FOOD AND CHEMICAL TOXICOLOGY
·
2021
·
IF 6.025
Ying Liu,Shiyu Zhang,Tingting Jiang,Rong Tan,Yao Fu,Xiaojing Yang,Bowen Gong,Ying Zou,Weiwei Li,Jiang Zheng
Bergamottin (BGM) is a major furanocoumarin constituent of grapefruit and is reported to have inhibitory effects on cytochrome P450 enzymes. This study investigated the chemical interactions between BGM and the enzyme CYP2C9 . BGM exhibited time-, concentration-, and NADPH-dependent inhibition of CYP2C9. Co-incubation with diclofenac, a reversible inhibitor of CYP2C9, attenuated the time-dependent enzyme inhibition. Exhaustive dialysis did not restore enzyme activity post-inhibition. Glutathione (GSH) and catalase/superoxide dismutase failed to reverse BGM-induced CYP2C9 inactivation. A GSH trapping study suggested that BGM was metabolized to an epoxide and/or γ-ketoenal that may have been responsible for the enzyme inactivation. In conclusion, BGM can be characterized as a mechanism-based inactivator of CYP2C9 acting via the formation of an epoxide and/or γ-ketoenal.
研究领域:
药理学
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生物化学
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毒理学
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DOI
PubMed
高影响因子
Science China-Life Sciences
·
2020
·Vol. 64
·期 3
·443-451
·
IF 4.611
Luan Dongrui,Zhao Zengteng,Xia Dandan,Zheng Qiuling,Gao Xiaonan,Xu Kehua,Tang Bo
Sodium selenite has alleviating effects on liver fibrosis; however, its therapeutic molecular mechanism remains unclear. Herein, hydrogen selenide, a major metabolite of Na 2 SeO 3 , was tested to uncouple the sulfilimine bond in collagen IV, the biomarker of liver fibrosis. A mouse model of liver fibrosis was constructed via a CCl 4 -induced method, followed by the administration of 0.2 mg kg −1 Na 2 SeO 3 via gavage three times per week for 4 weeks. Changes in H 2 Se, NADPH, and H 2 O 2 levels were monitored in real time by using NIR-H 2 Se, DCI-MQ-NADPH, and H 2 O 2 probes in vivo , respectively. H 2 Se continuously accumulated in the liver throughout the Na 2 SeO 3 treatment period, but the levels of NADPH and H 2 O 2 decreased. The expression of collagen IV was analyzed through Western blot and liquid chromatography-mass spectrometry. Results confirmed that the sulfilimine bond of collagen IV in the fibrotic mouse livers could be broken by H 2 Se with the Na 2 SeO 3 treatment. Therefore, the therapeutic effect of Na 2 SeO 3 on liver fibrosis could be mainly attributed to H 2 Se that uncoupled the sulfilimine bond to induce collagen IV degradation. This study provided a reasonable explanation for the molecular mechanism of the in vivo function of Na 2 SeO 3 and the prevention of liver fibrosis by administering inorganic selenium.
研究领域:
药理学
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生物化学
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肝病学
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2021
·
IF 6.953
Yanan Zhang,Xue Li,Di Xu,Mengyun Wu,Yinghui Dai,Mingyu Xia,Dong Wang
Bufadienolides are the main active ingredients of Venenum Bufonis, which is a widely used traditional Chinese medicine secreted from parotoid gland and skin glands of Bufo bufo gargarizans . According to the transcriptome analysis, “cholesterol-bile acid-bufadienolidies pathway” was proposed as animal-derived bufadienolides biosynthesis pathway by us previously. In this pathway 3 β -hydroxysteroid dehydrogenase (3 β HSD) and steroid 5 β -reductase (SRD5 β ) might be the key enzymes to convert the A/B ring to cis -configuration. Therefore, as the second report of our group, here we report the cloning of the full length of SRD5 β cDNA of B. bufo gargarizans ( Bbg - SRD5β ) from the parotoid gland of B. bufo gargarizans for the first time, and site-directed mutagenesis was used to explored the character of Bbg -SRD5 β . Bbg -SRD5 β had an open reading frame of 981 bp and encoded 326 amino acids residues. The expression conditions of the recombinant Bbg -SRD5 β in E. coli BL21 (DE3) harbored with pCold- Bbg -SRD5 β was optimized as induction for 10 h at 15 °C with 0.1 mM IPTG. With NADPH as a cofactor, Bbg -SRD5 β can reduce the Δ 4,5 double bonds of progesterone to generate dihydroprogesterone õwithout substrate inhibition effect. The catalytic rate of mutant type Bbg -SRD5 β -Y132G was 1.8 times higher than that of wild type Bbg -SRD5 β . Although Bbg -SRD5 β was almost unable to reduce the progesterone to dihydroprogesterone after mutation of V309, the affinity of enzyme with NADPH changed significantly. Bbg -SRD5 β is the key enzymes to convert the A/B ring of steroid to cis -configuration, and V309 is a key site affecting the binding affinity of enzyme with NADPH, and the mutation of Y132 can adjust the catalytic rate of Bbg -SRD5 β .
