JOURNAL OF PHYSICAL CHEMISTRY B
·
2020
·
IF 2.857
Simin Cao,Haoyang Li,Yangyi Liu,Mengyu Wang,Mengjie Zhang,Sanjun Zhang,Jinquan Chen,Jianhua Xu,Jay R. Knutson,Ludwig Brand
The fluorescence of dinucleotide NADH has been exploited for decades to determine the redox state of cells and tissues in vivo and in vitro. Particularly, nanosecond (ns) fluorescence lifetime imaging microscopy (FLIM) of NADH (in free vs bound forms) has recently offered a label-free readout of mitochondrial function and allowed the different “pools” of NADH to be distinguished in living cells. In this study, the ultrafast fluorescence dynamics of NADH–dehydrogenase (MDH/LDH) complexes have been investigated by using both a femtosecond (fs) upconversion spectrophotofluorometer and a picosecond (ps) time-correlated single photon counting (TCSPC) apparatus. With these enhanced time-resolved tools, a few-picosecond decay process with a signatory spectrum was indeed found for bound NADH, and it can best be ascribed to the solvent relaxation originating in “bulk water”. However, it is quite unlike our previously discovered ultrafast “dark” component (∼26 ps) that is prominent in free NADH (Chemical Physics Letters2019, 726, 18–21). For these two critical protein-bound NADH exemplars, the decay transients lack the ultrafast quenching that creates the “dark” subpopulation of free NADH. Therefore, we infer that the apparent ratio of free to bound NADH recovered by ordinary (>50 ps) FLIM methods may be low, since the “dark” molecule subpopulation (lifetime too short for conventional FLIM), which effectively hides about a quarter of free molecules, is not present in the dehydrogenase-bound state.
研究领域:
生物化学
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细胞生物学
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医学影像学
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产品关联
DOI
PubMed
高影响因子
BIOORGANIC CHEMISTRY
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2022
·
IF 5.307
Bin Huang,Kai Yang,Charles Amanze,Zhen Yan,Hongbo Zhou,Xueduan Liu,Guanzhou Qiu,Weimin Zeng
7β-Hydroxysteroid dehydrogenases (7β-HSDHs) have attracted increasing attention due to their crucial roles in the biosynthesis of ursodeoxycholic acid (UDCA). However, most published 7β-HSDHs are strictly NADPH-dependent oxidoreductases with poor activity and low productivity. Compared with NADPH, NADH is more stable and cheaper, making it the more popular cofactor for industrial applications of dehydrogenases. Herein, by using a sequence and structure-guided genome mining approach based on the structural information of conserved cofactor-binding motifs, we uncovered a novel NADH-dependent 7β-HSDH ( Cle 7β-HSDH). The Cle 7β-HSDH was overexpressed, purified, and characterized. It exhibited high specific activity (9.6 U/mg), good pH stability and thermostability, significant methanol tolerance, and showed excellent catalytic efficiencies ( k cat / K m ) towards 7-oxo-lithocholic acid (7-oxo-LCA) and NADH (70.8 mM -1 s −1 and 31.8 mM -1 s −1 , respectively). Molecular docking and mutational analyses revealed that Asp42 could play a considerable role in NADH binding and recognition. Coupling with a glucose dehydrogenase for NADH regeneration, up to 20 mM 7-oxo-LCA could be completely transformed to UDCA within 90 min by Cle 7β-HSDH. This study provides an efficient approach for mining promising enzymes from genomic databases for cost-effective biotechnological applications.
