INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2023
·Vol. 24
·期 9
·8265
·
IF 5.6
Jing Li,Jianchao Zhang,Yan Zhu,Lukman O. Afolabi,Liang Chen,Xuesong Feng
Triple-negative breast cancer (TNBC) has been clearly recognized as a heterogeneous tumor with the worst prognosis among the subtypes of breast cancer (BC). The advent and application of current small-molecule drugs for treating TNBC, as well as other novel inhibitors, among others, have made treatment options for TNBC more selective. However, there are still problems, such as poor patient tolerance, large administration doses, high dosing frequency, and toxic side effects, necessitating the development of more efficient and less toxic treatment strategies. High expression of Nrf2, a vital antioxidant transcription factor, often promotes tumor progression, and it is also one of the most effective targets in BC therapy. We found that in MDA-MB-231 cells and SUM159 cells, brusatol (BRU) combined with polydatin (PD) could significantly inhibit cell proliferation in vitro, significantly downregulate the expression of Nrf2 protein as well as the expression of downstream related target genesHeme Oxygenase-1 (HO-1)andNAD(P)H dehydrogenase, quinone 1 (NQO1), and promote reactive oxygen species (ROS) levels to further strengthen the anti-tumor effect. Furthermore, we discovered in our in vivo experiments that by reducing the drug dosage three times, we could significantly reduce tumor cell growth while avoiding toxic side effects, providing a treatment method with greater clinical application value for TNBC treatment.Keywords:breast cancer;brusatol;polydatin;Nrf2;HO-1;NQO1;ROS
研究领域:
肿瘤学
·
药理学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Biotechnology Journal
·
2024
·Vol. 19
·期 3
·2300744
·
IF 3.2
Shuping Zou,Bing Zhang,Yuyue Han,Jinlong Liu,Kuo Zhao,Yaping Xue,Yuguo Zheng
NAD(P)H-dependent oxidoreductases are crucial biocatalysts for synthesizing chiral compounds. Yet, the industrial implementation of enzymatic redox reactions is often hampered by an insufficient supply of expensive nicotinamide cofactors. Here, a cofactor self-sufficient whole-cell biocatalyst was developed for the enzymatic asymmetric reduction of 2-oxo-4-[(hydroxy)(-methyl)phosphinyl] butyric acid (PPO) to L-phosphinothricin (L-PPT). The endogenous NADP + pool was significantly enhanced by regulating Preiss-Handler pathway toward NAD(H) synthesis and, in the meantime, introducing NAD kinase to phosphorylate NAD(H) toward NADP + . The intracellular NADP(H) concentration displayed a 2.97-fold increase with the strategy compared with the wild-type strain. Furthermore, a recombinant multi-enzyme cascade biocatalytic system was constructed based on the Escherichia coli chassis. In order to balance multi-enzyme co-expression levels, the strategy of modulating rate-limiting enzyme Pm GluDH by RBS strengths regulation successfully increased the catalytic efficiency of PPO conversion. Finally, the cofactor self-sufficient whole-cell biocatalyst effectively converted 300 mM PPO to L-PPT in 2 h without the need to add exogenous cofactors, resulting in a 2.3-fold increase in PPO conversion (%) from 43% to 100%, with a high space-time yield of 706.2 g L −1 d −1 and 99.9% ee . Overall, this work demonstrates a technological example for constructing a cofactor self-sufficient system for NADPH-dependent redox biocatalysis.
