Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 8.448
Filter
1.
Chem Biol Drug Des ; 103(5): e14539, 2024 May.
Article in English | MEDLINE | ID: mdl-38760181

ABSTRACT

Tyrosinase is a copper-containing enzyme involved in the biosynthesis of melanin pigment. While the excess production of melanin causes hyperpigmentation of human skin, hypopigmentation results in medical conditions like vitiligo. Tyrosinase inhibitors could be used as efficient skin whitening agents and tyrosinase agonists could be used for enhanced melanin synthesis and skin protection from UV exposure. Among a wide range of tyrosinase-regulating compounds, natural and synthetic derivatives of furochromenones, such as 8-methoxypsoralen (8-MOP), are known to both activate and inhibit tyrosinase. We recently reported a synthetic approach to generate a variety of dihydrofuro[3,2-c]chromenones and furo[3,2-c]chromenones in a metal-free condition. In the present study, we investigated these compounds for their potential as antagonists or agonists of tyrosinase. Using fungal tyrosinase-based in vitro biochemical assay, we obtained one compound (3k) which could inhibit tyrosinase activity, and the other compound (4f) that stimulated tyrosinase activity. The kinetic studies revealed that compound 3k caused 'mixed' type tyrosinase inhibition and 4f stimulated the catalytic efficiency. Studying the mechanisms of these compounds may provide a basis for the development of new effective tyrosinase inhibitors or activators.


Subject(s)
Enzyme Inhibitors , Monophenol Monooxygenase , Monophenol Monooxygenase/antagonists & inhibitors , Monophenol Monooxygenase/metabolism , Enzyme Inhibitors/pharmacology , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/chemical synthesis , Kinetics , Humans , Methoxsalen/pharmacology , Methoxsalen/chemistry , Enzyme Activators/chemistry , Enzyme Activators/pharmacology
2.
Mar Drugs ; 22(5)2024 Apr 29.
Article in English | MEDLINE | ID: mdl-38786597

ABSTRACT

Abnormal melanogenesis can lead to hyperpigmentation. Tyrosinase (TYR), a key rate-limiting enzyme in melanin production, is an important therapeutic target for these disorders. We investigated the TYR inhibitory activity of hydrolysates extracted from the muscle tissue of Takifugu flavidus (TFMH). We used computer-aided virtual screening to identify a novel peptide that potently inhibited melanin synthesis, simulated its binding mode to TYR, and evaluated functional efficacy in vitro and in vivo. TFMH inhibited the diphenolase activities of mTYR, reducing TYR substrate binding activity and effectively inhibiting melanin synthesis. TFMH indirectly reduced cAMP response element-binding protein phosphorylation in vitro by downregulating melanocortin 1 receptor expression, thereby inhibiting expression of the microphthalmia-associated transcription factor, further decreasing TYR, tyrosinase related protein 1, and dopachrome tautomerase expression and ultimately impeding melanin synthesis. In zebrafish, TFMH significantly reduced black spot formation. TFMH (200 µg/mL) decreased zebrafish TYR activity by 43% and melanin content by 52%. Molecular dynamics simulations over 100 ns revealed that the FGFRSP (T-6) peptide stably binds mushroom TYR via hydrogen bonds and ionic interactions. T-6 (400 µmol/L) reduced melanin content in B16F10 melanoma cells by 71% and TYR activity by 79%. In zebrafish, T-6 (200 µmol/L) inhibited melanin production by 64%. TFMH and T-6 exhibit good potential for the development of natural skin-whitening cosmetic products.


Subject(s)
Melanins , Melanoma, Experimental , Monophenol Monooxygenase , Takifugu , Zebrafish , Animals , Melanins/biosynthesis , Takifugu/metabolism , Monophenol Monooxygenase/antagonists & inhibitors , Monophenol Monooxygenase/metabolism , Mice , Melanoma, Experimental/drug therapy , Melanoma, Experimental/metabolism , Cell Line, Tumor , Microphthalmia-Associated Transcription Factor/metabolism , Muscles/drug effects , Muscles/metabolism , Intramolecular Oxidoreductases/metabolism , Receptor, Melanocortin, Type 1/metabolism , Molecular Dynamics Simulation , Cyclic AMP Response Element-Binding Protein/metabolism
3.
Food Chem ; 451: 139409, 2024 Sep 01.
Article in English | MEDLINE | ID: mdl-38692236

