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Epimedin B exhibits pigmentation by increasing tyrosinase family proteins expression,activity,and stability

Journal of Pharmaceutical Analysis 2024年第1期14卷 69-85页

作者： Chen Hong Yifan Zhang Lili Yang Haoyang Xu Kang Cheng Zhi Lv Kaixian Chen Yiming Li Huali Wu Department of TCM Chemistry School of PharmacyShanghai University of Traditional Chinese MedicineShanghai200000China Department of Dermatology Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese MedicineShanghai200000China International Education College Shanghai University of Traditional Chinese MedicineShanghai200000China Shanghai Inoherb Cosmetics Co. Ltd.Shanghai200000China

Epimedin B(EB)is one of the main flavonoid ingredients present in Epimedium brevicornum Maxim.,a traditional herb widely used in China.Our previous study showed that EB was a stronger inducer of melanogenesis and an activator of tyrosinase(TYR).However,the role of EB in melanogenesis and the mechanism underlying the regulation remain unclear.Herein,as an extension to our previous investigation,we provide comprehensive evidence of EB-induced pigmentation in vivo and in vitro and elucidate the melanogenesis mechanism by assessing its effects on the TYR family of proteins(TYRs)in terms of expression,activity,and stability.The results showed that EB increased TYRs expression through microphthalmia-associated transcription factor-mediated p-Akt(referred to as protein kinase B(PKB))/glycogen synthase kinase 3β(GSK3β)/β-catenin,p-p70 S6 kinase cascades,and protein 38(p38)/mitogen-activated protein(MAP)kinase(MAPK)and extracellular regulated protein kinases(ERK)/MAPK pathways,after which EB increased the number of melanosomes and promoted their maturation for melanogenesis in melanoma cells and human primary melanocytes/skin tissues.Furthermore,EB exerted repigmentation by stimulating TYR activity in hydroquinone-and N-phenylthiourea-induced TYR inhibitive models,including melanoma cells,zebrafish,and mice.Finally,EB ameliorated monobenzone-induced depigmentation in vitro and in vivo through the enhancement of TYRs stability by inhibiting TYR misfolding,TYR-related protein 1 formation,and retention in the endoplasmic reticulum and then by downregulating the ubiquitination and proteolysis processes.These data conclude that EB can target TYRs and alter their expression,activity,and stability,thus stimulating their pigmentation function,which might provide a novel rational strategy for hypopigmentation treatment in the pharmaceutical and cosmetic industries.

关键词： Epimedin B Melanosome tyrosinase Melanogenesis

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Rapid detection of tyrosinase in shrimp by polyacid-based electrochemical sensors

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Tungsten 2023年第1期5卷 81-90页

作者： Xiao-Wen Chen Yu-Shan Chen Xiao-Mei Ding Li Wang Jing Liu Rui-Fang Wang College of Food and Biological Engineering Jimei UniversityXiamen 361021China School of Information Engineering Jimei UniversityXiamen 361021China

The increase through tyrosinase levels after shrimp death causes enzymatic browning,affecting the appearance and freshness of products,and it is essential to establish a simple and rapid method to detect tyrosinase in the aquatic product supply chain to control the market’s good quality.In this study,we used a constant potential deposition method to create a [(Polystyrene Sodium Sulfonate(PS S)/Polypyrrole(PPy))(P2Mo17Ni/PPy)5] polystyrene sodium sulfonate polymetallic oxide thin film electrodes that can realize the detection of tyrosinase(TYR) in shrimp samples.The method yielded a limit of detection(Signal-Noise ratio(S/N)=3) of 0.1028 U·mL^(-1) within the linear detection of 3.22-26.04 U·mL^(-1),and the recoveries of the samples ranged from 96.37% to 102.40%.The above data confirmed that the method showed some accuracy in the detection of TYR content in shrimps and provided a theoretical basis for the preparation of tyrosinase sensors for the application of polyoxometalates in aquatic products.

关键词： Polyoxometalates Electrochemical tyrosinase Sensors

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Comparison of structural and physicochemical properties of potato protein and potato flour modified with tyrosinase

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Journal of Integrative Agriculture 2022年第5期21卷 1513-1524页

作者： ZHU Yu YUAN Yu-han MEI Li-ping DING Shuang-kun GAO Yu-chen DU Xian-feng GUO Li Anhui Engineering Laboratory of Agro-products Processing Anhui Agricultural UniversityHefei 230036P.R.China State Key Laboratory of Biobased Material and Green Papermaking School of Food Sciences and EngineeringQilu University of Technology(Shandong Academy of Sciences)Jinan 250353P.R.China College of Life Science and Technology Xinjiang UniversityUrumchi 830046P.R.China

