EUROPEAN JOURNAL OF PHARMACOLOGY
·
2019
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IF 3.17
Jiamao Lin,Lijun Fang,Hui Li,Zhenxiang Li,Linmao Lyu,Haiyong Wang,Jun Xiao
Doxorubicin (DOX) is a classic anti-tumor chemotherapeutic used to treat a wide range of tumors. One major downfall of DOX treatment is it can induce fatal cardiotoxicity. Astragaloside IV (AS-IV) is one of the primary active ingredients that can be isolated from the traditional Chinese herbal medicine, Astragalus membranaceus . This study uses both in vitro and in vivo tools to investigate whether AS-IV alleviates DOX induced cardiomyopathy . We found that AS-IV supplementation alleviates body weight loss, myocardial injury , apoptosis of cardiomyocytes, cardiac fibrosis and cardiac dysfunction in DOX-treated mice. Also, DOX-induced cardiomyocyte injury and apoptosis were effectively improved by AS-IV treatment in vitro . NADPH oxidase (NOX) plays an important role in the progress of the oxidative signal transduction and DOX-induced cardiomyopathy. In this study, we found that AS-IV treatment relieves DOX-induced NOX2 and NOX4 expression and oxidative stress in cardiomyocytes. In conclusion, AS-IV, an antioxidant, attenuates DOX-induced cardiomyopathy through the suppression of NOX2 and NOX4.
研究领域:
药理学
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心脏病学
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肿瘤学
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分子生物学
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PubMed
高影响因子
CATALYSIS LETTERS
·
2018
·Vol. 148
·期 6
·1714-1722
·
IF 2.911
Luo Wei,Deng Xin-Xing,Huo Jiao,Ruan Tao,Gong Zhi-Wei,Yan Jia-Bao,Yang Zhong-Hua,Quan Can,Cui Zhi-Fang
Abstract Microalgae are excellent biocatalyst candidates for photo-biocatalytic-asymmetric reduction of prochiral ketones to produce enantiomer alcohols. In the biocatalysis asymmetric reduction of carbonyl group process, the cofactor, NADPH, plays a key role. So the current study focused on the effect of NADPH availability on the reduction. Photosynthesis is a main pathway for NADPH regeneration in microalgae, and the ferredoxin-NADP + oxidoreductase (FNR) is the key enzyme in this process. In this work, we constructed an engineered cyanobacterium Synechocystis sp. PCC6803:: Ω- P petE-petH to overexpress FNR gene petH to strengthen NADPH regeneration. The results show that the intracellular NADPH content was increased 80.8% compared to the wild type. Applying the Synechocystis sp. PCC6803:: Ω- P petE-petH to catalyze the model asymmetric reduction reaction (reduction of acetophenone to S -phenylethanol), the yield was improved 61.2% compared to the wild-type. This work makes the microalgal biocatalysis asymmetric reduction process become more preponderant via providing a new route to enhance the regeneration of cofactor NADPH. Graphical Abstract Putative metabolic process of nicotinamide adenine dinucleotide phosphate (NADPH) regeneration and the photo-biocatalytic-asymmetric reduction reaction of prochiral ketones in microalgal cells.
研究领域:
生物化学
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合成生物学
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DOI
高影响因子
ACS Sustainable Chemistry & Engineering
·
2019
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IF 6.97
Guochao Xu,Cheng Zhu,Aitao Li,Yan Ni,Ruizhi Han,Jieyu Zhou,Ye Ni
Cofactor regeneration using diols as “smart cosubstrates” is one of the most promising approaches, due to the thermodynamic preference and 0.5-equiv requirement. In order to establish an efficient NADPH regeneration system with 1,4-butanediol (1,4-BD), a NADP+-dependent alcohol dehydrogenase from Kluyveromyces polysporus (KpADH) was engineered to solve the kinetic imbalance. Several hotspots were identified through molecular dynamic simulation and subjected to saturation and combinatorial mutagenesis. Variant KpADHV84I/Y127M exhibited a lower KM of 15.1 mM and a higher kcat of 30.1 min–1 than WTKpADH. The oxidation of 1,4-BD to 4-hydroxybutanal was found to be the rate-limiting step, for which the kcat/KM value of double mutant KpADHV84I/Y127M was 2.00 min–1·mM–1, 11.6-fold higher than that of WTKpADH. KpADHV84I/Y127M preferred diols with a longer chain length (C5–C6). The ratio of kcat/KM toward 2-hydroxytetrahydrofuran (2-HTHF), in comparison to 1,4-BD, in KpADHV84I/Y127M was dramatically reduced by almost 100-fold compared to WTKpADH, which was advantageous for NADPH regeneration. As much as 100 mM phenylpyruvic acid could be reduced into d-phenylalanine with 99.2% conversion in 6 h using merely 0.5 equiv of 1,4-BD. Both the improved catalytic efficiency toward 1,4-BD and the balanced kcat/KM between 1,4-BD and 2-HTHF contributed to the higher NADPH regeneration efficiency. This study provides guidance for engineering alcohol dehydrogenases for cosubstrate specificity toward diols and its application in NADPH regeneration for the preparation of chiral compounds of pharmaceutical relevance.