ABSTRACT

Herein, nineteen buckwheat honey samples collected from 19 stations of different ecological zones of Kazakhstan were analysed for their pollen density, physicochemical properties, chemical composition, antioxidant, anticholinesterase, tyrosinase inhibitory, and urease inhibitory activities with chemometric approaches. Twelve phenolic compounds and fumaric acid were identified using HPLC-DAD, and mainly fumaric, p-hydroxybenzoic, p-coumaric, trans-2-hydroxy cinnamic acids, and chrysin were detected in all samples. The honey samples collected from the Northern zone exhibited best antioxidant activity in lipid peroxidation inhibitory (IC50:8.65 ± 0.50 mg/mL), DPPH• (IC50:17.07 ± 1.49 mg/mL), ABTS•+ (IC50:8.90 ± 0.65 mg/mL), CUPRAC (A0.50:7.51 ± 0.30 mg/mL) and metal chelating assay (IC50:10.39 ± 0.71 mg/mL). In contrast, South-eastern zone samples indicated better acetylcholinesterase (55.57 ± 0.83%), butyrylcholinesterase (49.59 ± 1.09%), tyrosinase (44.40 ± 1.21%), and moderate urease (24.57 ± 0.33%) inhibitory activities at 20 mg/mL. The chemometric classification of nineteen buckwheat honey was performed using PCA and HCA techniques. Both were supported by correlation analysis. Thirteen compounds contributed significantly to the clustering of buckwheat honey based on geographical origin.


Subject(s)
Antioxidants , Fagopyrum , Honey , Honey/analysis , Honey/classification , Fagopyrum/chemistry , Fagopyrum/classification , Antioxidants/chemistry , Antioxidants/analysis , Kazakhstan , Monophenol Monooxygenase/antagonists & inhibitors , Chemometrics , Phenols/analysis , Phenols/chemistry , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/pharmacology , Enzyme Inhibitors/analysis
4.
J Agric Food Chem ; 72(19): 10958-10969, 2024 May 15.
Article in English | MEDLINE | ID: mdl-38703118

ABSTRACT

Demand for the exploration of botanical pesticides continues to increase due to the detrimental effects of synthetic chemicals on human health and the environment and the development of resistance by pests. Under the guidance of a bioactivity-guided approach and HSQC-based DeepSAT, 16 coumarin derivatives were discovered from the leaves of Ailanthus altissima (Mill.) Swingle, including seven undescribed monoterpenoid coumarins, three undescribed monoterpenoid phenylpropanoids, and two new coumarin derivatives. The structure and configurations of these compounds were established and validated via extensive spectroscopic analysis, acetonide analysis, and quantum chemical calculations. Biologically, 5 exhibited significant antifeedant activity toward the Plutella xylostella. Moreover, tyrosinase being closely related to the growth and development of larva, the inhibitory potentials of 5 against tyrosinase was evaluated in vitro and in silico. The bioactivity evaluation results highlight the prospect of 5 as a novel category of botanical insecticide.


Subject(s)
Ailanthus , Coumarins , Insecticides , Plant Extracts , Plant Leaves , Plant Leaves/chemistry , Animals , Coumarins/pharmacology , Coumarins/chemistry , Ailanthus/chemistry , Plant Extracts/chemistry , Plant Extracts/pharmacology , Insecticides/chemistry , Insecticides/pharmacology , Molecular Structure , Larva/drug effects , Larva/growth & development , Moths/drug effects , Moths/growth & development , Monophenol Monooxygenase/antagonists & inhibitors , Monophenol Monooxygenase/metabolism , Biological Assay , Monoterpenes/pharmacology , Monoterpenes/chemistry , Feeding Behavior/drug effects , Enzyme Inhibitors/pharmacology , Enzyme Inhibitors/chemistry
5.
PLoS One ; 19(5): e0298502, 2024.
Article in English | MEDLINE | ID: mdl-38814922

ABSTRACT

The skin microbiome maintains healthy human skin, and disruption of the microbiome balance leads to inflammatory skin diseases such as folliculitis and atopic dermatitis. Staphylococcus aureus and Cutibacterium acnes are pathogenic bacteria that simultaneously inhabit the skin and cause inflammatory diseases of the skin through the activation of innate immune responses. Silkworms are useful invertebrate animal models for evaluating innate immune responses. In silkworms, phenoloxidase generates melanin as an indicator of innate immune activation upon the recognition of bacterial or fungal components. We hypothesized that S. aureus and C. acnes interact to increase the innate immunity-activating properties of S. aureus. In the present study, we showed that acidification is involved in the activation of silkworm hemolymph melanization by S. aureus. Autoclaved-killed S. aureus (S. aureus [AC]) alone does not greatly activate silkworm hemolymph melanization. On the other hand, applying S. aureus [AC] treated with C. acnes culture supernatant increased the silkworm hemolymph melanization. Adding C. acnes culture supernatant to the medium decreased the pH. S. aureus [AC] treated with propionic acid, acetic acid, or lactic acid induced higher silkworm hemolymph melanization activity than untreated S. aureus [AC]. S. aureus [AC] treated with hydrochloric acid also induced silkworm hemolymph melanization. The silkworm hemolymph melanization activity of S. aureus [AC] treated with hydrochloric acid was inhibited by protease treatment of S. aureus [AC]. These results suggest that acid treatment of S. aureus induces innate immune activation in silkworms and that S. aureus proteins are involved in the induction of innate immunity in silkworms.