The present study modified potato protein and flour with tyrosinase to promote the diversification of potato staple foods.The results indicated that tyrosinase treatment markedly altered the secondary structure of proteins.After tyrosinase treatment,the maximum decomposition temperature of potato protein and flour increased from 322.32 to 332.40℃ and from 294.24 to 299.61℃,respectively.tyrosinase treatment remarkably reduced the pasting viscosity of potato flour,that is,the peak viscosity,through reducing viscosity,breakdown,final viscosity,and setback by 32.50,60.98,13.04,68.24,and 74.31%,respectively.In contrast,tyrosinase treatment increased the shear resistance and hardness of the protein and flour gels;the maximum stress values of the protein and flour gels increased from 1.48 to 10.1% and from 6.87 to 14.8%,respectively.Furthermore,tyrosinase treatment promoted viscoelastic properties and structural stability of potato protein and flour.These results may provide an important foundation for the development of novel potato staple foods.

关键词： potato protein starch structural tyrosinase physicochemical features

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Engineering trienzyme cascade-triggered fluorescent immunosensor platform by sequentially integrating alkaline phosphatase, tyrosinase and horseradish peroxidase

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Chinese Chemical Letters 2023年第4期34卷 444-448页

作者： Yujie Sun Lei Wen Huili Ma Wenlin Ma Zhenqian Fu Yinhui Li Chengwu Zhang Lin Li Jinhua Liu Institute of Advanced Materials(IAM) Key Laboratory of Flexible Electronics(KLOFE)Nanjing Tech UniversityNanjing 211816China Key Laboratory for Green Organic Synthesis and Application of Hunan Province Key Laboratory of Environmentally Friendly ChemistryApplication of Ministry of EducationCollege of ChemistryXiangtan UniversityXiangtan 411105China School of Basic Medical Sciences Shanxi Medical UniversityTaiyuan 310003China

Mulit-enzyme cascades are a major type of chemical transformations and play a crucial role in biological signal transduction and metabolism. Herein, a trienzyme cascade-triggered fluorescent immunosensor platform was constructed by sequentially integrating alkaline phosphatase(ALP), tyrosinase(TYR)and horseradish peroxidase(HRP). The proposed platform was based on HRP-induced a rapid in situ fluorogenic reaction between dopamine(DA) and 1,5-dihydroxynaphthalene(DHA) to produce a strong yellow azamonardine fluorescent compound(AFC). The obtained AFC was clearly characterized by highresolution mass spectrum,1H NMR,^(13)C NMR and theoretical calculations. The integration of the twoenzyme system(TYR and HRP) or three-enzyme system(ALP, TYR and HRP) led to a maximum of 400.0-fold and 250.0-fold fluorescence enhancements, respectively. Using cardiac troponin I(c Tn I) as the model antigen, a trienzyme cascade-triggered fluorescent immunosensor platform was developed for quantitative detecting c Tn I in a wide linear range from 2 ng/m L to 150 ng/m L with a detection limit of 0.67ng/m L. In addition, the proposed platform was successfully applied in detection of c Tn I in serum of clinical patients. Overall, the developed fluorescent immunosensor performs powerful implications for researching enzyme cascade systems in the field of biomedicine.

关键词： Enzyme cascade Alkaline phosphatase tyrosinase Horseradish peroxidase Cardiac troponin I

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tyrosinase-modified carbon felt-based flow-biosensors:The role of ultra-sonication in shortening the enzyme immobilization time and improving the sensitivity for p-chlorophenol

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Journal of Environmental Sciences 2011年第6期23卷 1038-1043页

作者： Yasushi Hasebe Department of Material Science and Engineering Graduate School of EngineeringSaitama Institute of Technology1690 FusaijiFukayaSaitama 369-0293Japan Department of Life Science and Green Chemistry Saitama Institute of Technology1690 FusaijiFukayaSaitama 369-0293Japan

tyrosinase（TYR） was covalently immobilized onto amino-functionalized carbon felt surface via glutaraldehyde-coupling under ultrasonic treatment for 10 min.The resulting TYR-immobilized carbon felt was used as a working electrode unit of bioelectrocatalytic flow-through detector for TYR substrates（catechol,p-chlorophenol（p-CP）,p-cresol,phenol etc.）.Cathodic peak currents based on the electroreduction of enzymatically produced o-quinones were detected at-50 mV vs.Ag/AgCl.Compared with previous work in which TYR was immobilized onto amino-functionalized carbon felt for 16 hr without the ultrasonic treatment,we succeeded in（1） shortening the enzyme immobilization time from 16 hr to 10 min,（2） enhancing the sensitivity of p-CP,and（3） improving the operational stability of p-CP.The ultrasonic treatment during the TYR immobilization step would lead to certain changes in the structure of the immobilized TYR and the morphology of the immobilized TYR-layer on the carbon felt surface.