研究领域:
生物技术
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生物化学
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酶学
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药学
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产品关联
DOI
PubMed
高影响因子
INDUSTRIAL & ENGINEERING CHEMISTRY RESEARCH
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2019
·
IF 3.375
Biaobiao Ma,Shiyong Sun,Huichao He,Rui Lv,Jianjun Deng,Tingting Huo,Yulian Zhao,Haili Yu,Lei Zhou
Photocatalysis is a green and sustainable technique for converting solar energy into energy-storage molecules. A novel type of metal-free photocatalyst consisting of polydopamine (PDA) surface-functionalized graphitic carbon nitride (g-C3N4) was fabricated to enable the highly efficient visible-light-driven photocatalytic production of reduced nicotinamide adenine dinucleotide (NADH) through the photomediated reduction of NAD+. The photocatalytic performance and kinetic processes were investigated to evaluate the efficiency of NADH regeneration by the PDA/g-C3N4 photocatalysts. It was shown that the PDA layer thus formed could promote the charge carrier transfer of g-C3N4 to increase the solar-to-chemical conversion efficiency under visible light irradiation. A photocurrent density three times higher than that of pristine g-C3N4 was obtained. The photocatalytic activity of the PDA/g-C3N4 photocatalysts appeared to be dependent on the pH, with an increase in the pH from 3.0 to 9.5 substantially enhanced the NADH yield. The 20 wt % PDA/g-C3N4 exhibited remarkably improved photocatalytic NADH regeneration efficiency, achieving an NADH yield of 94.7% with an initial reaction rate of 2.4 mmol·h–1·g–1, which is approximately six times higher than that of pristine g-C3N4. Our study thus identifies a promising photocatalyst for enhancing the photocatalytic efficiency for nicotinamide cofactor regeneration.
研究领域:
材料科学
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化学
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可持续能源
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催化
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产品关联
DOI
高影响因子
MOLECULES
·
2024
·Vol. 29
·期 10
·2354
·
IF 4.2
Jia-Le Zhou,Hao-Han Chen,Jian Xu,Ming-Yu Huang,Jun-Feng Wang,Hao-Jie Shen,Sheng-Xiang Shen,Cheng-Xian Gao,Chao-Dong Qian
Background:Staphylococcus aureusis a common pathogenic microorganism in humans and animals. Type II NADH oxidoreductase (NDH-2) is the only NADH:quinone oxidoreductase present in this organism and represents a promising target for the development of anti-staphylococcal drugs. Recently, myricetin, a natural flavonoid from vegetables and fruits, was found to be a potential inhibitor of NDH-2 ofS. aureus. The objective of this study was to evaluate the inhibitory properties of myricetin against NDH-2 and its impact on the growth and expression of virulence factors inS. aureus. Results: A screening method was established to identify effective inhibitors of NDH-2, based on heterologously expressedS. aureusNDH-2. Myricetin was found to be an effective inhibitor of NDH-2 with a half maximal inhibitory concentration (IC50) of 2 μM. In silico predictions and enzyme inhibition kinetics further characterized myricetin as a competitive inhibitor of NDH-2 with respect to the substrate menadione (MK). The minimum inhibitory concentrations (MICs) of myricetin againstS. aureusstrains ranged from 64 to 128 μg/mL. Time–kill assays showed that myricetin was a bactericidal agent againstS. aureus. In line with being a competitive inhibitor of the NDH-2 substrate MK, the anti-staphylococcal activity of myricetin was antagonized by MK-4. In addition, myricetin was found to inhibit the gene expression of enterotoxin SeA and reduce the hemolytic activity induced byS. aureusculture on rabbit erythrocytes in a dose-dependent manner. Conclusions: Myricetin was newly discovered to be a competitive inhibitor ofS. aureusNDH-2 in relation to the substrate MK. This discovery offers a fresh perspective on the anti-staphylococcal activity of myricetin.
研究领域:
微生物学
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药理学
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生物化学
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DOI
PubMed
高影响因子
INDUSTRIAL & ENGINEERING CHEMISTRY RESEARCH
·
2017
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IF 2.843
Dong Yang,Hongjian Zou,Yizhou Wu,Jiafu Shi,Shaohua Zhang,Xiaodong Wang,Pingping Han,Zhenwei Tong,Zhongyi Jiang
NAD(P)H is a critical cofactor (biological source of hydrogen) that participates in many enzymatic hydrogenations for energy conversion and storage with resultant oxidation to NAD(P)+. Due to its high cost, the regeneration of NAD(P)H is a critical and feasible way to ensure the sustainability of these enzymatic hydrogenations. Intrigued by the photoreaction process in thylakoid membrane, we explored a two-dimensional (2D) isotype heterojunction photocatalyst, termed as quantum dots@flake graphitic carbon nitride (QDs@Flake g-C3N4), for visible-light-driven NAD(P)H regeneration. The catalyst was synthesized by one-step calcination using cyanamide-treated cyanuric acid–melamine (CM) complex as starting material, where cyanamide plays dual roles: (a) assisting the transformation of CM from bulky to stacked structure and further to flakes after calcination, and (b) acting as raw material for the generation of QDs on the flakes. To replicate both the functional and structural properties of the natural photoreaction system, QDs@Flake g-C3N4 exploited the two types of g-C3N4 (i.e., QDs and flake) and a heterojunction interface to, respectively, mimic the functional components of light-harvesting systems (LHSs, i.e., photosystem I and photosystem II) and electron transport chains (ETCs), and utilize the flake structure as the analogue of 2D thylakoid membrane. Therefore, QDs@Flake g-C3N4 showed remarkably improved capability in visible light harvesting and charge separation, and exhibited elevated performance in photocatalytic NADH regeneration with a regeneration yield of up to 40%. The NADH regeneration approach was then coupled with alcohol dehydrogenase catalyzed hydrogenation of formaldehyde, achieving continuous methanol production.