Subject(s)
Bombyx , Hemolymph , Melanins , Staphylococcus aureus , Animals , Hemolymph/metabolism , Hemolymph/microbiology , Hemolymph/immunology , Bombyx/microbiology , Bombyx/immunology , Staphylococcus aureus/immunology , Melanins/metabolism , Immunity, Innate , Hydrogen-Ion Concentration , Monophenol Monooxygenase/metabolism
6.
Chem Commun (Camb) ; 60(43): 5618-5621, 2024 May 23.
Article in English | MEDLINE | ID: mdl-38713525

ABSTRACT

A tyrosinase-activatable fluorescent probe with endoplasmic reticulum targetability was developed for the first time. It can ratiometrically fluoresce and hence be used to monitor refluxed tyrosinase into the endoplasmic reticulum.


Subject(s)
Endoplasmic Reticulum , Fluorescent Dyes , Monophenol Monooxygenase , Monophenol Monooxygenase/metabolism , Endoplasmic Reticulum/metabolism , Fluorescent Dyes/chemistry , Fluorescence , Humans , Spectrometry, Fluorescence
7.
J Enzyme Inhib Med Chem ; 39(1): 2357174, 2024 Dec.
Article in English | MEDLINE | ID: mdl-38814149

ABSTRACT

Tyrosinase, a pivotal enzyme in melanin synthesis, is a primary target for the development of depigmenting agents. In this work, in vitro and in silico techniques were employed to identify novel tyrosinase inhibitors from a set of 12 anilino-1,4-naphthoquinone derivatives. Results from the mushroom tyrosinase activity assay indicated that, among the 12 derivatives, three compounds (1, 5, and 10) demonstrated the most significant inhibitory activity against mushroom tyrosinase, surpassing the effectiveness of the kojic acid. Molecular docking revealed that all studied derivatives interacted with copper ions and amino acid residues at the enzyme active site. Molecular dynamics simulations provided insights into the stability of enzyme-inhibitor complexes, in which compounds 1, 5, and particularly 10 displayed greater stability, atomic contacts, and structural compactness than kojic acid. Drug likeness prediction further strengthens the potential of anilino-1,4-naphthoquinones as promising candidates for the development of novel tyrosinase inhibitors for the treatment of hyperpigmentation disorders.


Subject(s)
Agaricales , Dose-Response Relationship, Drug , Enzyme Inhibitors , Monophenol Monooxygenase , Naphthoquinones , Monophenol Monooxygenase/antagonists & inhibitors , Monophenol Monooxygenase/metabolism , Naphthoquinones/pharmacology , Naphthoquinones/chemistry , Naphthoquinones/chemical synthesis , Enzyme Inhibitors/pharmacology , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/chemical synthesis , Agaricales/enzymology , Structure-Activity Relationship , Molecular Structure , Molecular Docking Simulation , Molecular Dynamics Simulation
8.
Nat Microbiol ; 9(6): 1454-1466, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38806673

ABSTRACT

With rising global temperatures, permafrost carbon stores are vulnerable to microbial degradation. The enzyme latch theory states that polyphenols should accumulate in saturated peatlands due to diminished phenol oxidase activity, inhibiting resident microbes and promoting carbon stabilization. Pairing microbiome and geochemical measurements along a permafrost thaw-induced saturation gradient in Stordalen Mire, a model Arctic peatland, we confirmed a negative relationship between phenol oxidase expression and saturation but failed to support other trends predicted by the enzyme latch. To inventory alternative polyphenol removal strategies, we built CAMPER, a gene annotation tool leveraging polyphenol enzyme knowledge gleaned across microbial ecosystems. Applying CAMPER to genome-resolved metatranscriptomes, we identified genes for diverse polyphenol-active enzymes expressed by various microbial lineages under a range of redox conditions. This shifts the paradigm that polyphenols stabilize carbon in saturated soils and highlights the need to consider both oxic and anoxic polyphenol metabolisms to understand carbon cycling in changing ecosystems.


Subject(s)
Carbon Cycle , Microbiota , Permafrost , Polyphenols , Soil Microbiology , Polyphenols/metabolism , Permafrost/microbiology , Bacteria/metabolism , Bacteria/genetics , Bacteria/enzymology , Bacteria/classification , Carbon/metabolism , Oxidation-Reduction , Arctic Regions , Monophenol Monooxygenase/metabolism , Monophenol Monooxygenase/genetics , Soil/chemistry , Ecosystem
9.
Molecules ; 29(9)2024 Apr 23.
Article in English | MEDLINE | ID: mdl-38731413