关键词： enzyme immobilization under ultrasonic treatment flow-biosensor carbon felt tyrosinase phenolic compounds

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Targeted disruption of tyrosinase causes melanin reduction in Carassius auratus cuvieri and its hybrid progeny

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Science China(Life Sciences) 2019年第9期62卷 1194-1202页

作者： Qingfeng Liu Yanhua Qi Qiuli Liang Jia Song Junmei Liu Wuhui Li Yuqin Shu Min Tao Chun Zhang Qinbo Qin Jing Wang Shaojun Liu State Key Laboratory of Developmental Biology of Freshwater Fish Hunan Normal UniversityChangsha 410081China College of Life Sciences Hunan Normal UniversityChangsha 410081China

The white crucian carp(Carassius auratus cuvieri,WCC) not only is one of the most economically important fish in Asia,characterized by strong reproductive ability and rapid growth rates,but also represents a good germplasm to produce hybrid progenies with heterosis.Gene knockout technique provides a safe and acceptant way for fish breeding.Achieving gene knockout in WCC and its hybrid progeny will be of great importance for both genetic studies and hybridization breeding.tyrosinase(TYR) is a key enzyme in melanin synthesis.Depletion of tyr in zebrafish and mice results in mosaic pigmentation or total albinism.Here,we successfully used CRISPR-Cas9 to target tyr in WCC and its hybrid progeny(WR) derived from the cross of WCC(♀) and red crucian carp(Carassius auratus red var.,RCC,♂).The level of TYR protein was significantly reduced in mutant WCC.Both the mutant WCC and the mutant WR showed different degrees of melanin reduction compared with the wild-type sibling control fish,resulting from different mutation efficiency ranging from 60% to 90%.In addition,the transcriptional expression profiles of a series of pivotal pigment synthesis genes,i.***.tyrp1,mitfa,mitfb,dct and sox10,were down-regulated in tyr-CRISPR WCC,which ultimately caused a reduction in melanin synthesis.These results demonstrated that tyr plays a key role in melanin synthesis in WCC and WR,and CRISPR-Cas9 is an effective tool for modifying the genome of economical fish.Furthermore,the tyr-CRISPR models could be valuable in understanding fundamental mechanisms of pigment formation in non-model fish.

关键词： white crucian carp hybridization CRISPR-Cas9 tyrosinase pigmentation

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Catalysis-based specific detection and inhibition of tyrosinase and their application

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Journal of Pharmaceutical Analysis 2020年第5期10卷 414-425页

作者： Yunwei Qu Qing Zhan Shubo Du Yang Ding Bin Fang Wei Du Qiong Wu Haidong Yu Lin Li Wei Huang Key Laboratory of Flexible Electronics(KLOFE)&Institute of Advanced Materials(IAM) Nanjing Tech University(NanjingTech)NanjingPR China Department of Chemistry National University of SingaporeSingapore Frontiers Science Center for Flexible Electronics Xi’an Institute of Flexible Electronics(IFE)and Xi’an Institute of Biomedical Materials&EngineeringNorthwestern Polytechnical University127 West Youyi RoadXi’an710072China

tyrosinase is an important enzyme in controlling the formation of melanin in melanosome,and plays a key role in the pigmentation of hair and skin.The abnormal expression or activation of tyrosinase is associated with several diseases such as albinism,vitiligo,melanoma and Parkinson disease.Excessive deposition of melanin could cause diseases such as freckles and brown spots in the human body,and it is also closely related to browning of fruits and vegetables and insect molting.Detecting and inhibiting the activity of tyrosinase is of extraordinary value in the progress of diagnosis and treatment of these diseases.Therefore,many selective optical detection probes and small molecular inhibitors have been developed,and have made significant contributions to the basic and clinical research on these diseases.In this paper,the detection and inhibition of tyrosinase and their application in whitening products are reviewed,with special emphasis on development of fluorescent probes and inhibitors.Hopefully,this review will help design more efficient and sensitive tyrosinase probes and inhibitors,as well as shed light on novel treatment of diseases such as melanoma.

