MOLECULAR PHARMACEUTICS
·
2013
·
IF 4.57
Wenjing Li,Xinru Li,Yajie Gao,Yanxia Zhou,Shujin Ma,Yong Zhao,Jinwen Li,Yan Liu,Xinglin Wang,Dongdong Yin
The present study aimed to investigate the effect of monomethoxy poly(ethylene glycol)-block-poly(d,l-lactic acid) (mPEG-PLA) on the activity of P-glycoprotein (P-gp) in Caco-2 cells and further unravel the relationship between PLA chain length in mPEG-PLA and influence on P-gp efflux and the action mechanism. The transport results of rhodamine 123 (R123) across Caco-2 cell monolayers suggested that mPEG-PLA unimers were responsible for its P-gp inhibitory effect. Furthermore, transport studies of R123 revealed that the inhibitory potential of P-gp efflux by mPEG-PLA analogues was strongly correlated with their structural features and showed that the hydrophilic mPEG-PLA copolymers with an intermediate PLA chain length and 10.20 of hydrophilic–lipophilic balance were more effective at inhibiting P-gp efflux in Caco-2 cells. The fluorescence polarization measurement results ruled out the plasma membrane fluidization as a contributor for inhibition of P-gp by mPEG-PLA. Concurrently, mPEG-PLA inhibited neither basal P-gp ATPase (ATP is adenosine triphosphate) activity nor substrate stimulated P-gp ATPase activity, suggesting that mPEG-PLA seemed not to be a substrate of P-gp and a competitive inhibitor. No evident alteration in P-gp surface level was detected by flow cytometry upon exposure of the cells to mPEG-PLA. The depletion of intracellular ATP, which was likely to be a result of partial inhibition of cellular metabolism, was directly correlated with inhibitory potential for P-gp mediated efflux by mPEG-PLA analogues. Hence, intracellular ATP-depletion appeared to be possible explanation to the inhibition mechanism of P-gp by mPEG-PLA. Taken together, the establishment of a relationship between PLA chain length and impact on P-gp efflux activity and interpretation of action mechanism of mPEG-PLA on P-gp are of fundamental importance and will facilitate future development of mPEG-PLA in the drug delivery area.
研究领域:
药学
·
药理学
·
药物递送系统
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产品关联
DOI
PubMed
高影响因子
BIOORGANIC CHEMISTRY
·
2026
·
IF 4.7
Jinnian Wang,Haoming Zhao,Shijun Tong,Xingjian Zhang,Shuyu Ding,Han Zhang,Zhong-Hua Yang
The industrial production of pyridoxal 5′-phosphate (PLP)—an indispensable enzymatic cofactor—is hampered by inefficient multistep chemical syntheses and the economic infeasibility of ATP-dependent biocatalytic routes. To address these challenges, we report an ATP-autonomous biocatalytic cascade that couples pyridoxal kinase (PLK) with polyphosphate kinase (PPK) for sustainable PLP synthesis. This system harnesses inexpensive sodium hexametaphosphate (SHMP) as a phosphoryl donor to drive continuous ATP regeneration, thereby enabling efficient phosphorylation of pyridoxal to PLP without exogenous ATP input. Recombinant PLK and PPK were co-expressed in E. coli BL21(DE3), yielding specific activities of 50 and 145 U/g cell , respectively. The whole-cell biocatalyst achieved only about 25% conversion yield due to mass transfer limitations and intracellular PLP degradation. Notably, while the optimized cell-free system (45 °C, pH 6.5, PLK:PPK = 1:3, 25 mM SHMP) attained a remarkable 95% PLP yield. This 3.8-fold enhancement demonstrates the critical advantage of eliminating cellular barriers in complex multi-enzyme cascades. Our work establishes a robust, ATP-independent platform for high-efficiency PLP biosynthesis that seamlessly integrates cofactor regeneration with cascade catalysis, offering a scalable and economically viable route for industrial biomanufacturing. Moreover, it demonstrates the broader utility of polyphosphate-driven energy recycling in sustainable biocatalysis.
