RESUMO
Amino acid surfactants have gained significant importance in overcoming the limitations of conventional surfactants, notably, their low biocompatibility and biodegradability. However, the current amino acid surfactants lack multifunctional properties due to the nonreactivity of their aliphatic chains, necessitating the development of a new type of amino acid surfactant. A novel melanin-like amino acid surfactant and a biomimetic synthesis route were devised by mimicking the biosynthesis of melanin. Renewable natural polyphenol compounds with catechol moieties were utilized as building blocks for the hydrophobic group. In a proof-of-concept experiment, ethyl protocatechuate was oxidized to o-quinone and subsequently covalently linked to the amino group of lysine via Michael addition. The chemical structure was verified using liquid chromatography-tandem mass spectroscopy. The melanin-like amino acid surfactant exhibited excellent surface-active properties, with a critical micelle concentration of 1.59 mN m-1. Furthermore, it demonstrated remarkable emulsifying, foaming, solubilizing, dispersing, and wetting capabilities. Notably, it also possessed multifunctionality, including antibacterial activity, antioxidant activity, robustness, and mildness. These outstanding properties indicate significant potential for various applications. This strategy offers innovative insights and a versatile, modular toolbox for synthesizing multifunctional amino acid surfactants that mimic melanin. The approach allows for the easy interchange of o-quinone building blocks, which is akin to snap jewelry.
Assuntos
Joias , Tensoativos , Tensoativos/química , Melaninas , Aminoácidos , Biomimética , QuinonasRESUMO
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.
Assuntos
Levodopa , Tirosina 3-Mono-Oxigenase , Tirosina 3-Mono-Oxigenase/química , Levodopa/química , Carbono/química , Ferro/química , Porosidade , TaninosRESUMO
Endometriosis is a common disease of reproductive-age women and an important cause of dysmenorrhea and infertility. Information on endometriosis is complex and there is a lack of summarization of available results. The study aims to evaluate the overall distribution of publications related to endometriosis to provide a foundation for further research. The Web of Science Core Collection was searched for articles published in the field of endometriosis. Our survey revealed the structure, hotspots, and development trends of endometriosis-related research and publications.
Assuntos
Endometriose , Infertilidade , Humanos , Feminino , Bibliometria , Dismenorreia , ReproduçãoRESUMO
Hydroxytyrosol, one of the most powerful natural antioxidants, exhibits certificated benefits for human health. In this study, a biomimetic approach to synthesize hydroxytyrosol from the hydroxylation of tyrosol was established. EDTA-Fe2+ coordination complex served as an active center to simulate tyrosine hydroxylase. H2O2 and ascorbic acid were used as oxygen donor and hydrogen donor, respectively. Hydroxy radical and singlet oxygen contributed to active species. The biomimetic system displayed analogous component, structure, and activity with TyrH. Hydroxytyrosol titer of 21.59 mM, and productivity of 9985.92 mg·L-1·h-1 was achieved with 100 mM tyrosol as substrate. The proposed approach provided efficient and convenient route to quickly produce high amount of hydroxytyrosol.
Assuntos
Peróxido de Hidrogênio , Tirosina 3-Mono-Oxigenase , Humanos , BiomiméticaRESUMO
Tyrosinase plays an important role in melanin synthesis. Inhibition against tyrosinase activity has been extensively focused on for pharmaceutical, food, cosmetic, and agricultural purpose. The inhibitory mechanism of scutellarein on tyrosinase was elaborated by coupling enzyme kinetics, multi-spectroscopy and computational simulation. Scutellarein remarkably inhibited tyrosinase activity with an IC50 value of 91 µM. Scutellarein reversibly inhibited tyrosinase in a competitive manner. Fluorescence quenching validated that interaction of scutellarein with tyrosinase occurred to form a complex with a binding constant of 6.11 × 104 M-1. Thermodynamic parameters suggested that scutellarein spontaneously bound with tyrosinase via hydrogen bond and van der Waals force. Three-dimensional fluorescence spectra and circular dichroism spectra revealed that scutellarein induced an obvious conformational change in tyrosinase. Molecular docking result predicted that scutellarein mainly bound with tyrosinase via Arg268 residue. Scutellarein effectively controlled the enzymatic browning of apple slices during storage. This research could give theoretical guiding significance in various application for tyrosinase inhibitors.
