摘要
目的:对猪苓菌核表皮黑色素进行鉴定,为从分子水平研究猪苓菌核发育过程中黑色素的关键作用提供基础。方法:采用碱提酸沉法提取并纯化猪苓菌核表皮黑色素,利用紫外可见光谱和红外光谱鉴定表皮黑色素,并对其溶解性、结构特性、稳定性和抗氧化性进行进一步测定。结果:猪苓菌核表皮色素不溶于水和常见的有机溶剂,在pH3时完全沉淀,溶于碱性溶液;不具有鞣质和黄酮类物质的特性;紫外光谱结果表明,在200~800 nm,吸光值随波长增加呈指数型下降,以波长对吸光值取对数作图,斜率为-0.0024,样品放置一月后,斜率为-0.0036;红外光谱鉴定出—OH、脂肪族—CH、芳香环中C=C和C=N骨架、羧酸外C=O、COO—等官能团的存在;温度升高和pH增大对其具有增色效应,冷冻和光照对其具有减色效应;对羟自由基和超氧阴离子自由基具有显著地清除活性,具有一定的还原能力,1 g猪苓菌核表皮色素的多酚含量相当于(224.48±11.46)mg没食子酸。结论:首次鉴定了猪苓菌核表皮黑色素,对于猪苓的药理药效及发育过程研究提供了新的思路和基础。
引文
[1]许广波,傅伟杰,赵旭奎.我国猪苓研究的进展[J].菌物研究,2003(1):58-61.
[2]陈彩霞,王泽昊,FENG J,等.植物病原真菌的菌核研究进展[J].微生物学通报,DOI:10.13344/j.microbiol,china.180117.
[3]李雯瑞.猪苓优良菌株筛选与菌核生长发育过程的研究[D].陕西咸阳:西北农林科技大学,2013.
[4]徐锦堂,郭顺星,李灵玉,等.猪苓菌核生长发育规律观察[J].中国药学杂志,1991,26(12):714-716.
[5]李雯瑞,梁宗锁,陈德育,等.不同发育阶段猪苓菌核显微结构和成分含量的比较[J].西北林学院学报,2013,28(4):116-121.
[6]樊荣,徐小鸿,曹亚松,等.大丽轮枝菌黑色素合成相关基因与微菌核形成的关系[J].菌物学报,2017(12):1608-1615.
[7]江绍锋,王陈骄子,舒灿伟,等.水稻纹枯病菌黑色素形成的转录组分析[A]//彭友良,李向东.中国植物病理学会2017年学术年会论文集[C].北京:中国农业科学技术出版社,2017:35.
[8]曹志艳,杨胜勇,董金皋.植物病原真菌黑色素与致病性关系的研究进展[J].微生物学通报,2006,33(1):154-158.
[9]姚增玉.山杏种皮黑色素提取及其性质研究[D].陕西咸阳:西北农林科技大学,2007.
[10]戚建华.食品加工剩余物板栗壳的利用:Cu(Ⅱ)的生物吸附和天然色素的提取、分级与表征[D].陕西咸阳:西北农林科技大学,2010.
[11]凌关庭.抗氧化食品与健康[M].北京:化学工业出版社,2004.
[12]DUAN X,JIANG Y,SU X,et al.Antioxidant properties of anthocyanins extracted from litchi(Litchi chinensis,Sonn.)fruit pericarp tissues in relation to their role in the pericarp browning[J].Food Chemistry,2007,101(4):1365-1371.
[13]OYAIZU M.Studies on products of browning reaction.Antioxidative activities of products of browning reaction prepared from glucoseamine[J].Japanese Journal of Nutrition&Dietetics,1986,44(6):307-315.
[14]SPANOS G A,WROLSTAD R E.Influence of processing and storage on the phenolic composition of Thompson Seedless grape juice[J].Journal of Agricultural&Food Chemistry,1990,38(7):1565-1571.
[15]SAUER K.The genomics and proteomics of biofilm formation[J].Genome Biology,2003,4(6):219.
[16]LABBATE M,QUECK S Y,KOH K S,et al.Quorum sensing-controlled biofilm development in Serratia liquefaciens MG1[J].Journal of Bacteriology,2004,186(3):692.
[17]LIN W P,LAI H L,LIU Y L,et al.Effect of melanin produced by a recombinant Escherichia coli on antibacterial activity of antibiotics[J].Journal of Microbiology Immunology&Infection,2005,38(5):320-326.
[18]SHIMA S,MATSUOKA H,IWAMOTO T,et al.Antimicrobial action of epsilon-poly-L-lysine[J].Journal of Antibiotics,1984,37(11):1449.P39
[19]HARKI E,TALOU T,DARGENT R.Purification,characterisation and analysis of melanin extracted from Tuber melanosporum Vitt[J].Food Chemistry,1997,58(1-2):69-73.
[20]BILI?SKA B.On the structure of human hair melanin from an infrared spectroscopy analysis of their interactions with Cu2+,ions[J].Spectrochimica Acta Part A Molecular&Biomolecular Spectroscopy,2001,57(12):2525-2533.
[21]HONG L,SIMON J D.Insight into the Binding of Divalent Cations to Sepia Eumelanin from IR Absorption Spectroscopy[J].Photochemistry&Photobiology,2010,82(5):1265-1269.
[22]ELLIL A H A A.Sclerotial development,melanin production and lipid peroxidation by Sclerotium rolfsii[J].Folia Microbiologica,1999,44(2):181-186.
[23]李斌.黑木耳黑色素对细菌群体感应调控行为的抑制及其抗氧化活性的研究[D].南京:南京农业大学,2011.
[24]YUAN W,BURLEIGH S H,DAWSON J O.Melanin biosynthesis by Frankia strain CeI5[J].Physiologia Planta-(上接P33)rum,2010,131(2):180-190.
[25]KORYTOWSKI W,SARNA T.Bleaching of melanin pigments.Role of copper ions and hydrogen peroxide in autooxidation and photooxidation of synthetic dopa-melanin[J].Journal of Biological Chemistry,1990,265(21):12410-12416.
[26]HENSON J M,BUTLER M J,DAY A W.THE DARKSIDE OF THE MYCELIUM:Melanins of phytopathogenic fungi[J].Annual Review of Phytopathology,1999,37(4):447-471.
[27]BUTLER M,GARDINER R B.Degradation of melanin or inhibition of its synthesis:are these a significant approach as a biological control of phytopathogenic fungi[J].Biological Control,2005,32(2):326-336.
[28]ROSAS A L,CASADEVALL A.Melanization decreases the susceptibility of Cryptococcus neoformans to enzymatic degradation[J].Mycopathologia,2001,151(2):53-56.
[29]BECHINGER C,BASTMEYER M.Optical measurement of invasive forces exerted by appressoria of a plant pathogenic fungus[J].Science,1999,285(5435):1896-1899