Inhibition of Angiotensin-Converting Enzyme (ACE) L.

helveticus
and Novel Functional Ingredient Keep Normal Blood Pressure
GSLC Assignment

Functional Foods

Group 2

Name NIM
Evi : 1901526016
Giovina Stefanny : 1901460051
Ivan Yudhistira : 1901513884
Michelle Muliawidjaja : 1901502022
Ronald Horison : 1901492375
Vini Octaviani Puspita : 1901527574

Lecturer : Ir. Ingrid Suryanti Surono, M.Sc., Ph.D.

Department of Food Technology

2017
Background
Hypertension affects up to 30% of the adult population in most countries. Besides
pharmacological therapy, nutritional factors play a significant role in the prevention and treatment of
hypertension. Prevention of diseases may in the future be just as important as treatment of diseases.
Interest on the possible health-promoting properties of food is increasing and more and more research
is targeted at the search for new biologically active compounds in different food products. The most
studied bioactive peptides at present are those inhibiting angiotensin-converting enzyme (ACE). ACE,
as a part of the renin-angiotensin system (RAS), has an important role in the regulation of blood pressure
by converting angiotensin I to a potent vasoconstrictor, angiotensin II, which induces the release of
aldosterone and therefore increases the sodium concentration and blood pressure further (Jakala, 2010).

Definition of ACE
Angiotensin converting enzyme is a dipeptidyl carboxypeptidase (EC. 3.4.15.1) and was
originally isolated from horse blood (Skeggs et al., 1956). ACE plays a crucial role in the regulation
of blood pressure as it promotes the conversion of angiotensin-I to the potent vasoconstrictor
angiotensin-II as well as inactivates the vasodilator bradykinin, which has a depressor action in the
renin-angiotensin system. This potent vasoconstrictor is also involved in the release of a Na-retaining
steroid, aldosterone from the adrenal cortex, which has a tendency to increase blood pressure (Li et al.,
2003). Angiotensin converting enzyme (ACE) inhibitors are used in the management of hypertension
and congestive heart failure. The ACE inhibitors interfere with the conversion of angiotensin I to
angiotensin II. Ingestions involving small amounts of ACE inhibitors may result in limited or no toxic
effects (Spiller, 2014).

ACE Inhibitor Properties

ACE inhibitors can be classified on the basis of their interaction with the active site zinc (II)
centre. Inhibitors such as captopril, zofenopril, rentiapril, alacepril bind to the metal centre through the
thiol moiety, leading to the formation of zinc(II)-thiolates. Compounds such as enalarpril, lisinopril,
ramipril, spirapril, etc. interact through the carboxylate moiety and fosinopril interacts through the
phosphate group. In addition to the coordination with the active site metal centre, these inhibitors
interact with the binding pockets present at the active site of ACE. Therefore, the side chains and
stereochemistry of inhibitors play a crucial role (Bhuyan and Mugesh, 2011).

The enzyme consists of 27 helices (96% of the total amino acid residues) and six relatively
short -strands. The overall shape is ellipsoid (approx. dimension, 72 52 48 ) with a central groove
that extends for about 30 into the active site and divides the enzyme into two sub-domains. The cavity
is covered by four helices and a -strand. Three of these helices contain charged amino acid residues
and restrict the access of larger polypeptides to the active site cleft. Two chloride ions are bound to the
interior of the enzyme with a highly ordered active site containing zinc (II) ion (Bhuyan and Mugesh,
2011).

