Fonsecaea monophora黑素的理化性质及其合成途径的分析研究

中国真菌学杂志　 2017, Vol. 12 　Issue (3): 129-134.

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Fonsecaea monophora黑素的理化性质及其合成途径的分析研究

陈映丹¹, 陆春¹, 李美荣¹, 马寒¹, 尹颂超¹, 郑智心¹, 李海仪¹, 席丽艳², 冯佩英¹

1. 广州市中山大学附属第三医院皮肤科, 广州 510630;
2. 广州市中山大学孙逸仙纪念医院皮肤科, 广州 510120

收稿日期:2017-02-13 出版日期:2017-06-28 发布日期:2017-06-28
通讯作者: 陆春,E-mail:luliyuan@tom.com;冯佩英,E-mail:fengpeiying@medmail.com.cn E-mail:luliyuan@tom.com;fengpeiying@medmail.com.cn
作者简介:陈映丹,女(汉族),硕士研究生在读.E-mail:chenyingdan89@163.com
基金资助:
国家自然科学基金（81401650）

Detection of physiochemical characterization and synthesis pathway of melanin from Fonsecaea monophora

CHEN Ying-dan¹, LU Chun¹, LI Mei-rong¹, MA Han¹, YIN Song-chao¹, ZHENG Zhi-xin¹, LI Hai-yi¹, XI Li-yan², FENG Pei-ying¹

1. Department of Dermatology, Third Affiliated Hospital, Sun Yat-sen University, Guangzhou 510630, China;
2. Department of Dermatology, Sun Yat-sen memorial Hospital of Sun Yat-sen University, Guangzhou 510120, China

Received:2017-02-13 Online:2017-06-28 Published:2017-06-28

摘要/Abstract

摘要：

目的探讨Fonsecaea monophora黑素的理化性质及其合成途径。方法通过化学分析、紫外光谱、红外光谱和电子顺磁共振波谱等明确F.monophora黑素的理化性质；通过比对F.monophora菌株在基础培养基（马铃薯葡萄糖琼脂（PDA）培养基、L-DOPA PDA培养基）和含黑素抑制剂培养基（DOPA黑素抑制剂培养基、DHN黑素抑制剂培养基）的菌落生长情况，采用BioTek酶标仪EON定量分析其黑素合成，以明确F.monophora的黑素合成途径。结果 F.monophora黑素与合成L-DOPA黑素的理化性质相似；菌株在含L-DOPA培养基较PDA培养基产生更多的黑素，且在含DOPA黑素抑制剂叠氮化钠及DHN黑素抑制剂苯肽、三环唑培养基中其黑素合成均明显降低。结论 F.monophora黑素主要为L-DOPA黑素，可能共同存在DOPA黑素和DHN黑素合成途径。

关键词: Fonsecaea monophora, 黑素, 理化性质, 合成途径

Abstract:

Objective To study the melanin physiochemical characterization and melanin synthesis pathway of Fonsecaea monophora.Methods Pure melanin mass was extracted from the F.monophora isolate using the cell wall crude extract method.Then we used Ultravolet (Uv),Fourier transformed infrared (FT-IR) and electron paramagnetic resonance (EPR) spectra assay to evaluate the physiochemical characterization of melanin.Furthermore,we observed the pigment production of the colonies on different mediums (PDA,PDA with L-DOPA,PDA with DOPA-melanin inhibitors and PDA with DHN-melanin inhibitors),and the quantity of melanin produced on different above media were measured using BioTek Eon microplate reader.Results The melanin extracted from F.monophora shared similar physiochemical and spectroscopic properties with the synthetic L-DOPA melanin.The formation of melanin pigment statistically significant increased on the L-DOPA medium,and decreased on the DOPA-melanin inhibitor medium (sodium azide) and DHN-melanin inhibitors medium (phthalide and tricyclazole) compare to their blank contrast medium.Conclusions The melanin produced by F.monophora is mainly DOPA melanin,and it may synthesize simultaneously by DOPA-melanin and DHN-melanin pathway.

Key words: Fonsecaea monophora, melanin, physiochemical characterization, melanin synthesis pathway

中图分类号:

R379.9

陈映丹, 陆春, 李美荣, 马寒, 尹颂超, 郑智心, 李海仪, 席丽艳, 冯佩英. Fonsecaea monophora黑素的理化性质及其合成途径的分析研究[J]. 中国真菌学杂志, 2017, 12(3): 129-134.

CHEN Ying-dan, LU Chun, LI Mei-rong, MA Han, YIN Song-chao, ZHENG Zhi-xin, LI Hai-yi, XI Li-yan, FENG Pei-ying. Detection of physiochemical characterization and synthesis pathway of melanin from Fonsecaea monophora[J]. Chinese Journal of Mycology, 2017, 12(3): 129-134.

