摘要
利用液体培养基对冬虫夏草菌丝体进行培养 ,在 18～ 2 0℃ 160r min摇瓶中培养 72h ,可得到 2 66g L的湿菌体 ,并对天然冬虫夏草和冬虫夏草菌丝体的蛋白粗提液进行了蛋白含量 ,SDS PAGE和HPLC分析。结果表明天然冬虫夏草与冬虫夏草菌丝体中水溶性蛋白含量分别占其干重的 2 .0 0 %和 7.2 4% ;在 12 %SDS PAGE中低分子蛋白标准的回归方程为Y =5.0 1847 1.11588X ,其相关系数r = 0 .9913 1,由此推算出天然冬虫夏草与冬虫夏草菌丝体含有 4种相同的蛋白质亚基 ,其相对分子质量分别为 46971,510 87,86681和 95685;天然冬虫夏草中特有的蛋白质亚基的相对分子质量为 1714 2及小于 12 3 72的多肽 ,其余的蛋白质亚基稍有差异 ,在天然冬虫夏草中蛋白质亚基的相对分子质量为 2 8942 ,3 72 3 7和 580 91,而在冬虫夏草菌丝体中为 2 8658,3 4 92 0和 550 18。水溶性蛋白经HPLC分析 ,可以分离出不同的蛋白峰 。
Cultured at 18～20℃,160 r/min for about 72 hours in shakers using liquid media, 266g∕L wet weight Cordyceps mycelia was obtain d. Analysised proteins from natural Cordyceps sinensis and cultured Cordyceps my celia by Lowry's methed, the result sh'owed the hydrosoluble proteins in them were 2.00% and 7.24% re'spectively. 12%SDS PAGE indicated the linear regression formula of low molecular weight marker is Y=5.01847 1.11588X. The correlation coefficient is r= 0.99131. The relative molecular mass of the protein subunits could be calculated by it. They were 95685,86681,58097,51087,46971,37231,28942,17142 and 12372 from natural Cordyceps sinensis and 95685,86681,55018,51087,46971,34920 and 28658 from cultured Cordyceps mycelia. In HPLC, TSK GEL G2000SW(7.5 mm×300 mm )was used. The moble phase consisted of 20 mmol/L pH 8.0 Tris HCl with a flow rate of 1.0ml/min . HPLC expressed the proteins from cultured Cordyceps mycelia were simple and ea sily separative to those from natured Cordyceps sinensis.out 72hoursinshakersusingliquidme
出处
《药物生物技术》
CAS
CSCD
2003年第5期304-307,共4页
Pharmaceutical Biotechnology
