Antioxidant and angiotensin I converting enzyme inhibitory activity of Bambusae caulis in Liquamen

Bambusae caulis in Liquamen (BCL) is a nutritious liquid that can be extracted from heat-treated bamboo stems. It is also an important herbal medicine in Asia. In this study, antioxidant and angiotensin I converting enzyme (ACE) inhibitory activity of BCL were investigated. BCL significantly quenched DPPH and peroxyl radicals measured by electron spin resonance (ESR) spectroscopy, while IC₅₀ values were 79.85 and 28.85 μg/ml, respectively. The ability of BCL to inhibit the oxidative damage of DNA was assessed in vitro by measuring the conversion of supercoiled pBR322 plasmid DNA to the open circular form. It was found that BCL significantly protected hydroxyl radical-induced DNA damage in a dose-dependant manner, while also inhibiting apoptosis in hydrogen peroxide-induced PC-12 cells and angiotensin I converting enzyme (ACE) activity. These findings suggest that BCL may be a beneficial ingredient in functional foods and/or pharmaceuticals.     

Antioxidant and angiotension‐converting enzyme inhibition capacities of various parts of Benincasa hispida (wax gourd)

Vegetables and fruits have been shown to be good sources of antioxidants. Benincasa hispida (wax gourd) has been used in traditional Chinese medicine to treat hypertension and inflammation. The aims of this study were to investigate the abilities of antioxidation and inhibition of angiotensin‐converting enzyme (ACE) activity of wax gourd pulp, core, seed, and peel prepared by different extraction methods. The fresh weights required to reach 50% inhibition of linoleic acid oxidation were higher in fresh extracts, compared to other extraction methods. Fresh weights required to reach 50% inhibition were the lowest in seed. The seed had the lowest Cu²⁺‐induced low‐density lipoprotein (LDL) oxidation percentage and inhibition level of ACE activity among all parts. The higher antioxidant capacity of the seed may result from the higher total phenolics contents and superoxide dismutase activity. The abilities of antioxidation and ACE activity inhibition may provide protective effects against cardiovascular diseases and cancers.     

Identification of angiotensin converting enzyme inhibitory and antioxidant peptides in a whey protein concentrate hydrolysate produced at semi‐pilot scale

Antioxidant and angiotensin converting enzyme (ACE) inhibitory peptides were identified in a 5 kDa ultrafiltration permeate of a whey protein hydrolysate generated at semi‐pilot scale. Further laboratory scale ultrafiltration of this 5 kDa permeate resulted in a 0.65 kDa permeate with antioxidant, (1.11 ± 0.074 μmol TE per mg dry weight, oxygen radical absorbance capacity, ORAC) and ACE inhibitory (ACE IC₅₀ 0.215 ± 0.043 mg mL^?1) activities. Semi‐preparative (SP) reverse phase high‐performance liquid chromatography (RP‐HPLC) of the 0.65 kDa permeate resulted in a fraction (SP_F3) with a 4.4‐fold increase in ORAC activity (4.83 ± 0.45 μmol TE mg dry weight) and a 1.3‐fold increase in ACE inhibitory activity (84.35 ± 1.36% inhibition when assayed at 0.28 mg mL^?1). Peptides within SP_F3 were identified using UPLC‐ESI‐MS/MS. Met‐Pro‐Ile had the highest ORAC activity (205.75 ± 12.08 μmol TE per mmol peptide) while Met‐Ala‐Ala and Val‐Ala‐Gly‐Thr had the highest ACE inhibitory activities (IC₅₀:515.50 ± 1.11 and 610.30 ± 2.41 μm , respectively).     

Antioxidant enzyme activities and antioxidant capacity in longissimus muscle from bulls fed diets rich in polyunsaturated fatty acids

The effect of a treatment diet composed of grass silage and concentrate including rapeseed (with/without feeding restriction) was compared with a control diet of maize silage/grass silage (70:30) and concentrate including soybean, on the antioxidant enzyme activities of fresh longissimus muscle from German Simmental bulls. Additionally, the effect of diet on antioxidant capacity (AOC) of hydrophilic and lipophilic antioxidants was evaluated in fresh and stored beef muscle using the FRAP-ferric reducing ability and TEAC – Trolox-equivalent antioxidant capacity assays at different reaction times. Catalase and superoxide dismutase activities were significantly higher in the treatment diet groups, and glutathione peroxidase activity was not different. AOC was not affected by the diet. However, storage affected the values of FRAP and TEAC assays, and the results were time-depending. 30 min were found like a minimum reaction time for both assays. Generally, AOC values of the hydrophilic antioxidants were significantly higher than lipophilic values.     