Assuntos
Inibidores Enzimáticos , Monofenol Mono-Oxigenase , Monofenol Mono-Oxigenase/metabolismo , Simulação de Acoplamento Molecular , Cinética , Dicroísmo Circular , Inibidores Enzimáticos/farmacologia , Inibidores Enzimáticos/químicaRESUMO
Silver nanoparticles (AgNPs) attract a great deal of attention for potent antibacterial capacity, but their use is challenged by limited stability. Inspired by the adhesive and redox properties of the mussel foot proteins containing L-3,4-dihydroxyphenylalanine (DOPA), a facile strategy for in situ synthesis of AgNPs using DOPA-containing fibroin is developed. Tyrosine residues in fibroin are transformed into DOPA via biomimetic synthesis method with content of 0.55 mol%. In situ synthesis generates stable and small AgNPs through DOPA bound in fibroin as a reducing and stabilizing agent. Narrow size distribution with average diameter of 20 nm and excellent monodispersity are obtained. Cross-linking with lysine increases the content of ß-sheet to form hydrogel and achieves gradual release of silver. The material exhibits excellent antibacterial properties against Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli bacteria. It can be potentially applied in biological and medical fields to treat bacterial infections.
Assuntos
Fibroínas , Nanopartículas Metálicas , Fibroínas/farmacologia , Fibroínas/química , Prata/farmacologia , Prata/química , Di-Hidroxifenilalanina , Nanopartículas Metálicas/química , Testes de Sensibilidade Microbiana , Antibacterianos/química , Bactérias Gram-NegativasRESUMO
Tyrosinase is pivotal for melanin formation. Measuring monophenolase activity is of great importance for both fundamental research and industrial applications. For the first time, a backpropagation (BP) artificial neural network with three-dimensional fluorescence spectroscopy was applied for the real-time determination of tyrosinase monophenolase activity. Principal component analysis (PCA) was utilized for the dimension reduction of three-dimensional fluorescence data. The four principal components served as inputs for the neural network. Network parameters were optimized using a genetic algorithm (GA). BP learning algorithm was applied to train the network model to determine tyrosine levels in a binary mixture containing tyrosine and L-DOPA without any chemical separation. The time course of tyrosine consumption by monophenolase was determined to calculate the initial velocity of the enzymatic reaction. The limit of detection of the monophenolase assay was 0.0615 U·mL-1. This combined strategy of PCA, GAs, and BP artificial neural networks for three-dimensional fluorescence spectroscopy was efficient for the real-time and in-situ determination of monophenolase activity in a cascade reaction.
Assuntos
Monofenol Mono-Oxigenase , Redes Neurais de Computação , Espectrometria de Fluorescência , TirosinaRESUMO
L-3,4-dihydroxyphenylalanine (L-DOPA) is in high demand as the cornerstone for treatment of Parkinson's disease. The current production of L-DOPA is associated with poor productivity and long production period. Biomimetic system inspired from tyrosine hydroxylase was developed to achieve the production of L-DOPA from tyrosine with high reactivity, efficiency, and specificity. The biomimetic system owned close resemblance of component and structure in comparison with tyrosine hydroxylase, consisting of tyrosine as substrate, a redox complex of Fe2+ and EDTA as the catalyst to simulate the active center of the natural tyrosine hydroxylase, hydrogen peroxide as the oxidant, and ascorbic acid as the reductant. HPLC, HPLC-MS/MS, 1H NMR, and specific rotation identified L-DOPA was generated. The system showed high catalytic activity and regioselectivity for hydroxylation of tyrosine as equal to tyrosine hydroxylase. FeIVO2+ was formed as the major active species, and NIH shift was observed. EDTA accelerated the reaction by reducing the redox potential of Fe3+/Fe2+ couple. Density functional theory calculation suggested formation of FeIVO2+ was more thermodynamically favorable. The biomimetic system shared analogous catalytic mechanism with TyrH. Process parameters was optimized for maximum production of L-DOPA, namely 6.4 mM tyrosine, 1.6 mM Fe2+, 1.92 mM EDTA, 150 mM H2O2, and 35 mM ascorbic acid in 0.2 M glycine-HCl buffer at pH 4.5 and 60 °C. The yield, titer, and productivity were obtained as 52.01%, 3.22 mM, and 48,210.68 mg L-1 h-1, respectively. The proposed method exhibited an amazing productivity, might provide a promising strategy to industrialize L-DOPA production.
