摘要
为了解决传统肼解法操作过程中碳末端氨基酸难以分离的问题,基于传统肼解法提出一种改进的方法测定碳末端氨基酸。采用水合肼肼解不同水解度的酪蛋白酶解液,分别利用庚醛和苯甲醛去除氨基酸酰肼,通过薄层层析法、高效液相色谱法测定2种方法获得的碳末端氨基酸并进行t检验。结果表明:采用庚醛和苯甲醛去除氨基酸酰肼得到的碳末端氨基酸溶液中均含有赖氨酸、鸟氨酸,氨基酸含量随着水解度的增大而增加,而且不同方法得到的赖氨酸、鸟氨酸含量无显著性差异(p>0.05),验证了改进肼解法的可行性,确定胰蛋白酶对酪蛋白中精氨酸、赖氨酸具有定向酶解作用。
To solve the problem that the carbon-terminal amino acids were difficult to separate during the operation of the traditional hydrazine hydrolysis method,an improved method for the determination of carbon-terminal amino acids was proposed based on the traditional hydrazine hydrolysis method.Hydrazine hydrazide hydrate was used to hydrolyze casein digests with different degrees of hydrolysis,and amino acid hydrazide was removed by using heptanal and benzaldehyde,respectively,and carbon-terminal amino acids obtained by both methods were determined by thin-layer chromatography and high-performance liquid chromatography with a t-test.The results show that the carbon-terminal amino acid solutions obtained by using heptanal and benzaldehyde to remove amino acid hydrazide contain lysine and ornithine,and the amino acid content increases with the increase of hydrolysis degree,and there is no significant difference(p>0.05)in the content of lysine and ornithine obtained by the different methods,verifying the feasibility of the improved hydrazide hydrolysis method,and determining the directional enzyme enzymatic digestion of trypsin on arginines and lysines in tyrosine protein.
作者
韩文玉
李凯
魏旭
李宝瑞
王元秀
HAN Wenyu;LI Kai;WEI Xu;LI Baorui;WANG Yuanxiu(School of Biological Science and Technology,University of Jinan,Jinan 250022,Shandong,China;Shandong Fupai Ejiao Co.,Ltd.,Jinan 250400,Shandong,China;Shandong Academy of Agricultural Sciences,Jinan 250100,Shandong,China)
出处
《济南大学学报(自然科学版)》
北大核心
2025年第3期420-425,共6页
Journal of University of Jinan(Science and Technology)
基金
山东省重点研发计划项目(2022CXGC010602)。
关键词
肼解法
酪蛋白
碳末端氨基酸
胰蛋白酶
定向酶解
hydrazolysis
casein
carbon-terminal amino acid
trypsin
directed enzymatic hydrolysis
作者简介
第一作者:韩文玉(1997—),女,山东德州人。硕士研究生,研究方向为生物活性物质制备。E-mail:1769094755@qq.com。;通信作者:王元秀(1966—),女,山东济南人。教授,硕士,硕士生导师,研究方向为生物活性物质制备。E-mail:chm_wangyx@ujn.edu.cn。;通信作者:李凯(1974—),男,山东聊城人。高级工程师,博士,研究方向为大健康产品开发、天然活性产物的开发与应用、生物分析化学。E-mail:kevinli2011@qq.com。