Antioxidant,enzyme inhibitory and antiproliferative activity of polyphenolic‐rich fraction of commercial dry ginger powder

Polyphenolic‐rich fraction obtained from locally produced dry ginger powder in Brahmaputra valley, India, and commercially available dry ginger (Zingiber officinale) rhizome powder consisted of [6]‐gingerol (41.9%), [6]‐shogaol (24.3%), 1‐dehydro‐6‐gingerdione (8.6%), [8]‐gingerol (7.2%), [10]‐gingerol (5.1%), [6]‐paradol (5.9%) and [4]‐gingerol (3.6%). Traces of methyl‐[6]‐gingerol and methyl‐[8]‐gingerol (both at 1.8%) were also detected. The fraction exhibited high antioxidant capacity [total phenolics (TP), ferric reducing antioxidant power (FRAP), oxygen radical absorbance capacity (ORAC) and cellular antioxidant activity (CAA assay)], effectively inhibited isolated digestive enzymes (α‐glucosidase, pancreatic lipase and angiotensin converting enzyme) and inhibited the proliferation of colon (HT29; IC₅₀ of 1.06 ± 0.02 mg mL^?1) and gastric (AGS IC₅₀ of 1.29 ± 0.03 mg mL^?1) adenocarcinoma cells, without affecting the proliferation of their nontransformed counterparts (IC₅₀ > 2.0 mg mL^?1). This case study demonstrates that locally produced and commercially available dry ginger powder from Brahmaputra valley, India, retains numerous food components that may enhance human health.     

Effect of angiotensin I converting enzyme inhibitory peptide purified from skate skin hydrolysate

Our objective was to evaluate the angiotensin I converting enzyme (ACE) inhibitory activity of skate skin protein hydrolysates and its corresponding fractions. The skate skin hydrolysates were obtained by enzymatic hydrolysis using alcalase, α-chymotrypsin, neutrase, pepsin, papain, and trypsin. Amongst the six hydrolysates, the α-chymotrypsin hydrolysate had the highest ACE inhibitory activity compared to other hydrolysates. The amino acid sequences of the purified peptides were identified to be Pro–Gly–Pro–Leu–Gly–Leu–Thr–Gly–Pro (975.38 Da), and Gln–Leu–Gly–Phe–Leu–Gly–Pro–Arg (874.45 Da). The purified peptides from skate skin had an IC₅₀ value of 95 μM and 148 μM, respectively, and the Lineweaver–Burk plots suggest that they act as a non-competitive inhibitor against ACE. Our study suggested that novel ACE inhibitory peptides derived from skate skin protein may be beneficial as anti-hypertension compounds in functional foods.     

Purification and characterisation of angiotensin I converting enzyme inhibitory peptides from lysozyme hydrolysates

Hen egg white lysozyme (HEWL) was hydrolysed with trypsin, papain and a combination of the two. The prepared hydrolysates exhibited ACE inhibitory activity. The hydrolysates were fractionated using ultrafiltration and reverse phase-high performance liquid chromatography (RP-HPLC). Three fractions, which showed the highest ACE inhibitory activities, were purified by RP-HPLC. They were the F₇ (from papain-trypsin hydrolysate), F₈ (from papain hydrolysate) and F₃ (from trypsin hydrolysate) fractions. The IC₅₀ values were 0.03, 0.155 and 0.23 mg/ml for F₇, F₈ and F₃, respectively. The F₇ fraction was the most potent ACE inhibitor peptide, and was composed of 12 amino acids, Phe-Glu-Ser-Asn-Phe-Asn-Thr-Gln-Ala-Thr-Asn-Arg (MW: 1428.6 Da). Lineweaver-Burk plots suggest that the F₇ peptide acts as an uncompetitive inhibitor against ACE. The kinetic parameters (K_m, V_max, and K_i) for the F₇ peptide were measured and compared to the control.     

