草菇子实体多肽的分离纯化及其结构分析  

Purification and structural identification of polypeptides from fruiting bodies of Volvariella volvacea

作　　者：张芳艺 张瑜萍 罗小芳 刘培培 林晓彤 邓优锦[2] 江玉姬[1,2,3] 陈炳智 ZHANG Fangyi;ZHANG Yuping;LUO Xiaofang;LIU Peipei;LIN Xiaotong;DENG Youjin;JIANG Yuji;CHEN Bingzhi(College of Food Science,Fujian Agriculture and Forestry University,Fuzhou 350002,Fujian,China;Mycological Research Center,Fujian Agriculture and Forestry University,Fuzhou 350002,Fujian,China;Key Laboratory of Processing of Subtropical Characteristic Fruits,Vegetables and Edible Fungi(Co-construction by Ministry and Province),Ministry of Agriculture and Rural Affairs,Fuzhou 350002,Fujian,China)

机构地区：[1]福建农林大学食品科学学院,福建福州350002 [2]福建农林大学菌物研究中心,福建福州350002 [3]农业农村部亚热带特色果蔬菌加工重点实验室(部省共建),福建福州350002

出　　处：《菌物学报》2023年第8期1775-1784,共10页Mycosystema

基　　金：国家自然科学基金(31801920);福建省自然科学基金(2020J01557);福建农林大学杰出青年科研人才计划项目(xjq202113);国家食用菌产业技术体系(CARS-20)。

摘　　要：采用碱性蛋白酶酶解提取草菇子实体多肽,透析法对其进行分离纯化得到3−10、1–3及1 kDa以下3种多肽组分,比较3种组分的体外抗氧化活性。用液相色谱-质谱联用(LC-MS/MS)、紫外吸收光谱、傅里叶红外光谱和氨基酸的测定方法表征纯化后多肽结构。结果表明,1–3 kDa组分的抗氧化活性最强,其1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基清除率达到99.81%,铁离子自由基还原能力为1.47,2,2′-联氮-双-(3-乙基苯并噻唑啉-6-磺酸)[2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid),ABTS+]自由基和羟自由基的清除能力分别达到99.15%、99.34%。此分子量的草菇抗氧化肽的总蛋白含量为71.12%,灰分含量为12.75%,水分含量为8.44%,其他为7.69%。经鉴定表明草菇子实体多肽是一种优质蛋白质。The polypeptides from fruiting bodies of Volvariella volvacea were extracted by alkaline protease enzymatic hydrolysis,and purified by dialysis method to yield three different components(3 to 10,1 to 3,and less than 1 kDa).The antioxidant activities in vitro of the three components were compared.The structure of purified polypeptide was characterized by LC-MS/MS,ultraviolet absorption spectroscopy,Fourier transform infrared spectroscopy,and amino acid determination methods.The result showed that the antioxidant activity of 1 to 3 kDa was strongest.The 1,1-diphenyl-2-picrylhydrazyl(DPPH)free radical clearance rate was 99.81%,the ferric ion reducing ability was 1.47,and the clearance rates of 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)(ABTS+)radical and hydroxyl radical were 99.15%and 99.34%,respectively.The total protein content of the antioxidant peptide of V.volvacea at this molecular weight was 71.12%,ash content was 12.75%,moisture content was 8.44%and the others were 7.69%.The results of structural identification showed that the polypeptides from fruiting bodies of V.volvacea were high quality protein materials.

关 键 词：草菇子实体多肽 分离纯化 透析 抗氧化活性 氨基酸 

分 类 号：TS219[轻工技术与工程—粮食、油脂及植物蛋白工程]