TALANTA
·
2016
·
IF 4.035
Yun Guan,Lanjunzi Liu,Chao Chen,Xiuzhi Kang,Qingji Xie
The facile preparation of poly( l -DOPA)-tyrosinase (PD M -Tyr) composite and its application both in substrate (phenol) and inhibitor (atrazine) sensing is reported here for the first time. Effective immobilization of enzyme is realized via in-situ entrapping Tyr in poly( l -DOPA) (PD M ), which is formed by Tyr catalytic polymerization of l -DOPA. The Tyr modified electrode is simply prepared by dipping the PD M -Tyr composite on an Au electrode and then covered by Nafion. The thus-prepared Tyr-immobilized electrode exhibits excellent performance superior to most Tyr-based electrochemical biosensors, the sensitivity to phenol is as high as 5122 μA mM −1 in the linear range of 10 nM~1.25 μM, the apparent Michaelis-Menten constant ( K M app ) determined as low as 3.13 μM indicates strong substrate binding and high catalytic activity of the immobilized Tyr. The biosensor also works well in atrazine biosensing, with a linear detection range of 50 ppb~30 ppm and a low detection limit of 10 ppb obtained. In addition, the biosensor shows excellent stability, precision, high sensitivity and fabrication simplicity.
研究领域:
生物化学
·
环境科学
·
分析化学
✓ 已验证
DOI
高影响因子
ENZYME AND MICROBIAL TECHNOLOGY
·
2017
·
IF 2.502
Ren-Chao Zheng,Xiao-Ling Tang,Hui Suo,Li-Lin Feng,Xiao Liu,Jian Yang,Yu-Guo Zheng
Tyrosine phenol-lyase (TPL) catalyzes the reversible cleavage of l -tyrosine to phenol, pyruvate and ammonia. When pyrocatechol is substituted for phenol, l -dihydroxyphenylalanine ( l -DOPA) is produced. The TPL-catalyzed route was regarded as the most economic process for l -DOPA production. In this study, a novel TPL from Fusobacterium nucleatum ( Fn -TPL) was successfully overexpressed in Escherichia coli and screened for l -DOPA synthesis with a specific activity of 2.69 U mg −1 . Fn -TPL was found to be a tetramer, and the optimal temperature and pH for α, β-elimination of l -tyrosine was 60 °C and pH 8.5, respectively. The enzyme showed broad substrate specificity toward natural and synthetic l -amino acids. Kinetic analysis suggested that the k cat / K m value for l -tyrosine decomposition was much higher than that for l -DOPA decomposition, while Fn -TPL exhibited similar catalytic efficiency for synthesis of l -tyrosine and l -DOPA. With whole cells of recombinant E. coli as biocatalyst, l -DOPA yield reached 110 g L −1 with a pyrocatechol conversion of 95%, which was comparable to the reported highest level. The results demonstrated the great potential of Fn -TPL for industrial production of l -DOPA.
研究领域:
生物化学
·
酶学
·
微生物学
✓ 已验证
DOI
高影响因子
JOURNAL OF SEPARATION SCIENCE
·
2013
·
IF 2.591
Xinchun Li,Zuanguang Chen,Fan Yang,Jianbin Pan,Yinbao Li
L -3,4-dihydroxyphenylalanine (L-DOPA) is a well-recognized therapeutic compound to Parkinson's disease. Tyrosine is a precursor for the biosynthesis of L-DOPA, both of which are widely found in traditional medicinal material, Mucuna pruriens . In this paper, we described a validated novel analytical method based on microchip capillary electrophoresis with pulsed electrochemical detection for the simultaneous measurement of L-DOPA and tyrosine in M . pruriens . This protocol adopted end-channel amperometric detection using platinum disk electrode on a homemade glass/polydimethylsiloxane electrophoresis microchip. The background buffer consisted of 10 mM borate (pH 9.5) and 0.02 mM cetyltrimethylammonium bromide, which can produce an effective resolution for the two analytes. In the optimal condition, sufficient electrophoretic separation and sensitive detection for the target analytes can be realized within 60 s. Both tyrosine and L-DOPA yielded linear response in the concentration range of 5.0–400 μM ( R 2 > 0.99), and the LOD were 0.79 and 1.1 μM, respectively. The accuracy and precision of the established method were favorable. The present method shows several merits such as facile apparatus, high speed, low cost and minimal pollution, and provides a means for the pharmacologically active ingredients assay in M . pruriens .
