Angiotensin I-Converting Enzyme (ACE) Inhibitory Activity and ACE Inhibitory Peptides of Salmon (Salmo salar) Protein Hydrolysates Obtained by Human and Porcine Gastrointestinal Enzymes (original) (raw)
Related papers
2014
Abstract: The objectives of the present study were two-fold: first, to detect whether salmon protein fractions possess angiotensin I-converting enzyme (ACE) inhibitory properties and whether salmon proteins can release ACE inhibitory peptides during a sequential in vitro hydrolysis (with commercial porcine enzymes) and ex vivo digestion (with human gastrointestinal enzymes). Secondly, to evaluate the ACE inhibitory activity of generated hydrolysates. A two-step ex vivo and in vitro model digestion was performed to simulate the human digestion process. Salmon proteins were degraded more efficiently by porcine enzymes than by human gastrointestinal juices and sarcoplasmic proteins were digested/hydrolyzed more easily than myofibrillar proteins. The ex vivo digested myofibrillar and sarcoplasmic duodenal samples showed IC50 values (concentration required to decrease the ACE activity by 50%) of 1.06 and 2.16 mg/mL, respectively. The in vitro hydrolyzed myofibrillar and sarcoplasmic samp...
Malaysian Applied Biology
Blood cockle (Anadara granosa) is the most abundant and available bivalves in Malaysia. Blood cockles meat has high protein content and has potential to generate bioactive peptides. To date, no study has been reported on purification and identification of angiotensin I converting enzyme (ACE) inhibitory peptides from blood cockle meat. Thus, the objectives of this study were to purify and characterize ACE inhibitory peptide from blood cockle meat hydrolysate. ACE inhibitory peptides from blood cockle meat hydrolysate (CMH) were prepared by enzymatic protein hydrolysis using Protamex®. Crude CMH was characterized for its stability against gastrointestinal proteases, at varying pH (2–11) and temperature (4–90°C). Next, crude CMH was purified by ultrafiltration, ion exchange chromatography and reverse-phase chromatography and its amino acid sequence was identified. It was found that crude CMH was highly stable at low pH and temperature, and was resistant to gastrointestinal proteases (...
Journal of the Science of Food and Agriculture, 2005
In vitro gastrointestinal digestion of pea and whey protein produced high angiotensin I converting enzyme (ACE) inhibitory activity with IC 50 values of 0.070 and 0.041 mg protein ml −1 respectively. Ultrafiltration/centrifugation using a membrane with a molecular weight cut-off of 3000 Da decreased the IC 50 value to 0.055 mg protein ml −1 for pea permeate and 0.014 mg protein ml −1 for whey permeate. Further fractionation by reverse phase HPLC gave IC 50 values as low as 0.016 mg protein ml −1 for pea and 0.003 mg protein ml −1 for whey. Consequently, these purification steps enriched the ACE inhibitory activity of the pea digest more than four times and that of the whey digest more than 13 times. HPLC profiles after digestion and ultrafiltration indicate that high ACE inhibitory activity is due to short and more hydrophobic peptides. The results also suggest that potent ACE inhibitory peptides were present alongside low active peptides in whey hydrolysate, while all peptides had more or less the same ACE inhibitory activity in pea hydrolysate. In addition, the hydrolysates and enriched fractions will resist in vivo gastrointestinal digestion after oral administration. Hence these ACE inhibitory peptides, as part of functional foods, can play significant roles in the prevention and treatment of hypertension.
Angiotensin I Converting Enzyme Inhibitory Peptides from Fish By-products
CRC Press eBooks, 2013
Three peptides which inhibit angiotensin I-converting enzyme were isolated from maitake (Gnfola frondosa) water extract digested by pepsin. Among them, Lys-Tyr-Thr-Phe-Ala-Val-Thr-Thr-Val-Lys-Thr-Trp-Val had the strongest angiotensin I-converting enzyme inhibitory activity with an IC50 value of 2.6 uM, and the others Gly-Pro-Ser-Gly-Pro-Ser-Gly, and Tyr-Pro-Ser also showed inhibitory activity with IC50 values of 2160 and 570 uM, respectively!
