Wang,De-Xiang Xu,Zhu-Xia Tan
Accumulating data demonstrate that bleomycin (BLM), a clinically used antineoplastic drug, induces pulmonary interstitial fibrosis. However, the specific mechanism remains unclear. This study was carried out for the purpose of evaluating the effect of mitochondrial dysfunction-associated senescence on BLM-induced pulmonary fibrosis. Adult C57BL/6 J mice were intratracheally instilled with BLM (2.5 mg/kg). Pulmonary α-SMA and Vimentin, two indicators of epithelial-mesenchymal transition (EMT), were increased, and pulmonary collagen deposition was shown in BLM-treated mice. Pulmonary p16 and p21, two biomarkers of cell cycle arrest, were elevated, and pulmonary SASP indicators were up-regulated in BLM-treated mice. The reduction of mitochondrial area in BLM-treated mouse lungs was revealed by transmission electron microscopy. ATP content was reduced in BLM-treated mouse lungs. Pulmonary SOD2 and IDH2, two enzymes located in mitochondria, were decreased in BLM-treated mice. Sirtuin3 (SIRT3), an NAD + -dependent deacetylase located in mitochondria, was down-regulated in BLM-treated mouse lungs. Interestingly, Sirt3 gene knockout aggravated BLM-evoked mitochondrial dysfunction-associated senescence in mouse lungs. Sirt3 gene knockout exacerbated BLM-induced lung fibrosis. Conversely, nicotinamide mononucleotide (NMN), an NAD + precursor, weakened BLM-induced down-regulation of mitochondrial SIRT3 activity in mouse lungs. NMN pretreatment attenuated BLM-induced mitochondrial dysfunction-associated senescence in mouse lungs. Finally, NMN pretreatment alleviated BLM-induced EMT and lung fibrosis. These results indicate that mitochondrial dysfunction-associated senescence partially contributed to BLM-induced pulmonary fibrosis.
研究领域:
分子生物学
·
细胞生物学
·
呼吸病学
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药理学
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遗传学
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生物化学
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产品关联
DOI
PubMed
高影响因子
Catalysis Science & Technology
·
2024
·
IF 4.4
Jianhua Liu,Jiashu Li,Fangshu Xing,Jian Liu
Developing biological cofactor regeneration strategies is crucial for photo-enzyme coupling systems. Here, crystalline poly(triazine imide) (PTI) containing the cyano group (–CN) is designed for photocatalytic nicotinamide cofactor oxidation regeneration via π–π interaction. The π-conjugation degree and electron delocalization of PTI can be regulated through –CN modification. Such a unique extended π-conjugated system promotes the adsorption of the reduced-state nicotinamide cofactor (NADH) on the aromatic skeleton. Moreover, the enhanced charge transfer dynamics expedite the formation of intermediate active species. Accordingly, the optimized conversion rate of NADH to NAD+ reached 98.2% within 60 min. In contrast, pristine PTI and amorphous polymeric carbon nitride (PCN) gave oxidation yields of 88.1% and 56.8%, respectively. The exclusive selectivity of the oxidized product was further verified by the enzymatic method and nuclear magnetic resonance characterization. This study introduces an innovative method for modulating non-covalent interactions for photo-enzymatic catalysis via the extended π-conjugated system.
研究领域:
材料科学
·
光化学
·
生物催化
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产品关联
DOI
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2018
·
IF 3.412
Cun-Duo Tang,Hong-Ling Shi,Jian-He Xu,Zhu-Jin Jiao,Fei Liu,Peng-Ju Ding,Hong-Fei Shi,Lun-Guang Yao,Yun-Chao Kan
d-Mandelate dehydrogenase (DMDH) has the potential to convert d-mandelic acid to phenylglyoxylic acid (PGA), which is a key building block in the field of chemical synthesis and is widely used to synthesize pharmaceutical intermediates or food additives. A novel NAD+-dependent d-mandelate dehydrogenase was cloned from Lactobacillus harbinensi (LhDMDH) by genome mining and expressed in Escherichia coli BL21. After being purified to homogeneity, the oxidation activity of LhDMDH toward d-mandelic acid was approximately 1200 U·mg–1, which was close to four times the activity of the probe. Meanwhile, the kcat/Km value of LhDMDH was 28.80 S–1·mM–1, which was distinctly higher than the probe. By coculturing two E. coli strains expressing LhDMDH and LcLDH, we developed a system for the efficient synthesis of PGA, achieving a 60% theoretical yield and 99% purity without adding coenzyme or cosubstrate. Our data supports the implementation of a promising strategy for the chiral resolution of racemic mandelic acid and the biosynthesis of PGA.