研究领域:
生物化学
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分子生物学
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中医药学
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药理学
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DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2024
·
IF 5.7
Xue Bai,Shuling Wang,Qin Zhang,Yuhan Hu,Jiawei Zhou,Lianhui Men,Dengyu Li,Jing Ma,Qiuhui Wei,Mengdie Xu,Xiaopu Yin,Tianyuan Hu
Miltiradiene serves as a crucial precursor in the synthesis of various high-value abietane-type diterpenes, exhibiting diverse pharmacological activities. Previous efforts to enhance miltiradiene production have primarily focused on the mevalonate acetate (MVA) pathway. However, limited emphasis has been placed on optimizing the supply of acetyl-CoA and NADPH. In this study, we constructed a platform yeast strain for miltiradiene production by reinforcing the biosynthetic pathway of geranylgeranyl diphosphate (GGPP) and acetyl-CoA, and addressing the imbalance between the supply and demand of the redox cofactor NADPH within the cytoplasm, resulting in an increase in miltiradiene yield to 1.31 g/L. Furthermore, we conducted modifications to the miltiradiene synthase fusion protein tSmKSL1-CfTPS1. Finally, the comprehensive engineering strategies and protein modification strategies culminated in 1.43 g/L miltiradiene in the engineered yeast under shake flask culture conditions. Overall, our work established efficient yeast cell factories for miltiradiene production, providing a foothold for heterologous biosynthesis of abietane-type diterpenes.
研究领域:
生物技术
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代谢工程
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药物化学
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DOI
PubMed
高影响因子
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS
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2014
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IF 3.043
Chenxi Ning,Erzheng Su,Dongzhi Wei
Lodderomyces elongisporus LH703 isolated from soil samples contained three novel aldo–keto reductases (AKRs) (LEAKR 48, LEAKR 49, and LEAKR 50). The three enzymes were cloned, expressed, and purified to homogeneity for characterization. These three AKRs shared <40% amino acid identity with each other. LEAKR 50 was identified as a member of AKR3 family, whereas the other two LEAKRs were identified as members of two novel AKR families, respectively. All the three AKRs required nicotinamide adenine dinucleotide phosphate as a cofactor. However, they showed diverse characteristics, including optimum catalyzing conditions, resistance to adverse reaction conditions, and substrate specificity. LEAKR 50 was estimated to be a promising biocatalyst that could reduce ethyl 4-chloroacetoacetate with high enantiomeric excess (98% e. e.) and high activity residue under adverse conditions.
研究领域:
生物化学
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酶学
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微生物学
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DOI
PubMed
高影响因子
Reaction Chemistry & Engineering
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2020
·Vol. 5
·期 3
·531-538
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IF 3.441
Xiaolin Pei,Jiapao Wang,Haoteng Zheng,Pengfei Cheng,Yifeng Wu,Anming Wang,Weike Su
D-(−)-Pantolactone (D-PL) is the key chiral intermediate used to synthesize calcium D-pantothenate. In a previous study, a conjugated polyketone reductase (CduCPR) was discovered to efficiently catalyse ketopantolactone (KPL) to D-PL with a high stereoselectivity. Nevertheless, this method was limited due to the expensive coenzyme, nicotinamide adenine dinucleotide phosphate hydrate (NADPH), and the poor stability of substrate KPL in pure aqueous buffer. Here, to overcome these problems, we developed a whole-cell biotransformation process to produce D-PL in a biphasic reaction system. Recombinant CduCPR and glucose dehydrogenase (BsuGDH) were co-expressed in Escherichia coli to simultaneously achieve the synthesis of D-PL and the regeneration of NADPH. The biphasic reaction system contained 15% dichloromethane (v/v) which significantly inhibited the undesirable hydrolysis of KPL. Approximately, 2.8% of 100 mM KPL was spontaneously hydrolysed to ketopantoic acid in the biphasic system, which was lower than 54% of pure aqueous buffer. The biphasic reaction system was optimized and scaled up to prepare D-PL. In a fed-batch biphasic reaction, the substrate KPL was continuously fed into the reactor after a batch reaction. The D-PL concentration reached 0.77 mol L−1 in the reaction mixture at 7 h, and its enantiomeric excess was 99%. This study presented an alternative method to asymmetrically synthesize enantiopure D-PL for further industrial application.