研究领域:
分析化学
·
药学
·
植物化学
✓ 已验证
DOI
高影响因子
CONSTRUCTION AND BUILDING MATERIALS
·
2025
·
IF 8
Xue Wang,Tingjun Wang,Chengsheng Gui,Hanxiang Guo,Haili Chen
s Bamboo is a sustainable and renewable material. However, its high hydrophilicity and nutrient-rich composition make it highly susceptible to microbial colonization. In this study, a simple two-step strategy was employed to fabricate stable nano-silver particles (AgNPs) containing polymer composites on bamboo surfaces to enhance antibacterial and anti-mold properties. L-3,4-dihydroxyphenylalanine (L-DOPA) was modified onto the bamboo surface via self-polymerization, providing catechol groups capable of chelating and reducing Ag⁺ ions in situ to form AgNPs. The incorporation of polyethyleneimine (PEI) into the L-DOPA could further promote AgNPs formation and enhance stability of the composite coating. The resulting BT/L-D/PEI/Ag sample was systematically characterized by SEM, XRD, FTIR, XPS, EDS, and water contact angle measurements. The results confirmed that L-D/PEI/Ag polymer composites were successfully coated onto the bamboo surface, and the BT/L-D/PEI/Ag exhibited a significantly enhanced silver loading (6.68 wt%) compared to the BT/L-D/Ag (2.97 wt%), corresponding to a 125 % increase. The modified surface showed improved hydrophobicity with a contact angle of 94.7°, effectively inhibiting microbial adhesion. Antibacterial tests revealed that the BT/L-D/PEI/Ag achieved complete (100 %) inhibition against Escherichia coli ( E. coli ) and Staphylococcus aureus ( S. aureus ), as well as superior antifungal activity against Aspergillus niger ( A. niger ). The synergistic interaction between L-D, PEI and Ag formed an efficient antimicrobial coating, providing a promising strategy for the development of high-value, antimicrobial bamboo materials for sustainable construction and consumer products.
研究领域:
材料科学
·
表面工程
·
纳米技术
·
抗菌技术
·
可持续材料
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF MATERIALS SCIENCE
·
2020
·Vol. 55
·期 18
·7981-7997
·
IF 3.553
Zhang Xiaoyong,Liu Huihui,Yue Lipei,Bai Yongping,He Jinmei
When it comes to bonding in an aqueous environment, intertidal marine organisms such as mussels, sandcastle worms, barnacles and polychaetes are providing inspiration for the assembly of synthetic molecules into the polymeric adhesives. Although intense demands in various life or industrial applications and many mussel-inspired systems have been shown to develop coatings and hydrogels, exploring the mimetic mussel adhesive system through the control of composition and polymer molecular weight is demonstrated less often. In this report, we showed a simple approach to synthesize mussel-inspired adhesive, P( N -butyl acrylate- co - l -Dopa-methacrylate anhydride), which possesses characteristics of simple synthesis, easy-using, no extra-crosslinker need, and excellent adhesion behaviour on diverse substrates. Adhesion behaviour was examined using low- and high-surface-energy substrates (e.g. PTFE and steel, respectively). Furthermore, the fabricated adhesives exhibited excellent underwater adhesive behaviour for steel in various aqueous solutions, including ion ((IO 4 ) − and Mn 3+ ), acid and alkali solutions (pH = 4, 6.7 and 8, respectively). The outstanding fabricated adhesives take advantage of the catechol functionality and wettability of the resulting polymer. These insights should help us to design next-generation underwater adhesive systems as well as open up a number of future research designs for mussel-inspired adhesives with performance not accessed previously.