研究领域:
生物技术
·
生物化学
·
代谢工程
·
酶学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIORESOURCE TECHNOLOGY
·
2015
·
IF 4.494
Yu-Cai He,Dan-Ping Zhang,Zhi-Cheng Tao,Yun Lu,Yun Ding,Feng Liu,Zheng-Zhong Zhu,Huan Rui,Gao-Wei Zheng,Xian Zhang
To reduce dependence on the expensive cofactor and effectively biotransform ethyl 4-chloro-3-oxobutanoate, l -glutamine and glycine were found to enhance the content of intracellular NADH and the reductase activity. Adding the mixture of 200 mM of l -glutamine and 500 mM of glycine to the reaction media, a 1.67-fold of reductase activity was increased over the control without the addition of the two compounds. Moreover, β-cyclodextrin (0.4 mol β-cyclodextrin/mol ethyl 4-chloro-3-oxobutanoate) was also added into this reaction media, and the biocatalytic activity of the whole-cell biocatalyst of Escherichia coli CCZU-K14 was increased by 1.34-fold than that without β-cyclodextrin. In this β-cyclodextrin–water media containing l -glutamine (200 mM) plus glycine (500 mM), ethyl ( S )-4-chloro-3-hydroxybutanoate (>99% ee ) could be obtained from 3000 mM ethyl 4-chloro-3-oxobutanoate in the yield of 98.0% after 8 h. All the positive features demonstrate the potential applicability of the bioprocess for the large-scale production of ethyl ( S )-4-chloro-3-hydroxybutanoate.
研究领域:
生物化学
·
酶工程
·
有机合成
✓ 已验证
产品关联
DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2026
·
IF 12.5
Xueying An,Hanjie Zhang,Xujing Zhang,Minyi Cai,Yining Zhou,Hantao Cai,Tao Shen,Sheng zhao,Zhihong Xu,Hui Wei,Qing Jiang
A lycopene-loaded CeO₂ nanozyme (Ly@Ce) is developed to combat chondrocyte senescence for osteoarthritis therapy • Ly@Ce efficiently scavenges mitochondrial ROS, restores mitochondrial dynamics, and elevates NAD + levels • Ly@Ce reverses H₂O₂-induced dysregulation of the p53-p21 signaling axis • Ly@Ce suppresses SASP expression and shifts ECM metabolism toward an anabolic program • The nanozyme integrates redox/NAD + reprogramming with senescence pathway inhibition, offering a disease-modifying strategy for OA Osteoarthritis (OA) is strongly associated with chondrocyte senescence, driven by oxidative stress, mitochondrial dysfunction, and the ensuing senescence-associated secretory phenotype (SASP). Herein, we report a lycopene-loaded ceria nanozyme (Ly@Ce) designed to simultaneously address these interconnected pathogenic cues. Ly@Ce markedly reduced intracellular and mitochondrial reactive oxygen species (ROS), mitigated excessive mitochondrial fragmentation, and restored mitochondrial membrane potential in senescent chondrocytes, indicating robust recovery of mitochondrial fitness. Notably, Ly@Ce also elevated intracellular nicotinamide adenine dinucleotide (NAD + ) levels, making it an active metabolic modulator rather than a passive ROS scavenger. Transcriptomic profiling revealed that Ly@Ce reversed H 2 O 2 -induced dysregulation of the p53-p21 signaling pathway, accompanied by suppression of SASP-related genes and a shift of extracellular matrix metabolism toward an anabolic program, thus delaying OA progression. Collectively, these findings establish Ly@Ce as a bioinspired, catalytic antioxidant nanoplatform that combines redox and NAD + reprogramming with p53-p21 inhibition, offering a disease-modifying strategy for senescence-associated OA and a potential framework for targeting other age-related pathologies characterized by mitochondrial decline and cellular senescence.
✓ 已验证
DOI
高影响因子
FREE RADICAL BIOLOGY AND MEDICINE
·
2025
·
IF 8.2
Xiao-Yi Zhang,Yi-Hao Zhang,Nan-Nan Liang,Shan-Shan Xu,Qiang-Sheng Li,Ya-Ping Song,Wei-Wei Zhang,Bo Wang,Hua Wang,Yi-Chao Huang,De-Xiang Xu
Accumulating evidence indicates that arsenic (As) exposure causes a decline in sperm quality. This study aimed to investigate the impact of As exposure on spermatogonial senescence. GC-1 cells were exposed to NaAsO 2 (10 μM). RNA sequencing and ribosome profiling sequencing were performed to identify key regulators of cellular senescence. Methylated RNA immunoprecipitation-qPCR was used to determine N 6 -methyladenosine (m6A) modification. The results revealed that the differentially expressed genes were enriched in pathways related to cellular senescence. Several established senescence markers, β-galactosidase activity, γ-H2AX, and P16, were elevated in As-exposed GC-1 cells. Further analysis revealed that P21 and its transcription factor ETS2 were upregulated. ETS2 knockout prevented As-induced P21 upregulation and cell senescence. The multi-omics joint analysis indicated that As exposure elevated ETS2 translation efficiency by YTHDC2-dependent m6A modification. Mechanistically, As exposure induced mitochondrial dysfunction. Alpha-ketoglutarate (α-KG), a tricarboxylic acid cycle intermediate and RNA demethylase cofactor, was reduced in As-exposed GC-1 cells. Additional experiments showed that As exposure induced NAD + depletion and suppressed SIRT3 activity. Supplementation with nicotinamide mononucleotide, an NAD + precursor, attenuated As-evoked α-KG reduction and ETS2 upregulation. These findings suggest that As induces spermatogonial senescence via m6A modification-mediated upregulation of ETS2 translation and identify NAD + replenishment as a potential countermeasure.
