远东拟沙丁鱼酶解产物的降血压效果评价及其ACE抑制肽的分离纯化与鉴定  

Antihypertensive effect of enzymatic hydrolysate from Sardinops melanosticta muscle on spontaneously hypertensive rats and identification of its angiotensin-converting enzyme(ACE) inhibitory peptide

作　　者：王晶晶[1] 吉薇[1] 吉宏武[1,2,3,4] 苏伟明[1,2,3,4] 李亚会[1] 刘书成[1,2,3,4] 

机构地区：[1]广东海洋大学,食品科技学院,广东湛江524088 [2]广东省水产品加工与安全重点实验室,广东湛江524088 [3]水产品深加工广东普通高校重点实验室,广东湛江524088 [4]广东省海洋食品工程技术研究中心,广东湛江524088

出　　处：《食品与发酵工业》2016年第4期63-68,共6页Food and Fermentation Industries

基　　金：广东省高等院校学科建设项目(编号:2013CXZDA020);广东省产学研合作项目(编号:2013B090600155);广东海洋大学创新强校项目(编号:GDOU2013050314,GDOU2014050203)

摘　　要：以远东拟沙丁鱼肌肉为原料,采用动物蛋白水解酶和风味蛋白酶联合水解制备其酶解产物(简称为SMMH),SMMH通过截留分子量分别为10 k Da和3 k Da超滤膜进行分离,获得3个组分,分别称为A(MW>10k Da)、B(3 k Da<MW<10 k Da)和C(MW<3 k Da)。以体外血管紧张素转移酶(angiotensin-converting enzyme,ACE)抑制率为指标,评价其活性。结果显示:C的ACE抑制率最大,为71.4%,A的ACE抑制率最低,为34.6%。为了进一步评价组分C降血压效果,以原发性高血压大鼠为模型,采用灌胃方式,研究其短期(10 h)与长期(28 d)降血压功效,结果显示:灌胃3种剂量组分C对SHRs大鼠的收缩压有显著降低作用,并且随着剂量的增加,降血压作用越显著。为了确定C组分降血压肽的结构,采用凝胶色谱、离子色谱、反向高效液相色谱相结合对其进行了分离纯化,获得了2个较纯的组分,经电喷雾质谱鉴定,一种为Lys-Val-Glu-Pro-Leu-Pro,另一种为Pro-Ala-Leu,它们的IC50分别为22.9μmol/L和12.2μmol/L。Sardinops melanosticta muscle was hydrolyzed using a combination of animal protein hydrolysis enzyme and flavor enzyme to obtain its hydrolysate abbreviated as SMMH. SMMH was then ultrafiltered with 10 k Da and 3k Da cut-off membranes to yield 3 fractions named as A（ MW 〉10 k Da）,B（ 3 k Da〈 MW〈 10 k Da） and C（ MW〉3 k Da）. Results from measurement of their ACE inhibitory activity suggested that the fraction C exhibited the strongest ACE inhibitory activity in vitro. To further evaluate its antihypertensive effect,a single oral administration（ short term） or continuous administration for 28 d（ long term） of fraction C were carried out. Fraction C supplementation decreased the systolic blood pressure（ SBP） in a dose-dependent manner. With the increase of dose,the systolic blood pressure in SHRs significantly decreased. To identify novel ACE inhibitory peptide,the fraction C was then further separated by gel filtration,ion-exchange chromatography,and reversed phase high-performance liquid chromatography（ RP-HPLC）. Two novel ACE inhibitory peptides were isolated. The amino acid sequences were KVEPLP and PAL with IC50 value of 12. 2 μmol / L and 22. 9 μmol / L,respectively.

关 键 词：远东拟沙丁鱼 酶解产物 血管紧张素转移酶(ACE)抑制肽 原发性高血压大鼠 分离纯化 

分 类 号：TS254.1[轻工技术与工程—水产品加工及贮藏工程] TS201.4[轻工技术与工程—食品科学与工程]