Bioresources and Bioprocessing
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2021
·Vol. 8
·期 1
·1-9
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IF 4.578
Zhang Xiaowang,Tan Zhuotao,Li Chaojian,Qi Siyu,Xu Mengjiao,Li Ming,Xiong Wenlong,Zhuang Wei,Liu Dong,Zhu Chenjie,Ying Hanjie
Background Lactones are important compounds in the field of medicine, material and chemical industry. One of the promising accesses to these flexible scaffolds is NAD(P) + -dependent alcohol dehydrogenases-catalyzed oxidative lactonization of diols, which relies on the construction of an efficient NAD(P) + regeneration system. Results In this study, a novel system combining horse liver alcohol dehydrogenase (HLADH) with the synthetic bridged flavin cofactor was established for biosynthesis of lactones. The reaction conditions of this system were optimized and a variety of lactones including chiral lactones were efficiently obtained from various diols. Compared to the previously reported NAD(P) + -regeneration systems, this system showed better regeneration efficiency and product yield. A two-phase system was further applied to solve the problem of product inhibition, and 80% yield was obtained at the condition of 300 mM substrate. Conclusions This study provides an efficient method to synthesis of lactones from diols under mild conditions. We believe this system will be a promising alternative to promote the synthesis of other valuable compounds. Graphic abstract
研究领域:
生物催化
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化学生物学
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有机化学
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药物化学
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PubMed
高影响因子
TOXICOLOGY AND APPLIED PHARMACOLOGY
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2023
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IF 3.8
Hancheng Huang,Shuchun Wei,Xiaohan Wu,Mengke Zhang,Benhong Zhou,Debin Huang,Weiguo Dong
The pathophysiological mechanism of hepatic fibrosis (HF) is related to the excessive activation of the DNA repair enzyme poly ADP-ribose polymerase-1 (PARP-1). The drugs, targeting PARP-1, are scarce. Therefore, the lead compound, moderately inhibiting PARP-1, with anti-HF properties should be identified. This study screened dihydrokaempferol (DHK) from herbs based on preliminary studies to intervene in a CCl 4 -induced liver injury and HF model in mice. In vitro , the expression levels of PARP-1-regulated related proteins and phosphorylation were examined. The binding pattern of DHK and PARP-1 was analyzed using molecular docking and molecular dynamics platforms. The results showed that DHK could significantly attenuate CCl 4 -induced liver injury and HF in mice. Moreover, it could also attenuate the toxic effects of CCl 4 on HepG2 and inhibit α-SMA and Collagen 1/3 synthesis of LX-2 cells in-vitro . Molecular docking revealed that DHK could competitively bind to the Glu-988 and His-862 residues of the upstream DNA repair enzyme PARP-1, moderately inhibiting its overactivation. This led to maintaining NAD + levels and energy metabolism in hepatocytes and inhibiting the activation of PARP-1-regulated downstream signaling pathways (TGF-β1, etc. ), related proteins (p-Smd2/3, etc. ), and inflammatory mediators while acting indirectly. Thus, DHK could attenuate CCl 4 -induced liver injury and HF in mice in a different mechanism from those of the existing reported flavonoids. It was associated with inhibiting the expression of downstream pathways and related cytokines by competitively binding to PARP-1. This study might provide a basis and direction for the design and exploration of anti-HF lead compounds.
研究领域:
药理学
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肝病学
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分子生物学
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毒理学
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PubMed
高影响因子
Biomedicines
·
2025
·Vol. 13
·期 10
·2395
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IF 3.9
Seongsu Kang,Jiwon Park,Eunbyul Cho,Dohyun Kim,Sanghyun Ye,Eui Taek Jeong,Seung-Hyun Jun,Nae-Gyu Kang
Background/Objectives:Enhancement of cellular NAD+mediated by NMN has emerged as a pivotal strategy in modulating the aging process. This study aimed to systematically investigate the anti-aging effects of NMN on human skin fibroblasts, focusing on how the former contributes to the improvement of cellular health and function. This study elucidated the molecular and functional mechanisms by which NMN contributes to the attenuation of skin aging.Methods:We performed extensive in vitro and transcriptomic analyses. Human skin fibroblasts were treated with NMN, and the induced biological responses were observed under oxidative stress/photo-aging models.Results:Transcriptome analysis revealed distinct gene expression patterns for NAD+and its precursors (NMN, NR, and NAM), showing significant differences between NMN and other precursors (NR and NMN). NMN seemed to be significantly involved in cytokine and chemokine activity. It significantly elevated cellular NAD+levels, activated sirtuin and autophagy pathways, and enhanced mitochondrial function, collectively maintaining cellular homeostasis under stress. Furthermore, it suppressed cellular senescence, promoted cell proliferation, supported extracellular matrix integrity, and accelerated wound healing.Conclusions:The study provided essential mechanistic evidence supporting the anti-aging effects of NMN in skin cells and addressed the current lack of scientific validation of NMN-based topical applications. The findings established a solid academic background for future translational research and the development of NMN-based therapeutics and cosmeceuticals.
