RSC Advances
·
2025
·Vol. 15
·期 1
·420-427
·
IF 4.6
Zongmei Huang,Jing Li,Lu-Shuang Li
Levodopa (L-Dopa), a precursor drug for dopamine has been widely used to treat Parkinson's disease. However, excess accumulation of L-Dopa in the body may cause movement disorders and uncontrollable emotions. Therefore, it is vital to monitor L-Dopa levels in patients. In this study, a carbon dot (CD)-based fluorescence sensing system was developed for sensitive detection of L-Dopa. The CDs were prepared using a novel biomass, Pandanus amaryllifolius Roxb., as a carbon source via a simple hydrothermal method. Interestingly, it was found that ethylenediamine doping in the preparation system increased the quantum yield of CDs, as well as their fluorescence response sensitivity to L-Dopa. After optimizing the preparation and sensing conditions, the detection limit of L-Dopa decreased from 1.54 μM to 0.05 μM. A complete methodological validation was conducted and the probe was successfully applied to the determination of L-Dopa in fetal bovine serum with excellent precision (RSD ≤ 2.99%) and recoveries of 88.50–99.71%. Overall, this work provides an effective strategy for the regulation of properties of CDs derived from biomass and an innovative method for clinical L-Dopa monitoring.
研究领域:
医学诊断
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材料科学
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药物分析
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DOI
PubMed
高影响因子
Microbial Biotechnology
·
2022
·Vol. 15
·期 5
·1610-1621
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IF 6.575
Xing Liu,Xiao Han,Yuan Peng,Chunlin Tan,Jing Wang,Hongsong Xue,Ping Xu,Fei Tao
Summary 3, 4-Dihydroxyphenyl- l -alanine ( l -DOPA) is a compound of high medical value and is considered effective as a treatment for Parkinson’s disease. Currently, bioproduction of l -DOPA is mainly carried out by whole-cell catalysis mediated by recombinant Escherichia coli carrying heterogeneous tyrosine phenol lyase. Vibrio natriegens is increasingly attracting attention owing to its superiority, including extremely rapid growth and high soluble protein expression capacity. In this study, we attempt to develop an efficient whole-cell catalyst for l -DOPA production using V. natriegens as the chassis. The maximum soluble protein expression by V. natriegens was accomplished in 4 h at 37°C, which was equivalent to that achieved by E. coli in 16 h at 16°C. Furthermore, the maximum productivity reached over 10.0 g l −1 h −1 in the early stage of biocatalysis, nearly two-fold higher than previously reported. Approximately 54.0 g l −1 l -DOPA was obtained with a catechol conversion rate greater than 95%. In conclusion, V. natriegens displays advantages, including rapid protein expression and catalytic rate in the catalysis process for l -DOPA production. These findings strongly suggest that V. natriegens has remarkable potential as a whole-cell catalysis chassis for the production of valuable chemicals.
研究领域:
生物工程
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医学
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微生物学
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DOI
PubMed
高影响因子
NANOTECHNOLOGY
·
2022
·Vol. 33
·期 13
·135503
·
IF 3.953
Shuyang Hu,Qiuyan Shuai,Yulong Lin,Yan Fu,Meng Li
L-3,4-dihydroxy-phenylalanine (L-dopa) is the most widely used drug in Parkinson's disease treatment. However, development of cost-effective and high-throughput sensors to accurate enantioselective discrimination of L-dopa and D-dopa remains challenging to date. Herein, on the basis of the peroxidase-mimic activity of chiral FexCuySe nanoparticles, we demonstrated a novel colorimetric sensor for determination of chiral dopa. The surface chiral ligand, L/D-histidine (L/D-His), endowed the nanozymes with enantioselectivity in catalyzing the oxidation of dopa enantiomers. According to the values of kcat/Km, the efficiency of L-His modified nanoparticles (L-FexCuySe NPs) towards L-dopa was 1.56 times higher than that of D-dopa. While, D-His can facilely reverse the preference of the nanozyme to D-dopa. On the basis of high catalytic activity and enantioselectivity of L-FexCuySe NPs in oxidation of L-dopa, the L-FexCuySe NPs-based system can be utilized for detection of L-dopa. The linear ranges for L-dopa determination were 5 μM–0.125 mM and 0.125 mM–1 mM with a detection limit of 1.02 μM. Critically, the developed sensor has been successfully applied in the quality control of clinical used L-dopa tablets. Our work sheds light on developing simple and sensitive chiral nanomaterials-based sensors for drug analysis.