ABSTRACT

Ultraviolet radiation can heighten tyrosinase activity, stimulate melanocyte production, impede the metabolism of numerous melanocytes, and result in the accumulation of plaques on the skin surface. α-Arbutin, a bioactive substance extracted from the arbutin plant, has been widely used for skin whitening. In this study, the whitening effect of α-arbutin by inhibiting tyrosinase activity and alleviating the photoaging effect induced by UVB are investigated. The results indicate that α-arbutin can inhibit skin inflammation, and its effectiveness is positively correlated with concentration. Moreover, α-arbutin can reduce the skin epidermal thickness, decrease the number of inflammatory cells, and down-regulate the expression levels of IL-1ß, IL-6 and TNF-α, which are inflammatory factors. It also promotes the expression of COL-1 collagen, thus playing an important role in anti-inflammatory action. Network pharmacology, metabolomics and transcriptomics further confirm that α-arbutin is related to the L-tyrosine metabolic pathway and may interfere with various signaling pathways related to melanin and other photoaging by regulating metabolic changes. Therefore, α-arbutin has a potential inhibitory effect on UVB-induced photoaging and possesses a whitening effect as a cosmetic compound.


Subject(s)
Arbutin , Skin Aging , Ultraviolet Rays , Arbutin/pharmacology , Ultraviolet Rays/adverse effects , Animals , Skin Aging/drug effects , Skin Aging/radiation effects , Mice , Monophenol Monooxygenase/metabolism , Monophenol Monooxygenase/antagonists & inhibitors , Humans , Skin/radiation effects , Skin/drug effects , Skin/metabolism , Skin/pathology
10.
Bioorg Chem ; 147: 107397, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38691905

ABSTRACT

Phenolics, abundant in plants, constitute a significant portion of phytoconstituents consumed in the human diet. The phytochemical screening of the aerial parts of Centaurium spicatum led to the isolation of five phenolics. The anti-tyrosinase activities of the isolated compounds were assessed through a combination of in vitro experiments and multiple in silico approaches. Docking and molecular dynamics (MD) simulation techniques were utilized to figure out the binding interactions of the isolated phytochemicals with tyrosinase. The findings from molecular docking analysis revealed that the isolated phenolics were able to bind effectively to tyrosinase and potentially inhibit substrate binding, consequently diminishing the catalytic activity of tyrosinase. Among isolated compounds, cichoric acid displayed the lowest binding energy and the highest extent of polar interactions with the target enzyme. Analysis of MD simulation trajectories indicated that equilibrium was reached within 30 ns for all complexes of tyrosinase with the isolated phenolics. Among the five ligands studied, cichoric acid exhibited the lowest interaction energies, rendering its complex with tyrosinase the most stable. Considering these collective findings, cichoric acid emerges as a promising candidate for the design and development of a potential tyrosinase inhibitor. Furthermore, the in vitro anti-tyrosinase activity assay unveiled significant variations among the isolated compounds. Notably, cichoric acid exhibited the most potent inhibitory effect, as evidenced by the lowest IC50 value (7.92 ± 1.32 µg/ml), followed by isorhamnetin and gentiopicrin. In contrast, sinapic acid demonstrated the least inhibitory activity against tyrosinase, with the highest IC50 value. Moreover, cichoric acid exhibited a mixed inhibition mode against the hydrolysis of l-DOPA catalyzed by tyrosinase, with Ki value of 1.64. Remarkably, these experimental findings align well with the outcomes of docking and MD simulations, underscoring the consistency and reliability of our computational predictions with the actual inhibitory potential observed in vitro.


Subject(s)
Enzyme Inhibitors , Molecular Docking Simulation , Monophenol Monooxygenase , Phenols , Monophenol Monooxygenase/antagonists & inhibitors , Monophenol Monooxygenase/metabolism , Enzyme Inhibitors/pharmacology , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/isolation & purification , Phenols/chemistry , Phenols/pharmacology , Phenols/isolation & purification , Molecular Structure , Dose-Response Relationship, Drug , Structure-Activity Relationship , Molecular Dynamics Simulation , Agaricales/enzymology
11.
Exp Dermatol ; 33(5): e15101, 2024 May.
Article in English | MEDLINE | ID: mdl-38770555

ABSTRACT

Skin hyperpigmentation is mainly caused by excessive synthesis of melanin; however, there is still no safe and effective therapy for its removal. Here, we found that the dermal freezer was able to improve UVB-induced hyperpigmentation of guinea pigs without causing obvious epidermal damage. We also mimic freezing stimulation at the cellular level by rapid freezing and observed that freezing treatments <2.5 min could not decrease cell viability or induce cell apoptosis in B16F10 and Melan-A cells. Critically, melanin content and tyrosinase activity in two cells were greatly reduced after freezing treatments. The dramatic decrease in tyrosinase activity was associated with the downregulation of MITF, TYR, TRP-1 and TRP-2 protein expression in response to freezing treatments for two cells. Furthermore, our results first demonstrated that freezing treatments significantly reduced the levels of p-GSK3ß and ß-catenin and the nuclear accumulation of ß-catenin in B16F10 and Melan-A cells. Together, these data suggest that fast freezing treatments can inhibit melanogenesis-related gene expression in melanocytes by regulating the Wnt/ß-catenin signalling pathway. The inhibition of melanin production eventually contributed to the improvement in skin hyperpigmentation induced by UVB. Therefore, fast freezing treatments may be a new alternative of skin whitening in the clinic in the future.