关键词： tyrosinase Melanin Detection probe Inhibitors Melanoma

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Evaluation of a resoruf in-based fluorescent probe for tyrosinase detection in skin pigmentation disorders

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Bio-Design and Manufacturing 2021年第4期4卷 806-817页

作者： Yibo Hu Hongliang Zeng Jiaxing Jiang Sheng Yang Jinhua Huang Jing Chen Qinghai Zeng Department of Dermatology The Third Xiangya Hospital of Central South UniversityTongzipo Road No.138Yuelu DistrictChangsha 410013China Institute of Chinese Materia Medica Hunan Academy of Chinese MedicineChangsha 410006China School of Chemistry and Food Engineering Changsha University of Science and Technology(CSUST)Changsha 410114China

Purpose Skin pigmentation disorders,such as vitiligo and melasma,are difficult to diagnose in the early stages,but abnor-mal tyrosinase levels and tyrosinase activity are potential indicators.Some resorufin-based fluorescence probes(RBFPs)have been designed to detect tyrosinase in tumors,but they have not been used in skin pigmentation disorders.In this study,one of these RBFPs(synthesized by resorufin salt coupled with 3-(bromomethyl)phenol)was evaluated comprehensively.Methods The RBFP was tested in different kinds of mouse and human skin cells,as well as in in vivo models,including zebrafish,guinea pigs,and Sprague-Dawley rats.In addition,small interfering RNAs(siRNAs),kojic acid,and 1-phenyl-2-thiourea(PTU)were used to inhibit tyrosinase levels or tyrosinase activity.Results This probe successfully detected tyrosinase and emitted red fluorescence in melanoma cells and melanocytes.Fluorescence was also observable in zebrafish and on the skin of guinea pigs when using the RBFP.In mouse and human cells,the RBFP showed good selectivity to tyrosinase.Moreover,in the case of decreased tyrosinase levels or activity caused by siRNAs,kojic acid,or PTU,the probe was sensitive to these changes.Further,the RBFP showed no toxic effects at concentrations oftyrosinase detection,but suggest the need for further improvement of fluorescent probes in the diagnosis of skin pigmentation disorders.

关键词： tyrosinase Pigmentation disorder Diagnosis Fluorescence probe

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CuO/Cu2O nanowire array photoelectrochemical biosensor for ultrasensitive detection of tyrosinase

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Science China Chemistry 2020年第7期63卷 1012-1018页

作者： Xiaoxi Guo Jinghua Wu Lian Xia Meihao Xiang Fengli Qu Jinghong Li College of Chemistry and Chemical Engineering Qufu Normal UniversityQufu 273165China Department of Chemistry Key Laboratory of Bioorganic Phosphorus Chemistry&Chemical BiologyTsinghua UniversityBeijing 100084China

Photoelectrochemical(PEC) biosensors have shown great promise in bioanalysis and diagnostic applications in recent years. In this work, the CuO/Cu2O nanowire array(CuO/Cu2O Nanowire) supported on copper foam was prepared as a photocathode for detection of tyrosinase though quinone-chitosan conjugation chemistry method. The in-situ generated quinones that were the catalytic product of tyrosinase acted as electron acceptors, which were captured by the chitosan deposited on the surface of the electrode. Direct immobilization of electron acceptor on the electrode surface improved the photocurrent conversion efficiency and thus sensitivity. The as-prepared biosensor can realize a rapid response in a wide linear range of 0.05 U/mL to 10 U/mL with the detection limit as low as 0.016 U/mL of tyrosinase. The current work provides a new perspective to design and develop highly sensitive and selective PEC biosensor.

关键词： tyrosinase Cu0/Cu20 nanowire photoelectrochemical biosensor chitosan modified electrode quinones

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Phenol content,antioxidant and tyrosinase inhibitory activity of mangrove plants in Micronesia

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Asian Pacific Journal of Tropical Medicine 2014年第7期7卷 531-535页

作者： Sung-Suk Suh Jinik Hwang Mirye Park Heung-Sik Park Taek-Kyun Lee South Sea Environment Research Department.Korea Institute of Ocean Science and Technology Korea South Pacific Ocean Research Center Korea Institute of Ocean Science and Technology

Objective:To find out and compare the in vitro antioxidant and tyrosinase inhibitory activities of two species of mangrove plants.Methods:Mangrove samples were harvested at the shoreline on the island of Weno,Chuuk State in Micronesia.The phenol content,antioxidant activity(based on DPPH-free radical scavenging)and tyrosinase inhibitory activity in different tissues(leaves,barks and roots)of Rhizophora stylosa(R.stylosa)and Sonneratia alba(S.alba),collected from the island of Weno.Results:Total phenol content ranged from 4.87 to 11.96 mg per g of freeze dried samples.The highest antioxidant activity was observed in R.styiosa bark(85.5%).The highest tyrosinase inhibitory activity was found in S.alba bark.Also,total phenol content and antioxidant activity were higher in methanol extracts than in aqueous extracts.Conclusions:Taken together,the results of this study proved that mangroves can be excellent sources of antioxidant compounds.

关键词： Rhizophora stylosa Sonneratia alba Phebolics Antioxidant activity tyrosinase inhibitory activity

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