泾阳茯砖茶中“金花菌”的分离、鉴定及抗氧化酶活性
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摘要
目的分离鉴定泾阳茯砖茶中的金花菌,探讨其发酵产物的抗氧化酶活性。方法采用涂布平板法分离纯化泾阳茯砖茶中的"金花菌",通过其ITS序列分析进行分子鉴定。利用抗氧化酶试剂盒测定不同时期"金花菌"发酵液的抗氧化酶活性。结果从茶叶中分离出1株"金花菌"J17,通过ITS序列分析初步确定其为赤散囊菌(Eurotium rubrum)。在J17菌株发酵后第1、7、10、14和21天,其发酵液中过氧化物酶(POD)活性(U/mL)分别为0.22±0.11、1.22±0.13、2.46±1.64、7.35±2.77和45.19±1.54;过氧化氢酶(CAT)活性(U/mL)分别为0.06±0.00、0.45±0.00、1.52±0.20、2.35±0.20和2.75±0.37;总超氧化物歧化酶(T-SOD)活性(U/mL)分别为25.55±10.12、50.90±3.72、76.77±6.75、89.07±3.33和36.34±12.32。发酵14d左右,3种抗氧化酶活性均较高,与茶砖在发花车间的发花时间一致。茯砖茶沸水冲泡液和沸水冲洗液中POD和CAT活性差异无统计学意义,而沸水冲泡液的T-SOD活性显著高于沸水冲洗液。结论泾阳茯砖茶的抗氧化酶活性与茶砖发花期产生的赤散囊菌有关,泾阳茯砖茶适用于热水冲泡或煮沸后饮用。
Objective To isolate and identify Jinhuafungi from Jingyang Fuling(poria)tile tea and determine the activity of antioxidant enzyme in its fermentation products.Methods A strain of Jinhuafungi was isolated and purified from Jingyang Fuling tile tea by using spread-plate method and identified using ITS sequence analysis.The activities of antioxidant enzymes at different stages of fermentations were determined using antioxidant enzyme kits.Results The isolated Jinhua fungi was named J17 and identified as Eurotium rubrum.At the 1st,7th,10 th,14th and 21 st days of fermentation,the activities of peroxidase(POD)were0.22±0.11,1.22±0.13,2.46±1.64,7.35±2.77 and 45.19±1.54U/mL respectively;those of catalase(CAT)were 0.06±0.00,0.45±0.00,1.52±0.20,2.35±0.20 and 2.75±0.37 U/mL respectively;those of total superoxide dismutase(T-SOD)were 25.55±10.12,50.90±3.72,76.77±6.75,89.07±3.33 and 36.34±12.32U/mL respectively.After fermented for 14 days,the activities of the three antioxidant enzymes in the fermented broths maintained at high levels,corresponding to the time of fungal-fermentation process during the Fuling tile tea processing.The activities of POD and CAT in boiled water brewed tea and in boiled water washed tea were not significantly different,but the activity of T-SOD in brewed tea was significant higher than in washed tea.Conclusion The predominant fungus in Jingyang Fuling tile tea is Eurotium rubrum,the fermented broth of which has antioxidant activities.From antioxidant perspectives,the better way to make Jingyang Fuling tile tea is brewing it with boiled water.