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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DOI
高影响因子
PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY
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2019
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IF 1.117
Chuhong Zong,Xu Zhang,Fei Yang,Yafeng Zhou,Nan Chen,Zuisu Yang,Guofang Ding,Fangmiao Yu,Yunping Tang
(S)-1-(2, 6-dichloro-3-fluorophenyl) ethanol, the key chiral intermediate of crizotinib, was prepared from 1-(2, 6-dichloro-3-fluorophenyl) ethanone using the alcohol dehydrogenases from Lactobacillus kefir (ADH-LK) with a tetrad mutant (ADH-LKM, F147L/Y190P/V196L/A202W), coupled with glucose dehydrogenase (GDH). In the present study, ADH-LKM and GDH were successfully heterologous expressed in recombinant Escherichia coli. During the regeneration of NADPH with GDH, 150 g/L substrate was totally transformed into target chiral alcohol with an enantiomeric excess value of 99.9% after 12 h at 30 °C (pH 7.0). Our study demonstrates the potential for industrial green production of the key chiral intermediate of crizotinib.
研究领域:
生物催化
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药物化学
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酶工程
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产品关联
DOI
PubMed
高影响因子
Signal Transduction and Targeted Therapy
·
2022
·Vol. 7
·期 1
·1-17
·
IF 38.104
Zeng Hao,Pan Ting,Zhan Meiling,Hailiwu Renaguli,Liu Baolin,Yang Hua,Li Ping
Endothelial-to-mesenchymal transition (EndoMT), the process wherein endothelial cells lose endothelial identity and adopt mesenchymal-like phenotypes, constitutes a critical contributor to cardiac fibrosis. The phenotypic plasticity of endothelial cells can be intricately shaped by alteration of metabolic pathways, but how endothelial cells adjust cellular metabolism to drive EndoMT is incompletely understood. Here, we identified 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3) as a critical driver of EndoMT via triggering abnormal glycolysis and compromising mitochondrial respiration. Pharmacological suppression of PFKFB3 with salvianolic acid C (SAC), a phenolic compound derived from Salvia miltiorrhiza , attenuates EndoMT and fibrotic response. PFKFB3-haplodeficiency recapitulates the anti-EndoMT effect of SAC while PFKFB3-overexpression augments the magnitude of EndoMT and exacerbates cardiac fibrosis. Mechanistically, PFKFB3-driven glycolysis compromises cytoplasmic nicotinamide adenine dinucleotide phosphate (reduced form, NADPH) production via hijacking glucose flux from pentose phosphate pathway. Efflux of mitochondrial NADPH through isocitrate/α-ketoglutarate shuttle replenishes cytoplasmic NADPH pool but meanwhile impairs mitochondrial respiration by hampering mitochondrial iron-sulfur cluster biosynthesis. SAC disrupts PFKFB3 stability by accelerating its degradation and thus maintains metabolic homeostasis in endothelial cells, underlying its anti-EndoMT effects. These findings for the first time identify the critical role of PFKFB3 in triggering EndoMT by driving abnormal glycolysis in endothelial cells, and also highlight the therapeutic potential for pharmacological intervention of PFKFB3 (with SAC or other PFKFB3 inhibitors) to combat EndoMT-associated fibrotic responses via metabolic regulation.
