金钱菇中降血压肽的纯化和磁性脂质体分离鉴定  

Purification of anti-hypertensive peptide and separation identification of magnetic liposome from Lentinus edodes

作　　者：陆苑[1] 陆玉婷[1] 杨思敏 冯学珍[1] 伍善广[1] LU Yuan;LU Yuting;YANG Simin;FENG Xuezhen;WU Shanguang(Medical School,Guangxi University of Science and Technology,Liuzhou 545005,China)

机构地区：[1]广西科技大学医学部,广西柳州545005

出　　处：《广西科技大学学报》2023年第3期116-122,138,共8页Journal of Guangxi University of Science and Technology

基　　金：国家自然科学基金项目(21766003);广西高校中青年教师科研基础能力提升项目(2021KY0342);广西自然科学基金项目(2019GXNSFBA245098)资助。

摘　　要：为了从金钱菇中快速分离出降血压肽,利用超声细胞破碎法提取金钱菇蛋白,采用透析、超滤的方法对提取物进行纯化,以固定化金属亲和磁性脂质体(metal affinity-immobilied magnetic liposome,MA-IML)作为吸附介质,以金钱菇提取物为吸附对象,吸附分离其中的降血压肽,并解析其结构,利用分子对接分析其结合血管紧张素I转化酶(angiotensin-I converting enzyme,ACE)的能力。结果表明:对金钱菇提取液用透析、超滤的方法进行纯化后,蛋白含量由37.67%提高至51.18%,多糖含量由26.80%降低至6.90%,ACE抑制率由61.83%提高至79.77%,纯化后更有利于分离降血压肽;用MA-IML粒子吸附分离出金钱菇纯化液中的降血压活性组分,经反相高效液相色谱法分析其为单一组分,该组分为五肽,其氨基酸序列为苯丙氨酸-丙氨酸-色氨酸-丙氨酸-酪氨酸(Phe-Ala-Trp-Ala-Tyr,FAWAY),IC50为179μmol/L。分子对接模拟表明,FAWAY能与ACE自发发生结合并形成稳定的构象。该研究制备的MA-IML可作为从金钱菇中分离降血压肽的有效工具。To separate anti-hypertensive peptide from Lentinus edodes,the Lentinus edodes proteins were extracted by using an ultrasonic cell disrupter and purified by dialysis and ultra-filtration.The metal affinity-immobilized magnetic liposome(MA-IML)particles were prepared and used as absorption medium to separate anti-hypertensive peptide from Lentinus edodes.The amino acid sequence of the separated peptide was analyzed and verified.The angiotensin-I converting enzyme(ACE)binding energy of the peptides was also explored by molecular docking.The results showed that after purification by dialysis and ultra-filtration,the protein content of Lentinus edodes extracts increased from 37.67%to 51.18%,the polysaccharide content decreased from 26.80%to 6.90%,and the ACE inhibition rate increased from 61.83%to 79.77%,which helped to separate anti-hypertensive peptides.The anti-hypertensive component was separated by MA-IML adsorption and identified as a single peptide by reverse phase high-performance liquid chromatography.The amino acid sequence was identified to be Phe-Ala-Trp-Ala-Tyr(FAWAY).The IC50 of FAWAY was 179μmol/L.Molecular docking results showed FAWAY could spontaneously bind to ACE,forming a stable conformation.The prepared MA-IML can be used as an effective tool for separating anti-hypertensive peptide from Lentinus edodes.

关 键 词：磁性脂质体 金钱菇 降血压肽 血管紧张素I转化酶(ACE) 分离鉴定 分子对接 

分 类 号：R284.2[医药卫生—中药学]