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产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2016
·
IF 4.758
Chun An Shi,Xin Zhang,Hong Jun Yin,Hui Fang,Yong Mei Zhao,Luo Liu,Zheng Long Wu,Hai Jun Xu
Adenosine triphosphate (ATP) is an important small biological molecule in all living organisms and the level of it is a vital physiological index associated with many diseases. In this study, a convenient, sensitive and precise method is proposed to quantify ATP that combines glucose phosphorylation with the surface-enhanced Raman scattering (SERS) technique. First, a portion of glucose was consumed by ATP in the glucose phosphorylation reaction. Then, the quantitative determination of the residual glucose was realized by utilizing the SERS technique instead of classical spectrophotometry to analyze the charge transfer complex (CTC) of a chromogen. The ATP concentration of a sample can be determined by subtracting the residual glucose from the original amount. We have demonstrate that ATP is detectable and quantifiable from a glucose concentration 1–1000 times that of the ATP. The detection limit of ATP is as low as 4.69 × 10 −7 M, which is sufficient for the quantification of ATP.
研究领域:
生物化学
·
分析化学
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DOI
高影响因子
BIOCHEMICAL ENGINEERING JOURNAL
·
2026
·
IF 3.6
Xiangcao Cui,Xiaosen Liu,Huiling Zhang,Kun Zhou,Cunzhuo Tang,Chen Deng,Xu Li,Yongjun Qiu,Liqiang Fan,Liming Zhao
A Whole-cell method co-displaying ACT and PPK2 enables low-cost butyrolactam biosynthesis. • Co-display of ACT and PPK2 increases butyrolactam productivity by 46.4% against ACT display alone. • Fed-batch fermentation achieves 39.13 g/L butyrolactam with 94.7% yield, showing industrial potential. Butyrolactam is a high value-added monomer and an important intermediate for various physiologically active substances, and has been widely used in the pharmaceutical, pesticide, and chemical industries. Though the biosynthesis of butyrolactam is gradually advancing, challenges such as the requirement for expensive cofactor adenosine triphosphate (ATP) and the barrier of substrate transmembrane transport in whole-cell synthesis hinder its large-scale industrial application. In this study, an integrated whole-cell catalytic strategy combining ice nucleation protein (INP)-mediated surface display and ATP regeneration was developed. Compared with the butyrolactam productivity of cells intracellularly expressing β-alanine CoA transferase (ACT), co-expressing polyphosphate kinase 2 (PPK2) to regenerate ATP and display ACT on the cell surface only increased it by 23.1% and 25.0%, respectively. Co-displaying ACT and PPK2 further increased the butyrolactam productivity, which was 46.4% more than that of only ACT displayed cells. Using fed-batch fermentation strategy, a final butyrolactam titer of 39.13 g/L and a yield of 94.7% were achieved in the strain co-displaying ACT and PPK2. Our findings demonstrate a potentially attractive strategy for the industrial butyrolactam production.
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DOI
高影响因子
JOURNAL OF CHEMICAL TECHNOLOGY AND BIOTECHNOLOGY
·
2023
·Vol. 98
·期 4
·1025-1031
·
IF 3.4
Jie Zhang,Chenlei Sun,Qingshi Wen,Rongxin Miao,Bingyun Zhang,Ziyi Yan,Hanjie Ying,Dong Liu,Junzhi Wang
BACKGROUND Acetyl phosphate/acetate kinase system is a cost-saving and efficient ATP regeneration system. However, it suffers from accumulation of salts during catalytic reactions. It inhibits the activity of acetate kinase (ACK), a crucial enzyme of this process, and hence decreases the rate of product accumulation and yield of CTP. RESULTS A new halotolerant ACK enzyme based ATP regeneration platform was established in this research by applying the concept of rational design to optimize its building blocks. Selection of residues for rational design followed several rules. First, the less conserved residue positions on the protein surface were considered. Second, avoiding significant residue interactions, suchas salt bridges. Third, following the feature of nature occurring halotolerant enzymes, the basic and neutral surface residues were replaced with acidic ones. Our mutants represent high catalytic activities and high IC50 value (inhibit activity by 50%) at high salt concentrations. Also, when the halotolerant ACK was applied to catalyze CTP production, the maximum titer was 213.5 ± 1.6 mM (which is 12.3-fold higher than that of the traditional process), the productivity was 13.3 ± 0.1 mM L −1 h −1 , and the molar yield to 5′-cytidylate monophosphate (CMP) and utilization efficiency of energy were 92.4% and 72.7%, respectively. CONCLUSION This strategy can be applied to modify other enzymes in the industry and gives insight into resolving salt accumulation during biocatalytic processes. In addition, the halotolerant Adenosine Triphosphate (ATP) regeneration system built in this work can be applied to the synthesis of several other high energy compounds. © 2023 Society of Chemical Industry (SCI).