Objective To isolate and identify Jinhuafungi from Jingyang Fuling(poria)tile tea and determine the activity of antioxidant enzyme in its fermentation products.Methods A strain of Jinhuafungi was isolated and purified from Jingyang Fuling tile tea by using spread-plate method and identified using ITS sequence analysis.The activities of antioxidant enzymes at different stages of fermentations were determined using antioxidant enzyme kits.Results The isolated Jinhua fungi was named J17 and identified as Eurotium rubrum.At the 1st,7th,10 th,14th and 21 st days of fermentation,the activities of peroxidase(POD)were0.22±0.11,1.22±0.13,2.46±1.64,7.35±2.77 and 45.19±1.54U/mL respectively;those of catalase(CAT)were 0.06±0.00,0.45±0.00,1.52±0.20,2.35±0.20 and 2.75±0.37 U/mL respectively;those of total superoxide dismutase(T-SOD)were 25.55±10.12,50.90±3.72,76.77±6.75,89.07±3.33 and 36.34±12.32U/mL respectively.After fermented for 14 days,the activities of the three antioxidant enzymes in the fermented broths maintained at high levels,corresponding to the time of fungal-fermentation process during the Fuling tile tea processing.The activities of POD and CAT in boiled water brewed tea and in boiled water washed tea were not significantly different,but the activity of T-SOD in brewed tea was significant higher than in washed tea.Conclusion The predominant fungus in Jingyang Fuling tile tea is Eurotium rubrum,the fermented broth of which has antioxidant activities.From antioxidant perspectives,the better way to make Jingyang Fuling tile tea is brewing it with boiled water.
引文
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[13] HUANG Yan,SHI Rui,SU Erzheng.Progress in the research and application of the Eurotium cristatum[J].Chin J Bioprocess Eng,2017,15(1):49-56.(in Chinese)黄彦,石瑞,苏二正.冠突散囊菌的研究与应用进展[J].生物加工过程,2017,15(1):49-56.
[14] SONG Jiale,ZHOU Yanyuan,ZHAO Chaochao,et al.Effect of Liubao Tea total flavonoid on antioxidant function in aged mice[J].Food Res Dev,2016,37(20):173-176.(in Chinese)宋家乐,周燕园,赵超超,等.六堡茶总黄酮对老龄小鼠抗氧化功能的影响[J].食品研究与开发,2016,37(20):173-176.
[15] Fan W,Chen Y,Sun J,et al.Effects of tea polyphenol on quality and shelf life of pork sausages[J].J Food Sci Technol,2014,51(16):191-195.
[16] Giroux HJ,GrandpréGD,Fustier P,et al.Production and characterization of Cheddar-type cheese enriched with green tea extract[J].Dairy Sci Technol,2013,93(3):241-254.
[17] Chen Z,Lin Z.Tea and human health:Biomedical functions of tea active components and current issues[J].J Zhejiang UnivSci B,2015,16(2):87-102.
[18] YANG Fulin,DENG Fangming,ZHAO Lingyan,et al.Development of Black Tea microbiology[J].J Microbiol,2006,26(1):81-84.(in Chinese)杨抚林,邓放明,赵玲艳,等.黑茶微生物学研究进展[J].微生物学杂,2006,26(1):81-84.
[2] ZHAO Longfei,XU Yajun,ZHOU Hongjie.Study on Aspergillium niger′s growth property during Pile-fermentation process[J].Food Res Dev,2007,28(10):1-3.(in Chinese)赵龙飞,徐亚军,周红杰.黑曲霉在普洱茶发酵过程中生长特性的研究[J].食品研究与开发,2007,28(10):1-3.
[3]邵元元,肖建中,李飞鸣,等.桑叶茯砖茶“金花”菌的分离鉴定[J].中国蚕业,2017,38(3):11-14.
[4] PENG Xiaoyun,LIANG Faliang,LI Dongli,et al.Secondary metabolites of Eurotium cristatumfrom Fu Brick Tea and their biological activities[J].Chin Tradit Herb Drugs,2013,44(14):1881-1886.(in Chinese)彭晓赟,梁法亮,李冬利,等.茯砖茶中冠突散囊菌的次级代谢产物及其生物活性研究[J].中草药,2013,44(14):1881-1886.
[5] DING Ting,LYU Jiali.Biological characteristics analysis of Jinhua Fungi from Fuzhuan Brick Tea in Shaanxi[J].Food Ind,2012(1):104-106.(in Chinese)丁婷,吕嘉枥.陕西茯砖茶中“金花”菌的生物学特性分析[J].食品工业,2012(1):104-106.