Subject(s)
Freezing , Hyperpigmentation , Melanins , Melanocytes , Monophenol Monooxygenase , Ultraviolet Rays , Wnt Signaling Pathway , beta Catenin , Animals , Melanins/biosynthesis , Melanins/metabolism , Melanocytes/metabolism , Mice , Hyperpigmentation/metabolism , beta Catenin/metabolism , Monophenol Monooxygenase/metabolism , Guinea Pigs , Microphthalmia-Associated Transcription Factor/metabolism , Cell Survival , Intramolecular Oxidoreductases/metabolism , Glycogen Synthase Kinase 3 beta/metabolism , Apoptosis , Oxidoreductases/metabolism , Interferon Type I , Pregnancy Proteins
12.
Front Biosci (Landmark Ed) ; 29(5): 194, 2024 May 20.
Article in English | MEDLINE | ID: mdl-38812330

ABSTRACT

BACKGROUNDS: Melanogenesis, regulated by genetic, hormonal, and environmental factors, occurs in melanocytes in the basal layer of the epidermis. Dysregulation of this process can lead to various skin disorders, such as hyperpigmentation and hypopigmentation. Therefore, the present study investigated the effect of ultrasonic-assisted ethanol extract (SHUE) from Sargassum horneri (S. horneri), brown seaweed against melanogenesis in α-melanocyte-stimulating hormone (MSH)-stimulated B16F10 murine melanocytes. METHODS: Firstly, yield and proximate compositional analysis of the samples were conducted. The effect of SHUE on cell viability has been evaluated by using 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. After that, the melanin content and cellular tyrosinase activity in α-MSH-stimulated B16F10 murine melanocytes were examined. Western blot analysis was carried out to investigate the protein expression levels of microphthalmia-associated transcription factor (MITF), tyrosinase, tyrosinase-related protein-1 (TRP1), and tyrosinase-related protein-2 (TRP2). In addition, the effect of extracellular signal-regulated kinase (ERK) on the melanogenesis process was assessed via Western blotting. RESULTS: As per the analysis, SHUE contained the highest average yield on a dry basis at 28.70 ± 3.21%. The findings showed that SHUE reduced the melanin content and cellular tyrosinase activity in α-MSH-stimulated B16F10 murine melanocytes. Additionally, the expression levels of MITF, TRP1, and TRP2 protein were significantly downregulated by SHUE treatment in α-MSH-stimulated B16F10 murine melanocytes. Moreover, SHUE upregulated the phosphorylation of ERK and AKT in α-MSH-stimulated B16F10 murine melanocytes. In addition, experiments conducted using the ERK inhibitor (PD98059) revealed that the activity of SHUE depends on the ERK signaling cascade. CONCLUSION: These results suggest that SHUE has an anti-melanogenic effect and can be used as a material in the formulation of cosmetics related to whitening and lightening.


Subject(s)
Ethanol , Melanins , Melanocytes , Monophenol Monooxygenase , Sargassum , Animals , Sargassum/chemistry , Melanins/biosynthesis , Melanins/metabolism , Monophenol Monooxygenase/metabolism , Monophenol Monooxygenase/antagonists & inhibitors , Melanocytes/drug effects , Melanocytes/metabolism , Mice , Ethanol/chemistry , Microphthalmia-Associated Transcription Factor/metabolism , alpha-MSH/metabolism , Plant Extracts/pharmacology , Plant Extracts/chemistry , Cell Survival/drug effects , Melanoma, Experimental/metabolism , Cell Line, Tumor , Intramolecular Oxidoreductases/metabolism
13.
J Agric Food Chem ; 72(15): 8742-8748, 2024 Apr 17.
Article in English | MEDLINE | ID: mdl-38564658

ABSTRACT

Tyrosinase is capable of oxidizing tyrosine residues in proteins, leading to intermolecular protein cross-linking, which could modify the protein network of food and improve the texture of food. To obtain the recombinant tyrosinase with microbial cell factory instead of isolation tyrosinase from the mushroom Agaricus bisporus, a TYR expression cassette was constructed in this study. The expression cassette was electroporated into Trichoderma reesei Rut-C30 and integrated into its genome, resulting in a recombinant strain C30-TYR. After induction with microcrystalline cellulose for 7 days, recombinant tyrosinase could be successfully expressed and secreted by C30-TYR, corresponding to approximately 2.16 g/L tyrosinase in shake-flask cultures. The recombinant TYR was purified by ammonium sulfate precipitation and gel filtration, and the biological activity of purified TYR was 45.6 U/mL. The purified TYR could catalyze the cross-linking of glycinin, and the emulsion stability index of TYR-treated glycinin emulsion was increased by 30.6% compared with the untreated one. The cross-linking of soy glycinin by TYR resulted in altered properties of oil-in-water emulsions compared to emulsions stabilized by native glycinin. Therefore, cross-linking with this recombinant tyrosinase is a feasible approach to improve the properties of protein-stabilized emulsions and gels.


