干酪乳杆菌LC-15发酵乳血管紧张素转换酶体外抑制活性的研究

血管紧张素转换酶抑制剂筛选模型是一个用于抗高血压药物和功能性食品研究的有效模型。通过考察乳酸菌蛋白质水解能力与ACE抑制活性的关系，以一株具有较高ACE抑制活性的干酪乳杆菌LC-15为研究对象，研究了发酵时间、添加酪蛋白、酶水解处理和模拟胃肠消化对LC-15发酵乳的ACE抑制活性的影响。结果表明，乳酸菌对牛乳蛋白质的水解能力与ACE抑制活性呈现一定的正相关关系；干酪乳杆菌LC-15在复原脱脂乳中发酵至42h时，发酵乳的ACE抑制活性达到最高，此时ACE抑制率为66％；通过在复原脱脂乳中添加4％酪蛋白可以使LC-15发酵乳的ACE抑制活性提高到81％；控制复原脱脂乳水解度为10％，然后再利用LC-15进行发酵得到的发酵乳的ACE抑制活性达到85％；在模拟胃肠环境消化条件下LC-15发酵乳的ACE抑制活力从最初的85％下降到78％。     

具有ACE抑制活性乳酸菌菌株的筛选与鉴定

以ACE抑制活性和蛋白水解活性为检测指标,选择138株乳酸菌为出发菌株,筛选出具有强ACE抑制活性的乳酸菌菌株.结果表明,筛选出具有强ACE抑制活性的4株乳酸菌,其中3株菌为瑞士乳杆菌,1株菌为干酪乳杆菌.瑞士乳杆菌KLDS1.0485和干酪乳杆菌KLDS1.0486比例为1:1,制成的发酵乳ACE抑制活性可达到61.55%.因此,组合瑞士乳杆菌KLDS1.0485和干酪乳杆菌KLDS1.0486可作为制备乳源ACE抑制肽的优良菌株.     

乳酸菌发酵乳血管紧张素转化酶抑制活力的比较研究

以18种共计61株乳酸菌为研究对象,比较了乳酸菌发酵脱脂乳的血管紧张素转化酶抑制活力(ACE抑制活力),并探讨了ACE抑制活力与pH值、滴定酸度、蛋白水解度的关系。结果表明,乳杆菌ACE抑制活力最强,双歧杆菌次之,链球菌、乳球菌、明串珠菌和片球菌ACE抑制活力很低。ACE抑制活力最高的是Lacto- bacillus helveticus 6024,发酵28 h后ACEI%可达到80%以上,其OPA指数可达到0.800以上,而球菌的ACE抑制活力和蛋白水解能力都很低,甚至检测不到。乳酸菌产酸能力做为一个单一因素与ACE抑制率没有明显相关性(Spearman相关系数r_3=0.526),乳酸菌的蛋白水解能力与ACE抑制率具有显著的正相关性(Spearman相关系数r_3=0.938)。通过对2株产ACE抑制活力高的L.helveticus 6024和L.helveticus T16的研究发现,其共同特点是产酸能力和蛋白水解能力都很强。     

产ACE抑制肽乳酸菌的筛选、益生特性评定及应用

目的:筛选出富产血管紧张素转化酶(Angiotensin converting enzyme,ACE)抑制肽的乳酸菌并评定其益生特性.方法:依据蛋白水解度及ACE抑制率对乳酸菌进行初筛,然后依据模拟胃肠消化后的ACE抑制率最终筛出2株乳酸菌ZJUIDS09和ZJUIDS11,进一步评价其耐酸、耐胆盐、抗生素耐药性和抑菌...     

