Journal of Pharmaceutical Analysis
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2024
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IF 6.1
Yang Xu,Shujing Lv,Xiang Li,Chuanjia Zhai,Yulian Shi,Xuejiao Li,Zhiyang Feng,Gan Luo,Ying Wang,Xiaoyan Gao
Sennoside A (SA), a typical prodrug, exerts its laxative effect only after its transformation into rheinanthrone catalyzed by gut microbial hydrolases and reductases. Hydrolases have been identified, but reductases remain unknown. By linking a photoreactive group to the SA scaffold, we synthesized a photoaffinity probe to covalently label SA reductases and identified SA reductases using activity-based protein profiling. From lysates of an active strain, Bifidobacterium pseudocatenulatum ( B . pseudocatenulatum ), 397 proteins were enriched and subsequently identified using mass spectrometry. Among these proteins, chromate reductase/nicotinamide adenine dinucleotide phosphate (NADPH)-dependent flavin mononucleotide (FMN) reductase/oxygen-insensitive NADPH nitroreductase (nfrA) was identified as a potent SA reductase through further bioinformatic analysis and Uniprot database screening. We also determined that recombinant nfrA could reduce SA. Our study contributes to further illuminating mechanisms of SA transformation to rheinanthrone and simultaneously offers an effective method to identify gut bacterial reductases.
研究领域:
微生物学
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药理学
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蛋白质组学
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产品关联
DOI
PubMed
高影响因子
Redox Biology
·
2018
·
IF 7.126
Dong Hou,Zhaojian Liu,Xiuhua Xu,Qiao Liu,Xiyu Zhang,Beihua Kong,Jian-Jun Wei,Yaoqin Gong,Changshun Shao
PARP inhibitors have been widely tested in clinical trials, especially for the treatment of breast cancer and ovarian cancer, and were shown to be highly successful. Because PARP primarily functions in sensing and repairing DNA strand breaks, the therapeutic effect of PARP inhibition is generally believed to be attributed to impaired DNA repair. We here report that oxidative stress is also increased by PARP inhibition and mediates the antitumor effect. We showed that PARP1 is highly expressed in specimens of high grade serous ovarian carcinoma and its activity is required for unperturbed proliferation of ovarian cancer cells. Inhibition or depletion of PARP leads to not only an increase in DNA damage, but also an elevation in the levels of reactive oxygen species (ROS). Importantly, antioxidant N-acetylcysteine (NAC) significantly attenuated the induction of DNA damage and the perturbation of proliferation by PARP inhibition or depletion. We further showed that NADPH oxidases 1 and 4 were significantly upregulated by PARP inhibition and were partially responsible for the induction of oxidative stress. Depletion of NOX1 and NOX4 partially rescued the growth inhibition of PARP1-deficient tumor xenografts. Our findings suggest that in addition to compromising the repair of DNA damage, PARP inhibition or depletion may exert extra antitumor effect by elevating oxidative stress in ovarian cancer cells.
研究领域:
肿瘤学
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分子生物学
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癌症治疗
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产品关联
DOI
PubMed
高影响因子
EUROPEAN JOURNAL OF PHARMACOLOGY
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2022
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IF 5.195
Hongxiao Yang,Juncai Bai,Chengchuang zhan,Shuang Liu,Yunan Gao,Lihua Zhong,Yajing Lv,Jing Chi,Jiaren Liu,Xinrui Yang,Wei Yang
Cardiac hypertrophy occurs as a result of high levels of thyroid hormone, which may contribute to heart failure and is closely related to oxidative stress . Hydrogen is a good antioxidant. In this study, we found that intragastric levothyroxine administration for two weeks caused obvious cardiac hypertrophy without reduced systolic function. Additionally, hydrogen inhalation ameliorated the levothyroxine-induced metabolic increase and cardiac hypertrophy in rats. Serum brain natriuretic peptide expression was also attenuated by hydrogen treatment. However, hydrogen had no significant effect on levothyroxine -induced serum troponin I and serum thyroid hormone changes. Hydrogen treatment also reduced the levothyroxine-induced increase in cardiac malondialdehyde , 8-hydroxy-2-deoxyguanosine and serum hydrogen peroxide levels and upregulated superoxide dismutase and glutathione peroxidase activity. Additionally, western blotting results showed that hydrogen inhalation inhibited the expression of cardiac nicotinamide adenine dinucleotide phosphate oxidase 2 (NOX2), angiotensin II type 1 receptor, sarcoplasmic reticulum Ca 2+ -ATPase (SERCA2), phospho-phospholamban and α-myosin heavy chain proteins. In conclusion, the present study revealed a protective effect of hydrogen on levothyroxine -induced cardiac hypertrophy by regulating angiotensin II type 1 receptors and NOX2-mediated oxidative stress in rats.