参考文献

[1] Queiroz-Telles F,de Hoog S,Santos D W,et al.Chromoblastomycosis[J].Clin Microbiol Rev,2017,30(1):233-276.
[2] Najafzadeh M J,Vicente V A,Sun J,et al.Fonsecaea multimorphosa sp.nov,a new species of Chaetothyriales isolated from a feline cerebral abscess[J].Fungal Biol,2011,115(10):1066-1076.
[3] Xi L,Sun J,Lu C,et al.Molecular diversity of Fonsecaea (Chaetothyriales) causing chromoblastomycosis in southern China[J].Med Mycol,2009,47(1):27-33.
[4] Fransisca C,He Y,Chen Z,et al.Molecular identification of chromoblastomycosis clinical isolates in Guangdong[J].Med Mycol,2017.
[5] Franzen A J,de Souza W,Farina M,et al.Morphometric and densitometric study of the biogenesis of electron-dense granules in Fonsecaea pedrosoi[J].FEMS Microbiol Lett,1999,173(2):395-402.
[6] Franzen AJ,Cunha MM,Miranda K,et al.Ultrastructural characterization of melanosomes of the human pathogenic fungus Fonsecaea pedrosoi[J].J Struct Biol,2008,162(1):75-84.
[7] Alviano DS,Franzen AJ,Travassos LR,et al.Melanin from Fonsecaea pedrosoi induces production of human antifungal antibodies and enhances the antimicrobial efficacy of phagocytes[J].Infection and Immunity,2004,72(1):229-237.
[8] Cunha MM,Franzen AJ,Seabra SH,et al.Melanin in Fonsecaea pedrosoi:a trap for oxidative radicals[J].BMC Microbiol,2010,10(1):80.
[9] Pal AK,Gajjar DU,Vasavada AR.DOPA and DHN pathway orchestrate melanin synthesis in Aspergillus species[J].Med Mycol,2014,52(1):10-18.
[10] Xi L,Lu C,Sun J,et al.Chromoblastomycosis caused by a meristematic mutant of Fonsecaea monophora[J].Med Mycol,2009,47(1):77-80.
[11] Teixeira PA,De Castro RA,Ferreira FR,et al.L-DOPA accessibility in culture medium increases melanin expression and virulence of Sporothrix schenckii yeast cells[J].Med Mycol,2010,48(5):687-695.
[12] Kumar CG,Mongolla P,Pombala S,et al.Physicochemical characterization and antioxidant activity of melanin from a novel strain of Aspergillus bridgeri ICTF-201[J].Lett Appl Microbiol,2011,53(3):350-358.
[13] Sajjan S,Kulkarni G,Yaligara V,et al.Purification and physiochemical characterization of melanin pigment from Klebsiella sp.GSK[J].J Microbiol Biotechnol,2010,20(11):1513-1520.
[14] Liu D,Wei L,Guo T,et al.Detection of DOPA-melanin in the dimorphic fungal pathogen Penicillium marneffei and its effect on macrophage phagocytosis in vitro[J].PLoS One,2014,9(3):e92610.
[15] 陈映丹,陆春,冯佩英.黑素与真菌抗药性的研究进展[J].中国真菌学杂志,2016,11(4):248-251.
[16] de A S R,Angluster J,de Souza W,et al.Carbohydrate and lipid components of hyphae and conidia of human pathogen Fonsecaea pedrosoi[J].Mycopathologia,1995,132(2):71-77.
[17] Corbellini VA,Scroferneker ML,Carissimi M,et al.Lipolytic activity of chromoblastomycosis agents measured by infrared spectroscopy and chemometric methods[J].Med Mycol,2009,47(1):63-69.
[18] Gessler NN,Egorova AS,Belozerskaia TA.Melanin pigments of fungi under extreme environmental conditions (review)[J].Prikl Biokhim Mikrobiol,2014,50(2):125-134.
[19] Li XQ,Guo BL,Cai WY,et al.The role of melanin pathways in extremotolerance and virulence of Fonsecaea revealed by de novo assembly transcriptomics using illumina paired-end sequencing[J].Stud Mycol,2016,83:1-18.
[20] Langfelder K,Streibel M,Jahn B,et al.Biosynthesis of fungal melanins and their importance for human pathogenic fungi[J].Fungal Genet Biol,2003,38(2):143-158.
[21] Franzen AJ,Cunha MM,Batista EJ,et al.Effects of tricyclazole (5-methyl-1,2,4-triazol benzothiazole),a specific DHN-melanin inhibitor,on the morphology of Fonsecaea pedrosoi conidia and sclerotic cells[J].Microsc Res Tech,2006,69(9):729-737.
[22] Revankar SG,Sutton DA.Melanized fungi in human disease[J].Clin Microbiol Rev,2010,23(4):884-928.
[23] Schmaler-Ripcke J,Sugareva V,Gebhardt P,et al.Production of pyomelanin,a second type of melanin,via the tyrosine degradation pathway in Aspergillus fumigatus[J].Appl Environ Microbiol,2009,75(2):493-503.
[24] Alviano D S,Franzen A J,Travassos L R,et al.Melanin from Fonsecaea pedrosoi induces production of human antifungal antibodies and enhances the antimicrobial efficacy of phagocytes[J].Infect Immun,2004,72(1):229-237.
[25] Almeida-Paes R,Frases S,Araujo G D S,et al.Biosynthesis and functions of a melanoid pigment produced by species of the Sporothrix complex in the presence of L-tyrosine[J].Appl Environ Microbiol,2012,78(24):8623-8630.
[26] Frases S,Salazar A,Dadachova E,et al.Cryptococcus neoformans can utilize the bacterial melanin precursor homogentisic acid for fungal melanogenesis[J].Appl Environ Microbiol,2007,73(2):615-621.
[27] Romero-Martinez R,Wheeler M,Guerrero-Plata A,et al.Biosynthesis and functions of melanin in Sporothrix schenckii[J].Infect Immun,2000,68(6):3696-3703.
[28] van de Sande WW,de Kat J,Coppens J,et al.Melanin biosynthesis in Madurella mycetomatis and its effect on susceptibility to itraconazole and ketoconazole[J].Microbes Infect,2007,9(9):1114-1123.

Fonsecaea monophora黑素的理化性质及其合成途径的分析研究

Detection of physiochemical characterization and synthesis pathway of melanin from Fonsecaea monophora

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