研究领域:
生物化学
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医学诊断
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纳米技术
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DOI
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Shiming Tang,Zhilin Ouyang,Ying Huo,Daocheng Liao,Zhiteng Zhang,Ying Lin,Suiping Zheng
Hydroxytyrosol has been proven beneficial to human health. However, the process involving the conversion of L-DOPA to 3,4-dihydroxyphenylacetaldehyde (3,4-DHPAA) in hydroxytyrosol biosynthesis typically required the simultaneous use of decarboxylase and oxidative deaminase. In addition, phenylacetaldehyde reductase from Solanum lycopersicum (SlPAR) in hydroxytyrosol biosynthesis exhibits poor thermal stability. In this study, we unexpectedly discovered that L-DOPA decarboxylase from Pseudomonas putida (PpDODC) exhibits weak dual-functional activity for both decarboxylation and oxidative deamination of L-DOPA. Through a dual-function reshaping strategy, the best dual-functional mutant PpDODC/Y79F/Y324F achieved an enzyme activity of 0.95 U/mg and exhibited a 256.8-fold increase in activity compared to the wild-type. Through rational design of SlPAR, the optimal mutant SlPAR/G52D/V188I/N234W/Q286P (SlPAR-M4) maintained stability after 12 h' treatment at 40 °C. Based on these mutants, we established a simplified cascade to synthesize hydroxytyrosol from L-DOPA, achieving a hydroxytyrosol yield of 31.4 mM from 32 mM L-DOPA in a 5-hour reaction. This process achieved the highest molar conversion rate (98.2 %) currently reported for the synthesis of hydroxytyrosol from L-DOPA. This study provides a novel solution for hydroxytyrosol synthesis from a new perspective, and the mutants hold promise for widespread application in biotransformation studies of hydroxytyrosol and other structurally-similar compounds.
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
MICROCHIMICA ACTA
·
2020
·Vol. 187
·期 7
·1-8
·
IF 6.232
Li Shuhuai,Pang Chaohai,Ma Xionghui,Zhao Min,Li Haibo,Wang Mingyue,Li Jianping,Luo Jinhui
A luminescent double recognition nanoprobe is described as a new strategy for the selective determination of chiral molecules. C-dots/Ir/Au fluorescent nanoparticles, synthesised under hydrothermal conditions, are used as a high-performance probe in combination with a molecularly imprinted polymer (MIP) and calix[6]arene as a double recognition element. Thiolated calix[6]arene is grafted on C-dots/Ir/Au as the first recognition element, which then forms a host–guest complex with the target molecule levodopa (L-DOPA). Subsequently, an MIP is prepared on the C-dots/Ir/Au (MIP/C-dots/Ir/Au) by chemical polymerisation. After the removal of L-DOPA, double recognition imprinting cavities are formed. The fluorescence intensity at 478 nm of the nanoprobe is effectively quenched by adsorption of L-DOPA on MIP/C-dots/Ir/Au, which provides a method for L-DOPA determination. Owing to the double recognition strategy, this method has excellent selectivity which can effectively avoid interference from enantiomer D-DOPA, and a imprinting factor of 7.1 is obtained for L-DOPA. This accurate and reliable method, with a wide linear range (5 × 10 −10 to 1.2 × 10 −7 mol L −1 ) and a low limit of detection (1.45 × 10 −10 mol L −1 ), was successfully applied to the determination of L-DOPA in real samples, giving standard recoveries of 89.7–110.0%. Thus, the proposed sensing method provides a viable approach for the determination of a single enantiomer.