研究领域:
生物化学
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代谢工程
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合成生物学
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DOI
高影响因子
ENZYME AND MICROBIAL TECHNOLOGY
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2025
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IF 3.7
Yingying Zhu,Dawei Ni,Zeyu Li,Zhebin Hao,Liang Wang,Wanmeng Mu
Indigo is an important blue pigment widely used in textile, food, and medicine industries. Biological production of indigo attracts increasing attention recently. Cell factory production of indigo encounters the problem of the toxicity of the precursor indole. Enzymatic production is the alternative biological approach, however, NADPH regeneration should be solved. In this study, flavin-containing monooxygenase from Methylophaga aminisulfidivorans was used for enzymatic production of indigo from indole and formate dehydrogenase from Pseudomonas sp. 101 was co-expressed for NADPH regeneration. Indigo production was enhanced by combination of molecular modification, promoter engineering, and translation initiation region engineering. Finally, 0.183 g/L of indigo was produced from 0.5 g/L of indole and 0.5 mM of sodium formate, with the conversion ratio of 32.5 %. This study demonstrates a feasible and effective strategy for enzymatic production of indigo.
研究领域:
生物催化
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酶促合成
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代谢工程
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工业微生物学
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应用生物化学
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DOI
PubMed
高影响因子
CHINESE JOURNAL OF CHEMISTRY
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2022
·Vol. 41
·期 1
·57-63
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IF 5.56
Ziwei Su,Xiangxiang Zhang,Weichen Wang,Mingdong Dong,Xiaojun Han
Comprehensive Summary The bottom-up construction of self-powered artificial cells is significant to understand the energy supply and metabolism of nature cells. Here, we demonstrate an efficient manner to build thylakoid-containing artificial cells, which continuously convert light energy into chemical energy to supply adenosine 5’-triphosphate (ATP) under light illumination. The production of ATP supplies energy to promote the biological enzyme cascade reactions, where glucose is transformed into glucose 6-phosphate (G6P) under the catalysis of hexokinase (HK). G6P was further converted to gluconolactone 6-phosphate (PG) in the presence of 6-phosphate dehydrogenase (G6PDH), meanwhile NADP + was converted to nicotinamide adenine dinucleotide phosphate (NADPH). The self-powered artificial cells were demonstrated to generate ATP and NADPH successively, which provided a way for building more complicated artificial cells.
研究领域:
生物化学
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细胞生物学
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合成生物学
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能量代谢
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DOI
高影响因子
FREE RADICAL BIOLOGY AND MEDICINE
·
2025
·
IF 8.2
Yuqian Li,Lijingzhe Hou,Kang Cheng,Guangbo He,Jau-Shyong Hong,Sheng Li,Jie Zhao,Yanjie Guo
Inflammatory bowel disease (IBD), including Crohn's disease and ulcerative colitis, is a chronic inflammatory condition of the gastrointestinal tract with limited treatment options and no definitive cure. Emerging evidence implicates NADPH oxidase (Nox) in IBD pathogenesis, but the specific subtype involved and its therapeutic potential remain unclear. To assess the efficacy of dextromethorphan (DM), a potent Nox2 inhibitor, we employed a dextran sulfate sodium (DSS)-induced chronic colitis model in mice. Mice exposed to intermittent DSS for 34 days developed IBD-like symptoms, including weight loss, elevated disease activity index (DAI), colon shortening, and epithelial barrier disruption. DM exhibited a bimodal protective effect: both standard-dose DM (10 mg/kg/day) and ultralow-dose DM (ULDM, 10 ng/kg/day) significantly reduced colitis severity, with ULDM proving more effective and selected for further studies. Notably, ULDM remained effective when post-administered after DSS exposure. Histological analyses revealed that ULDM reduced immune cell infiltration, crypt damage, and tissue disruption. It also suppressed pro-inflammatory cytokine expression (MCP-1, IL-6, IL-1β, TNF-α) and oxidative stress markers (myeloperoxidase, malondialdehyde, 8-hydroxydeoxyguanosine), while enhancing anti-inflammatory cytokines (IL-4, IL-10) and antioxidant enzymes (catalase, superoxide dismutase). Importantly, ULDM protected Nox1 KO but not Nox2 KO mice, indicating a Nox2-dependent mechanism. In vitro, ULDM inhibited Nox2 activation in primary macrophages and RAW 264.7 cells by blocking the membrane translocation of p47 phox . Furthermore, our study suggested a feed-forward inflammatory cycle between epithelial cell death and macrophage overactivation that exacerbated colitis. Together, these findings demonstrated that Nox2 played a central role in DSS-induced chronic colitis and identified ultralow-dose dextromethorphan as a promising, mechanism-based therapeutic candidate for IBD.
