蚕丝蛋白水解液中多肽相对分子质量分布的测定
Determination of molecular weight distribution of peptides in silk protein hydrolyzate
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摘要: 蚕丝经脱胶后用硫酸进行水解处理,然后经过一系列的中和、过滤、沉淀脱色等过程制备得到丝素肽。水解过程采用正交实验优化水解工艺条件。然后利用高效液相色谱法,测定了蚕丝蛋白水解液中多肽组分相对分子质量的分布情况。根据胰岛素、酵母核糖核酸、杆菌肽和还原性谷胱甘肽四种标准品的相对分子质量对数与色谱峰的保留时间的关系,绘制出标准相对分子质量分布的标准曲线。结果表明:蚕丝蛋白在固液比为1∶40,硫酸浓度为30%,水解温度为100℃,水解时间8h的条件下进行水解其丝素肽得率最高。蚕丝蛋白水解液中多肽组分的相对分子质量基本都分布在3500u以下,且用此方法得到的色谱峰的分离效果也很好。该方法简便快速且灵敏度高,结果的准确性好,为丝素肽的研究提供了更有效的方法。Abstract: The degummed silk was hydrolyzed using sulfuric acid and then after a series of decoloration, filtration, sedimentation, etc.to prepare fibroin peptide. Hydrolysis process conditions were optimized by using orthogonal test L 9 ( 34) . Then the molecular weight distribution of the silk protein hydrolyzate was determined by high-performance liquid chromatography. The molecular weight distribution curve was obtained as the relationship between molecular weight of standard and elution time.The results showed that silk fibroin peptide yield was the highest when silk protein was hydrolyzed under the conditions of solid-liquid ratio of 1∶40, the sulfuric acid concentration of 40%, hydrolysis temperature of 80℃, hydrolysis time 8h and the molecular weight distribution less than 3500u.The repetition of result was good. In conclusion, it suggested that it was an efficient way to analysis distribution of polypeptide compounds.It provided a new way in the study of silk peptide.