微生物发酵法制备大豆ACE抑制肽菌种的筛选

以大豆分离蛋白为底物,以血管紧张素转化酶(ACE)抑制率和肽含量为检测指标,选择5株乳酸菌、3株霉菌和1株枯草芽孢杆菌为出发菌种,通过微生物发酵筛选出具有高ACE抑制活性的发酵菌株。结果表明:乳酸菌中具有强ACE抑制活性的菌株为保加利亚乳杆菌,ACE抑制率和肽含量分别为57.93%和3.27mg/mL;霉菌中具有强ACE抑制活性的菌株为米曲霉,ACE抑制率和肽含量分别为41.23%和2.17mg/mL;枯草芽孢杆菌ACE抑制率和肽含量分别为45.02%和2.25mg/mL。综合比较9种菌株的发酵特性,选用保加利亚乳杆菌作为制备大豆抗高血压活性肽优良菌株。     

酵母菌与乳酸菌发酵马乳产ACE抑制肽

通过使用从传统酸马乳中分离纯化和筛选出的乳酸菌与酵母菌共发酵马乳,获得具有血管紧张素转换酶(angiotensin-converting enzyme,ACE)抑制活性的生物活性肽.经过发酵性能和生长性能测试后筛选到发酵马乳ACE抑制率为38.67％的乳酸乳球菌(Lactococcus lactis)L8和ACE抑制率...     

产ACE抑制肽菌株的筛选及其在模拟消化道环境中的活性分析

为了筛选出对血管紧张素转换酶（angiotensin-converting enzyme,ACE）抑制活性较高并且具有较强的耐酸耐胆盐和耐盐的乳酸菌,对实验室保存的27株乳酸菌进行了产酸能力、蛋白水解活力、ACE抑制活性、模拟胃肠道消化及酸、胆盐和盐耐受性的测定。结果显示,菌株M3的ACE抑制率可达71.94%±1.39%,具有较好的蛋白水解活力和产酸能力,蛋白水解活力为（86.66±3.51）μg/mL亮氨酸,滴定酸度为（71.67±2.86）°T。菌株M3经人工胃肠液分别处理3 h后,ACE抑制率为74.96%±1.73%;在pH3的环境中3 h,耐受性为14.34%±1.21%,活菌数能达到7 lg CFU mL?1以上;在含有0.3%胆盐的培养基中3 h,耐受性为37.50%±2.47%;在含有4% NaCl的培养基中24 h,耐受性为37.32%±1.84%。该菌经16S rDNA鉴定为Lactobacillus (Lb.) paracasei subsp. paracasei M3。因此,菌株Lb. paracasei subsp. paracasei M3可用作发酵牛乳富产ACE抑制肽且能耐受消化道环境具有益生菌潜力的菌株。     

杏仁蛋白水解物对血管紧张素转化酶抑制作用的研究

分别采用Protamex、Alcalase、Neutrase、Flavourzyme、Proleather FG-F、木瓜蛋白酶水解杏仁蛋白,利用高效液相色谱法测定水解物对血管紧张素转化酶(ACE)的抑制活性,以水解度(DH)和水解产物对ACE的抑制率为指标对酶解过程进行分析,并研究水解物的体外消化稳定性。结果表明,Proleather FG-F 和Alcalase 对杏仁蛋白有较好的水解效果,其水解物对ACE 抑制率较高,IC50 分别为1.24mg/ml 和0.98mg/ml。模拟胃肠消化实验结果表明,在消化酶的作用下杏仁蛋白水解物仍具有较强的ACE 抑制活性。     

具有ACE抑制活性乳酸菌的特性研究

选择了3株Lactobacillus helveticus(H—1，H—2，LH)和Lactobacillus casei(C，LC)为实验对象，分别制作发酵乳，并测定其蛋白水解活性、生长特性及ACE抑制活性(ACEI)。结果表明，发酵乳中脯氨酸及含有脯氨酸的多肽越多，其ACE抑制活性越强；而且具有较大滴定酸度的菌种很可能具有较高的ACEI活性。经筛选，H—1的A—CEI活性最高。该菌经30℃培养18h后具有最大约ACE抑制活性。     