研究领域:
生物化学
·
工业酶学
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DOI
高影响因子
Theranostics
·
2018
·
IF 8.537
Zhanwei Zhou,Qingyan Zhang,Minghua Zhang,Huipeng Li,Gang Chen,Chenggen Qian,David Oupicky,Minjie Sun
Stimuli-responsive polycations have been developed for improved nucleic acid transfection and enhanced therapeutic efficacy. The most reported mechanisms for controlled release of siRNA are based on polyelectrolyte exchange reactions in the cytoplasm and the degradation of polycations initiated by specific triggers. However, the degradation strategy has not always been sufficient due to unsatisfactory kinetics and binding of cationic fragments to siRNA, which limits the gene silencing effect. In this study, a new strategy that combines degradation and charge reversal is proposed. Methods: We prepared a polycation (CrossPPA) by crosslinking of phenylboronic acid (PBA)-grafted 1.8k PEI with alginate. It was compared with 25k PEI, 1.8k PEI and 1.8k PEI-PBA on siRNA encapsulation, ATP-responsive behavior and mechanism, cytotoxicity, cell uptake, siRNA transfection, in vivo biodistribution and in vivo anti-tumor efficacy. The in vitro and in vivo experiments were performed on 4T1 murine breast cancer cells and 4T1 tumor model separately. Results: The crosslinking strategy obviously improve the siRNA loading ability of 1.8k PEI. We validated that intracellular levels of ATP could trigger CrossPPA disassembly and charge reversal, which resulted in efficient and rapid siRNA release due to electrostatic repulsion. Besides, CrossPPA/siRNA showed strong cell uptake in 4T1 cells compared with 1.8k PEI/siRNA. Notably, the cytotoxicity of CrossPPA was pretty low, which was owing to its biodegradability. Furthermore, the crosslinked polyplexes significantly enhanced siRNA transfection and improved tumor accumulation. The high gene silencing ability of CrossPPA polyplex led to strong anti-tumor efficacy when using Bcl2-targeted siRNA. Conclusion: These results indicated that the ATP-triggered disassembly and charge reversal strategy provided a new way for developing stimuli-responsive siRNA carriers and showed potential for nucleic acid delivery in the treatment of cancer.
研究领域:
生物医学工程
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药学
·
癌症研究
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纳米技术
·
基因治疗
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DOI
PubMed
高影响因子
ACS Applied Bio Materials
·
2022
·
IF 0
Juan Qiao,Qian Ma,Yuying Song,Li Qi
It is necessary to develop reliable chemiluminescence strategies for determination of intracellular adenosine triphosphate (ATP), which is vital in life science and clinical diagnosis. However, the current chemiluminescence methods based on firefly luciferase suffered from low delivery efficiency, unsatisfied targeting performance, and autohydrolysis in living biosystem. To circumvent these drawbacks, a thermoresponsive polymer nanocomposite modified with firefly luciferase and ATP aptamer (PFLNC@aptamer) was fabricated, which targeted ATP and determined the intracellular ATP levels via measuring the chemiluminescence signals at different temperatures. The PFLNC@aptamer exhibited capability for the enzymolysis efficiency regulation, increased 21.0% with temperature change from 37.0 to 25.0 °C. The ATP detection limit was 3.3 nM with a linear relationship from 10.0 nM to 0.1 mM. Moreover, the thermoresponsive nanocomposite could also effectively avoid the interference during delivering firefly luciferase into the living cells and effectively discriminate ATP via the immobilized ATP aptamer, which further confirmed its reliability for practical applications. It paves a specific avenue for effective intracellular ATP monitoring in fundamental and applied research.