研究领域:
毒理学
·
分子生物学
·
表观遗传学
·
生殖生物学
·
生物信息学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
PROTEIN EXPRESSION AND PURIFICATION
·
2025
·
IF 1.2
Shuping Fei,Jiatong Zhao,Wenwen Wu,Chaoqun Lian,Wanggang Tang
Histidinol dehydrogenase (HisD, E.C. 1.1.1.23), encoded by the hisD gene and catalyzing the final two steps in the l -histidine biosynthesis, has emerged as a promising antibacterial target for several human pathogens, such as Brucella suis and Mycobacterium tuberculosis . Herein, biochemical properties on recombinant HisD from the human pathogen Neisseria gonorrhoeae ( Ng HisD) were characterized in detail. SDS-PAGE and size-exclusion chromatography revealed that recombinant Ng HisD is a homodimer (∼105 kDa native size; ∼47 kDa subunit). Kinetic analysis identified this protein as a highly selective NAD + -dependent enzyme (specific activity: 17.3 ± 0.4 U mg −1 ), exhibiting minimal activity with NADP + . The K M values for NAD + and l -histidinol were 0.9 ± 0.3 mM and 45 ± 6 μM, respectively. Maximal activity occurred at pH 9.5 and 45 °C in the presence of Mn 2+ . Heat-inactivation experiment showed that rapid inactivation began to occur following incubation at 45 °C for 30 min. The metal ions coordinated by the enzyme were not sequestered by 10 mM EDTA but were efficiently chelated by 1,10-phenanthroline. The recombinant Ng HisD lost 85 % activity in the presence of 1 mM 1,10-phenantroline. Our study not only establishes a foundation for investigating the physiological function of Ng HisD in vivo but also uncovers the absence of the canonical histidine biosynthesis pathway in Neisseria cinerea .
研究领域:
微生物学
·
分子生物学
·
生物化学
·
感染性疾病
·
抗菌药物靶点
✓ 已验证
产品关联
DOI
PubMed
高影响因子
CATALYSIS LETTERS
·
2021
·Vol. 151
·期 9
·2527-2536
·
IF 3.186
Du Hui-Jun,Luo Wei,Appiah Bright,Zou Zhi-Cheng,Yang Zhong-Hua,Zeng Rong,Luo Li
Abstract The intracellular NADPH insufficient supply is the main bottleneck to the synthesis of chiral alcohols by asymmetric reduction with whole-cell catalysis. Herein, we provide a novel strategy to strengthen intracellular NADPH supply through introducing an NADP + -dependent glyceraldehyde 3-phosphate dehydrogenase ( gap B from Bacillus subtilis 168) into the Embden-Meyerhof pathway and a NAD kinase ( yfj B from E. coli MG1655) to further enhance the NADP(H) pool. A recombinant E. coli ( E. coli BL21 (DE3)/pETDuet-1- gap B- yue D&pET28a- yfj B) was constructed to co-express gap B and yfj B with a carbonyl reductase gene yue D together. The result showed that the intracellular NADPH amount increased by 134.4% with the strategy. To the model reaction (asymmetric reduction of acetophenone to S -phenyl ethanol), the yield was 3.7-fold with this strategy compared to the control. This provides a technological route for strengthening the intracellular NADPH supply in E. coli for biocatalysis and biosynthesis. Graphic Abstract
研究领域:
生物催化
·
代谢工程
·
微生物生物技术
✓ 已验证
DOI
高影响因子
BRAIN RESEARCH BULLETIN
·
2024
·
IF 3.5
Cong Chen,Tong Wang,Tong-Yao Gao,Ya-Ling Chen,Yun-Bi Lu,Wei-Ping Zhang