Subject(s)
Cross-Linking Reagents , Gene Expression , Globulins , Hypocreales , Monophenol Monooxygenase , Recombinant Proteins , Soybean Proteins , Monophenol Monooxygenase/biosynthesis , Monophenol Monooxygenase/genetics , Monophenol Monooxygenase/isolation & purification , Monophenol Monooxygenase/metabolism , Cross-Linking Reagents/isolation & purification , Cross-Linking Reagents/metabolism , Hypocreales/classification , Hypocreales/genetics , Hypocreales/growth & development , Hypocreales/metabolism , Globulins/chemistry , Globulins/metabolism , Soybean Proteins/chemistry , Soybean Proteins/metabolism , Electroporation , Cellulose , Ammonium Sulfate , Chromatography, Gel , Fractional Precipitation , Emulsions/chemistry , Emulsions/metabolism , Recombinant Proteins/biosynthesis , Recombinant Proteins/genetics , Recombinant Proteins/isolation & purification , Recombinant Proteins/metabolism , Protein Stability , Endoplasmic Reticulum/metabolism , Protein Sorting Signals , Oils/chemistry , Water/chemistry
14.
Int J Mol Sci ; 25(8)2024 Apr 17.
Article in English | MEDLINE | ID: mdl-38674004

ABSTRACT

Phenolic compounds, originating from industrial, agricultural, and urban sources, can leach into flowing waters, adversely affecting aquatic life, biodiversity, and compromising the quality of drinking water, posing potential health hazards to humans. Thus, monitoring and mitigating the presence of phenolic compounds in flowing waters are essential for preserving ecosystem integrity and safeguarding public health. This study explores the development and performance of an innovative sensor based on screen-printed electrode (SPE) modified with graphene (GPH), poly(3,4-ethylenedioxythiophene) (PEDOT), and tyrosinase (Ty), designed for water analysis, focusing on the manufacturing process and the obtained electroanalytical results. The proposed biosensor (SPE/GPH/PEDOT/Ty) was designed to achieve a high level of precision and sensitivity, as well as to allow efficient analytical recoveries. Special attention was given to the manufacturing process and optimization of the modifying elements' composition. This study highlights the potential of the biosensor as an efficient and reliable solution for water analysis. Modification with graphene, the synthesis and electropolymerization deposition of the PEDOT polymer, and tyrosinase immobilization contributed to obtaining a high-performance and robust biosensor, presenting promising perspectives in monitoring the quality of the aquatic environment. Regarding the electroanalytical experimental results, the detection limits (LODs) obtained with this biosensor are extremely low for all phenolic compounds (8.63 × 10-10 M for catechol, 7.72 × 10-10 M for 3-methoxycatechol, and 9.56 × 10-10 M for 4-methylcatechol), emphasizing its ability to accurately measure even subtle variations in the trace compound parameters. The enhanced sensitivity of the biosensor facilitates detection and quantification in river water samples. Analytical recovery is also an essential aspect, and the biosensor presents consistent and reproducible results. This feature significantly improves the reliability and usefulness of the biosensor in practical applications, making it suitable for monitoring industrial or river water.


Subject(s)
Biosensing Techniques , Bridged Bicyclo Compounds, Heterocyclic , Graphite , Monophenol Monooxygenase , Phenols , Polymers , Rivers , Water Pollutants, Chemical , Biosensing Techniques/methods , Graphite/chemistry , Rivers/chemistry , Polymers/chemistry , Phenols/analysis , Water Pollutants, Chemical/analysis , Bridged Bicyclo Compounds, Heterocyclic/chemistry , Enzymes, Immobilized/chemistry , Electrochemical Techniques/methods , Electrodes , Limit of Detection
15.
Biosensors (Basel) ; 14(4)2024 Apr 19.
Article in English | MEDLINE | ID: mdl-38667195

ABSTRACT

Tyrosinase (TYR) emerges as a key enzyme that exerts a regulatory influence on the synthesis of melanin, thereby assuming the role of a critical biomarker for the detection of melanoma. Detecting the authentic concentration of TYR in the skin remains a primary challenge. Distinguished from ex vivo detection methods, this study introduces a novel sensor platform that integrates a microneedle (MN) biosensor with surface-enhanced Raman spectroscopy (SERS) technology for the in situ detection of TYR in human skin. The platform utilized dopamine (DA)-functionalized gold nanoparticles (Au NPs) as the capturing substrate and 4-mercaptophenylboronic acid (4-MPBA)-modified silver nanoparticles (Ag NPs) acting as the SERS probe. Here, the Au NPs were functionalized with mercaptosuccinic acid (MSA) for DA capture. In the presence of TYR, DA immobilized on the MN is preferentially oxidized to dopamine quinone (DQ), a process that results in a decreased density of SERS probes on the platform. TYR concentration was detected through variations in the signal intensity emitted by the phenylboronic acid. The detection system was able to evaluate TYR concentrations within a linear range of 0.05 U/mL to 200 U/mL and showed robust anti-interference capabilities. The proposed platform, integrating MN-based in situ sensing, SERS technology, and TYR responsiveness, holds significant importance for diagnosing cutaneous melanoma.