研究领域:
材料科学
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生物化学
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光催化
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产品关联
DOI
高影响因子
TALANTA
·
2021
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IF 6.057
Bo-Wen Liu,Pengcheng Huang,Fang-Ying Wu
Hypoxia refers to the lack of oxygen supply to cells or tissues. The overexpression of nitroreductase has been shown to be closely related to the degree of hypoxia, which leads to the level of nitroreductase (NTR) being used as an indicator of hypoxia. We reported a facile visual detection of NTR based on the aggregation of gold and silver alloy nanoparticles . Compared with gold nanoparticles (AuNPs), the aggregation behavior of Au 80 Ag 20 NPs caused a more prominent color change. Copper ions (Cu 2+ ) can be rapidly reduced by nicotinamide adenine dinucleotide (NADH) under the catalysis of Au 80 Ag 20 NPs. But NADH is consumed as an electron donor during the catalytic reduction reaction of p-nitrophenol (pNP) by NTR. A decrease of NADH amount results in the aggregation of Au 80 Ag 20 NPs by the excess Cu 2+ and different aggregation degrees of Au 80 Ag 20 NPs lead to observable color change. A linear correlation of A 600 /A 505 = 0.0285 [NTR]+0.361 (R 2 = 0.980) was obtained with a limit of detection (LOD) of 0.23 μg/mL for UV–vis spectrophotometer. For visual detection, the values of R/B against the concentration of NTR obtains a calibration curve of R/B = −0.031 [NTR]+ 1.54 (R 2 = 0.985) with a LOD of 0.76 μg/mL, which is of the same order of magnitude as the UV–vis spectrophotometer analysis. As a comparison, Au 80 Ag 20 NPs was replaced by several different composition nanoparticles (Au NPs, Au 70 Ag 30 NPs, Au 50 Ag 50 NPs) to be a chromogenic substrate, and the results suggest the Au 80 Ag 20 NPs is the most sensitive substrate in our assay. Selectivity tests showed that the detection system did not respond to other common substances, and the reaction mechanism was verified by inhibitor research. Finally, the assay was used on the human serum samples with spiking NTR, and the recovery rates of this assay with UV–vis spectrophotometer were basically consistent with RGB analysis.
研究领域:
生物化学
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医学检测
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纳米技术
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产品关联
DOI
PubMed
高影响因子
GREEN CHEMISTRY
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2025
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IF 9.2
yang Zhou,LingLong Huang,Yuan Tao,ChangQing Luo,Jian-Miao Xu,Zhi-Qiang Liu,Yu-Guo Zheng
The efficient regeneration of nicotinamide adenine dinucleotide (NADH), a key coenzyme in biocatalytic redox reactions, is essential for achieving sustainable chemical synthesis. However, existing 1,4-NADH regeneration methods generally suffer from high cost and poor stability, which limit their potential for industrial-scale applications.We proposed an innovative green electrochemical catalytic system for efficient regeneration of 1,4-NADH, which achieving a coenzyme conversion rate exceeding 99.0% through optimization of electrode materials and reaction conditions. In this study, cyclic voltammetry was used to investigate the effective potentials of multiple metal electrodes in the NAD+ reduction process, and the key factors affecting the NAD+ reduction efficiency were deeply resolved, revealing the complex mechanisms of the role of different electrode materials in the electron transfer and protonation processes. It was found that the introduction of foam metal and carbon nanoparticles could significantly enhance the electron transfer efficiency and increase the conversion of 1,4-NADH to 99.3%. In addition, the metal-carbon composite electrode prepared by sintering technique further enhanced the electron transfer efficiency and excellent stability. This study provides a new idea for the recycling of 1,4-NADH and contributes important insights for the efficiency improvement of electrochemical catalytic systems. Through material innovation and reaction optimization, this study demonstrates the key pathways to achieve sustainable chemical processes and opens up new directions for green chemical synthesis. In addition, it has the potential to advance biocatalytic technology in industrial applications and shows promising applications in biocatalysis and energy conversion.