研究领域:
分子生物学
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皮肤科学
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抗衰老研究
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细胞生物学
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转录组学
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代谢调控
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再生医学
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药理学
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DOI
PubMed
高影响因子
EUROPEAN JOURNAL OF PHARMACOLOGY
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2024
·
IF 4.2
Ya-Ling Chen,Yi Wang,Qiu-Yu Fang,Tong Wang,Cong Chen,Tong-Yao Gao,Ming Wu,Wei-Ping Zhang,Yun-Bi Lu
Poly (ADP-ribose) polymerase-1 (PARP-1) activity significantly increases during cerebral ischemia/reperfusion. PARP-1 is an NAD + -consumption enzyme. PARP-1 hyperactivity causes intracellular NAD + deficiency and bioenergetic collapse, contributing to neuronal death. Besides, the powerful trigger of PARP-1 causes the catalyzation of poly (ADP-ribosyl)ation (PARylation), a posttranslational modification of proteins. Here, we found that PARP-1 was activated in the ischemic brain tissue during middle-cerebral-artery occlusion and reperfusion (MCAO/R) for 24 h, and PAR accumulated in the neurons in mice. Using immunoprecipitation, Western blotting, liquid chromatography-mass spectrometry, and 3D-modeling analysis, we revealed that the activation of PARP-1 caused PARylation of hexokinase-1 and lactate dehydrogenase-B, which, therefore, caused the inhibition of these enzyme activities and the resulting cell energy metabolism collapse. PARP-1 inhibition significantly reversed the activity of hexokinase and lactate dehydrogenase, decreased infarct volume, and improved neuronal deficiency. PARP-1 inhibitor combined with pyruvate further alleviated MCAO/R-induced ischemic brain injury in mice. As such, we conclude that PARP-1 inhibitor alleviates neuronal death partly by inhibiting the PARylation of metabolic-related enzymes and reversing metabolism reprogramming during cerebral ischemia/reperfusion injury in mice. PARP-1 inhibitor combined with pyruvate might be a promising therapeutic approach against brain ischemia/reperfusion injury.
研究领域:
神经科学
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药理学
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分子生物学
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PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2025
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IF 6.2
Hailiang Guo,Wenjin Xia,Changyang Chen,Song Yang,Dongmei Sun,Jizu Qiao,Jianya Su
Nicotinamidase (Naam), which is essential for insect NAD synthesis but absent in mammals, represents a potential pesticide target. Virtual screening identified 1-[4-(trifluoromethyl)-3-pyridinyl]-ethanone (4-TFPE) as a potent inhibitor of Myzus persicae (IC50 = 4.156 μM) and Drosophila melanogaster (IC50 = 2.855 μM) Naam. While 4-TFPE reduced pupal weight, development, and longevity in both species, it showed no lethal toxicity. Crucially, Naam expression was upregulated by nicotinamide accumulation, revealing a feedback regulation mechanism that compromises its pesticidal potential. Furthermore, Flumetnicam’s neurotoxic effects were found to be Naam-independent. These findings highlight how target compensation mechanisms can undermine pesticide development focused on single metabolic enzymes, suggesting the need for multitarget strategies.