Nicotinamide phosphoribosyltransferase (NAMPT) is the key enzyme in the salvaging synthesize pathway of nicotinamide adenine dinucleotide (NAD). The neuroprotective roles of NAMPT on neurodegeneration have been explored in aging brain and Alzheimer’s Disease. However, its roles in Parkinson’s Disease (PD) remain to be elucidated. We found that the dopaminergic neurons in substantia nigra expressed higher levels of NAMPT than the other types of neurons. Using conditional knockout of the Nampt gene in dopaminergic neurons and utilizing a NAMPT inhibitor in the substantia nigra of mice, we found that the NAMPT deficiency triggered the time-dependent loss of dopaminergic neurons, the impairment of the dopamine nigrostriatal pathway, and the development of PD-like motor dysfunction. In the rotenone-induced PD mouse model, nicotinamide ribose (NR), a precursor of NAD, rescued the loss of dopaminergic neurons, the impairment of dopamine nigrostriatal pathway, and mitigated PD-like motor dysfunction. In SH-SY5Y cells, NAD suppression induced the accumulation of reactive oxygen species (ROS), mitochondrial impairment, and cell death, which was reversed by N-acetyl cysteine, an antioxidant and ROS scavenger. Rotenone decreased NAD level, induced the accumulation of ROS and the impairment of mitochondria, which was reversed by NR. In summary, our findings show that the ablation of NAMPT in dopaminergic neurons leads to neurodegeneration and contributes to the development of PD. The NAD precursors have the potential to protect the degeneration of dopaminergic neurons, and offering a therapeutic approach for the treatment of PD.
研究领域:
神经科学
·
分子生物学
·
神经退行性疾病
✓ 已验证
产品关联
DOI
PubMed
高影响因子
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
·
2016
·Vol. 182
·期 3
·898-909
·
IF 1.606
Xu Jian-Miao,Cheng Feng,Fu Fang-Tian,Hu Hai-Feng,Zheng Yu-Guo
L -2-aminobutyric acid ( L -ABA) as a precursor for the anticonvulsant and the antituberculotic is a key intermediate in the chemical and pharmaceutical industries. Recently, leucine dehydrogenase (LeuDH) with NAD + regeneration was developed for L -ABA production on a large scale. Previously, the L -ABA yield was improved by optimizing conversion conditions, including cofactor regeneration and enzyme immobilization but not protein engineering on LeuDH due to lacking an applicable high-throughput screening (HTS) method. Recently, an HTS assay was developed by us, which enables researchers to engineer LeuDH in a relatively short period of time. Herein, a semirational engineering was performed on LeuDH to increase the catalytic efficiency of BcLeuDH. Firstly, the structure of wild-type (WT) BcLeuDH was modeled and seven potentially beneficial positions were selected for mutation. Five beneficial variants were then identified from the seven site-saturation mutagenesis (SSM) libraries by HTS and confirmed by rescreening via amino acid analyzer. The “best” variant M5 (WT + Q358N) showed 44.5-fold higher catalytic efficiency ( k
cat / K
M ) than BcLeuDH WT, which suggested that BcLeuDH M5 is an attractive candidate for L -ABA production on a large scale. Furthermore, the structure-functional relationship was investigated based on the docking and kinetic results.