Subject(s)
Biosensing Techniques , Early Detection of Cancer , Melanoma , Monophenol Monooxygenase , Spectrum Analysis, Raman , Biosensing Techniques/instrumentation , Biosensing Techniques/methods , Monophenol Monooxygenase/analysis , Monophenol Monooxygenase/metabolism , Skin/enzymology , Animals , Mice , Melanoma/diagnosis , Melanoma/enzymology , Metal Nanoparticles/chemistry , Gold/chemistry , Needles/standards , Enzyme-Linked Immunosorbent Assay , Silver/chemistry , Sensitivity and Specificity , Early Detection of Cancer/instrumentation , Early Detection of Cancer/methods
16.
J Med Chem ; 67(9): 7260-7275, 2024 May 09.
Article in English | MEDLINE | ID: mdl-38651218

ABSTRACT

Artificial intelligence (AI) de novo molecular generation is a highly promising strategy in the drug discovery, with deep reinforcement learning (RL) models emerging as powerful tools. This study introduces a fragment-by-fragment growth RL forward molecular generation and optimization strategy based on a low activity lead compound. This process integrates fragment growth-based reaction templates, while target docking and drug-likeness prediction were simultaneously performed. This comprehensive approach considers molecular similarity, internal diversity, synthesizability, and effectiveness, thereby enhancing the quality and efficiency of molecular generation. Finally, a series of tyrosinase inhibitors were generated and synthesized. Most compounds exhibited more improved activity than lead, with an optimal candidate compound surpassing the effects of kojic acid and demonstrating significant antipigmentation activity in a zebrafish model. Furthermore, metabolic stability studies indicated susceptibility to hepatic metabolism. The proposed AI structural optimization strategies will play a promising role in accelerating the drug discovery and improving traditional efficiency.


Subject(s)
Artificial Intelligence , Enzyme Inhibitors , Monophenol Monooxygenase , Zebrafish , Animals , Monophenol Monooxygenase/antagonists & inhibitors , Monophenol Monooxygenase/metabolism , Enzyme Inhibitors/pharmacology , Enzyme Inhibitors/chemistry , Enzyme Inhibitors/chemical synthesis , Molecular Docking Simulation , Structure-Activity Relationship , Molecular Structure , Humans , Drug Discovery
17.
Int J Biol Macromol ; 267(Pt 2): 131513, 2024 May.
Article in English | MEDLINE | ID: mdl-38608979

ABSTRACT

Tyrosinase is a copper oxidase enzyme which catalyzes the first two steps in the melanogenesis pathway, L-tyrosine to L-dopa conversion and, then, to o-dopaquinone and dopachrome. Hypopigmentation and, above all, hyperpigmentation issues can be originated depending on their activity. This enzyme also promotes the browning of fruits and vegetables. Therefore, control of their activity by regulators is research topic of great relevance. In this work, we consider the use of inhibitors of monophenolase and diphenolase activities of the enzyme in order to accomplish such control. An experimental design and data analysis which allow the accurate calculation of the degree of inhibition of monophenolase activity (iM) and diphenolase activity (iD) are proposed. The IC50 values (amount of inhibitor that causes 50 % inhibition at a fixed substrate concentration) can be calculated for the two activities and from the values of IC50M (monophenolase) and IC50D(diphenolase). Additionally, the strength and type of inhibition can be deduced from these values. The data analysis from these IC50D values allows to obtain the values of [Formula: see text] or [Formula: see text] , or and [Formula: see text] from the values of IC50M. In all cases, the values of the different must satisfy their relationship with IC50M and IC50D.


Subject(s)
Enzyme Inhibitors , Monophenol Monooxygenase , Monophenol Monooxygenase/antagonists & inhibitors , Monophenol Monooxygenase/metabolism , Monophenol Monooxygenase/chemistry , Enzyme Inhibitors/pharmacology , Enzyme Inhibitors/chemistry , Inhibitory Concentration 50 , Kinetics , Oxidoreductases/antagonists & inhibitors , Oxidoreductases/metabolism , Humans
18.
Protein Expr Purif ; 219: 106483, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38609025

ABSTRACT

Mussel foot proteins (Mfps) possess unique binding properties to various surfaces due to the presence of L-3,4-dihydroxyphenylalanine (DOPA). Mytilus edulis foot protein-3 (Mefp-3) is one of several proteins in the byssal adhesive plaque. Its localization at the plaque-substrate interface approved that Mefp-3 plays a key role in adhesion. Therefore, the protein is suitable for the development of innovative bio-based binders. However, recombinant Mfp-3s are mainly purified from inclusion bodies under denaturing conditions. Here, we describe a robust and reproducible protocol for obtaining soluble and tag-free Mefp-3 using the SUMO-fusion technology. Additionally, a microbial tyrosinase from Verrucomicrobium spinosum was used for the in vitro hydroxylation of peptide-bound tyrosines in Mefp-3 for the first time. The highly hydroxylated Mefp-3, confirmed by MALDI-TOF-MS, exhibited excellent adhesive properties comparable to a commercial glue. These results demonstrate a concerted and simplified high yield production process for recombinant soluble and tag-free Mfp3-based proteins with on demand DOPA modification.