研究领域:
昆虫学
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生物化学
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药理学
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分子生物学
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农业科学
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DOI
PubMed
高影响因子
JOURNAL OF PHYSICAL CHEMISTRY B
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2026
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IF 3.2
Xiaonan Li,Xirong Huang
Abstract An oil-in-water microemulsion is an excellent candidate medium for the electrochemical regeneration of the coenzyme NADH and its subsequent coupled conversion application due to its good electrical conductivity and solubilization capacity for hydrophobic substrates. In this work, the reduced methyl viologen (MV+•)-mediated electrochemical reduction of coenzyme NAD+ was studied in a Triton X-100 (TX-100)-stabilized methyl tert-butyl ether (MTBE)-in-water microemulsion. The effects of the surfactant concentration and the oil content on the electrochemical behavior of the MV2+/MV+• couple and the activity of diaphorase (DH) were systematically investigated. The results show that in the microemulsion, the couple exhibits excellent reversibility. Compared with the aqueous buffer system, the couple in the microemulsion system has a more positive formal potential and a lower redox peak current, depending on the levels of both the surfactant and the oil. The positive shift of the formal potential is attributed to the stabilizing effect of the surfactant aggregates on the relatively hydrophobic MV+• species, and the decrease of the redox currents is ascribed to the decrease of the electroactive area of the electrode due to the adsorption of the surfactant and the oil as well as the decrease of the diffusion coefficients of MV species associated with or solubilized in the surfactant aggregates. The activity of DH in the microemulsion system also depends on the TX-100 concentration and the MTBE content. Increases in both [TX-100] and [MTBE] result in a decrease in DH activity. This phenomenon could be explained based on the [TX-100]/[MTBE]-dependent changes in the local acidity and the local concentration of the substrate DCPIP in the microemulsions. Based on cyclic voltammetry, the MV+•-mediated electroreduction of NAD+ in the TX-100-stabilized MTBE/H2O (0.1 M PB, pH = 7.0) microemulsion was studied. For the microemulsion system containing 0.5 mM MV2+, 5 mM NAD+, and 26.6 μM DH, when a scan rate of 2 mV s–1 was applied, a typical steady-state S-shaped CV curve was obtained. UV–vis spectroscopy confirms that the NADH generated by the mediated reaction is bioactive. By coupling the MV+•-mediated NADH electrochemical regeneration reaction with the horse liver alcohol dehydrogenase (HLADH)-catalyzed reduction of the hydrophobic substrate 4-phenyl-2-butanone, the enantiomerically pure (S)-4-phenyl-2-butanol can be selectively obtained. After 12 h, the yield of the product in the microemulsion is approximately 10 times that in the DMSO-buffer mixed homogeneous system. This difference in yield is caused by the differences in enzyme activity and stability, as well as in the substrate local concentration and reactivity in the two systems. The present study indicates that the TX-100-stabilized MTBE-in-water microemulsion is a suitable medium for the electrochemical regeneration of NADH and the electroenzymatic conversion of water-insoluble substrates catalyzed by NADH-dependent oxidoreductases.
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高影响因子
Small
·
2024
·
IF 13
Zining Hao,Shu Guo,Wenkang Tu,Qiang Wang,Jing Wang,Xuwu Zhang,Yuchu He,Dawei Gao
Cellular senescence, a vulnerable state of growth arrest, has been regarded as a potential strategy to weaken the resistance of tumor cells, leading to dramatic improvements in treatment efficacy. However, a selective and efficient strategy for inducing local tumor cellular senescence has not yet been reported. Herein, piezoelectric catalysis is utilized to reduce intracellular NAD + to NADH for local tumor cell senescence for the first time. In detail, a biocompatible nanomedicine (BTO/Rh-D@M) is constructed by wrapping the piezoelectric BaTiO 3 /(Cp * RhCl 2 ) 2 (BTO/Rh) and doxorubicin (DOX) in the homologous cytomembrane with tumor target. After tumors are stimulated by ultrasound, negative and positive charges are generated on the BTO/Rh by piezoelectric catalysis, which reduce the intracellular NAD + to NADH for cellular senescence and oxidize H 2 O to reactive oxygen species (ROS) for mitochondrial damage. Thus, the therapeutic efficacy of tumor immunogenic cell death-induced chemo-immunotherapy is boosted by combining cellular senescence, DOX, and ROS. The results indicate that 23.9% of the piezoelectric catalysis-treated tumor cells senesced, and solid tumors in mice disappeared completely after therapy. Collectively, this study highlights a novel strategy to realize cellular senescence utilizing piezoelectric catalysis and the significance of inducing tumor cellular senescence to improve therapeutic efficacy.