研究领域:
药理学
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心脏病学
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内分泌学
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分子生物学
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DOI
PubMed
高影响因子
JOURNAL OF CATALYSIS
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2024
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IF 6.5
Chun-Xian Cai,Zi-Wen Zhou,Chun-Xiu Liu,Yun-Jie Wei,Xiao-Qi Yu,Li-Hong Zhou,Na Wang
Photo-enzyme catalysis featuring sustainability and selectivity is attractive, but remains challenges from efficiency and enzyme inactivation. This work provides a compatible and efficient photocatalyst-enzyme hybrid system for chiral alcohol synthesis. A covalent organic framework (COF) possessing broad visible light absorption edge to 554 nm was used for photocatalytic NADPH regeneration, achieving 82 % yield in 90 min. Then the influence of photocatalysis on coupled enzyme Lb ADH was unveiled by fluorescence spectra and circular dichroism. Further a facile and mild strategy of entrapping Lb ADH into Ca 2+ /Mn 2+ doped alginate gel was carried out. Impressively, enzyme immobilized strategy not only enhances 3.2 times activity, but also protects Lb ADH from diverse harsh environments, thereby improving photo-enzyme catalytic yield from 26 % to 82 %. To our delight, this one-pot photo-enzyme catalytic system achieves highly yield (up to 99 %) with excellent enantioselectivity (90–99 ee %) for various chiral alcohols, and fulfills semipreparative scale preparation of ( R )-1-[3,5-Bis(trifluoromethyl)phenyl] ethanol, prochiral intermediate of Aprepitant. This work sheds light on the new synthetic route of chiral compounds and development of photo-enzyme catalytic field.
研究领域:
生物化学
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催化化学
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产品关联
DOI
高影响因子
Biotechnology for Biofuels
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2018
·Vol. 11
·期 1
·1-14
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IF 5.497
Wang Xiang,Dong Hong-Po,Wei Wei,Balamurugan Srinivasan,Yang Wei-Dong,Liu Jie-Sheng,Li Hong-Ye
Background Metabolic engineering has emerged as a potential strategy for improving microalgal lipid content through targeted changes to lipid metabolic networks. However, the intricate nature of lipogenesis has impeded metabolic engineering. Therefore, it is very important to identify the crucial metabolic nodes and develop strategies to exploit multiple genes for transgenesis. In an attempt to unravel the microalgal triacylglycerol (TAG) pathway, we overexpressed two key lipogenic genes, glycerol-3-phosphate acyltransferase (GPAT1) and lysophosphatidic acid acyltransferase (LPAT1), in oleaginous Phaeodactylum tricornutum and determined their roles in microalgal lipogenesis. Results Engineered P. tricornutum strains showed enhanced growth and photosynthetic efficiency compared with that of the wild-type during the growth phase of the cultivation period. However, both the cell types reached stationary phase on day 7. Overexpression of GPAT1 and LPAT1 increased the TAG content by 2.3-fold under nitrogen-replete conditions without compromising cell growth, and they also orchestrated the expression of other key genes involved in TAG synthesis. The transgenic expression of GPAT1 and LPAT1 influenced the expression of malic enzyme and glucose 6-phosphate dehydrogenase, which enhanced the levels of lipogenic NADPH in the transgenic lines. In addition, GPAT1 and LPAT1 preferred C16 over C18 at the sn-2 position of the glycerol backbone. Conclusion Overexpression of GPAT1 together with LPAT1 significantly enhanced lipid content without affecting growth and photosynthetic efficiency, and they orchestrated the expression of other key photosynthetic and lipogenic genes. The lipid profile for elevated fatty acid content (C16-CoA) demonstrated the involvement of the prokaryotic TAG pathway in marine diatoms. The results suggested that engineering dual metabolic nodes should be possible in microalgal lipid metabolism. This study also provides the first demonstration of the role of the prokaryotic TAG biosynthetic pathway in lipid overproduction and indicates that the fatty acid profile can be tailored to improve lipid production.