Subject(s)
Dihydroxyphenylalanine , Mytilus edulis , Animals , Dihydroxyphenylalanine/chemistry , Dihydroxyphenylalanine/metabolism , Mytilus edulis/genetics , Mytilus edulis/chemistry , Mytilus edulis/metabolism , Recombinant Proteins/genetics , Recombinant Proteins/biosynthesis , Recombinant Proteins/isolation & purification , Recombinant Proteins/chemistry , Recombinant Proteins/metabolism , Verrucomicrobia/genetics , Verrucomicrobia/metabolism , Monophenol Monooxygenase/genetics , Monophenol Monooxygenase/metabolism , Monophenol Monooxygenase/chemistry , Proteins/genetics , Proteins/chemistry , Proteins/isolation & purification , Hydroxylation , Escherichia coli/genetics , Escherichia coli/metabolism
19.
Food Chem ; 450: 139392, 2024 Aug 30.
Article in English | MEDLINE | ID: mdl-38640546

ABSTRACT

The combinational effects of kojic acid and lauroyl arginine ethyl ester hydrochloride (ELAH) on fresh-cut potatoes were investigated. Kojic acid of 0.6% (w/w) effectively inhibited the browning of fresh-cut potatoes and displayed antimicrobial capacity. The color difference value of samples was decreased from 175 to 26 by kojic acid. In contrast, ELAH could not effectively bind with the active sites of tyrosinase and catechol oxidase at molecular level. Although 0.5% (w/w) of ELAH prominently inhibited the microbial growth, it promoted the browning of samples. However, combining kojic acid and ELAH effectively inhibited the browning of samples and microbial growth during the storage and the color difference value of samples was decreased to 52. This amount of kojic acid inhibited enzyme activities toward phenolic compounds. The results indicated that combination of kojic acid and ELAH could provide a potential strategy to extend the shelf life of fresh-cut products.


Subject(s)
Arginine , Monophenol Monooxygenase , Pyrones , Solanum tuberosum , Pyrones/pharmacology , Pyrones/chemistry , Arginine/chemistry , Arginine/analogs & derivatives , Arginine/pharmacology , Solanum tuberosum/chemistry , Solanum tuberosum/growth & development , Monophenol Monooxygenase/metabolism , Food Preservation/methods , Catechol Oxidase/metabolism , Food Preservatives/pharmacology , Food Preservatives/chemistry , Bacteria/drug effects , Bacteria/genetics
20.
Peptides ; 177: 171215, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38608837

ABSTRACT

Melasma is a common skin disease induced by an increase in the content of melanin in the skin, which also causes serious physical and mental harm to patients. In this research, a novel peptide (Nigrocin-OA27) from Odorrana andersonii is shown to exert a whitening effect on C57 mice pigmentation model. The peptide also demonstrated non-toxic and antioxidant capacity, and can significantly reduce melanin content in B16 cells. Topical application effectively delivered Nigrocin-OA27 to skin's epidermal and dermal layers and exhibited significant preventive and whitening effects on the UVB-induced ear pigmentation model in C57 mice. The whitening mechanism of Nigrocin-OA27 may be related to reduced levels of the microphthalmia-associated transcription factor and the key enzyme for melanogenesis-tyrosinase (TYR). Nigrocin-OA27 also inhibited the catalytic activity by adhering to the active core of TYR, thereby reducing melanin formation and deposition. In conclusion, Nigrocin-OA27 may be a potentially effective external agent to treat melasma by inhibiting aberrant skin melanin synthesis.


Subject(s)
Melanins , Microphthalmia-Associated Transcription Factor , Monophenol Monooxygenase , Ultraviolet Rays , Animals , Melanins/metabolism , Melanins/biosynthesis , Microphthalmia-Associated Transcription Factor/metabolism , Microphthalmia-Associated Transcription Factor/genetics , Mice , Monophenol Monooxygenase/metabolism , Ultraviolet Rays/adverse effects , Peptides/pharmacology , Peptides/chemistry , Skin Pigmentation/drug effects , Skin Pigmentation/radiation effects , Mice, Inbred C57BL , Skin/drug effects , Skin/metabolism , Skin/radiation effects , Skin/pathology , Signal Transduction/drug effects
SELECTION OF CITATIONS
SEARCH DETAIL
...