研究领域:
心脏病学
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细胞代谢
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药理学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2024
·
IF 13.3
Mei Xue,Zhiwen Ma,Yali Pan,Sihui Mao,Yingqiang Sun,Yupeng Yuan
Using photoluminescent (PL) graphitic carbon nitride (g-C 3 N 4 ) to convert ultraviolet (UV) light into chloroplast absorbed blue or near infrared (NR) is an attractive methodology to facilitate sunlight energy storage and carbon sequestration by microalgae. However, most researches focus on inhibiting water bloom by g-C 3 N 4 , applying PL g-C 3 N 4 to promote growth of microalgae is rarely reported. Accordingly, PL g-C 3 N 4 is synthesized by a microwave heating method and co–cultured with Chlorella vulgaris ( C. vulgaris ) with concentration of 0, 50, and 250 mg/L, named as C. vulgaris -0, C. vulgaris -50, and C. vulgaris -250, respectively, to study effect of PL g-C 3 N 4 on growth of C. vulgaris . Results indicate that there is no significant effect of microwave synthesized g-C 3 N 4 on morphology of microalgae cells, while cell viability and growth rate of C. vulgaris -50 is obviously enhanced. Further analysis indicates that, on the one hand, PL fluorescence at 615 nm enlarges light absorption of microalgae; on the other hand, photogenerated electrons by g-C 3 N 4 accelerate electron transfer in photosystem, subsequently enhancing formation of NADPH and ATP, which is favorable to growth of microalgae. Metabolism analysis illustrates that glucose, fructose, and galactose, involved in glycolysis and photosynthesis, are all up-regulated in C. vulgaris -50 group. This work reveals low concentration of PL g-C 3 N 4 facilitates protein synthesis and energy metabolism through enhancing light absorption and electron transfers, providing a biochemical strategy to enhance carbon sequestration.
研究领域:
环境科学
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微生物学
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生物技术
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生物化学
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产品关联
DOI
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Yuqian Chi,Xiaoliang Zhu,Yaxuan Chen,Xin Li,Ziyi Jiang,Xiaoyang Jian,Mengyuan Lian,Xiaodi Wu,Lei Wang,Mengmeng Sun,Xiaowei Shi
Piperlonguminine (PLG) is a major alkaloid found in Piper longum fruits. It has been shown to possess a variety of biological activities, including anti-tumor, anti-hyperlipidemic, anti-renal fibrosis and anti-inflammatory properties. Previous studies have reported that PLG inhibits various CYP450 enzymes. The main objective of this study was to identify reactive metabolites of PLG in vitro and assess its ability to inhibit CYP450. In rat and human liver microsomal incubation systems exposed to PLG, two oxidized metabolites (M1 and M2) were detected. Additionally, in microsomes where N -acetylcysteine was used as a trapping agent, N -acetylcysteine conjugates (M3, M4, M5 and M6) of four isomeric O -quinone-derived reactive metabolites were found. The formation of metabolites was dependent on NADPH. Inhibition and recombinant CYP450 enzyme incubation experiments showed that CYP3A4 was the primary enzyme responsible for the metabolic activation of PLG. This study characterized the O -dealkylated metabolite (M1) through chemical synthesis. The IC 50 shift assay showed time-dependent inhibition of CYP3A4, 2C9, 2E1, 2C8 and 2D6 by PLG. This research contributes to the understanding of PLG-induced enzyme inhibition and bioactivation.
研究领域:
药理学
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毒理学
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生物化学
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产品关联
DOI
PubMed
高影响因子
BIORESOURCE TECHNOLOGY
·
2024
·
IF 9.7
Guofu Zhao,Ming Chen,Jingwen Liu,Shang Wang,Dongmei Fu,Chunzhi Zhang
Antioxidant addition is an effective strategy to achieve docosahexaenoic acid (DHA) overproduction in oleaginous microorganisms. Nevertheless, antioxidants like phenolic compounds sometimes exert pro-oxidant activity. In this work, effects of proanthocyanidins (PAs) on fermentation performance and oxidative stress in Schizochytrium sp. were investigated. Low PAs addition (5 mg/L) reduced reactive oxygen species and enhanced lipogenic enzymes activities and NADPH, resulting in significant increase in lipid (20.3 g/L) by 33.6 % and DHA yield (9.8 g/L) by 53.4 %. In contrast, high PAs addition (500 mg/L) exerted pro-oxidant effects, aggravated oxidative damage and lipid peroxidation, leading to sharp decrease in biomass (21.3 g/L) by 35.1 %, lipid (8.2 g/L) by 46.0 %, and DHA (2.9 g/L) by 54.8 %. Therefore, the antioxidant concentration is especially crucial in DHA production. This study is the first to report concentration-dependant dual roles of PAs in oxidative stress and DHA production in Schizochytrium sp., providing new insights into microbial DHA production.
研究领域:
微生物学
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生物化学
·
生物技术
✓ 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
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生物化学
·
细胞生物学
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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
危险品化学品经营许可证（带存储）