研究领域:
生物化学
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分析化学
·
医学诊断
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产品关联
DOI
PubMed
Water
·
2024
·Vol. 16
·期 7
·921
·
IF 3
Teng-Zhou Li,Cheng-Zhuang Chen,Shao-Ying Xing,Ling Liu,Ping Li,Zhi-Hua Li
Triphenyltin (TPT) is an organotin pollutant widely found in the aquatic environment. It has endocrine-disrupting and osmotic pressure toxicity. In this study, the physiological and biochemical effects of TPT and various salinities were investigated in different tissues (gut, gill, and brain) of marine medaka. The exposure experiments were conducted for 42 days in different salinities (0, 15, and 30 ppt) without TPT exposure and in different salinity groups with TPT exposure concentrations of 100 ng/L, respectively. The results showed that the Na+-K+-ATPase (NKA) and Ca2+ATPase activity had significant tissue-specific differences, with the highest activity observed in the gills, indicating their major contribution to osmoregulation. Changes in salinity also resulted in significant alterations in the ion concentrations of the gut and gill tissues in the 0-C and 15-C groups. While the changes in Na+and Cl−were relatively stable, the presence of TPT disrupted the regulation of Ca2+and K+. In conclusion, substantial variations were observed in the osmoregulatory capacity of marine medaka tissues. Environmental concentrations of TPT had little effect on osmotic enzyme activity but interfered with the regulation of Ca2+and K+concentrations in the tissues. This study provides valuable insights into the osmotic toxicity of TPT in aquatic environments with different salinities.
研究领域:
环境科学
·
水生毒理学
·
海洋生物学
·
生理学
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产品关联
DOI
高影响因子
ACS Omega
·
2021
·
IF 3.512
Yun Wang,Puling Li,Xue Zhang,Lingyun Li,Mengjiao Liu,Xiaoqing Li,Yejia Dai,Cun Zhang,Shaojing Li
The processing method for Chinese traditional herbal medicine is “Pao Zhi” in Chinese. This study examined the efficacy of the Pao Zhi on the preparations of Gardeniae Fructus (GF) on a mitochondrial respiratory function in rats. To determine the efficacy of Pao Zhi, we investigated the effects of GF heat processing on mitochondrial respiratory function. To test the GF components, the rats were randomly divided into a geniposide-alone group, crocin-alone group, and combination groups and treated with geniposide and crocin at different ratios. The results showed that a high dose, raw GF was more effective in improving the neurological function, mitochondrial respiratory function, and activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase than the preparations that underwent heating. Moreover, mitochondrial ROS production was the lowest in the raw GF-treated group. In addition, treatments with crocin and GC3 were more effective than geniposide in improving the functional deficit in MCAO rats. In conclusion, our results suggest that raw GF is the most suitable preparation for the treatment of cerebral ischemia, and its underlying mechanisms may be associated with the improvement of mitochondrial respiratory function, increased activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase, and reduced oxidative stress in mitochondria. Our findings suggest that raw GF, especially crocin, could be an ideal therapeutic agent for ischemic stroke.
研究领域:
药理学
·
神经科学
·
中医药学
·
线粒体生物学
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产品关联
DOI
PubMed
高影响因子
Aquaculture Reports
·
2021
·
IF 3.216
Dongyong Fu,Yadong Hu,Peng Chu,Tao Wang,Mingxu Chu,Yonghai Shi,Shaowu Yin,Yongxiang Zhu,Yaohui Wang,Zhenglong Guo
Copper nanoparticles (Cu-NPs) are water pollutants, which has significant toxic environmental effects. However, the effects of Cu-NPs on Takifugu fasciatus are still relatively unknown. Compared with the control (no add CuSO 4 and Cu-NPs). This study aims to describe the pathological changes in the gill and liver of pufferfish exposed to CuSO 4 (Cu 2+ for short) and Cu-NPs dissolved at a concentration of either 20 or 100 μg/L, in a semi-static waterborne exposure regime for 28 days. All Cu treatments resulted in organ injuries, and the pathologies observed following exposure to Cu-NPs were broadly of the same type as those observed following exposure to Cu 2+ . In the gill, these included lamellar fusion, clubbed tips, and mucocytes swollen, and in the liver included oedema, aggregation of blood cells, and necrosis. Furthermore, enzyme activity assays revealed that Ca 2+ -ATP activity significantly increased under the influence of Cu, whilst Na + -K + -ATPase activity was significantly inhibited. In this study, the CRT in T. fasciatus (referred to as Tf CRT) was identified and full-length cDNA sequences were characterized. Subsequently, RT-PCR and western blot analysis were carried out to assess the expressions of Tf CRT in the gill and liver at both the mRNA and protein levels. Under both the Cu-NPs and Cu 2+ treatments, both the mRNA and protein expressions of Tf CRT were considerably higher in the gill and liver compared to that of the control treatment. Furthermore, there were no significant differences between Cu-NPs and Cu 2+ treatment (at the same treatment concentration) in the expressions of Tf CRT at the mRNA or protein levels. Overall, both Cu 2+ and Cu-NPs had negative effects on T. fasciatus , and Cu-NPs had the toxic effects similar to dissolved Cu. Our results provide the ecological safety data service of Cu-NPs in water, and provide a reference for the health T. fasciatus aquaculture.