ACE Inhibitor Sources and Examples

Inhibition of ACE is considered to be a useful therapeutic approach in the treatment of
hypertension. Therefore, in the development of drugs to control high blood pressure, ACE inhibition
has become an important activity. Many studies have been attempted in the synthesis of ACE inhibitors
such as captopril, enalapril, alcacepril and lisinopril, which are currently used in the treatment of
essential hypertension and heart failure in humans (Ondetti, 1977). Here are some example of drugs
that used as Captopril, Enalapril, Fosinopril, Lisinopril, Perindopril, Quinapril, Ramipril, Trandolapril.
However, these synthetic drugs are believed to have certain side effects such as cough, taste
disturbances, skin rashes or angioneurotic edema all of which might be intrinsically linked to synthetic
ACE inhibitors (Atkinson, 1979).
Many research groups have combed for novel ACE inhibitors from natural products
(Maruyama, 1989), microbial sources (Demain,1989) and food proteins (Fujita, 2000). Marine by-
products generated during marine food processing, have low commercial value, but the detection of in
vitro ACE inhibitory activity in these products might provide significant environmental and cost
benefits. Marine-derived bioactive peptides have been shown to possess many physiological functions,
including antihypertensive or ACE inhibition (Yokoyama, 1992). A marine-derived antihypertensive
peptides have shown potent ACE inhibitory activities (Table 1). The potency of these marine-derived
peptides to inhibit ACE activity has been expressed as an IC50 value, which is the ACE inhibitor
concentration, leading to 50% inhibition of ACE activity.

. ACE inhibitory peptides, derived from a multitude of plant and animal proteins such as milk,
soy or fish, represent sources of health-enhancing components. These ACE inhibitory peptides can be
enzymatically released from precursor proteins in vitro and in vivo, respectively during food processing
and gastrointestinal digestion (Leo, et al, 2009) .

Target Group and Target Health Benefit

Angiotensin Converting Enzyme (ACE) inhibitors are used in the management of hypertension
and congestive heart failure. ACE inhibitors used to lower blood pressure in people with high blood
pressure (hypertension), because it have an ability to relax constricted blood vessels. A recent study
looked at patients at high risk for having heart attacks or developing heart failure (people with known
blockages in arteries, diabetes, or hypertension) and found that treatment with an ACE inhibitor
prevented future heart attacks and deaths from heart disease in these groups of people. ACE inhibitors
seem to be particularly effective drugs for reducing heart attacks in patients with diabetes. These drugs
can also protect kidney function in patients with diabetes and patients with other forms of mild kidney
disease, especially those with protein in the urine (Sweitzer, 2003).

Renal Angiotensin Converting Enzyme

ACE is produced in large quantities by the kidneys. In particular, renal ACE and locally
generated angiotensin II will affect several key process in sodium transport and the induction of sodium
and water retention resulting in the elevation of blood pressure. Angiotensin II, generated in the renal
tubular lumen, may enhance NaCl transport across macula densa cells and thus increase the basal
sensitivity of tubuloglomerular feedback . The angiotensin II is a master regulator of nephron sodium
transport and plays a key role in experimental hypertension (Bernstein, et al., 2014).

Mechanism of ACE Inhibition (Bioactivity)

These compound inhibit competitively the activity of ACE (also termed kininase II) to prevent
formation of the active octapeptide, angiotensin II, from the inactive decapeptide, angiotensin I. This
occurs in blood and tissues including kidney, heart, blood vessels, adrenal gland and brain. Angiotensin
II is a potent vasoconstrictor, promotes aldosterone release, facilities sympathetic activity and has other
potentially harmful effects on the cardiovascular system. Reduction in blood pressure secondary to
vasodilatation following ACE inhibition is greatest when the renin-angiotensin system is stimulated
(e.g. following diuretic therapy or in renal artery stenosis) but ACE inhibitors also lower blood pressure
when there is normal or low activity of the renin-angiotensin system. Inhibition of ACE (kininase II)
also leads to accumulation of kinins including bradykinin which promotes vasodilator activity and may
contribute to the overall effectiveness of ACE inhibitors (BHS, 2008). The direct application of an ACE
inhibitor in the proximal tubular lumen significantly reduces proximal tubular sodium reabsorption.
This is probably as a result of Na+/H+ exchanger 3 (NHE3) modulation, as this is the predominant
mechanism for proximal tubular sodium absorption.