研究领域:
微生物学
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生物技术
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代谢工程
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植物科学
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DOI
PubMed
高影响因子
BIOCHEMICAL ENGINEERING JOURNAL
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2015
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IF 2.467
Qin Xu,Wei-Yi Tao,He Huang,Shuang Li
An NADPH-dependent carbonyl reductase (YlCR2) from Yarrowia lipolytica was discovered by genome mining, overexpressed in Escherichia coli BL21 and purified to homogeneity. To efficiently synthesize ethyl ( S )-4-chloro-3-hydroxybutanoate (( S )-CHBE) (99%, e.e), the highly stereoselective bioreduction of ethyl 4-chloro-3-oxobutanoate (COBE) into ( S )-CHBE with the recombinant E. coli BL21/pETYlCR2 was successfully demonstrated in an n -butyl acetate-water biphasic system (1:1, v/v) with NADPH self-regeneration by substrate-coupled system using sorbitol or mannitol as co-substrate. The optimum reaction condition for the biotransformation of COBE in the biphasic system were 3000 mM COBE, (1.2 or 1.3 mmol/mmol COBE) mannitol or sorbitol, 0.2 mM or 0.4 mM NADP + , 0.12 or 0.14 g (wet weight)/ml cell dosage, pH 5.0, 30 °C; ( S )-CHBE with yield of 90% and e.e of 99% was obtained after 10 h reaction. Furthermore, 3000 mM COBE could also be completely biotransformed after 20 h without addition of expensive cofactor NADP + . Significantly, E. coli BL21/pETYlCR2 shows the high potential in the industrial production of ( S )-CHBE.
研究领域:
生物化学
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合成生物学
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酶工程
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DOI
高影响因子
JOURNAL OF PLANT PHYSIOLOGY
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2023
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IF 4.3
Lin Xiao,Wenna Ma,Jiarong Zhang,Xiaojun Pu,Zed Rengel,Zhongbang Song,Qi Chen
Flavonols are well-known antioxidants that prevent stomatal closure via interfering with ROS signaling. Phytomelatonin regulates stomatal closure, but the signaling pathways are still largely unknown. Here, we investigated the role of flavonols in phytomelatonin-mediated stomatal closure in tobacco plants. The application of melatonin induced stomatal closure through NADPH oxidase-mediated ROS production. Transgenic tobacco plants overexpressing soybean GmSNAT1 (coding for serotonin N -acetyltransferase that catalyzes the penultimate step in phytomelatonin biosynthesis) had higher phytomelatonin concentration, accumulated more ROS in guard cells and were more sensitive to melatonin-induced stomatal closure than the wild-type plants, which was associated with the higher expression of PMTR1 -homologous genes. Exogenous melatonin decreased flavonol concentrations in guard cells and the expression of flavonoid-related genes in wild-type and transgenic tobacco plants, and these inhibitory effects were more obvious in GmSNAT1 -overexpressing plants than the wild type. However, the melatonin-mediated stomatal closure and ROS production were diminished by the application of kaempferol (a type of flavonol). Additionally, transgenic tobacco plants with increased expression of NtFLS (encoding flavonol synthase) were less sensitive to melatonin-induced stomatal closure. In conclusion, phytomelatonin hampers the biosynthesis of flavonols in guard cells, which results in high concentration of ROS and induces stomatal closure in tobacco plants.
研究领域:
植物生理学
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分子生物学
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植物生物化学
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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
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代谢工程
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微生物生物技术
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DOI
高影响因子
Bioresources and Bioprocessing
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2022
·Vol. 9
·期 1
·1-14
·
IF 4.983
Liu Kun,Wang Feng-Qing,Liu Ke,Zhao Yunqiu,Gao Bei,Tao Xinyi,Wei Dongzhi
Progesterone is one of the classical hormone drugs used in medicine for maintaining pregnancy. However, its manufacturing process, coupled with organic reagents and poisonous catalysts, causes irreversible environmental pollution. Recent advances in synthetic biology have demonstrated that the microbial biosynthesis of natural products, especially difficult-to-synthesize compounds, from building blocks is a promising strategy. Herein, overcoming the heterologous cytochrome P450 enzyme interdependency in Mycolicibacterium neoaurum successfully constructed the CYP11A1 running module to realize metabolic conversion from waste phytosterols to progesterone. Subsequently, progesterone yield was improved through strategies involving electron transfer and NADPH regeneration. Mutant CYP11A1 (mCYP11A1) and adrenodoxin reductase (ADR) were connected by a flexible linker (L) to form the chimera mCYP11A1-L-ADR to enhance electron transfer. The chimera mCYP11A1-L-ADR, adrenodoxin (ADX), and ADR-related homolog ARH1 were expressed in M. neoaurum, showed positive activity and produced 45 mg/L progesterone. This electron transfer strategy increased progesterone production by 3.95-fold compared with M. neoaurum expressing mCYP11A1, ADR, and ADX. Significantly, a novel inorganic–biological hybrid system was assembled by combining engineered M. neoaurum and InP nanoparticles to regenerate NADPH, which was increased 84-fold from the initial progesterone titer to 235 ± 50 mg/L. In summary, this work highlights the green and sustainable potential of obtaining synthetic progesterone from sterols in M. neoaurum.