益生菌发酵藜麦制备ACE抑制肽

目的:探究利用益生菌发酵藜麦制备血管紧张素转换酶(ACE)抑制发酵液潜在的能力及发酵工艺对其的影响。方法:以藜麦为原料,ACE抑制率为指标,对18株乳酸菌和12株酵母制备具有ACE抑制能力的发酵液进行了筛选,并对接种量、发酵温度、发酵时间进行了正交优化;对发酵液中的肽进行液质鉴定,并对筛选出的肽进行体内试验评价降压效果。结果:30株菌中Lactobacillus paracasei L2(副干酪乳杆菌)作用效果最好,发酵藜麦制备的发酵液ACE抑制率高达(86.50±0.25)%;Lactobacillus paracasei L2菌株发酵藜麦制备具有ACE抑制活性的发酵液最佳发酵条件为接种量5%,发酵时间48 h,发酵温度34℃,该条件下ACE抑制率最高,为(90.41±0.16)%;通过对液质鉴定结果比对数据库得到两条ACE抑制效果较好的两条肽NIFRPFAPEL和AALEAPRILNL,它们均能够在自发性高血压大鼠(SHR)体内显现出显著的降血压作用,均在灌胃4 h时达到最佳降压效果。收缩压(SBP)分别降低(2.67±0.21),(3.46±0.01) kPa,舒张压(DBP)分...     

Initial proteolysis of milk proteins and its effect on formation of ACE-inhibitory peptides during gastrointestinal proteolysis: a bioinformatic, in silico, approach

Angiotensin-I-converting enzyme (ACE)-inhibitory peptides are encrypted in milk protein sequences. They may express an antihypertensive effect if they are released by proteolysis in foods and/or during gastrointestinal digestion. A bioinformatic, in silico, approach was developed to evaluate how systematic initial proteolysis, i.e. cleavage after one specific type of amino acid (C-end) at a time in milk proteins, influence the formation of ACE-inhibitory peptides by subsequent gastrointestinal proteolysis. Computer simulation was done and a peptide QSAR model was used to estimate the combined ACE inhibition by digested proteins. Initial proteolytic cleavage at the C-end of amino acids isoleucine and proline gave, based on calculations, increased effect of ACE-inhibitory peptides after gastrointestinal proteolysis of milk proteins. Cleavage after most other amino acid residues had little or no effect. Results indicate that initial proteolysis in foods have to be specific in order to increase formation of bioavailable ACE-inhibitory peptides during gastrointestinal digestion.     

Screening of bacteria from the cattle gastrointestinal tract for inhibitory activity against enterohemorrhagic Escherichia coli O157:H7, O111:H-, and O26:H11

A quick and reproducible microgel plate assay was adapted to screen bacteria from cattle gastrointestinal tracts for production of compounds inhibitory to the growth of three enterohemorrhagic Escherichia coli (EHEC) serotypes: O157:H7, O111:H-, and O26:H11. The inhibitory activity of 309 bacteria, isolated on several agar media, was assessed by a microgel assay performed in 96-well microtiter plates. Fifty-three isolates secreted inhibitory compounds with a molecular weight of less than 1,000. In 12 isolates, the inhibitory activity was attributable to compounds other than lactic or acetic acid. These compounds were highly heat tolerant, with varying sensitivity to digestion by proteolytic enzymes. The inhibitory isolates were identified as lactic acid-producing bacteria on the basis of a combination of analyses, including 16S-rDNA restriction fragment length polymorphisms, 16S-rDNA gene sequences, and fermentation end products. The lactic acid bacteria of ruminants may contain antibacterial compounds not yet described. Naturally occurring populations of lactic acid bacteria may have potential as probiotics, to reduce the carriage of EHEC in the gastrointestinal tract of ruminants.     

Activity and bioavailability of food protein-derived angiotensin-I-converting enzyme–inhibitory peptides

Angiotensin-I-converting enzyme (ACE) inhibitory peptides are able to inhibit the activity of ACE, which is the key enzymatic factor mediating systemic hypertension. ACE-inhibitory peptides can be obtained from edible proteins and have the function of antihypertension. The amino acid sequences and the secondary structures of ACE-inhibitory peptides determine the inhibitory activities and stability. The resistance of ACE-inhibitory peptides to digestive enzymes and peptidase affect their antihypertensive bioactivity in vivo. In this paper, the mechanism of ACE-inhibition, sources of the inhibitory peptides, structure–activity relationships, stability during digestion, absorption and transportation of ACE-inhibitory peptides, and consumption of ACE-inhibitory peptides are reviewed, which provide guidance to the development of new functional foods and production of antihypertensive nutraceuticals and pharmaceuticals.     