研究领域:
生物技术
·
生物化工
·
酶学
·
药学
✓ 已验证
DOI
PubMed
高影响因子
Synthetic and Systems Biotechnology
·
2025
·
IF 4.4
Haizhao Xue,Yinghan Hu,Aabid Manzoor Shah,Xueying Wang,Xiaojia Guo,Yanzhe Huang,Zongbao K. Zhao
The nicotinamide cofactors including nicotinamide adenine dinucleotide (NAD) and its phosphate (NADP) play important roles in facilitating redox reactions for energy metabolism and biosynthesis. To expand the cofactor menu, a non-natural cofactor nicotinamide cytosine dinucleotide (NCD) has been introduced recently. The reduced forms of these cofactors carry reducing equivalents that are essential for cellular metabolism. However, there is a long standing challenge to rationally transfer reducing equivalent from one cofactor to another, albeit such process is highly demanding in metabolic engineering. This study develops a new approach based on malic enzyme (ME)-mediated transhydrogenation to enable reducing equivalents exchange among different cofactors. We used wild-type ME, MaeB and an engineered ME* that favors NAD, NADP and NCD, respectively, to demonstrate such conversions. When an in vitro system initiated with equal amount of NADH and NCD in the presence of ME, ME* and excess amount of pyruvate was held for 2 h, up to 65% NADH was consumed and 57% NCDH was generated. When implemented into NCD self-sufficient Escherichia coli cells, the system directed reducing equivalents toward NCDH-linked formation of lactate. Overall, this work offers an effective strategy to regulate intracellular reducing equivalents that may serve as a novel tool for metabolic engineering and synthetic biology.
研究领域:
生物化学
·
分子生物学
·
代谢工程
·
合成生物学
·
微生物学
✓ 已验证
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2025
·
IF 8.5
Yuangui Tang,Chenyun Dai,Huihui Yang,Xian Yang,Ziyu Chen,Jinxiu Dou,Liuxu Zhang,Tao Bai,Junfang Zheng
Non-histone proteins localized in membrane, cytosol and nucleus are lactylated to promote tumor progression. However, whether mitochondrial proteins undergo lactylation and contribute to tumor progression remains unexplored. Here, we identified multiple lactylated mitochondrial proteins in human renal cell carcinoma (RCC) cells using lactylome profiling. Among these, malate dehydrogenase 2 (MDH2)–the only lactylated protein in the tricarboxylic acid (TCA) cycle–emerged as a key target, with K239 as its lactylation site. MDH2 K239la levels are regulated by KAT8 and SIRT3. Under low-glucose, high-lactate conditions mimicking the tumor microenvironment, MDH2 K239la elevated the NADH/NAD + ratio to drive ATP production and boosted NADPH generation to reduce ROS. This enables cells to alleviate oxidative stress, sustain mitochondrial function, and promote RCC malignancy in vitro and in vivo . Mechanistically, MDH2 K239la enhances MDH2 enzymatic activity and strengthens its interaction with the citrate transporter SLC25A1. This facilitates citrate efflux, fueling IDH1-dependent NADPH production when lactate serves as an energy source. Collectively, we unveil a lactylation-dependent mechanism that reprograms mitochondrial metabolism to confer oxidative stress resistance and drive RCC progression. Targeting mitochondrial proteins lactylation, exemplified by MDH2 K239la , represents a promising therapeutic strategy for RCC.
研究领域:
癌症生物学
·
生物化学
·
细胞生物学
·
分子生物学
·
代谢工程
·
肿瘤学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2024
·
IF 8.5
Yicheng Ren,Quan Ma,Dong Li,Yanpei Chen,Qinyang Cheng,Zisheng Luo
Fluctuations in energy status are critical physiological factors influencing postharvest fruit quality. However, the role of autophagy, a stress-resistant biological process, in regulating postharvest fruit energy status remains unclear. In this study, we treated strawberry fruit with the autophagy inhibitor hydroxychloroquine (HCQ) to investigate the role of autophagy in maintaining energy status. The results demonstrated that HCQ treatment inhibited autophagosome formation, reducing relative autophagic activity by 73 % on day 6. HCQ treatment also reduced the energy charge and the NADH/NAD + ratio by 5 % and 61 %, respectively, on day 2. Corresponding to the decline in energy status, HCQ treatment led to a reduction in cytochrome C oxidase (CCO) and succinate dehydrogenase (SDH) activities by 47.48 % and 56.34 %, respectively, on day 3. Additionally, fluctuations in energy charge led to abnormalities in ripening and decreased stress resistance in autophagy-inhibited fruit. In conclusion, this study provides evidence that autophagy-mediated energy charge sustainability mitigates postharvest quality deterioration in fruit.
研究领域:
植物生理学
·
采后生物学
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）