研究领域:
环境毒理学
·
水生毒理学
·
分子生物学
·
鱼类生理学
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DOI
高影响因子
EUROPEAN JOURNAL OF PHARMACOLOGY
·
2017
·
IF 2.896
Cheng-yuan Li,Mei Su,Yun-yi Yan,Lin Zhou,Lu-yao Ao,Wei-rong Fang,Yun-man Li
1-(5-(1H-indol-5-yl)-1-(pyridin-3-ylsulfonyl)-1H-pyrrol-3-yl)-N-methylmethanamine (KFP-H008)，a novel and potent potassium-competitive acid blocker for the treatment of acid secretion related diseases, has not been reported previously. In this study, we demonstrated that KFP-H008 inhibits basal acid secretion, 2-deoxy- D -glucose- (2DG-) stimulated gastric acid secretion in rats. KFP-H008 blocked histamine-stimulated acid secretion in rats and heidenhain pouch dogs and reversed acid output in isolated gastric perfusion under histamine stimulation. In all the animal experiments, KFP-H008 exerted a more effective, potent and longer-lasting inhibitory action in comparison with lansoprazole , a proton pump inhibitor (PPI) commonly used in clinic. KFP-H008 inhibited H + -K + -ATPase activity both at pH 6.5 and pH 7.5, and was unaffected by pH. The inhibitory action was reversible and was achieved in a K + -competitive manner. Furthermore, KFP-H008 did not affect Na + -K + -ATPase activity, thus exhibiting high selectivity, which is different from PPIs. In all, KFP-H008, a novel potassium-competitive acid blocker, may provide new option for the patients with acid-related diseases and provide longer-lasting inhibitory action than drugs commonly used in clinical treatment.
研究领域:
药理学
·
胃肠病学
·
药物开发
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产品关联
DOI
PubMed
高影响因子
MARINE ENVIRONMENTAL RESEARCH
·
2022
·
IF 3.737
Shaoying Xing,Ping Li,Shuwen He,Zhihan Cao,Xu Wang,Xuqian Cao,Bin Liu,Chengzhuang Chen,Hong You,Zhi-Hua Li
Triphenyltin (TPT) has attracted considerable attention owing to its vitality, bioaccumulation , and lurking damage. TPT widely exists in complex salinity areas such as estuaries and coastal regions. However, there are few studies on the toxicological behavior of TPT under different salinity. In the study, juvenile Nile tilapia ( Oreochromis niloticus ) were utilized as model animals to investigate the effects of environmental relevant TPT exposure on the osmoregulation and energy metabolism in gill under different salinity. The results showed that salinity and TPT single or combined exposure affected the morphology of the gill tissue. After TPT exposure, Na + -K + -ATPase (NKA) activity significantly decreased at 0 ppt, while NKA and Ca 2+ -Mg 2+ -ATPase (CMA) activities significantly increased at 15 ppt. In addition, significantly higher succinate dehydrogenase (SDH) and lactate dehydrogenase (LDH) activities were found in the control fish compared to the TPT-exposed ones at 15 ppt. Quantitative real-time PCR results showed that TPT exposure affected the expression of osmoregulation and energy metabolism-related genes under different salinity. Overall, TPT exposure interfered with osmoregulation and energy metabolism under different salinity. The study will provide reference data for assessing the toxicity of organotin compounds in complex-salinity areas.
研究领域:
水生毒理学
·
环境科学
·
鱼类生理学
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DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）