ACE inhibitors vary in efficiency and duration. Because of the effect, some ACE-Inhibitors is
not suitable for once - daily dosing. Captopril has the shortest duration of action and has to be
administered twice or thrice daily to provide blood pressure lowering over 24 hours. Although many
ACE inhibitors are recommended for once daily dosing, for some, such as enalapril, a more consistent
response is achieved by twice daily administration. Captopril can be alone or with other agents in the
management of hypertension. block the conversion of angiotensin I to the vasoconstrictor angiotensin
II. ACE inhibitors also prevent the degradation of bradykinin and other vasodilatory prostaglandins.
ACE inhibitors also increase plasma renin levels and reduce aldosterone levels. Aldosterone, is essential
for sodium conservation in the kidney, salivary glands, sweat glands and colon. By reducing the
aldosterone, it will help reducing the sodium concentration (Davis, 2015). Since duration of blood
pressure reduction is dose dependent, with most ACE inhibitors, smooth blood pressure lowering over
24 hours is only achieved at the maximum recommended dose. To avoid precipitous initial fall in blood
pressure or decline in renal function, it is generally advised to start therapy with low dosages in the
elderly, and in patients with compromised renal function or heart failure (BHS, 2008).

Lactobacillus helveticus: the importance, the characteristics, the correlations with inhibition of
ACE
Lactobacillus helveticus is a homofermentative, Gram-positive, rod-shaped thermophilic
microorganism belonging to lactic acid bacteria (LAB). These microorganisms are used in the dairy
industry as a starter predominantly employed to ferment milk in manufacturing of several
cheeses (Skrzypczak et al., 2015). Lactobacillus helveticus is gaining also importance as health-
promoting culture in probiotic and nutraceutic food products. It has the potential to produce bioactive
peptides or bacteriocins, and exert synbiotic effect when associated with prebiotics in fermented dairy
products. Lactobacillus helveticus can therefore be considered as a multifunctional LAB with increasing
importance in the food industry (Giraffa, 2014).

Scientific reports have demonstrated evidence that the L. helveticus species strains exhibit
health-promoting properties (Taverniti & Guglielmetti, 2012). L. helveticus is able to stimulate the
immune system, increases defense against pathogens, has an influence on the composition of the
intestinal microbiota, and prevent gastrointestinal infections. L. helveticus was also demonstrated to
establish synergistic interactions with other bacterial strains to antagonize pathogens (Gareau et al.,
2010). Various desired health-promoting sequences with antimicrobial, immunostimulating, opioid,
mineral binding, and antihypertensive activities have been isolated from products fermented with L.
helveticus (Griffiths & Tellez, 2013).

L. helveticus is among the most nutritionally fastidious LAB, requiring 14 exogenous amino
acids (Chopin, 1993). To assure its nutritional requirements when grown in milk, L. helveticus relies on
a potent proteolytic system capable of producing short peptides and liberating amino acids from casein
(Callanan et al., 2008). This explains why it has higher proteolytic activity than most other lactobacilli.
The proteolytic system of L. helveticus consists mainly of envelope-associated proteinases (CEPs)
which initially cleave caseins to large peptides, intracellular peptidases further degrading peptides to
small peptides and amino acids, and specific transport proteins which transfer amino acids and peptides
across the cytoplasmic membrane (Slattery et al., 2010).
 Some L. helveticus strains have been used in the production of antihypertensive-fermented milk
foods (Lpez-Fandio et al., 2006). The ability of individuals L. helveticus to produce antihypertensive
peptides is most likely related to the completeness and efficiency of their proteolytic system (Griffiths
and Tellez, 2013). Because of having a high proteolytic activity, L. helveticus is deemed as a good
candidate for producing fermented milks with high angiotensin-converting enzyme inhibitors (ACEI)
activity (Yamamoto et al., 1994; Fuglsang et al., 2003).

A large number of studies proved that among LAB, L. helveticus has the highest capacity to
form ACE inhibitory peptides from food proteins as a consequence of the marked activity of its cell
envelope-associated proteinases (CEPs) (Yamamoto et al., 1993). The first observations of the ability
of L. helveticus to generate ACE inhibitory peptides showed that the peptides liberated from casein by
the CEP activity of L. helveticus CP790 were able to suppress ACEs (Yamamoto et al., 1994). It was
also found that the ACE inhibitory activity was higher in the whey fractions of milk fermented with L.
helveticus than in milk fermented with other LAB, including L. delbrueckii, L. casei, Streptococcus
thermophilus, Lactococcus lactis and, L. acidophilus (Yamamoto et al., 1994).

