沈阳地区北虫草野生与市售菌株人工培育子实体的蛋白质组学比较  

Proteomics comparison of cultivated fruiting bodies of wild and commercial strains of Cordyceps militaris in Shenyang

作　　者：齐贺[1,2] 王千慧 刘颖[1] 卜桐 黄竹青[1] 付赵军 张帆 郑剑玲[1] QI He;WANG Qianhui;LIU Ying;BU Tong;HUANG Zhuqing;FU Zhaojun;ZHANG Fan;ZHENG Jianling(Liaoning Institute of Basic Medicine,Shenyang,Liaoning 110101,China;Liaoning University of Traditional Chinese Medicine,Shenyang,Liaoning 110032,China)

机构地区：[1]辽宁省基础医学研究所,辽宁沈阳110101 [2]辽宁中医药大学,辽宁沈阳110032

出　　处：《微生物学通报》2021年第8期2619-2631,共13页Microbiology China

基　　金：辽宁省自然科学基金(20170540486);辽宁省科学技术研究项目(L201901)。

摘　　要：【背景】北虫草作为冬虫夏草的代用品,具有与冬虫夏草类似的药理活性,其富含的蛋白质和氨基酸通常作为衡量真菌营养价值的重要指标,从中分离纯化具有潜在临床应用价值的蛋白质或多肽,已成为一个研究热点。【目的】检测沈阳北虫草野生与市售菌株人工培育子实体的蛋白质组成,分析相同培育条件下获得的蛋白种类、数量及其功能的差异,为深入研究鉴定沈阳地区北虫草药用蛋白和针对性驯化提供了蛋白质组学数据基础。【方法】采集沈阳棋盘山野生北虫草菌株,与市售人工栽培北虫草菌株同期分别经组织分离、液体发酵后培育获得子实体,通过蛋白提取、胰酶酶解后,采用非标定量技术液相色谱-质谱联用方法,对野生和市售来源培育的子实体样本进行定量蛋白组的研究。【结果】共鉴定到9233条特异性肽段和1923个蛋白,其中含有1163个可定量蛋白,野生来源培育子实体有214个蛋白表达发生上调,181个蛋白表达发生下调,对这些差异蛋白进行功能富集分析发现,其主要参与能量生产/转换、氨基酸转运/代谢、抗氧化功能。在相同的营养摄取条件下,野生来源培育菌种在各个能量代谢、氨基酸代谢功能中的相关蛋白表达量高于市售来源培育的菌种。野生来源培育菌种的一种抗氧化重要蛋白(Gene Name:ISF02112)表达量远远高于(Fold Change>9)市售来源培育菌种。同时与抗氧化和代谢功能相关的差异蛋白有22个。【结论】沈阳地区北虫草野生菌株经适当人工培育会保留部分优良的生物学特性,2种来源菌株培育的子实体具有丰富及优异抗氧化功能的蛋白,子实体蛋白的抗氧化能力与其整体代谢能力相关。本研究结果为深入研究鉴定北虫草药用蛋白和针对性驯化提供蛋白质组学数据基础。[Background]As a substitute of Cordyceps sinensis,Cordyceps militaris has similar pharmacological activities with Cordyceps sinensis.It is rich in protein and amino acids,which is usually used as an important index to measure the nutritional value of fungi.It has become a research focus to isolate and purify proteins or peptides with potential clinical application value from Cordyceps militaris.[Objective]The protein composition of fruiting bodies of wild and commercial strains of Cordyceps militaris in Shenyang were detected,and the differences of protein types,quantities and functions under the same cultivation conditions were analyzed,which provided the proteomic data basis for further study and identification of protein for medicinal use and targeted domestication of northern caterpillar in Shenyang area.[Methods]Wild Cordyceps militaris strains were collected from Qipan Mountain of Shenyang.Fruiting bodies of wild and artificially cultivated Cordyceps militaris strains were obtained by tissue isolation and liquid fermentation respectively at the same time.The quantitative proteome of fruit body samples from wild and commercial sources was studied by non-standard quantitative technology liquid chromatography-mass spectrometry after protein extraction and trypsin enzymolysis.[Results]A total of 9233 specific peptide fragments and 1923 proteins were identified,including 1163 quantifiable proteins.214 proteins were up-regulated and 181 proteins were down-regulated in fruit bodies cultured from wild sources.After functional enrichment analysis,these differential proteins were mainly involved in energy production/conversion,amino acid transport/metabolism and antioxidant function.Under the same nutritional conditions,the expression of protein related to energy metabolism and amino acid metabolism of wild-type strains was higher than that of commercial strains,and the expression of an important antioxidant protein(Gene Name:ISF02112)from wild cultivated strains was much higher than that in commercial strains(fold c

关 键 词：北虫草 子实体 蛋白质组学 

分 类 号：S567.3[农业科学—中草药栽培]