Design and production of genetically modified soybean protein with anti‐hypertensive activity by incorporating potent analogue of ovokinin(2–7)
Design and production of genetically modified soybean protein with anti‐hypertensive activity by incorporating potent analogue of ovokinin(2–7)
复制标题
通过掺入有效的卵激肽类似物设计和生产具有抗高血压活性的转基因大豆蛋白(2–7)
DOI：
10.1016/s0014-5793(01)02434-6
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发表时间：
2001
期刊：
影响因子：
3.5
通讯作者：
M. Yoshikawa
中科院分区：
文献类型：
--
作者：
N. Matoba;N. Doyama;Yuko Yamada;N. Maruyama;S. Utsumi;M. Yoshikawa
有效的抗高血压肽，RPLKPW，是根据卵激肽的结构设计的（2-7）。将编码该肽的序列插入大豆β-伴大豆球蛋白α′亚基基因的3个同源位点。天然α′亚基以及修饰的、含有RPLKPW的α′亚基在大肠杆菌中表达，从可溶性级分中回收，然后通过离子交换色谱纯化。重组蛋白经胰蛋白酶和糜蛋白酶体外消化后，释放出RPLKPW肽段。此外，与天然α′亚基不同，以10 mg/kg的剂量经口给予未消化的含RPLKPW的α′亚基在自发性高血压大鼠中发挥了抗高血压作用。这些结果首次提供了通过定点诱变引入食物蛋白质中的生理活性肽即使在低剂量下也可以在体内实际发挥作用的证据。
The potent anti-hypertensive peptide, RPLKPW, has been designed based on the structure of ovokinin(2–7). The sequence encoding this peptide was introduced into three homologous sites in the gene for soybean β-conglycinin α′ subunit. The native α′ subunit as well as the modified, RPLKPW-containing α′ subunit were expressed in Escherichia coli, recovered from the soluble fraction and then purified by ion-exchange chromatography. The RPLKPW peptide was released from recombinant RPLKPW-containing α′ subunit after in vitro digestion by trypsin and chymotrypsin. Moreover, the undigested RPLKPW-containing α′ subunit given orally at a dose of 10 mg/kg exerted an anti-hypertensive effect in spontaneously hypertensive rats, unlike the native α′ subunit. These results provide evidence for the first time that a physiologically active peptide introduced into a food protein by site-directed mutagenesis could practically function in vivo even at a low dose.