研究领域:
免疫学
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药理学
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胃肠病学
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分子生物学
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炎症性肠病研究
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2019
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IF 4.784
Fei-Long Li,Qiang Zhou,Wei Wei,Jian Gao,Ye-Wang Zhang
Enzymatic NADP + regeneration is a promising approach to produce valuable chemicals under economic conditions. Among all the enzymatic routes, using water-forming NADH oxidase is an ideal one because there is no by-product. However, most NADH oxidases have a low specific activity to NADPH. In this work, a thermostable NADH oxidase from Lactobacillus rhamnosus (LrNox) was rationally engineered to switch its specificity from NADH to NADPH. The results show that mutants D177A, G178R, D177A/G178R, D177A/G178R/L179S improved the NADPH activity by a factor of 4–6. The highest NADPH catalytic efficiency ( K cat /K m 223.71 S −1 μm −1 , 47.6-fold higher than wild-type LrNox) and 51% of NADH activity retention were achieved by replacing the single amino acid Leu179 for serine (L179S) in LrNox. Modeling of L179S-NADPH complex reveals that the phosphate group of NADPH interacts with the hydroxyl of Ser179 with a strong hydrogen bond and several shorter hydrogen bonds with the amino group of Lys185 could stabilize the binding of NADPH in the L179S mutant. This work provides an efficient method for converting NAD(P)H specificity and shows that L179S mutant is a potential and efficient auxiliary enzyme for NADP + regeneration.
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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DOI
PubMed
高影响因子
ACS Synthetic Biology
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2025
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IF 3.9
Jiaying Ai,Ziyang Yin,Jikai Gao,Wenjing Wang,Fuping Lu,Hui-Min Qin,Shuhong Mao
P450 enzymes are promising biocatalysts and play an important role in the field of drug synthesis due to their high catalytic activity and stereoselectivity. CYP109E1 from Bacillus megaterium was used to convert VD3 for the production of 25(OH)VD3. However, the industrial production was still limited due to the low catalytic performance of CYP109E1. To overcome this, we constructed an engineered strain containing a modified CYP109E1 coupled with an efficient electron transfer chain and NADPH regeneration system. First, Adx4–108T69E-Fpr was identified as the most compatible redox partner for the enzyme based on in-silico analysis. Then, targeted mutations were introduced at the substrate channel of CYP109E1, resulting in higher production efficiency. Next, the production of 25(OH)VD3 was increased by 13.1% after introducing a double Adx4–108T69E expression cassette. Finally, an NADPH regeneration system was introduced by overexpressing zwf, which increased the yield of 25(OH)VD3 48.7%. These results demonstrate that recombinant Escherichia coli BL21 (DE3) coexpressing CYP109E1_R70A-ZWF and 2Adx4–108T69Es-Fpr is an efficient whole-cell biocatalyst for the synthesis of 25(OH)VD3, illustrating an attractive strategy for improving the catalytic efficiency of P450 enzymes.
研究领域:
生物化学
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药物合成
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酶工程
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DOI
PubMed
高影响因子
JOURNAL OF BIOSCIENCE AND BIOENGINEERING
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2022
·
IF 3.185
Lin Yuan,Yan-Li Qin,Zhi-Cheng Zou,Bright Appiah,Hao Huang,Zhong-Hua Yang,Can Qun
The intracellular NAD(P)H insufficiency is the key factor which limits the reduced product (such as chiral alcohols) synthesis by whole cell biocatalysis or microbial cell factory. In this paper, we reported a novel solution to increase NADPH supply through strengthening the pentose phosphate pathway (PPP) flux with overexpression of extra zwf (gene for glucose 6-phosphatedehydrogenase) and glk (gene for glucokinase) by recombinant Escherichia coli BL21(DE3)/pETDuet-1- glk - zwf and pET28a- bccr containing a carbonyl reductase gene bccr . The amount of intracellular NADPH was significantly increased from 150.3 μmol/L to 681.8 μmol/L after strengthening the PPP flux, which was 4.5-fold to that of the control. It was applied to improve the asymmetric reduction of 4-chloroacetoacetate to ethyl S -4-chloro-3-hydroxybutylate catalyzed by the Bc CR, which increased the reaction yield 2.8-fold to the control. This strategy provides a new way to increase NADPH supply in E. coli cell factories.