研究领域:
生物化学
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电化学
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可持续化学合成
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DOI
高影响因子
Energies
·
2022
·Vol. 15
·期 12
·4242
·
IF 3.252
Xue-Yong Li,Jia-Ying Huang,Qiang Zhou,Yuan-Yuan Xu,Ponnandy Prabhu,Ye-Wang Zhang
Acetoin, a four-carbon hydroxyl-keto compound, is used in the food, pharmaceutical, and chemical industries. The cascade enzymatic production is considered a promising and efficient method to produce acetoin. However, the stability and compatibility of the enzymes under the same catalytic conditions are challenges that need to be resolved. In this work, alcohol dehydrogenase, acetaldehyde lyase, and NADH oxidase were selected to work at the same conditions to efficiently convert ethanol into acetoin. These three enzymes were immobilized on epoxy-modified magnetic nanomaterials to obtain highly stable biocatalysts. The stability and the immobilization conditions, including temperature, pH, enzyme–carrier ratio, and immobilization time, were optimized to obtain the immobilized enzymes with a high catalytic activity. The cascade reactions catalyzed by the immobilized enzymes yielded a high conversion of 90%, suggesting that the use of immobilized enzymes is a promising way to produce acetoin.Keywords:immobilization;NADH oxidase;acetaldehyde lyase;alcohol dehydrogenase;acetoin;cascade enzymatic reactions
研究领域:
生物技术
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酶学
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化学工程
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产品关联
DOI
高影响因子
Catalysis Science & Technology
·
2022
·Vol. 12
·期 12
·4057-4065
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IF 6.177
Huichao Lin,Yang Liu,Chonghui Yang,Gaozhen Zhao,Jiaao Song,Taiyi Zhang,Xiaowen Huang
Artificial photosynthesis (APS) is an effective way to convert solar energy into chemical energy by simulating the natural photosynthesis system. Currently, APS plays an important role in the regeneration of coenzymes and synthesis of organic matter. Nevertheless, the photocatalytic materials and oxidoreductases used in most bulk reactors are simply distributed in the reaction solution, which results in problems such as poor stability, high price and they cannot be reused. Moreover, the photocatalytic system also has a certain influence on the catalytic activity of oxidoreductase. In this work, we report an efficient and stable microfluidic artificial photosynthetic system for reduced nicotinamide adenine dinucleotide (NADH) regeneration and L-glutamate synthesis under visible light. We fabricate glass capillary and polydimethylsiloxane (PDMS) microfluidic reactors, followed by immobilization of few-layer g-C3N4 and glutamate dehydrogenase in light and dark reactions zones, respectively. The system possesses advantages over the traditional method in terms of its high efficiency, low cost, and sustainability. The experimental results show that the NADH regeneration rate of the system reached 56.03%, the maximum production rate of L-glutamate was 98.3%, and the reaction conversion rate of immobilized glutamate dehydrogenase still reaches 53.2% after ten times of reuse. All these results prove that our system is applicable for synthesizing high value-added compounds by simulating the natural photosynthesis system, which may be a new feasible scheme to alleviate the energy and food crisis.