研究领域:
材料科学
·
生物仿生学
✓ 已验证
DOI
高影响因子
ACS Applied Materials & Interfaces
·
2015
·
IF 6.723
Mengzhen Dai,Lingen Sun,Long Chao,Yueming Tan,Yingchun Fu,Chao Chen,Qingji Xie
Electrochemical/chemical oxidative synthesis and biosensing/biofuel cell applications of poly(L-DOPA) (PD) are studied versus polydopamine (PDA) as a recent hotspot biomaterial. The enzyme electrode developed by coelectrodeposition of PD and glucose oxidase (GOx), uricase, or tyrosinase shows biosensing performance superior to that of the corresponding PDA-based enzyme electrode. The chemical oxidative polymerization of L-DOPA (PDC) by NaAuCl4 in GOx-containing neutral aqueous solution is used to immobilize GOx and gold nanoparticles (AuNPs). The thus-prepared chitosan (CS)/GOx-PDC-AuNPs/Auplate/Au electrode working in the first-generation biosensing mode responds linearly to glucose concentration with a sensitivity of 152 μA mM–1 cm–2, which is larger than those of the CS/GOx-PDAC-AuNPs/Auplate/Au electrode, the CS/GOx-poly(3-anilineboronic acid) (PABA)-AuNPs/Auplate/Au electrode, and the most reported GOx-based enzyme electrodes. This PDC-based enzyme electrode also works well in the second-generation biosensing mode and as an excellent bioanode in biofuel cell construction, probably because PD as an amino acid polymer has the higher biocompatibility and the more favorable affinity to the enzyme than PDA. The PD material of great convenience in synthesis, outstanding biocompatibility for preparing high-performance bionanocomposites, and strong capability of multifunctional coatings on many surfaces may find wide applications in diversified fields including biotechnology and surface-coating.
研究领域:
生物技术
·
材料科学
·
电化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
TALANTA
·
2025
·
IF 6.1
Shuojiang Liu,Ying Chen,Xingliang Cheng,Gongke Li,Yuling Hu
Surface-enhanced Raman scattering (SERS) in conjunction with nanozymes has emerged as a powerful technique for bioanalysis due to their signal conversion and amplification effect. However, clinical applications continue to encounter challenges such as inadequate selectivity. Herein, we proposed a novel approach for the selective SERS detection of L-DOPA in human serum with dual enzyme-mimicking MOF-919(Fe–Cu). This material exhibits both catechol oxidase-like and peroxidase-like activities, enabling it to catalyze catecholic compounds to o -quinones and subsequently oxidize 3,3′,5,5′-tetramethylbenzidine (TMB) molecules into SERS-active ox-TMB. Ultrasonic-assisted synthesis ensures uniform and dense loading of AgNPs within MOF-919(Fe–Cu), thereby enhancing SERS activity. A sensitive and selective method for SERS detection of L-DOPA was developed. The catechol oxidase-like activity of MOF-919(Fe–Cu) provides ideal selectivity and anti-interference to L-DOPA, while the cascade catalytic system transforms weak SERS signals of L-DOPA into amplified SERS signals of ox-TMB. The mechanism of dual enzyme-mimicking activity was further discussed. The detection process was optimized to achieve high sensitivity towards L-DOPA with a linear range 0.5–100 μmol/L and low limit of detection (LOD) of 0.16 μmol/L. Furthermore, it has been applied in serum sample of Parkinson's patients, demonstrating excellent recovery and low relative error compared to HPLC-UV. This approach offers an economical, efficient and accurate strategy for the detection of L-DOPA and provides valuable insights for the integration of selective multifunctional nanozyme and sensitive SERS detection method.
研究领域:
生物化学
·
纳米医学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF ELECTROANALYTICAL CHEMISTRY
·
2025
·
IF 4.1
Xiaofeng Wang,Xiao Liu,Lixue Wu,Gang Hu,Zhaohui Fang
Chiral enantiomers of drugs often exhibit distinct pharmacological activities despite their similar physical and chemical properties. Developing efficient methods for their distinguishment is crucial for pharmaceutical quality control. Here, we presented a novel approach using a CTT (Chlorite-Tetramethylammonium Hydroxide-Tetrathionate) clock reaction system for kinetic chiral enantiomer recognition. DOPA enantiomers were selected for this purpose, in which L-DOPA is therapeutic efficacy in treating Parkinson's disease by precursoring dopamine while D-DOPA lacks such a function. By introducing equal amounts of L-DOPA and D-DOPA into the CTT clock system, we observed a more prolonged induction period for L-DOPA compared to D-DOPA, enabling their quick differentiation. Moreover, a linear relationship between the induction period and the concentration (1.190 × 10 −6 mol/L to 4.5238 × 10 −6 mol/L) of L-DOPA or D-DOPA allowed quantitative detection of L-DOPA or D-DOPA samples. This method could also be applied to analyze varying concentration ratios of L-DOPA and D-DOPA mixtures at a fixed total concentration. Study on mechanism indicates that both L-DOPA and D-DOPA react with ClO 2 · radical to finally generate DHICA through oxidation, with their kinetic disparities evident during this process. These kinetic disparities between L-DOPA and D-DOPA oxidation arises from stereospecific hydrogen-bonding interactions in the cyclodopa intermediate. The weaker hydrogen bond in L-DOPA-derived cyclodopa likely facilitates its cleavage during oxidation, thereby accounting for the faster reaction kinetics observed in the clock system compared to D-DOPA. This makes L-DOPA exhibit more fast consumption of chlorine dioxide than D-DOPA, and resulting larger the extension in the induction period with L-DOPA than that with D-DOPA. Such a method coupling CCT clock demonstrates its versatility and potential for broader applications in chiral drug analysis.