研究领域:
合成生物学
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生物技术
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环境科学
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药学
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微生物学
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产品关联
DOI
PubMed
高影响因子
BIORESOURCE TECHNOLOGY
·
2021
·
IF 9.642
Yu Zhang,Minhua Wei,Guihong Zhao,Wenjie Zhang,Yingzi Li,Beibei Lin,Yanjun Li,Qingyang Xu,Ning Chen,Chenglin Zhang
l -Homoserine is a valuable non-proteinogenic amino acid used in the synthesis of various important compounds. Microbial fermentation has potential value for producing l -homoserine on a large scale, but suffers from a low yield and the need for expensive additives. In this study, a non-induced, non-auxotrophic, plasmid-free Escherichia coli chassis for the high-efficiency production of l -homoserine was constructed. Initially, the l -homoserine degradation pathway was dynamically attenuated. Subsequently, systems metabolic engineering strategies were employed, including reinforcing the synthetic flux, improving NADPH generation, and elevating l -homoserine efflux. The constructed strain HOM-14, produced 60.1 g/L l -homoserine without additional supplements or inducers, which achieved the highest fermentative production efficiency of l -homoserine till date. Moreover, common byproducts, such as acetate, did not accumulate. The strategies presented here can be applied in the further engineering of chassis for the scale-up production of l -homoserine and derivatives.
研究领域:
生物技术
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代谢工程
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微生物学
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产品关联
DOI
PubMed
高影响因子
Frontiers in Cell and Developmental Biology
·
2020
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IF 5.186
Xu Huanhuan,Hu Lihong,Liu Titi,Chen Fei,Li Jin,Xu Jing,Jiang Li,Xiang Zemin,Wang Xuanjun,Sheng Jun
Glucose-6-phosphate dehydrogenase (G6PDH) is the rate-limiting enzyme in the pentose phosphate pathway (PPP) and plays a crucial role in the maintenance of redox homeostasis by producing nicotinamide adenine dinucleotide phosphate (NADPH), the major intracellular reductant. G6PDH has been shown to be a biomarker and potential therapeutic target for renal cell carcinoma (RCC). Here, we report a previously unknown biochemical mechanism through which caffeine, a well-known natural small molecule, regulates G6PDH activity to disrupt cellular redox homeostasis and suppress RCC development and progression. We found that caffeine can inhibit G6PDH enzymatic activity. Mechanistically, caffeine directly binds to G6PDH with high affinity (KD = 0.1923 μM) and competes with the coenzyme NADP+ for G6PDH binding, as demonstrated by the decreased binding affinities of G6PDH for its coenzyme and substrate. Molecular docking studies revealed that caffeine binds to G6PDH at the structural NADP+ binding site, and chemical cross-linking analysis demonstrated that caffeine inhibits the formation of dimeric G6PDH. G6PDH inhibition abrogated the inhibitory effects of caffeine on RCC cell growth. Moreover, inhibition of G6PDH activity by caffeine led to a reduction in the intracellular levels of NADPH and reactive oxygen species (ROS), and altered the expression of redox-related proteins in RCC cells. Accordingly, caffeine could inhibit tumor growth through inhibition of G6PDH activity in vivo. Taken together, these results demonstrated that caffeine can target G6PDH to disrupt redox homeostasis and inhibit RCC tumor growth, and has potential as a therapeutic agent for the treatment of RCC.
研究领域:
肿瘤学
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生物化学
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药理学
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产品关联
DOI
PubMed
高影响因子
PROCESS BIOCHEMISTRY
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2022
·
IF 4.885
Wenhao Qu,Yajun Bai,Tai-Ping Fan,Xiaohui Zheng,Yujie Cai
Lactobacillus reuteri DSM20016 has several aldo-keto reductases (AKRs), and they were heterologously expressed in E. coli BL21 (DE3) in this study. Subsequently, these AKRs were purified, and their enzymatic properties were investigated. It was found that these enzymes used NADPH as coenzyme and did not depend on metal ions. The optimum pH of these AKRs was a slightly acidic (AKR-1 and AKR-6: 5.5; AKR-3 and AKR-8: 6; AKR-2 and AKR-4: 6.5). These enzymes had a broad substrate spectrum and could convert a variety of aldehydes to alcohols; however, they did not catalyze the reverse reactions. All results showed that Lactobacillus reuteri had the ability to produce various volatile compounds, which allowed Lactobacillus reuteri to impart complex aroma to something.
研究领域:
微生物学
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生物化学
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酶学
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食品科学
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DOI
高影响因子
危险品化学品经营许可证（带存储）