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.
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。