Journal of Agricultural and Food Chemistry, 2010
Pacific hake fish protein hydrolysate (FPH) with promising chemical assay based antioxidative capacity was studied for in vitro angiotensin-I-converting enzyme (ACE)-inhibitory potential, intestinal cell permeability characteristics, and intracellular antioxidative potential using the Caco-2 cell model system. FPH showed substrate-type inhibition of ACE with IC 50 of 161 μg of peptides/mL. HPLC analysis revealed that different peptides were responsible for antioxidative and ACE-inhibitory activity. FPH inhibited 2,2 0 -azobis(2-amidinopropane) dihydrochloride-induced oxidation in Caco-2 cells at noncytotoxic concentrations. In vitro simulated gastrointestinal digestion increased (P < 0.05) antioxidative capacity; ACE-inhibitory activity of FPH remained unchanged, although individual peptide fractions showed decreased or no activity after digestion. Some FPH peptides passed through Caco-2 cells: the permeates showed 2,2 0 -azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activity but no ACE-inhibitory activity. These results suggest the potential for application of Pacific hake FPH to reduce oxidative processes in vivo. Further studies are needed to assess prospective antihypertensive effects.
Current Pharmaceutical Design, 2009
The existence of endogenous bioactive protein or peptide with angiotensin-converting enzyme (ACE) inhibitory activity in snakehead fish fillet is promising to be investigated. The purposes of this research were to extract ACE inhibitory endogenous protein or peptide from snakehead fish fillet and to fractionate the active compounds using ultrafiltration. The extraction employed two solvents, i.e. aquadest and 50% ethanol. Fractionation was conducted using ultrafiltration membranes of 10,000; 5,000 and 3,000 Molecular W eight Cut Off (MW CO) to separate the protein or peptide into the sizes of >10 kDa, 5-10 kDa, 3-5 kDa and <3 kDa. The parameters observed were protein and peptide content, ACE inhibitory activity (in vitro) and also protein and peptide profiles. The result revealed that the snakehead fish fillet contained ACE inhibitory endogenous bioactive protein or peptide. The 50% ethanol was more effective in extracting peptide of <10 kDa than the aquadest. Yet, the aquadest was better in extracting higher molecular weight protein of >10 kDa than the 50% ethanol. The fraction of <3 kDa by aquadest had the highest ACE inhibitor activity per g protein (7.85% inhibition of ACE per g protein). Thus, the fraction of <3 kDa aquadest is the most promising option for further research and development of natural anti-hypertension compound. From the result, snakehead fish fillet was potential to be utilized as a functional food as well as functional ingredient to fight hypertension.
European Food Research and Technology, 2010
In this study, smooth hound protein hydrolysates (SHPHs), obtained by treatment with various gastrointestinal proteases, were analyzed for their angiotensin I-converting enzyme (ACE) inhibitory activities. Protein hydrolysates were obtained by treatment with crude alkaline enzyme extract, low molecular weight (LMW) alkaline protease, trypsin-like protease and pepsin from Mustelus mustelus, and bovine trypsin. All hydrolysates exhibited inhibitory activity toward ACE. Hydrolysate generated with alkaline protease extract displayed the highest ACE inhibitory activity, and the higher inhibition activity (82.6% at 2 mg/mL) was obtained with a hydrolysis degree of 18.8%. This hydrolysate was then fractionated by size exclusion chromatography on a Sephadex G-25 into five major fractions (P1–P5). ACE inhibitory activities of all fractions were assayed, and P3 was found to display a high ACE inhibitory activity (62.24% at 1 mg/mL). P3 was then fractionated by reversed-phase high-performance liquid chromatography (RP-HPLC) and ten fractions of ACE inhibitors were found (F1–F10). Sub-fraction F3 showed the strongest ACE inhibitory activity, being able to suppress more than 60% of initial enzyme activity at a concentration of 100 μg/mL. The amino acid sequence of peptide F3 was determined by ESI/MS and ESI–MS/MS as Ala-Gly-Ser, and the IC50 value for ACE inhibitory activity was 0.13 ± 0.03 mg/mL. Further, purified peptide F3 maintained inhibitory activity even after in vitro digestion with gastrointestinal proteases in order to demonstrate gastrointestinal stability digestion to enable oral application. These results indicate that smooth hound protein hydrolysate possesses potent antihypertensive activity.
