利用毕赤酵母表达系统生产具有抑制血管紧张素 I 转换酶活性的功能肽。
Production of functional peptides with inhibition ability against angiotensin I-Converting enzyme using P. pastoris expression system.
机构信息
Nugen Bioscience (Taiwan) Co., Ltd, 4F., No.35, Keya Rd., Daya Dist., Taichung City, 428, Taiwan.
Department of Seafood Science, National Kaohsiung University of Science and Technology, No.142 Hai-Chuan Rd. Nan-Tzu, Kaohsiung, 81143, Taiwan.
出版信息
J Food Drug Anal. 2018 Jul;26(3):1097-1104. doi: 10.1016/j.jfda.2018.02.001. Epub 2018 Feb 21.
To obtain the angiotension-I converting enzyme inhibitor (ACEI), a fusion ACEI polypeptide encoded with 8 DNA sequences of GPL, GPM, IKW, IVY, IRPVQ, IWHHT, IYPRY and IAPG, which were selected and designed and cloned into pGAPZαC and then transformed into Pichia pastoris SMD1168H. After 3 days induction, the fraction with highest ACEI activity was expressed and purified using a Ni Sepharose™ 6 Fast Flow. The IC of recombinant ACEI polypeptide was 88.2 μM. A 128-fold increase of ACEI activity (0.69 μM) was obtained after pepsin digestion, which was equivalent to 0.022 μM of captopril. Reverse phase HPLC indicated all the 8 peptides contained in ACEI-hydrolysate after pepsin digestion.
为了获得血管紧张素转化酶抑制剂（ACEI），我们设计并克隆了 8 个 DNA 序列 GPL、GPM、IKW、IVY、IRPVQ、IWHHT、IYPRY 和 IAPG 编码的融合 ACEI 多肽到 pGAPZαC 中，然后转化到毕赤酵母 SMD1168H 中。诱导 3 天后，使用 Ni Sepharose™ 6 Fast Flow 表达和纯化具有最高 ACEI 活性的部分。重组 ACEI 多肽的 IC 为 88.2 μM。胃蛋白酶消化后，ACEI 活性增加了 128 倍（0.69 μM），相当于卡托普利的 0.022 μM。反相 HPLC 表明胃蛋白酶消化后的 ACEI 水解物中含有所有 8 种肽。
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