Production and transepithelial transportation of angiotensin-I-converting enzyme (ACE)-inhibitory peptides from whey protein hydrolyzed by immobilized Lactobacillus helveticus proteinase
Production and transepithelial transportation of angiotensin-I-converting enzyme (ACE)-inhibitory peptides from whey protein hydrolyzed by immobilized Lactobacillus helveticus proteinase
复制标题
利用固定化瑞士乳杆菌蛋白酶水解的乳清蛋白生产和跨上皮运输血管紧张素-I 转换酶 (ACE) 抑制肽。
DOI：
10.3168/jds.2018-14899
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发表时间：
2019-02-01
影响因子：
3.5
通讯作者：
Pan, Daodong
中科院分区：
文献类型：
--
作者：
Guo, Yuxing;Jiang, Xiaoxiao;Pan, Daodong
用海藻酸钠固定化helveticus乳杆菌LB 10蛋白酶水解乳清蛋白，制备血管紧张素i转换酶（ACE）抑制肽。所生成的水解产物进行了ace抑制活性和跨Caco-2细胞单层运输能力的测试。采用响应面法确定了7.55 mg/mL的蛋白酶、2.03 g/100 mL的海藻酸钠、0.39%的戊二醛为最佳固定条件。与游离蛋白酶相比，固定化蛋白酶具有显著的pH、热稳定性和储存稳定性以及可重复使用性。在人肠道吸收模型中，乳清蛋白水解物通过凝胶过滤层析分离，ace抑制肽混合物通过Caco-2细胞单层运输。采用超高效液相色谱-串联质谱法在cco -2细胞基底外侧检测二肽和三肽KA、EN、DIS、EVD、LF、AIV和VFK（半数最大抑制浓度（平均±标准差）分别为1.24±0.01、1.43±0.04、1.59±0.27、1.32±0.05、1.60±0.39、2.66±0.02和1.76±0.09 mmol/L）。这些结果表明，乳清蛋白水解物通过Caco-2细胞膜运输后，acace抑制肽存在于Caco-2细胞模型的基底外侧。
Lactobacillus helveticus LB 10 proteinases immobilized with sodium alginate were used to hydrolyze whey protein to produce angiotensin-I-converting enzyme (ACE)-inhibitory peptides. The generated hydrolysates were tested for ACE-inhibitory activity and for their ability to be transported across Caco-2 cell monolayers. Using a response surface method, we determined that a proteinase concentration of 7.55 mg/mL, sodium alginate concentration of 2.03 g/100 mL, and glutaraldehyde concentration of 0.39% were found to be the optimal immobilization conditions. Compared with free proteinase, the immobilized proteinase had significantly higher pH, thermal and storage stability, and reusability. Whey protein hydrolysates were fractionated by gel filtration chromatography and ACE-inhibitory peptide mixtures were transported across Caco-2 cell monolayers in a human intestinal-absorption model. The di- and tripeptides KA, EN, DIS, EVD, LF, AIV, and VFK (half-maximal inhibitory concentrations (mean ± standard deviation) of 1.24 ± 0.01, 1.43 ± 0.04, 1.59 ± 0.27, 1.32 ± 0.05, 1.60 ± 0.39, 2.66 ± 0.02, and 1.76 ± 0.09 mmol/L, respectively) were detected on the basolateral side of the Caco-2 cell monolayer using ultra-performance liquid chromatography-tandem mass spectrometry. These results highlight that ACE-inhibitory peptides are present on the basolateral side of the Caco-2 cell model after transportation of whey protein hydrolysate across the Caco-2 cell membrane.