Enhancement of angiotensin I converting enzyme inhibitory activity and improvement of the emulsifying and foaming properties of corn gluten hydrolysate using ultrafiltration membranes

In this study, we attempted to enhance angiotensin I converting enzyme (ACE) inhibitory activity using an ultrafiltration (UF) membrane and to improve the emulsifying and foaming properties of fractions. ACE inhibitory activities of the corn gluten hydrolysate prepared by Flavourzyme had the same trend as protein contents and the IC₅₀ value was 0.18 mg solid after 8 h hydrolysis. The hydrolysate was separated by using two kinds of UF membrane (10 and 5 kDa cut-off membranes) and three fractions, >10 kDa, 5–10 kDa, and <5 kDa, were obtained. The yields of these three fractions were 58.3%, 27.2% and 14.5% as dry matter, respectively. The IC₅₀ value (the concentration of ACE inhibitor required to inhibit 50% of the ACE activity under specific conditions) of the <5 kDa fraction was 0.05 mg solid and was approximately fourfold that of the original, 5–10, and >10 kDa fractions. The average hydrophobicity of the <5 kDa fraction was slightly lower than that of the others according to the hydrophobicity index of Nozaki and Tanford, but according to the Krigbaum and Meirovitch index, that of the <5 kDa fraction was slightly higher than those of the others, showing there might be no significant differences among the fractions. The emulsifying activity and emulsion stability in the <5 kDa fraction was higher than those in the other fractions. Two fractions, <5 kDa and >10 kDa, showed poor foaming capacity at below pH 6.0. The foam stabilities of two fractions (<5 kDa and >10 kDa) increased with increasing pH, but that of 5–10 kDa decreased with increasing pH. The UF treatment was an effective method for the enhancement of ACE inhibitory activity with improvement of functional properties.     

具有血管紧张素转化酶抑制活性的益生菌筛选鉴定及其发酵特性的研究

目的 筛选具有血管紧张素转化酶(ACE)抑制作用的微生物, 提供适用于降高血压发酵产品的菌株。方法 从7种发酵食品中分离菌株后, 利用体外ACE酶抑制率法筛选ACE抑制率大于80%的菌株, 再通过耐盐和耐NaNO2实验, 筛选出目的菌, 并对其进行生物学特性测定和分子生物学鉴定。结果 从发酵食品中分离得到98株菌中, 筛选出F2和D2两个目的菌, F2和D2的ACE酶抑制率分别为83.13%和98.00%, 且均能耐受6%的NaCl及100 mg/kg的NaNO2。结论 筛选出的F2菌株为地衣芽孢杆菌, 对金黄色葡萄球菌抑菌效果显著; D2为球形芽孢杆菌, 具有脂肪酶活性; 且F2和D2菌均具有蛋白酶活性。     

Production of angiotensin I converting enzyme inhibitory (ACE-I) peptides during milk fermentation and their role in reducing hypertension

Fermented milk is a potential source of various biologically active peptides with specific health benefits. Angiotensin converting enzyme inhibitory (ACE-I) peptides are one of the most studied bioactive peptides produced during milk fermentation. The presence of these peptides is reported in various fermented milk products such as, yoghurt, cheese, sour milk, etc., which are also available as commercial products. Many of the ACE-I peptides formed during milk fermentation are resistant to gastrointestinal digestion and inhibit angiotensin converting enzyme (ACE) in the rennin angiotension system (RAS). There are various factors, which affect the formation ACE-I peptides and their ability to reach the target tissue in active form, which includes type of starters (lactic acid bacteria (LAB), yeast, etc.), substrate composition (casein type, whey protein, etc.), composition of ACE-I peptide, pre and post-fermentation treatments, and its stability during gastrointestinal digestion. The antihypertensive effect of fermented milk products has also been proved by various in vitro and in vivo (animal and human trials) experiments. This paper reviews the literature on fermented milk products as a source of ACE-I peptides and various factors affecting the production and activity of ACE-I peptides.     

Antioxidant and angiotensin converting enzyme (ACE) inhibitory activities of cocoa (Theobroma cacao L.) autolysates

The present study investigated antioxidant and angiotensin converting enzyme (ACE) inhibitory activities of cocoa autolysates. After removal of cocoa fat, alkaloids and polyphenols, the remaining proteinous powder was autolyzed at pH 3.5 and 5.2. At similar concentrations, autolysates produced at pH 3.5 indicated the highest reducing power and ACE inhibition activity. However, those generated at pH 5.2 showed the highest antioxidant activity based on ??-carotene bleaching assay. The results displayed a dose-dependent trend. Based on amino acids composition, slight differences were detected between autolysates, and as it was found, they were rich in hydrophobic amino acids. Qualitative and quantitative tests were applied to assure that the results from the assays were not due to the polyphenols of cocoa autolysates. Based on the results no polyphenols could be detected from cocoa autolysates. It can be indicated that among other useful substances of cocoa, its peptides and amino acids could contribute to its health-promoting properties. Furthermore, these bioactive substances can be exploited into functional foods or used as a source of nutraceuticals.     