Schematic presentation of the MIP/C-dots/Ir/Au for L-DOPA detection. A fluorescence double chiral recognition nanoprobe is prepared of C-dots/Ir/Au nanoparticles as signal probe, and a molecularly imprinted polymer (MIP) and calix[6]arene as a double recognition element. Owing to the double recognition strategy, this method has strong specificity and can effectively avoid interference from enantiomers and racemates.
研究领域:
分析化学
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药物化学
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材料科学
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DOI
PubMed
高影响因子
ChemistrySelect
·
2023
·Vol. 8
·期 2
·e202203646
·
IF 2.1
Siying Ren,Wen-Chao Geng,Xuexian Cui,Bian Wu,Zhe Zheng
Graphical We present a robust supramolecular tandem assay to monitor L-DOPA decarboxylase activity and screen enzyme inhibitors using cucurbit[7]uril and acridine orange reporter pair. L–DOPA decarboxylase (DDC) deficiency causes serious progressive neurodegenerative disorders. Developing a sensitive enzyme activity monitoring and inhibitors or activators screening approach is highly on demand for DDC deficiency diagnosis. Here, we report a simple and sensitive fluorescence supramolecular tandem assay based on reporter pair of cucurbit[7]uril and acridine orange for DDC activity monitoring continuously. Furthermore, this strategy demonstrates the potential of STA to screen inhibitors of DDC.
研究领域:
生物化学
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医学诊断
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药物开发
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DOI
高影响因子
MOLECULES
·
2022
·Vol. 27
·期 18
·6088
·
IF 4.927
Ke Shun Hu,Chong Le Chen,Huan Ru Ding,Tian Yu Wang,Qin Zhu,Yi Chen Zhou,Jia Min Chen,Jia Qi Mei,Sheng Hu,Jun Huang,Wei Rui Zhao,Le He Mei
Salvianic acid A (SAA), as the main bioactive component of the traditional Chinese herbSalvia miltiorrhiza, has important application value in the treatment of cardiovascular diseases. In this study, a two-step bioprocess for the preparation of SAA froml-DOPA was developed. In the first step,l-DOPA was transformed to 3,4-dihydroxyphenylalanine (DHPPA) using engineeredEscherichia colicells expressing membrane-bound L-amino acid deaminase fromProteus vulgaris.After that, the unpurified DHPPA was directly converted into SAA by permeabilized recombinantE. colicells co-expressingd-lactate dehydrogenase fromPediococcus acidilacticiand formate dehydrogenase fromMycobacterium vaccaeN10. Under optimized conditions, 48.3 mM of SAA could be prepared from 50 mM ofl-DOPA, with a yield of 96.6%. Therefore, the bioprocess developed here was not only environmentally friendly, but also exhibited excellent production efficiency and, thus, is promising for industrial SAA production.Keywords:salvianic acid A;l-DOPA;membrane-boundl-amino acid deaminases;biocatalysis;molecular biology;biological engineering
研究领域:
生物技术
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药学
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生物化工
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DOI
PubMed
高影响因子
Analytical Methods
·
2014
·Vol. 7
·期 4
·1387-1394
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IF 1.938
Lu Lin,Hui-Ting Lian,Xiang-Ying Sun,Ya-Ming Yu,Bin Liu
A molecularly imprinted polymer (MIP) based sensor for detecting L-dopa was fabricated based on the composite of graphene (GR) and chitosan (CS). The microstructure and composition of the imprinted films characterized by scanning electron microscopy (SEM), Fourier transform infrared (FTIR), Raman spectroscopy and electrochemical impedance spectroscopy (EIS) indicated that the modified film of GR-MIPs had been formed successfully. Electrochemical evaluation results showed that the sensor exhibited a satisfactory selectivity for molecular binding with an imprinting factor (IF) of 6.2 and a wide linear range from 0.4 μM to 100 μM with a low detection limit of 0.012 μM. This electrochemical sensor exhibited great promise for accurate determination of L-dopa in pharmaceutical preparations and human blood serum for recoveries ranging from 95.0 to 108.0%. Furthermore, compared to imprinted films without graphene doping, GR-MIPs showed a superior chiral sensing for L-dopa in the presence of D-dopa. The results reveal that the insertion of graphene brought about an enhanced sensitivity for chiral sensing and has the potential for application in chiral recognition.
