摘要
以水解度(DH)和多肽得率(TCA-YSP)为指标,筛选碱性蛋白酶、中性蛋白酶、木瓜蛋白酶、菠萝蛋白酶和复合蛋白酶等,采用超滤、Sephadex-G15、Sephadex-G50和半制备液相色谱等对腰果粗寡肽进行分离纯化。通过分析型液相色谱和红外光谱鉴定腰果寡肽的纯度和结构。结果显示:碱性蛋白酶得到的酶解液DH和TCA-YSP为(17.06±1.52)%,(26.28±0.13)%,均明显高于其他蛋白酶的,为最适蛋白酶;经分离纯化得到纯度高的腰果寡肽,其氨基酸序列为Ile-Ile-Ala-Ile-Pro-Ala-GIy-Val-Ala-His(IIAIPAGVAH);肥大细胞脱颗粒程度测定结果表明,200μg/mL的腰果寡肽具有显著抑制肥大细胞组胺和β-氨基己糖甘氨酶(β-HEX)释放及磷脂酰丝氨酸外翻的作用,显示腰果寡肽具有优异的抗过敏效应。
The proteases such as alkaline protease,neutral protease,papain,bromelain,and complex protease were screened using the degree of hydrolysis(DH)and peptide yield(TCA-YSP)as indicators.The crude oligopeptide of cashew nut was isolated by ultrafiltration,Sephadex-G15,Sephadex-G50 and semi-preparative liquid chromatography.The purity and structure of cashew nut oligopeptide were identified through analytical liquid chromatography and infrared spectra.The results showed that DH and TCA-YSP were(17.06±1.52)%and(26.28±0.13)%respectively,which were significantly higher than those of other proteases.Therefore,alkaline protease was the most suitable protease.The sequence of cashew nut oligopeptide was Ile-Ile-Ala-Pro-Ala-Val-Ala-His(IIAIPAGVAH).The result of antiallergic effect of cashew nut oligopeptide showed cashew oligopeptide at 200μg/mL significantly inhibited the release of histamine,the expression ofβ-aminohexose glycinase(β-HEX),and phosphatidylserine eversion in mast cell,indicating that cashew oligopeptide extracted from cashew nut owned outstanding antiallergic effect.
作者
陈笛
姜珂
周鑫
曹献英
CHEN Di;JIANG Ke;ZHOU Xin;CAO Xian-ying(School of Food Science and Technology,Hainan University,Haikou,Hainan 570228,China)
出处
《食品与机械》
北大核心
2020年第4期26-31,55,共7页
Food and Machinery
基金
海南自然科学基金创新研究团队项目(编号:2017CXTD001)。
关键词
腰果
寡肽
酶解
分离纯化
结构鉴定
抗过敏效应
cashew
oligopeptide
enzymolysis
isolation and purification
structural identification
anti-allergic effect