研究领域:
生物医学工程
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肿瘤学
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纳米技术
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免疫疗法
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DOI
PubMed
高影响因子
Small
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2022
·Vol. 19
·期 13
·2201305
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IF 15.153
Boyu Yang,Shubin Li,Wei Mu,Zhao Wang,Xiaojun Han
The bottom-up constructed artificial cells help to understand the cell working mechanism and provide the evolution clues for organisms. The energy supply and metabolism mimicry are the key issues in the field of artificial cells. Herein, an artificial cell containing cyanobacteria capable of light harvesting and carbon dioxide fixation is demonstrated to produce glucose molecules by converting light energy into chemical energy. Two downstream “metabolic” pathways starting from glucose molecules are investigated. One involves enzyme cascade reaction to produce H 2 O 2 (assisted by glucose oxidase) first, followed by converting Amplex red to resorufin (assisted by horseradish peroxidase). The other pathway is more biologically relevant. Glucose molecules are dehydrogenated to transfer hydrogens to nicotinamide adenine dinucleotide (NAD + ) for the production of nicotinamide adenine dinucleotide hydride (NADH) molecules in the presence of glucose dehydrogenase. Further, NADH molecules are oxidized into NAD + by pyruvate catalyzed by lactate dehydrogenase, meanwhile, lactate is obtained. Therefore, the cascade cycling of NADH/NAD + is built. The artificial cells built here pave the way for investigating more complicated energy-supplied metabolism inside artificial cells.
研究领域:
生物技术
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生物化学
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合成生物学
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细胞工程
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DOI
PubMed
高影响因子
SHOCK
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2024
·
IF 2.7
Liu Lijuan,Wang Ruoyu,Pu Xuexue,Zha Yutao,Yang Xiao,Fang Xiao,Liu Yong,Shao Min,Zhu Ling,Ren Xiaoou,Deng Guoqing,Yang Ke
Background Sepsis, a systemic inflammation syndrome initiated by infection, poses significant challenges due to its intricate pathophysiology. T cells play a crucial role in combating infections during sepsis. Despite previous observations indicating T cell dysfunction in sepsis, reliable in-vitro detection methods were lacking, and the factors influencing these impairments remained unclear. Methods We developed a novel method using the D 4 -Chip to assess sepsis T cell migration function. This microfluidic platform enabled precise analysis of migration function under controlled conditions. Additionally, We explored the impact of the plasma microenvironment on T cell behavior, along with the redox environment in sepsis, and assessed the potential efficacy of Mitoquinone mesylate (MitoQ), a mitochondrial-targeted drug. Results Our findings revealed impaired migration function in sepsis T cells compared to healthy controls. Interestingly, sepsis plasma enhanced the migration of healthy T cells, yet incubation with healthy plasma did not fully restore migration impairments in sepsis T cells. Subsequent investigations uncovered a significant increase in NADH/NAD+ levels in sepsis T cells, with healthy T cells exposed to various sepsis plasma conditions also showing elevated NADH/NAD+ levels. Importantly, MitoQ normalized abnormal intracellular NADH/NAD+ levels and enhanced the migration ability of T cells. Conclusions Short-term incubation with sepsis plasma does not directly inhibit T cell migration but instead affects T cell function by disrupting the intracellular redox environment. Improving the intracellular redox environment of sepsis patients contributes to restoring impaired migration and proliferation, with MitoQ demonstrating therapeutic potential.
研究领域:
免疫学
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传染病学
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药理学
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细胞生物学
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DOI
高影响因子
JOURNAL OF MEDICINAL CHEMISTRY
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2023
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IF 7.3
Yixian Fu,Yahui Huang,Chenchen Zhou,Xinge Li,Guoqiang Dong,Min Huang,Jian Ding,Chunquan Sheng
Nicotinamide adenine dinucleotide (NAD) is essentially involved in many biological processes of cancer cells, yet chemical intervention of NAD biosynthesis failed to obtain an optimal therapeutic benefit. We herein developed a new strategy to induce catastrophic NAD depletion by concurrently impairing NAD synthesis and promoting NAD consumption. We designed a series of new compounds that conjugate an inhibitor of nicotinamide phosphoribosyltransferase (NAMPT), a rate-limiting enzyme in the NAD salvage pathway, with a DNA-alkylating agent. Among them, compound 11b exhibited potent anticancer efficacy in cancer cell lines and mouse tumor models with intrinsic resistance to the parent compound FK866 or chlorambucil. Compound 11b caused catastrophic NAD depletion via a synergistic effect between the NAD salvage pathway blockade and DNA damage-triggered NAD consumption. Our findings suggest a new intervention strategy for causing catastrophic NAD depletion in cancer cells and provide basis for the development of new inhibitors targeting NAD metabolism.