Fractionation of angiotensin I converting enzyme inhibitory activity from pea and whey protein in vitro gastrointestinal digests

In vitro gastrointestinal digestion of pea and whey protein produced high angiotensin I converting enzyme (ACE) inhibitory activity with IC₅₀ 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 IC₅₀ 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 IC₅₀ 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. Copyright © 2004 Society of Chemical Industry     

Studies on the inhibitory effect of Graptopetalum paraguayense E. Walther extracts on the angiotensin converting enzyme

This study was aimed at evaluating the kinetic properties and capacities of water (GWE), 50% ethanolic (GE50) and 95% ethanolic (GE95) extracts from Graptopetalum paraguayense for the potential to inhibit angiotensin converting enzyme (ACE). The results showed that GWE, GE50 and GE95 showed potent inhibitory effects on ACE. It was found that the ACE inhibitory activities of all the tested extracts increased with the increase of their concentrations. In addition, the ACE inhibition of the tested extracts of G. paraguayense were significantly reduced after the addition of 1.5 mM ZnCl₂, suggesting the inhibitory action of the extracts may have resulted from the chelation of the ACE zinc cofactor. The inhibition kinetics, analyzed by Lineweaver–Burk plots, revealed that G. paraguayense extracts showed a mixed-type inhibition. A comparison of the 50% inhibition concentration (IC₅₀) and K_i values showed that the ethanolic extracts, including GE50 and GE95 exhibited the more effective ACE inhibitory activity than the water extracts of G. paraguayense.     

Free radical scavenging capacity and antioxidant enzyme activity in deerberry (Vaccinium stamineum L.)

Fruit from three genotypes (‘B-76’, ‘B-59’ and ‘SHF-3A’) of deerberry [Vaccinium stamineum L.] were evaluated for fruit quality, total anthocyanin and phenolic contents, antioxidants, antioxidant capacity, and antioxidant enzyme activity. The fruit soluble solids, titratable acids, total anthocyanins, and total phenolic contents varied with genotypes. Cyanidin 3-galactoside and cyanidin 3-arabinoside were the two predominant anthocyanins. Resveratrol was also found in deerberries. Among the three genotypes, ‘B-76’ had higher amount of anthocyanins, phenolic compounds and resveratrol than ‘B-59’ and ‘SHF-3A’. Deerberries contained potent free radical scavenging activities for 2,2-Di (4-tert-octylphenyl)-1-picrylhydrazyl (DPPH), 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS⁺), peroxyl radical (ROO), superoxide radicals (), hydrogen peroxide (H₂O₂), hydroxyl radicals (OH), and singlet oxygen (¹O₂) radicals and also had high activities of antioxidant enzymes including glutathione-peroxidase (GSH-POD), glutathione reductase (GR), superoxide dismutase (SOD), ascorbate peroxidase (AsA-POD), guaiacol peroxidase (G-POD), monodehydroascorbate radical reductase (MDAR), dehydroascorbate reductase (DHAR), glutathione reductase (GR)] and non-enzyme antioxidants [ascorbic acid (ASA) and reduced glutathione (GSH)]. Antioxidant capacities were highly correlated to antioxidant enzymes activities. Among the three genotypes, ‘B-76’ had the highest level of antioxidants and antioxidant enzyme activity.     

The antioxidant, angiotensin converting enzyme inhibition activity, and phenolic compounds of bamboo shoot extracts

This study was undertaken to evaluate the functional properties of two of the most popular species of edible bamboo shoots in Korea (Phyllostachys pubescens and Phyllostachys nigra). Powdered bamboo shoots were extracted with methanol and an aqueous suspension of the obtained methanol extract was partitioned successively with chloroform, ethyl acetate, and butanol, leaving a residual water extract. All obtained extracts were evaluated for their antioxidant capacity and antimicrobial activity, angiotensin converting enzyme (ACE) inhibition activity, and ascorbic acid and phenolic compound content. Methanol and water fractions showed a particularly high ascorbic acid contents. The ethyl acetate fraction contained a high concentration of phenolic compounds. Among all extracts, the ethyl acetate and butanol fractions showed particularly high antioxidant activity. Methanol extract had a significantly higher ACE inhibitory activity than other extracts. None of the extracts inhibited the tested bacteria.     