Assuntos
Di-Hidroxifenilalanina , Tirosina 3-Mono-Oxigenase , Ácido Ascórbico , Biomimética , Ácido Edético , Peróxido de Hidrogênio , Espectrometria de Massas em Tandem , Tirosina/química , Tirosina 3-Mono-Oxigenase/químicaRESUMO
A real-time assay for multiple enzyme activities in cascade reactions is required for research on metabolism and bioengineering. Tyrosinase has the bifunctional activity of monophenolase and diphenolase. A combined strategy of three-way calibration with excitation-emission matrix (EEM) fluorescence was developed for real-time and simultaneous determination of monophenolase and diphenolase activity with tyrosine as a substrate. Mathematical separation and second-order advantage were utilized to solve spectral overlapping and uncalibrated interferents during complex dynamic enzymatic processes. Kinetic evolution profiles of EEM were monitored to stack a fusion three-way data array together with static samples. Using a parallel factor analysis (PARAFAC) algorithm, pseudo-univariate calibration curves with limits of detection (LODs) of 3.00 µM and 0.85 µM were established to simultaneously and real-time measure tyrosine and DOPA. Progress curves for tyrosine consumption by monophenolase and DOPA consumption by diphenolase were obtained using the law of mass conservation to calculate the initial velocity. The LODs for monophenolase and diphenolase were 0.0232 Uâ mL-1 and 0.0316 Uâ mL-1. The method achieved real-time and simultaneous assays of multiple enzyme activities in cascade reactions. It showed potential application in the metabolic pathway and biochemical industry.
Assuntos
Monofenol Mono-Oxigenase , Oxirredutases , Calibragem , Catálise , Cinética , Monofenol Mono-Oxigenase/metabolismo , Oxirredutases/análiseRESUMO
Tyrosinase plays an essential role in melanin biosynthesis and inherently exhibits both monophenolase and diphenolase activity. A first derivative synchronous fluorometric assay was established for directly monitoring monophenolase activity. The zero-crossing point at 322 nm for the first-derivative under synchronous fluorescence with Δλ = 67 nm was utilized to selectively quantify tyrosine in the presence of the reaction product dihydroxyphenylalanine (DOPA). The limit of detection (LOD) for tyrosine was 0.54 µM. The fluorescence intensity of tyrosine was monitored at intervals of 30 s to establish the time course of tyrosine consumption. The LOD for the monophenolase activity was 0.0706 Uâ mL-1. The Michaelis-Menten e constant and maximum speed were 21.83 µM and 1.12 µM min-1, respectively. Zinc ions competitively inhibited the monophenolase activity, with an IC50 value of 14.36 µM. This assay is easily and rapidly executed and is of great significance for analyzing the kinetics of enzymatic reactions and in fundamental research on monophenolase. This approach has potential applications in the discovery of tyrosinase inhibitors for medicine and cosmetics, as well as in the industrial synthesis of substituted o-diphenol intermediates.
Assuntos
Monofenol Mono-Oxigenase , Oxirredutases , Monofenol Mono-Oxigenase/metabolismo , Oxirredução , Oxirredutases/metabolismo , Tirosina/metabolismoRESUMO
Tyrosinase is the key enzyme for melanogenesis with both monophenolase activity and diphenolase activity, which catalyzes the hydroxylation of tyrosine to L-DOPA and the further oxidation of DOPA, respectively. A continuous assay method was developed to directly monitor the real monophenolase activity using synchronous fluorescence. Complexation with borate to quench the native fluorescence of DOPA could selectively quantified the tyrosine in the binary mixture of tyrosine and DOPA under the wavelength difference Δλ = 67 nm for synchronous fluorescence. The limit of detection (LOD) for tyrosine were estimated to be 0.49 µM. Borate was used as a trapping agent for DOPA to abolish diphenolase activity, while hydroxylamine was used as a reducing agent to restore the catalytic cycle. The time course for consumption of tyrosine was established by monitoring the tyrosine fluorescence intensity at discrete intervals of 30 s. Calibration curve between monophenolase activity and tyrosinase concentration with range from 0.1830 U·mL-1 to 1.7034 U·mL-1, and LOD of 0.0721 U·mL-1. Using the proposed method, the Km and υmax for monophenolase was determined with values of 20.73 µM and 1.10 µM·min-1, respectively. Zinc ion was demonstrated to inhibit the monophenolase activity by competitive inhibition manner with IC50 of 14.36 µM. The assay method displayed a powerful application in kinetics and inhibitor screening for monophenolase.