印度尼西亚传统发酵牛肉（Cangkuk）中血管紧张素转换酶抑制肽的纯化及其产生的降压作用

Purification of angiotensin converting enzyme inhibitory peptides and antihypertensive effect generated from Indonesian traditional fermented beef (Cangkuk).

作者信息

Mirdhayati Irdha, Zain Wieda Nurwidada Haritsah, Fatah Abdul, Yokoyama Issei, Arihara Keizo

机构信息

Department of Animal Science, Faculty of Agriculture and Animal Science, Universitas Islam Negeri Sultan Syarif Kasim Riau, Pekanbaru, 28293, Indonesia.

Department of Animal Science, School of Veterinary Medicine, Kitasato University, Towada-shi, 034-8628, Japan.

出版信息

Anim Biosci. 2024 Oct;37(10):1799-1808. doi: 10.5713/ab.23.0433. Epub 2024 May 7.

DOI:10.5713/ab.23.0433

PMID:38754854

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11366520/

Abstract

OBJECTIVE

Traditional fermented meat products can be considered a source of bioactive peptides. Cangkuk, a traditional Indonesian fermented beef product is one source of angiotensin converting enzyme (ACE) inhibitory peptides. This study aimed to identify ACE-inhibitory peptides from Cangkuk and analyze their antihypertensive effects.

METHODS

The water-soluble fraction of Cangkuk was fractionated to obtain ACE-inhibitory peptides using an ethanol solvent at several concentrations and solid-phase extraction with an OASIS HLB cartridge followed by purification with reversed-phase high-performance liquid chromatography (RP-HPLC). HPLC-MS was used to identify target peptides, followed by automatic protein sequencer analysis to detect peptide sequences. Antihypertensive effects were analyzed on the water-soluble fraction and synthesized peptides. The animal model comprised 14-16-week-old male spontaneously hypertensive rats (SHRs) (~320 g average body weight) with mean systolic blood pressures (SBPs) higher than 190 mmHg. All oral doses of peptides were 1 mL in volume. Distilled water was used as a control. The antihypertensive activities of the sample and control were observed by measuring the SBP at 0, 2, 4, 6, 8 and 24 h after oral administration.

RESULTS

Two sequences of ACE inhibitory peptides were found, EAPLNPKANR (IC50 value of 44.6 μmol/L) and IVG (IC50 value of 97.3 μmol/L). The water-soluble fraction demonstrated an antihypertensive effect on SHRs after oral administration at 100 mg/kg body weight, maximally lowering the SBP by 14.9 mmHg 8 h after administration. The tripeptide IVG showed the highest reduction of SBP, 24.76±2.1 mmHg 8 h after administration. The decapeptide EAPLNPKANR showed the highest reduction of SBP, 21.0±1.9 mmHg, 8 h after administration. All the samples differed significantly from the control (p<0.01).

CONCLUSION

Cangkuk has potential as a functional food ingredient acting as an antihypertensive agent.

摘要

目的

传统发酵肉制品可被视为生物活性肽的来源。仓谷（Cangkuk），一种印度尼西亚传统发酵牛肉制品，是血管紧张素转换酶（ACE）抑制肽的来源之一。本研究旨在从仓谷中鉴定出ACE抑制肽，并分析其降压效果。

方法

采用不同浓度的乙醇溶剂对仓谷的水溶性部分进行分级分离，以获得ACE抑制肽，然后使用OASIS HLB柱进行固相萃取，接着用反相高效液相色谱（RP-HPLC）进行纯化。用HPLC-MS鉴定目标肽，随后用自动蛋白质测序仪分析以检测肽序列。对水溶性部分和合成肽的降压效果进行分析。动物模型为14 - 16周龄雄性自发性高血压大鼠（SHRs）（平均体重约320 g），平均收缩压（SBP）高于190 mmHg。所有肽的口服剂量体积均为1 mL。蒸馏水用作对照。在口服给药后0、2、4、6、8和24小时测量SBP，观察样品和对照的降压活性。

结果

发现了两种ACE抑制肽序列，EAPLNPKANR（IC50值为44.6 μmol/L）和IVG（IC50值为97.3 μmol/L）。水溶性部分以100 mg/kg体重口服给药后对SHRs显示出降压作用，给药后8小时最大程度降低SBP达14.9 mmHg。三肽IVG在给药后8小时显示出最高的SBP降低值，为24.76±2.1 mmHg。十肽EAPLNPKANR在给药后8小时显示出最高的SBP降低值，为21.0±1.9 mmHg。所有样品与对照均有显著差异（p<0.01）。