研究领域:
生物化学
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生物技术
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酶学
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微生物学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL BIOCHEMISTRY
·
2024
·
IF 2.6
Mengyu Jiang,Li Li,Yuan Jin,Liuliu Lu,Zhenchen Lu,Wangjie Lv,Xiaoqun Wang,Lei Di,Zhicheng Liu
Sepsis is a life-threatening condition characterized by organ dysfunction resulting from a dysregulated host response to infection. Dysregulated tryptophan (TRP) metabolites serve as significant indicators for endogenous immune turnovers and abnormal metabolism in the intestinal microbiota during sepsis. Therefore, a high coverage determination of TRP and its metabolites in sepsis is beneficial for the diagnosis and prognosis of sepsis, as well as for understanding the underlying mechanism of sepsis development. However, similar structures in TRP metabolites make it challenging for separation and metabolite identification. Here, high-performance liquid chromatography coupled with a diode array detector (HPLC-DAD) was developed to determine TRP metabolites in rat serum. The first-order derivative spectrophotometry of targeted metabolites in the serum was investigated and proved to be promising for chromatographic peak annotation across different columns and systems. The established method separating the targeted metabolites was optimized and validated to be sensitive and accurate. Application of the method revealed dysregulated TRP metabolites, associated with immune disorders and NAD + metabolism in both the host and gut flora in septic rats. Our findings indicate that the derivative spectrophotometry-assisted method enhances metabolite identifications for the chromatographic systems based on DAD detectors and holds promise for precision medicine in sepsis.
研究领域:
分析化学
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免疫学
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代谢组学
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微生物学
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产品关联
DOI
PubMed
高影响因子
BIOORGANIC CHEMISTRY
·
2023
·
IF 5.1
Zhishang Zhang,Wenqing Li,Haorui Wu,Zhuangfeng Liu,Huaiyi Huang
The challenge of antibiotic resistance worldwide has brought an urgent need to explore novel drugs for bacterial infections. Antimicrobial photodynamic therapy has been proven to be a potential antimicrobial modality but is limited by biofilms. In this study, we synthesized three cationic photosensitizers with strong photoinduced antimicrobial and antibiofilm activities toward gram-positive Staphylococcus aureus . The indole–pyridine compounds illustrated multiple type I/II photosensitization and coenzyme NAD(P)H photocatalytic activity upon excitation. A mechanistic study showed that intracellular reactive oxygen generation and NAD(P)H oxidation caused membrane damage, leading to protein/nucleus acid leakage. This research provides insights into the development of novel chemotherapeutics with synergetic photodynamic and photocatalytic reactivity.
研究领域:
药学
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微生物学
·
光化学
·
抗菌研究
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产品关联
DOI
PubMed
高影响因子
Antibiotics-Basel
·
2024
·Vol. 13
·期 6
·528
·
IF 4.3
Muchen Zhang,Huangwei Song,Siyuan Yang,Yan Zhang,Yunrui Tian,Yang Wang,Dejun Liu
The emergence of carbapenem-resistant Gram-negative pathogens presents a clinical challenge in infection treatment, prompting the repurposing of existing drugs as an essential strategy to address this crisis. Although the anticancer drug 5-fluorouracil (5-FU) has been recognized for its antibacterial properties, its mechanisms are not fully understood. Here, we found that the minimal inhibitory concentration (MIC) of 5-FU againstEscherichia coliwas 32–64 µg/mL, including strains carryingblaNDM-5, which confers resistance to carbapenems. We further elucidated the antibacterial mechanism of 5-FU againstE. coliby using genetic and biochemical analyses. We revealed that the mutation of uracil phosphoribosyltransferase-encoding geneuppincreased the MIC of 5-FU againstE. coliby 32-fold, indicating the role of theuppgene in 5-FU resistance. Additionally, transcriptomic analysis ofE. colitreated with 5-FU at 8 µg/mL and 32 µg/mL identified 602 and 1082 differentially expressed genes involved in carbon and nucleic acid metabolism, DNA replication, and repair pathways. The biochemical assays showed that 5-FU induced bacterial DNA damage, significantly increased intracellular ATP levels and the NAD+/NADH ratio, and promoted reactive oxygen species (ROS) production. These findings suggested that 5-FU may exert antibacterial effects onE. colithrough multiple pathways, laying the groundwork for its further development as a therapeutic candidate against carbapenem-resistant bacterial infections.
研究领域:
微生物学
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药理学
·
遗传学
·
生物化学
·
传染病学
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产品关联
DOI
PubMed
高影响因子
Redox Biology
·
2024
·
IF 10.7
Bi-Xin Xu,Tian-Yu Hu,Jin-Biao Du,Tao Xie,Ya-Wen Xu,Xin Jin,Si-Tao Xu,Hao-Wen Jin,Guangji Wang,Jiankun Wang,Le Zhen
Redox-responsive hydropersulfide prodrugs are designed to enable a more controllable and efficient hydropersulfide (RSSH) supply and to thoroughly explore their biological and therapeutic applications in oxidative damage. To obtain novel activation patterns triggered by redox signaling, we focused on NAD(P)H: quinone acceptor oxidoreductase 1 (NQO1), a canonical antioxidant enzyme, and designed NQO1-activated RSSH prodrugs. We also performed a head-to-head comparison of two mainstream structural scaffolds with solid quantitative analysis of prodrugs, RSSH, and metabolic by-products by LC-MS/MS, confirming that the perthiocarbamate scaffold was more effective in intracellular prodrug uptake and RSSH production. The prodrug was highly potent in oxidative stress management against cisplatin-induced nephrotoxicity. Strikingly, this prodrug possessed potential feedback activation properties by which the delivered RSSH can further escalate the prodrug activation via NQO1 upregulation. Our strategy pushed RSSH prodrugs one step further in the pursuit of efficient release in biological matrices and improved druggability against oxidative stress.
研究领域:
药学
·
生物化学
·
分子生物学
·
毒理学
✓ 已验证
DOI
PubMed
高影响因子
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