Peptides derived from sodium caseinate Hydrolysates produced by Lactobacillus helveticus NCC 2765
Peptides derived from sodium caseinate Hydrolysates produced by Lactobacillus helveticus NCC 2765
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DOI：
10.1021/jf049510t
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发表时间：
2004-11-17
影响因子：
6.1
通讯作者：
Juillerat, MA
中科院分区：
文献类型：
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作者：
Robert, MC;Razaname, A;Juillerat, MA
研究了乳酸菌发酵乳蛋白对血管紧张素转化酶（ACE）的抑制作用。helveticus NCC 2765（雀巢培养物保藏中心，Vers-chez-les-Blanc，瑞士）。水解酪蛋白酸钠1和2小时抑制ACE活性，通过体外ACE抑制试验测定。通过凝胶渗透色谱法分离具有最高ACE抑制潜力的水解产物，并收集其低分子量组分。随后通过反相高压液相色谱法对这些馏分进行亚分级。几个疏水亚组分显示出高的ACE抑制潜力，并使用离子阱质谱仪与电喷雾电离源，其肽的组成进行了测定。低分子量组分的分析鉴定了14种具有已知抗高血压活性的肽和1种具有先前描述的阿片样活性的肽。基于活性亚组分的肽组成，定义了两个潜在活性的新序列，并合成了以下合成肽：FVAPFPEVFG（α（S1）39-48），ENLLRFFVAPFPEVFG（α（S1）33-48），NENLLRFFVAPFPEVFG（alpha（S1）32-48）、LNENLLRFFVAPFPEVFG（alpha（S1）31-48）、NLHLPLPLL（beta 147-155）、ENLHLPLPLL（beta 146-155）和VENLHLPLPLL（beta 145-155）。评估这些合成肽的ACE抑制潜力，并测定IC 50值。NLHLPLPLL（β 147-155）是酪蛋白酸钠水解产物中也存在的唯一合成肽，NENLLRFFVAPFPEVFG（β 32-48）显示出对ACE活性的最高抑制，IC 50值分别为15和55 μ M。此外，使用模拟胃消化的体外模型评估所有合成肽的稳定性。β-酪蛋白衍生肽在胃蛋白酶和胰酶连续水解后保持完整，而α（S1）-酪蛋白衍生肽被胃蛋白酶降解。
Angiotensin-I-converting enzyme (ACE) inhibitory activity was identified in milk proteins fermented with Lactobacillus (Lb.) helveticus NCC 2765 (Nestle Culture Collection, Vers-chez-les-Blanc, Switzerland). Hydrolyzing sodium caseinate for 1 and 2 h inhibited ACE activity, as measured by an in vitro ACE inhibition test. The hydrolysates with the highest ACE inhibitory potential were fractionated by gel permeation chromatography and their low molecular weight fractions collected. These fractions were subsequently subfractionated by reverse-phase high-pressure liquid chromatography. Several hydrophobic subfractions showed high ACE inhibitory potential, and their peptide composition was determined using an ion trap mass spectrometer equipped with an elctrospray ionization source. Analysis of the low molecular weight fraction identified 14 peptides with known anti hypertensive activity and 1 with previously described opioid activity. On the basis of the peptide composition of active subfractions, two potentially active novel sequences were defined, and the following synthetic peptides were synthesized: FVAPFPEVFG (alpha(S1) 39-48), ENLLRFFVAPFPEVFG (alpha(S1) 33-48), NENLLRFFVAPFPEVFG (alpha(S1) 32-48), LNENLLRFFVAPFPEVFG (alpha(S1) 31-48), NLHLPLPLL (beta 147-155), ENLHLPLPLL (beta 146-155), and VENLHLIPLIPLL (beta 145-155). The ACE inhibitory potential of these synthetic peptides was assessed, and IC50 values were determined. NLHLPLPLL (beta 147-155), which was the only synthetic peptide also present in the sodium caseinate hydrolysates, and NENLLRFFVAPFPEVFG (as, 32-48) showed the highest inhibition of ACE activity, with IC50 values of 15 and 55 PM, respectively. Furthermore, the stability of all synthetic peptides was assessed using an in vitro model simulating gastric digestion. The beta-casein-derived peptides remained intact following the successive hydrolysis by pepsin and pancreatin, whereas alpha(S1)-casein-derived peptides were degraded by pepsin.