Rapid purification and characterization of angiotensin I-converting enzyme from porcine lung

IF 1.4 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Protein expression and purification Pub Date : 2025-07-17 DOI:10.1016/j.pep.2025.106780

Meng Li , Pan-Pan Xiao , Wan-Yu Li , Ling-Jing Zhang , Yu-Lei Chen , Tengchuan Jin , Min-Jie Cao

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引用次数: 0

Abstract

Angiotensin I-converting enzyme (ACE) is a key target for screening hypertension medicines. However, traditional method for ACE preparation is time-consuming, and commercial ACE is expensive. In this study, a simple and effective method for ACE purification was proposed. ACE was purified to homogeneity from porcine lungs using acid precipitation, ammonium sulfate fractionation, and chromatography on a HiTrap Q HP column. 2D-PAGE showed that the molecular weight of ACE was 180 kDa, and the pI was 5.7, its sequence was verified by LC-MS/MS. The thermal denaturation temperature of ACE was 58.8 ± 0.4 °C and it is a glycoprotein as confirmed by PAS staining. Circular dichroism and endogenous fluorescence spectroscopy showed that the addition of zinc ions led to changes in the structure of ACE, thereby inhibiting its activity. Compared with commercial ACE, the enzyme prepared in the present study exhibited higher purity and 1.3-fold higher specific activity. As ACE is widely required for studies on anti-hypertensive functional foods, effective preparation of high-purity ACE will be valuable for the investigation of potential functional foods and especially its inhibitory peptides.

查看原文本刊更多论文

猪肺血管紧张素i转化酶的快速纯化及特性研究。

血管紧张素i转换酶（ACE）是筛选高血压药物的关键靶点。然而，传统的ACE制备方法耗时长，且商业化的ACE价格昂贵。本研究提出了一种简单有效的ACE纯化方法。采用酸沉淀、硫酸铵分馏、HiTrap Q HP色谱柱纯化猪肺中ACE至均匀性。2D-PAGE显示ACE分子量为180 kDa， pI为5.7，其序列经LC-MS/MS验证。ACE热变性温度为58.8±0.4℃，经PAS染色证实为糖蛋白。圆二色性和内源荧光光谱显示，锌离子的加入导致ACE结构发生变化，从而抑制其活性。与商用ACE相比，本研究制备的酶纯度更高，比活性提高1.3倍。由于ACE在抗高血压功能食品的研究中被广泛需要，因此高效制备高纯度ACE对潜在功能食品尤其是其抑制肽的研究具有重要意义。

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来源期刊

Protein expression and purification 生物-生化研究方法

CiteScore

3.70

自引率

6.20%

发文量

120

审稿时长

32 days

期刊介绍： Protein Expression and Purification is an international journal providing a forum for the dissemination of new information on protein expression, extraction, purification, characterization, and/or applications using conventional biochemical and/or modern molecular biological approaches and methods, which are of broad interest to the field. The journal does not typically publish repetitive examples of protein expression and purification involving standard, well-established, methods. However, exceptions might include studies on important and/or difficult to express and/or purify proteins and/or studies that include extensive protein characterization, which provide new, previously unpublished information.