[6] HUANG Hao,ZHENG Hongfa,ZHAO Xi,et al.Identification and aflatoxin production of′Golden Flora′Fungi isolated from Fu Tea produced from different kinds of tea[J].Food Sci,2017,38(8):49-55.(in Chinese)黄浩,郑红发,赵熙,等.不同茶类发花茯茶中“金花”菌的分离、鉴定及产黄曲霉毒素分析[J].食品科学,2017,38(8):49-55.
[7] HU Zhiyuan,LIU Suchun,ZHAO Yunlin,et al.Dynamic microbial changes of Fuzhuan Brick Tea during processing and identification of dominant Fungi[J].Food Sci,2012,33(19):244-248.(in Chinese)胡志远,刘素纯,赵运林,等.茯砖茶生产过程中微生物动态变化及优势菌鉴定[J].食品科学,2012,33(19):244-248.
[8] DING Jinxiang,HUANG Yun,LI Yong,et al.Study on extraction technology of antioxidant bio-active components from Fuzhuan Tea[J].Food Res Dev,2013,34(16):12-14.(in Chinese)丁金祥,黄昀,李勇,等.茯砖茶抗氧化活性物质的提取工艺研究[J].食品研究与开发,2013,34(16):12-14.
[9] OU Huisuan,DENG Xuming,ZHANG Lingzhi,et al.Isolation and identification of Jinhua fungus in Liupao tea[J].J South Agric,2017,48(4):658-662.(in Chinese)欧惠算,邓旭铭,张灵枝,等.六堡茶中金花菌的分离与鉴定[J].南方农业学报,2017,48(4):658-662.
[10]刘作易,秦京.“金花”菌与茯砖茶品质[J].贵州农学院学报,1991,10(1):79-82.
[11] LIU Shiquan,ZHAO Yunlin,LEI Cunxi,et al.A Preliminary research on separation and identification of the Fungi form XIANG-YI Fuzhuan Tea[J].Hubei Agric Sci,2011,50(9):1765-1769.(in Chinese)刘石泉,赵云林,雷存喜,等.“湘益”牌茯砖茶真菌的分离及鉴定研究[J].湖北农业科学,2011,50(9):1765-1769.
[12]王志刚,童哲,程苏云.茯砖茶中散囊菌的种类组成和鉴定[J].茶叶科学,1991,11(2):157-161.
[13] HUANG Yan,SHI Rui,SU Erzheng.Progress in the research and application of the Eurotium cristatum[J].Chin J Bioprocess Eng,2017,15(1):49-56.(in Chinese)黄彦,石瑞,苏二正.冠突散囊菌的研究与应用进展[J].生物加工过程,2017,15(1):49-56.
[14] SONG Jiale,ZHOU Yanyuan,ZHAO Chaochao,et al.Effect of Liubao Tea total flavonoid on antioxidant function in aged mice[J].Food Res Dev,2016,37(20):173-176.(in Chinese)宋家乐,周燕园,赵超超,等.六堡茶总黄酮对老龄小鼠抗氧化功能的影响[J].食品研究与开发,2016,37(20):173-176.
[15] Fan W,Chen Y,Sun J,et al.Effects of tea polyphenol on quality and shelf life of pork sausages[J].J Food Sci Technol,2014,51(16):191-195.
[16] Giroux HJ,GrandpréGD,Fustier P,et al.Production and characterization of Cheddar-type cheese enriched with green tea extract[J].Dairy Sci Technol,2013,93(3):241-254.
[17] Chen Z,Lin Z.Tea and human health:Biomedical functions of tea active components and current issues[J].J Zhejiang UnivSci B,2015,16(2):87-102.
[18] YANG Fulin,DENG Fangming,ZHAO Lingyan,et al.Development of Black Tea microbiology[J].J Microbiol,2006,26(1):81-84.(in Chinese)杨抚林,邓放明,赵玲艳,等.黑茶微生物学研究进展[J].微生物学杂,2006,26(1):81-84.