研究领域:
分析化学
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材料科学
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生物医学工程
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DOI
高影响因子
TALANTA
·
2015
·
IF 3.545
Mengzhen Dai,Ting Huang,Long Chao,Qingji Xie,Yueming Tan,Chao Chen,Wenhua Meng
Horseradish peroxidase (HRP)-catalyzed polymerization of l -DOPA (vs. dopamine) in the presence of H 2 O 2 (and uricase (UOx)) was exploited to immobilize mono-/bi-enzymes for hydroquinone-mediated amperometric biosensing of H 2 O 2 and uric acid (UA). The relevant polymeric biocomposites (PBCs) were prepared in phosphate buffer solution containing HRP and l -DOPA (or plus UOx) after adding H 2 O 2 . The mono-/bi-enzyme amperometric biosensors were prepared simply by casting some of the PBCs on Au-plated Au (Au plate /Au) electrodes, followed by coating with an outer-layer chitosan (CS) film for each. UV–vis spectrophotometry , scanning electron microscopy, cyclic voltammetry and electrochemical impedance spectroscopy were used for film characterization and/or process monitoring. The HRP immobilized by enzyme catalysis well preserved its bioactivity, as confirmed by UV–vis spectrophotometry . Under optimized conditions, the monoenzyme CS/HRP-poly( l -DOPA) (PD)/Au plate /Au electrode potentiostated at −0.1 V responded linearly to H 2 O 2 concentration from 0.001 to 1.25 mM with a sensitivity of 700 μA mM −1 cm −2 and a limit of detection (LOD) of 0.1 μM, and the bienzyme CS/UOx–HRP–PD/Au plate /Au electrode at −0.1 V responded linearly to UA concentration from 0.001 to 0.4 mM with a sensitivity of 349 μA mM −1 cm −2 and a LOD of 0.1 μM. The mono-/bi-enzyme biosensors based on biosynthesized PD performed better than many reported analogues and those based on similarly biosynthesized polydopamine.
研究领域:
生物化学
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分析化学
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生物医学工程
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产品关联
DOI
PubMed
高影响因子
ENVIRONMENTAL POLLUTION
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2024
·
IF 7.6
Yue Niu,Yifan Pan,Yaqi Wang,Yongqi Fu,Zhangwu Zhao,Le Kang
Parkinson's disease (PD) is one of the fastest-growing neurodegenerative diseases and has been linked to the exposure to numerous environmental neurotoxins. Although lead (Pb) exposure has been related to the development of PD, the molecular target of Pb to cause the onset of PD is insufficiently investigated. Herein, we explored the effects of Pb exposure on behavior, pathophysiology, and gene expression of wild-type (WT) fly ( Drosophila melanogaster ) by comparison with its PD model. After exposure to Pb, the WT flies showed PD-like locomotor impairments and selective loss of dopaminergic (DAergic) neurons, displaying similar phenotypes to fly PD model (PINK1). Transcriptomic analysis showed the similarity in gene expression profiles between Pb treatment WT flies and PINK1 mutant flies. Moreover, Pb exposure resulted in endogenous dopamine deficits in WT flies. Analyses of gene expression and enzyme activity confirmed that Pb exposure reduced tyrosine hydroxylase (TH) activity and led to failure of dopamine synthesis. Furthermore, molecular dynamics simulation confirmed that Pb was adsorbed by TH and subsequently inhibited the enzymatic activity. Exogenous injection of L-dopa and melatonin could partially rescue the pathological phenotypes of Pb-exposed flies and PD fly model. Antagonist injection of microRNA-133, which negatively regulated the expression of TH gene, ultimately rescued in the manifestation of PD phenotypes in flies. Involvement of TH overexpression mutants of fly strongly promoted the resistance to Pb exposure and rescued both behavior and the number of DAergic neurons. Therefore, our study elucidates the Pb molecular target in dopamine pathway and mechanism underlying the risks of Pb exposure on the occurrence of PD at environmentally-relevant concentrations.