研究领域:
生物化学
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生物技术
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代谢工程
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DOI
PubMed
高影响因子
American Journal of Translational Research
·
2017
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IF 2.829
Yuanfeng Zhu,Shijun Fan,Ning Wang,Xiaoli Chen,Yongjun Yang,Yongling Lu,Qian Chen,Jiang Zheng,Xin Liu
Activation of NADPH oxidase 2 (NOX2) triggers reactive oxygen species (ROS) generation, both of which are essential for robust microbial clearance by phagocytes. However, it is unknown whether inhibition of NOX2 activation or ROS generation affects cellular phagocytosis. Here, we found that the classic NOX2 inhibitor diphenyleneiodonium (DPI) induced uptake of E. coli by murine peritoneal macrophages through enhancing phagocytosis, and this effect was temperature-sensitive and attenuated by cytochalasin D as well as chemical inhibition of Syk and PLCγ, two downstream kinases involved in actin polymerization during phagocytosis. DPI also decreased the production of TNF-α and IL-6 resulting from E. coli stimulation. The DPI-induced enhancement of phagocytosis was independent of NOX2 inhibition or ROS generation but depended on increased intracellular calcium and activation of the p38 MAPK signaling pathway. Furthermore, DPI enhanced bacterial elimination and ameliorated inflammation in E. coli -infected mice, leading to improved survival. Our results demonstrate that DPI facilitates ROS-independent bacterial phagocytosis by macrophages through activation of calcium and p38 MAPK signaling pathways.
研究领域:
免疫学
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分子生物学
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医学
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DOI
PubMed
高影响因子
BIORESOURCE TECHNOLOGY
·
2020
·
IF 7.539
Yuan-Yuan Li,Qing Li,Peng-Qi Zhang,Cui-Luan Ma,Jian-He Xu,Yu-Cai He
In this study, tin-loaded sulfonated zeolite (Sn-zeolite) catalyst was synthesized for catalysis of raw corncob (75.0 g/L) to 103.0 mM furfural at 52.3% yield in water (pH 1.0) at 170 °C. This corncob-derived furfural was subsequently biotransformed with recombinant E. coli CG-19 cells coexpressing NADPH-dependent reductase and glucose dehydrogenase at 35 °C by supplementary of glucose (1.5 mol glucose/mol furfural), sodium dodecyl sulfate (0.50 mM) and NADP + (1.0 μmol NADP + /mmol furfural) in the aqueous catalytic media (pH 7.5). Both sodium dodecyl sulfate (0.50 mM) and Sn 4+ (1.0 mM) could promote reductase activity by 1.4-folds. Within 3 h, furfural was wholly catalyzed into furfuryl alcohol. By combining chemical catalysis with Sn-zeolite and biocatalysis with CG-19 cells in one-pot, an effective and sustainable process was established for tandemly catalyzing renewable biomass into furfuryl alcohol under environmentally-friendly way.
研究领域:
生物化学
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催化化学
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可再生资源转化
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF PHARMACY AND PHARMACOLOGY
·
2021
·Vol. 74
·期 4
·596-604
·
IF 3.765
Sun Liyun,Shang Huoli,Wu Yuzhen,Xin Xiujuan
ObjectivesThe combination of gemcitabine (Gem) and hypericin (HY) enhances the apoptosis of Capan-2 cells, providing a promising option for the treatment of pancreatic cancer. Our study further explored the cytotoxic mechanism of HY combined with chemotherapy drugs on pancreatic cancer.MethodsThe proliferation rate of the cells assayed with the MTT method. The ROS (reactive oxygen species) levels of each treatment were evaluated by DCFH-DA oxidisation methods. The activity of glutathione reductase and the levels of reduced glutathione (GSH) and oxidised glutathione (GSSG) were assessed using assay kits. The expression levels of apoptosis-related proteins were analysed by western blotting.Key findingsThe activity of glucose-6-phosphate dehydrogenase (G6PDH), a key enzyme of the pentose phosphate pathway, significantly decreased in Gem + HY groups, however, the ROS level enhanced accompanying with GSH depleting, mitochondrial membrane depolarisation and cytochrome C release. Gem + HY inhibits the expression of Bcl-2 but stimulates Bax level, triggering caspase activation and PARP cleavage and thus promoted apoptosis of Capan-2 cells.ConclusionsWe demonstrated that Gem combined HY-PDT could inhibit the proliferation of Capan-2 cells and induce cell apoptosis. HY-PDT combined with Gem had a great potential on pancreatic cancer treatment clinically.
研究领域:
肿瘤学
·
药理学
·
细胞生物学
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DOI
PubMed
高影响因子
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