研究领域:
肿瘤学
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药理学
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分子生物学
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2018
·
IF 3.909
Di Xu,Mengyun Wu,Xue Li,Mingyu Xia,Dongchun Liu,Yinghui Dai,Qing Yu,Bin Wu,Dong Wang
Bufadienolides, one kind of steroids, are the major active component secreted by ear-side gland of Bufo species. Preliminary studies on high-throughput transcriptome sequencing about B. bufo gargarizans showed that the expression of 3 β -Hydroxysteroid dehydrogenase (3 β HSD) in ear-side gland was nearly 20 times higher than that in liver. The enzyme 3 β HSD is an essential step in the biosynthesis of steroid such as progesterone, estrogens and androgens in steroidogenic tissues. Accordingly, 3 β HSD is probably an important enzyme involved in the biosynthesis of bufadienolides. In this study, Bbg-3βHSD cDNA was cloned from the ear-side gland of B. bufo gargarizans . Genetic engineering techniques were used to construct a recombinant prokaryotic fusion expression plasmid pCOLD- Bbg3βHSD which was introduced into E. coli BL21 for prokaryotic expression. Bbg- 3 β HSD has an open reading frame (ORF) of 1134 bp and encodes 377 amino acid residues. The speculated protein molecular weight is 42.8 kDa and its theoretical isoelectric point is 8.68. Amino acid sequence homologous analysis showed that Bbg -3 β HSD was highly homologous to the 3 β HSD protein of other species. Phylogenetic tree showed the highest similarity between Bbg -3 β HSD and 3 β HSD from Rana rugosa . The optimized expression of recombinant Bbg -3 β HSD were achieved by inducing with 0.1 mmol L −1 IPTG at 15 °C for 20 h. Enzymatic activity in vitro shows that pregnenolone and dehydroepiandroesterone could be 3 β -oxidized by Bbg -3 β HSD when NAD + was used as the coenzyme. Enzymatic properties showed that the optimum reaction temperature of recombinant Bbg- 3 β HSD was 40 °C, the optimum pH was 8.5, and the optimum coenzyme concentration was 1.5 mmol L −1 .
研究领域:
生物化学
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分子生物学
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酶学
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药理学
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DOI
PubMed
高影响因子
BIOORGANIC CHEMISTRY
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2026
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IF 5.1
Nianci Chen,He Zhu,Chuhang Chi,Yilin Xu,Yunbing Tang,Yingfen Ying,Ren-shan Ge
Benzalkonium chloride (BAC) disinfectants are widely used, but their potential to disrupt reproductive endocrine systems remains known. Human placental 3β-hydroxysteroid dehydrogenase 1 (h3β-HSD1) plays a key role in progesterone biosynthesis and is the potential target. This study investigates the inhibitory effects of 9 BACs on h3β-HSD1 and rat placental (r3β-HSD4) homolog. Kinetic analyses revealed h3β-HSD1 exhibited a Michaelis constant Km of 0.261 μM, while r3β-HSD4 showed similar Km (0.254 μM). Screening identified C12-C18 BACs as potent inhibitors (IC 50 : 1.15–20.43 μM for h3β-HSD1; 1.90–20.75 μM for r3β-HSD4), with mixed/noncompetitive inhibition confirmed by Lineweaver-Burk plots. A BAC mixture demonstrated antagonistic effects. Functional cell assays in JAr cells showed that C12-C18 suppressed progesterone biosynthesis, with potency influenced by membrane permeability and inhibitory strength. Structure-activity relationship (SAR) and 3D-QSAR models highlighted hydrophobicity, alkyl chain length as well as steric features as key determinants, validated by pharmacophore mapping. Molecular docking revealed conserved NAD + -binding interactions, with C18 exhibiting the highest affinity via hydrogen bonds and hydrophobic contacts as well as van der Waals. Molecular dynamics simulations confirmed stable enzyme-ligand complexes. Pharmacokinetics prediction profiling indicated variable solubility (C12 > C14 > C16 = C18). These findings elucidate BAC-mediated endocrine-disrupting mechanisms and support structure-based optimization of inhibitors.
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PubMed
高影响因子
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