Formation of ACE and Suitable Technology For Functional Foods

There are several ways of forming Angiotensin Converting Enzyme Inhibitory such
as enzymatic hydrolysis with digestive enzymes and the other way is by fermentation of milk using
proteolytic starter cultures and action of enzymes that derived from these microorganism. The bacteria
proteolytic enzymes hydrolyze the proteins to release peptides into the hydrolysate. The hydrolysates
are then tested in vitro for a biological activity. If the hydrolysates show good bioactivity, they are then
confirmed through in vivo testing. The biologically active hydrolysate could then be developed into a
functional food. The factors that affect the inhibiting activity compound are the source of enzyme, the
enzyme concentration and the hydrolysis time. The good source for bioactive peptides ingredients is
bovine casein in milk. Meanwhile, ACE inhibitory is difficult to generate from whey protein because
the -Lactoglobulin in whey have rigid structure that have resistance to digestive enzymes (Unal and
Akalin, 2006). Another source of production ACE inhibitory that reported by Wu, et al (2015) is from
silkworm pupa (Bombyx mori) protein that was hydrolyzed using pepsin, trypsin and -chymotrypsin
and the hydrolysate was purified.

The source of making antihypertensive peptide, Lys-Val-Leu-Pro-Val-Pro-Gln, was purified

and identified from casein hydrolysate produced by a proteinase from L. helveticus CP790. Nakamura
et al (1955) identified two ACE inhibitory peptides (Val-Pro-Pro, Ile- Pro-Pro) in milk fermented with
a starter culture composed of L. helveticus and Saccharomyces cerevisiae. In the fermentation process
of milk, bioactive peptides can be generated by dairy starter cultures, which show high proteolytic
activity. Lactic acid bacteria utilize milk protein, especially casein, as a source of amino acids for
growth. For example, Lactobacillus helveticus and Lactococcus lactis strains have been shown to
produce bioactive peptides.

Lactic acid bacteria that used as a culture starter in dairy product having extracellular proteinase
that can release bioactive peptides from casein. The transport system that provided by lactic acid
bacteria allowing specific amino acid to be transported including di- and tripeptides and oligopeptides
of up to 18 amino acids. Meanwhile some longer oligopeptides that cannot be transported will be the
source of liberation of bioactive peptides that will degraded by peptidases in intracellular that happen
after the cell lysis (Korhonen and Pihlanto, 2003).

These are some examples of the production food products that have ACE inhibitory. One of
them is the new developed cheese by Ryhanen et al. (2001). This cheese has probiotic bacteria as starter
culture which uses combination of Lactobacillus acidophilus and L. bifidobacteria. In the analyses, the
cheese was found contain bioactive peptides with ACE inhibitory. Leppala et al. (1998) studied the
potential formation of ACE inhibitory peptides from cheese whey and caseins during fermentation, with
various commercial lactic acid starters used in the manufacture of yoghurt and sour whole milk.
Yamamoto (1999) obtained antihypertensive peptides from both casein and whey proteins by
extracellular proteinase from different strains of (CPN4, CP790) Lactobacillus helveticus.

Some peptides can only be found while using fermentation methods. Recently, an
antihypertensive effect related to angiotensin I-converting enzyme inhibitory peptides was reported for
sour milk produced by a starter containing Lactobacillus helveticus and Saccharomyces cerevisiae. The
substances were purified from sour milk fermented until the pH reaches around 3.3. These two
substances were identified as Val-Pro-Pro and Ile-Pro-Pro by analysis of amino acid composition and
amino acid sequences. The amino acid sequence of Val- Pro-Pro and Ile-Pro-Pro were found in the
primary structure of bovine b-casein and b-casein and j-casein. These peptides were produced during
fermentation but were not found in the hydrolysate of casein by an extracellular proteinase of L.
helveticus. This tell us some peptides seemed to be processed by intracellular proteinase that followed
by the peptidase action by fermentation process (Yamamoto and Takano, 1999).