Effect of simulated gastrointestinal digestion on the antihypertensive properties of synthetic beta-lactoglobulin peptide sequences

In this study, the antihypertensive activity in spontaneously hypertensive rats of two peptides isolated from beta-lactoglobulin hydrolysates with thermolysin was evaluated. These peptides, with sequences LLF [beta-lg f(103-105)] and LQKW [beta-lg f(58-61)], showed potent in vitro ACE-inhibitory activity. Two hours after administration, both sequences caused a clear and significant decrease in the blood pressure of these rats. The impact of a simulated gastrointestinal digestion on ACE-inhibitory and antihypertensive activities of these peptides was also studied. The results showed that both fragments were susceptible to proteolytic degradation after incubation with pepsin and Corolase PP. In addition, their in vitro ACE-inhibitory activity decreased after the simulated digestion. It is likely that fragment LQK was the active end product of the gastrointestinal digestion of peptide LQKW. The fragment LL, observed after digestion of peptide LLF, probably exert its antihypertensive effect through a mechanism of action different than ACE-inhibition.     

Angiotensin I-Converting Enzyme Inhibitory Activity of Soy Protein Digests in a Dynamic Model System Simulating the Upper Gastrointestinal Tract

ABSTRACT: The objective of this study was to investigate whether peptides with inhibitory activity against angiotensin I-converting enzyme (ACE) would be produced by digestion of isolated soy protein (ISP) in a dynamic model system simulating gastrointestinal conditions. Using the model system, 5% ISP solution was pumped into the stomach reactor containing pepsin and HCl. The peptic digest was continuously pumped into the duodenum reactor containing pancreatin and Oxgall bile. The effects of blanching (100°C, 10 min) followed by pasteurization (75°C, 15 s) or sterilization (121°C, 20 min) of ISP before digestion on the inhibitory activity were also investigated. During the first 30 min of digestion, significantly higher ( P < 0.05) ACE-inhibitory activity was generated from unheated ISP after sequential digestion in both reactors compared with after peptic digestion only in the stomach reactor. However, at 90 min, subsequent digestion by pancreatin of unheated and blanched-sterilized ISP decreased ACE-inhibitory activity compared with peptic digestion alone. The IC₅₀ values at the end of 90 min digestion in both reactors were 0.38 ±0.01, 0.37 ± 0.02, and 0.44 ± 0.02 mg/mL for unheated, blanched-pasteurized, and blanched-sterilized ISP, respectively. The results suggest the potential production of peptides with ACE-inhibitory activity upon physiological digestion of soy protein, including products that have been subjected to heat processing. Although clinical trials would be required to provide final evidence of efficacy of the soy peptides, the present findings support the application of soy protein as an ingredient for functional foods.     

Angiotensin converting enzyme-inhibitory activity of peptides isolated from Manchego cheese. Stability under simulated gastrointestinal digestion

In this study, several peptides, which had previously been identified in active HPLC fractions from Manchego cheese, were synthesised and their angiotensin converting enzyme (ACE)-inhibitory activities were measured. From 11 peptides, which were selected based on their structures, only two, VRYL and KKYNVPQL, showed considerable ACE-inhibitory activity with IC₅₀ values of 24.1 and 77.1 μ , respectively. Subsequently, the impact of the gastrointestinal digestion on ACE-inhibitory activity was evaluated. Some of the peptides selected were resistant to the incubation with pepsin followed by hydrolysis with a pancreatic extract. The ACE-inhibitory activity after simulated digestion did not change drastically except for peptide _s2-CN f(195-204) (TQPKTNAIPY) that exhibited an activity 6 times greater after simulated digestion. In contrast, after simulated digestion, the activities of peptides VRYL and KKYNVPQL decreased. The peptides not hydrolysed by gastrointestinal enzymes and peptide VRYL, which was only partly hydrolysed, were incubated with ACE and were found to be true inhibitors of the enzyme and to have a competitive inhibition pattern.