Biochimie, 2004
Angiotensin I converting enzyme (ACE) inhibitory peptides can induce antihypertensive effects after oral administration. By means of an ACE inhibitory peptide database, containing about 500 reported sequences and their IC 50 values, the different proteins in pea and whey were quantitatively evaluated as precursors for ACE inhibitory peptides. This analysis was combined with experimental data from the evolution in ACE inhibitory activity and protein degradation during in vitro gastrointestinal digestion. Pea proteins produced similar in silico scores and were degraded early in the in vitro digestion. High ACE inhibitory activity was observed after the simulated stomach phase and augmented slightly in the simulated small intestine phase. The major whey protein b-lactoglobulin obtained the highest in silico scores, which corresponded with the fact that degradation of this protein in vitro only occurred from the simulated small intestine phase on and resulted in a 10-fold increase in the ACE inhibitory activity. Whey protein obtained total in silico scores of about 124 ml/mg, compared to 46 ml/mg for pea protein, indicating that whey protein would be a richer source of ACE inhibitory peptides than pea protein. Although b-lactoglobulin is only partially digested, a higher ACE inhibitory activity was indeed found in the whey (IC 50 = 0.048 mg/ml) compared to the pea digest (IC 50 = 0.076 mg/ml). In silico gastrointestinal digestion of the highest scoring proteins in pea and whey, vicilin and albumin PA2, and b-lactoglobulin, respectively, directly released a number of potent ACE inhibitory peptides. Several other ACE inhibitory sequences resisted in silico digestion by gastrointestinal proteases. Briefly, the quantitative in silico analysis will facilitate the study of precursor proteins on a large scale and the specific release of bioactive peptides.
Food Research, 2023
In the fish processing sector, fish have been processed in large quantities, are generated and discarded into the sea or dumped into the oceans. However, by employing effective strategies, this fishery waste can be utilized and converted into fish protein hydrolysate (FPH). FPH is a rich source of amino acids and peptides that have biological properties such as angiotensin-converting enzyme (ACE) inhibition, antimicrobial, antioxidant, and anticancer activity. The FPH has been used as a functional food perspective related to improving human health, primarily by maintaining blood pressure and normal heart function as measured by antihypertensive activity. FPH can be produced by various fish species, parts, and hydrolysis methods. Several studies have been published on the acceptability of FPH in obtaining bioactive properties from various fish, each using a different method to obtain bioactive properties. FPH is commonly produced by fish species such as Atlantic Cod, Lizard Fish, Atlantic Chub Mackerel, Atlantic Horse Mackerel, and Kawakawa (Mackerel Tuna). Furthermore, we summarized the various methodologies used by various researchers based on raw material data collection, method of production, ACE-inhibitory assay, ACE-inhibitory activity, degree of hydrolysis, molecular weight, purification method, and ACE-I peptide structure in this review.
Polish Journal of Food and Nutrition Sciences, 2005, (55)2, 133–138, 2005
Enzymatic hydrolysate (Alcalase catalysed) was obtained from cracklings (CEH) and acid hydrolysates were prepared from cracklings (CAH) and chicken feathers (FAH). The degree of hydrolysis (DH) of CEH, CAH, and FAH were 14.0%, 53.8%, and 46.2%, respectively. The results of the SE-HPLC confirmed that small molecular weight peptides composed a considerable fraction of all hydrolysates. The crude hydrolysates exhibited angiotensin I-converting enzyme (ACE) inhibitory activity as determined using hippuryl-His-Leu as the substrate. The inhibition of ACE by CEH, CAH, and FAH was 72.3%, 50.0%, and 49.6% under identical assay conditions.