Effect of enzyme type and hydrolysis conditions on the in vitro angiotensin I‐converting enzyme inhibitory activity and ash content of hydrolysed whey protein isolate

Removal of salts from protein hydrolysate mixture on large scale is very difficult and relatively inefficient. Selecting practical proteinase system and hydrolysis conditions for the production of whey protein isolate (WPI) enzymatic hydrolysates with high angiotensin I‐converting enzyme (ACE) inhibitory activity and low ash content is very useful. The effect of alcalase, neutrase, trypsin and their combined system, i.e. alcalase‐neutrase and trypsin‐neutrase, under two different hydrolysis conditions, i.e. pH‐controlled and pH‐spontaneous drop, on the formation of ACE‐inhibitory peptides and the characteristics of WPI hydrolysate was investigated. Results showed that the ACE‐inhibitory activity of WPI hydrolysate obtained with alcalase was significantly higher than that of its trypsin or neutrase hydrolysate obtained at the same hydrolysis time by both pH‐controlled and pH‐spontaneous drop method (P < 0.05). The WPI hydrolysate obtained after 3 h incubation with alcalase plus 2 h with neutrase under pH‐spontaneous drop condition possessed the highest ACE‐inhibitory activity of 54.30% and the lowest ash content of 2.95%. This is practical as a functional ingredient in the food industry because of its high ACE‐inhibitory capability, commercial availability in large supply of alcalase and neutrase and no needing for additional desalting process.     

Angiotensin converting enzyme inhibitory activity of proteolytic digests of peanut (Arachis hypogaea L.) flour

Defatted raw and roasted peanut flour were hydrolyzed with alcalase or sequentially with pepsin and pancreatin, and then the hydrolyzates were fractionated by RP-HPLC and tested for hypotensive potential. This research revealed that proteolytic peanut digests have an inhibitory effect on the activity of angiotensin converting enzyme (ACE). Three fractions from the hydrophobic end of the chromatogram of each hydrolyzate were the most potent for inhibiting ACE activity in comparison to seven other fractions. These potentially potent fractions were then assayed for IC₅₀. Fractions from the alcalase digestion of raw peanut exhibited IC₅₀ values of 8.7-122 μg/ml, and those from roasted flour exhibited values of 12-235 μg/ml. IC₅₀ values of 7.9-65.9 μg/ml, and 11-36 μg/ml for raw and roasted peanut, respectively, from the pepsin-pancreatin system were observed. These values compare to the IC₅₀ value of 0.36 μg/ml of a known commercial ACE inhibitor (pGlu-Trp-Pro-Arg-Pro-Gln-Ile-Pro-Pro).     

Fermentation with Bacillus spp. as a bioprocess to enhance anthocyanin content,the angiotensin converting enzyme inhibitory effect,and the reducing activity of black soybeans

Food possessing anthocyanins, angiotensin converting enzyme (ACE) inhibitory activity or reducing activity show beneficial effect on human health. To develop healthy food, black soybeans were fermented with either Bacillus subtilis BCRC 14715 or Bacillus sp. CN11, or a mixture of both Bacillus spp. in the present study. The anthocyanin content, the ACE inhibitory activity and the reducing power of the fermented black soybean were then examined. It was found that the ACE inhibitory activity of the extracts of bean and viscous material from the fermented black soybeans varied with extraction solvents and starter organism, yet increased as the fermentation period was extended, regardless of starter organism. After 18 h of fermentation, the water extract of bean showed less ACE inhibitory activity than did the respective 80% ethanol extract. While the water extract of viscous material showed a higher ACE inhibitory activity than the respective ethanol extract. With respect to extraction yield, it was found that the ACE inhibitor in the fermented black soybean could be extracted more efficiently with water than 80% ethanol. Fermentation with B. subtilis BCRC 14715 was also found to increase the anthocyanin content of black soybean and the reducing activity of the extracts. Finally, the 80% ethanol extract showed a higher reducing activity than the water extract.