研究领域:
分析化学
·
手性药物分析
·
药学
·
化学动力学
·
生物化学
✓ 已验证
产品关联
DOI
高影响因子
BIOPROCESS AND BIOSYSTEMS ENGINEERING
·
2018
·Vol. 41
·期 9
·1347-1354
·
IF 2.139
Tang Xiao-Ling,Liu Xiao,Suo Hui,Wang Zhi-Chao,Zheng Ren-Chao,Zheng Yu-Guo
The tyrosine phenol lyase (TPL) catalyzed synthesis of L-DOPA was regarded as one of the most economic route for L-DOPA synthesis. In our previous study, a novel TPL from Fusobacterium nucleatum ( Fn -TPL) was exploited for efficient biosynthesis of L-DOPA. However, the catalytic efficiency decreased when the reaction system expanded from 100 mL to 1 L. As such, the bioprocess for scale-up production of L-DOPA was developed in this study. To increase the stability of substrate and product, as well as decrease the by-product formation, the optimum temperature and pH were determined to be 15 °C and pH 8.0, respectively. The initial concentration of pyrocatechol, pyruvate and ammonium acetate was fixed at 8, 5 and 77 g/L and a fed-batch approach was applied with sodium pyruvate, pyrocatechol and ammonium acetate fed in a concentration of 5, 5 and 3.5 g/L, respectively. In addition, L-DOPA crystals were exogenously added to inhibit cell encapsulation by the precipitated product. The final L-DOPA concentration reached higher than 120 g/L with pyrocatechol conversion more than 96% in a 15-L stirred tank, demonstrating the great potential of Fn -TPL for industrial production of L-DOPA.
研究领域:
生物化学
·
微生物学
·
制药工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Journal of Materials Chemistry B
·
2023
·Vol. 11
·期 47
·11235-11250
·
IF 7
Chan Chen,Haisheng Ren,Weikang Tang,Mengqi Han,Qinfei Chen,Hong Zhou,Jiadong Chen,Yuyue Gao,Wenbin Liu
L-3,4-Dihydroxyphenylalanine (L-DOPA) is widely used in Parkinson's disease treatment and is therefore in high demand. Development of an efficient method for the production of L-DOPA is urgently required. Nanozymes emulating tyrosine hydroxylase have attracted enormous attention for biomimetic synthesis of L-DOPA, but suffered from heterogeneity. Herein, a spherical porous iron–nitrogen–carbon nanozyme was developed for production of L-DOPA. Tannic acid chelated with ferrous ions to form a tannin-iron coordination framework as a carbon precursor. Iron and nitrogen co-doped carbon nanospheres were assembled via an evaporation-induced self-assembly process using urea as a nitrogen source, F127 as a soft template, and formaldehyde as a crosslinker. The nanozyme was obtained after carbonization and acid etching. The nanozyme possessed a dispersive iron atom anchored in the Fe–N coordination structure as an active site to mimic the active center of tyrosine hydroxylase. The material showed spherical morphology, uniform size, high specific surface area, a mesoporous structure and easy magnetic separation. The structural properties could promote the density and accessibility of active sites and facilitate mass transport and electron transfer. The nanozyme exhibited high activity to catalyze the hydroxylation of tyrosine to L-DOPA as tyrosine hydroxylase in the presence of ascorbic acid and hydrogen peroxide. The titer of DOPA reached 1.2 mM. The nanozyme showed good reusability and comparable enzyme kinetics to tyrosine hydroxylase with a Michaelis–Menten constant of 2.3 mM. The major active species was the hydroxyl radical. Biomimetic simulation of tyrosine hydroxylase using a nanozyme with a fine structure provided a new route for the efficient production of L-DOPA.
研究领域:
生物化学
·
材料科学
·
医学
✓ 已验证
DOI
PubMed
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
·
生物技术
·
酶学
✓ 已验证
产品关联
DOI
PubMed
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
·
药学
·
生物化工
✓ 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）