研究领域:
神经科学
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毒理学
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分子生物学
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遗传学
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
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2024
·
IF 8
Jiaxue Liang,Hang Qi,Feng Zhu,Shuqi Chen,Bohua Liu,Chongling Sun,Yanyan Wang
Real-time monitoring the drugs penetration into brain is crucial for clinical therapy of neurodegenerative diseases. In this study, a Blood-Brain Barrier-electrochemical sensing (BBB-ES) platform combining BBB on chip model and electrochemical sensing system was proposed, which facilitated stable BBB growth under fluidic conditions and realized real-time detection of L-Dopa across the BBB. Based on this system, the effects of fluid force on the cell morphology, skeleton arrangement and tight junction protein ZO-1 of BBB growth were evaluated, it was verified that the fluid force promote cell rearrangement. Additionally, L-Dopa detecting performance was optimized by modifying SWNTs-AuNPs-PPy/Tyr(Single-Walled Carbon Nanotubes-Au Nanoparticles-Polypyrrole/Tyrosinase) on electrochemical electrode, which showed a wide linear response range of 10–500 μM and excellent selectivity toward L-Dopa. The integrated BBB-ES system was finally applied to real-time monitor the penetration process of L-Dopa through the BBB, it was proved that the penetration of L-Dopa increased with the increase of flow rate and time, and finding that L-Dopa was delivered by passive diffusion without disrupting the integrity of BBB. Based on these findings, the system can be used to monitor the real-time process of different drugs penetrating BBB and even different organ barriers, which also provide the possibility to solve the clinical demand for drugs screen.
研究领域:
神经科学
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生物医学工程
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药学
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电化学
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DOI
高影响因子
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
·
2020
·Vol. 192
·期 1
·71-84
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IF 2.277
Zhu Hang-Qin,Tang Xiao-Ling,Zheng Ren-Chao,Zheng Yu-Guo
Tyrosine phenol-lyase (TPL) is a valuable and cost-effective biocatalyst for the biosynthesis of L-tyrosine and its derivatives, which are valuable intermediates in the pharmaceutical industry. A TPL from Morganella morganii ( Mm -TPL) was overexpressed in Escherichia coli and characterized. Mm -TPL was determined as a homotetramer with molecular weight of 52 kDa per subunit. Its optimal temperature and pH for β-elimination of L-tyrosine were 45 °C and pH 8.5, respectively. Mm -TPL manifested strict substrate specificity for the reverse reaction of β-elimination and ortho - and meta -substituted phenols with small steric size were preferred substrates. The enzyme showed excellent catalytic performance for synthesis of L-tyrosine, 3-fluoro-L-tyrosine, and L-DOPA with a yield of 98.1%, 95.1%, and 87.2%, respectively. Furthermore, the fed-batch bioprocess displayed space-time yields of 9.6 g L −1 h −1 for L-tyrosine and 4.2 g L −1 h −1 for 3-fluoro-L-tyrosine with a yield of 67.4 g L −1 and 29.5 g L −1 , respectively. These results demonstrated the great potential of Mm -TPL for industrial application.
研究领域:
生物化学
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生物技术
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药学
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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）