研究领域:
化学工程
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生物技术
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材料科学
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环境科学
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产品关联
DOI
高影响因子
ANALYST
·
2025
·
IF 3.3
Liping Lu,kang chunyu
Nicotinamide adenine dinucleotide (NADH) is a crucial biomolecule that serves as an essential cofactor in numerous biocatalytic reactions and a pivotal regulator of mitochondrial function and cellular physiology. This study developed a SECM platform utilizing a double-mediator system for the real-time, in situ, and non-invasive detection of intracellular NADH at the single living cell level. The system employs 1-methoxy-5-methylphenazinium methyl sulfate (mPMS), which permeates the cell membrane to oxidize intracellular NADH, and ferricyanide ([Fe(CN)₆]³⁻), which shuttles electrons from the reduced mPMS to the SECM tip. This methodology was applied to investigate mitochondrial impairment in human hepatoma (HepG2) cells induced by polycyclic aromatic sulfur heterocycles (PASHs). The metabolic response was corroborated by analyzing poly(ADP-ribose) polymerase (PARP) activity, adenosine triphosphate (ATP) content, and mitochondrial membrane potential (MMP). Results demonstrated that exposure to dibenzothiophene (DBT) activated PARP, leading to severe depletion of NADH and ATP and a significant decline in MMP, thereby confirming comprehensive mitochondrial dysfunction. Comparative analysis with its non-sulfur analog fluorene (Flu) revealed that the sulfur atom in DBT's structure is a critical determinant of its enhanced cytotoxicity. This work establishes a reliable and innovative analytical approach for assessing the subcellular metabolic effects of environmental pollutants, providing a new platform for in-depth cytotoxicity mechanistic studies.
研究领域:
分析生物医学
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环境毒理学
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细胞代谢
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分子毒理学
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肿瘤学
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生物电化学
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药理学
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产品关联
DOI
PubMed
高影响因子
Analytical Methods
·
2021
·Vol. 13
·期 19
·2223-2228
·
IF 2.896
Bo-Wen Liu,Peng-Cheng Huang,Fang-Ying Wu
A novel and efficient light-controlled colorimetric assay for the quantification and detection of nitroreductase (NTR) was constructed based on p-aminophenol (pAP)-catalyzed and nicotinamide adenine dinucleotide (NADH)-mediated generation of AgNPs. Due to the hydrolysis of p-nitrophenol by NTR in the presence of NADH, the hydrolysis product can be used as a catalyst to catalyze the reduction of Ag+ by NADH under the light. As the concentration of NTR increases, the value of absorbance at ca. 400 nm (A400) decreases and the color of the solution turns from brown to bright yellow. A linear correlation was obtained between A400 and the NTR concentration in the range from 1–50 μg mL−1 and the limit of detection (LOD) is 0.27 μg mL−1. The detection system does not respond to other common biological molecules due to the specificity of enzymes and the effect of the nitroreductase inhibitor on the NTR activity was also tested. Finally, we applied the assay to determine NTR in human serum samples by spiking different concentrations of NTR with a recovery of 85.2%–92.5%.
研究领域:
生物化学
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医学检测
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纳米技术
✓ 已验证
DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2025
·
IF 13.2
Wenjin Li,Lei Wan,Yan Dong,Hang Luo,Wangzhe Li,Tin Pou Lai,Jian Zhao,Guozheng Liu,Yanxin Gao,Yida Deng,Wenbin Hu,Liyun Zhang
The coupling of photocatalytic assemblies with enzyme-catalyzed reactions offers a promising strategy for storing light energy in the form of chemical bonds, producing clean fuels and high-value chemicals. Such systems require high performance and robustness of all components, as well as effective coupling between inorganic and enzymatic modules to achieve efficient light harvesting and high product yields. In this work, a composite photocatalytic module, consisting of a heterostructure formed by silver nanoclusters (AgNCs) and graphitic carbon nitride (g-C 3 N 4 ), was coupled with an amino-containing Rh-based complex ([Cp*Rh(bpy(NH₂)₂)H₂O] 2+ ) for more efficient electron delivery and NADH recycling. The regenerated NADH was subsequently utilized by the enzymatic module, where the metal-organic framework (MOF) ZIF-90 was employed as a protective compartmentalization matrix for NADH-dependent L-glutamate dehydrogenase (GluDH). The resulting system achieved efficient conversion of NAD + to NADH, along with a notable L-glutamate production rate of 570 μmol g cat −1 h −1 , underscoring the potential of composite photosynthetic systems and paves the way for future improvements through enhanced integration and optimization of key photocatalytic modules.
研究领域:
生物化学
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光催化技术
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酶工程
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产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）