Functional Ingredients to Keep Blood Pressure Normal

Food derived ACE-inhibitory peptides have been used as an alternative to synthetic drugs and
are considered as the best known class of bioactive peptides. Several studies have reported that food
originating peptides could be used as an alternative ACE inhibitor with their low IC50 value to synthetic
drugs that mean more effective than drugs (Dikmen, et al, 2017).

Different types of plants have been used to obtain ACE-inhibitory peptides such as wheat, peas,
mushrooms, soybeans, walnuts, date seed flour, bitter melon seeds and spinach. Milk and dairy products
have been concluded to be functional foods. A number of plant-based peptides have been investigated
for their potential ACE inhibitor activities by using in vitro and in vivo assays. These plant-based
peptides can be obtained by solvent extraction, enzymatic hydrolysis with or without novel food
processing methods, and fermentation. Antihypertensive peptides have been found in processed dairy
products (cheese, milk) without any intentional functional role. Cheese whey is one of the examples
functional ingredients that can be consumed to keep blood pressure normal. whey compounds exhibit a
number of functional, physiological and nutritional features that make them potentially useful for a wide
range of applications in meal. Whey which is high perishable can be converted into lactose- free powder,
condensed whey, whey protein concentrates and whey protein isolates. In whey protein components, -
lactalbumin and -lactoglobulin, were also shown to contain bioactive sequences. Peptides showing
opioid and angiotensin I-converting enzyme (ACE) inhibitory activity were found in -lactalbumin and
-lactoglobulin (Leppl, 2000). Another sources of ingredients that have ACE-inhibitory is bovine
milk . Bovine milk contains about 32 g/l protein of which 80% are caseins and 20% whey proteins.
Caseins can be divided into -, - and -casein. The whey fraction contains -lactalbumin, -
lactoglobulin and various other proteins (e.g., immunoglobulins, lactoferrin) in smaller quantities.
Antihypertensive peptide sequences have been detected both from casein and whey fractions.
Lactotripeptides isoleucine-proline-proline (Ile-Pro-Pro) and valine-proline-proline (Val-Pro-Pro) have
been found from sour milk. Whey fraction of a yoghurt-like product was found to contain a dipeptide
Tyr-Pro (Jakala, 2010).
According to the literature, there are certain egg proteins are a source of antuhypertensive
peptides. Hoppe (2010) identified a peptide with confirmed ACE inhibitory activity (YAEERYPIL) in
ovalbumin pepsin hydrolysate. Other peptides with the ACE inhibitory bioactivity or antihypertensive
effects found in the enzymatic hydrolysates of egg white proteins had the sequences RADHPF (derived
from ovalbumin and showing antihypertensive activity in SHR) and RVPSL (from ovotransferrin as the
ACE inhibitor) (Hoppe, 2010).

Marine organisms are rich sources of structurally diverse bioactive compounds. Hence, a great
interest has been developed nowadays to isolate bioactive compounds, which act as ACE inhibitors
from marine resources because of their numerous health beneficial effects. According to the researches,
it has reported that marine-derived bioactive peptides, chitooligosaccharide derivatives (COS) and
phlorotannins have potent ACE inhibitory activity (Je, et al, 2008). Marine-derived bioactive peptides
have been obtained widely by enzymatic hydrolysis of marine proteins (Je, 2008). In fermented marine
 food sauces such as blue mussel sauce and oyster sauce, enzymatic hydrolysis has already been done
by microorganisms, and bioactive peptides can be purified without further hydrolysis (Je, 2005). In
addition, several bioactive peptides have been isolated from marine processing by-products or wastes
(Kim, 2000). Moreover, some of these bioactive peptides have identified to possess nutraceutical
potentials that are beneficial in human health promotion (Defelice, 1995) and recently the possible roles
of food-derived bioactive peptides in reducing the risk of cardiovascular diseases has been shown
(Erdmann, 2008).

References
Atkinson, A.B Robertson, J.I.S. 1979. Captopril in the Treatment of Clinical Hypertension and Cardiac Failure. Lancet 2,
836839. 8.
Bernstein, Kenneth E. Jorge F. Giani, Xiao Z. Shen, and Romer A. Gonzalez-Villalobos. 2014. Renal Angiotensin-
Converting Enzyme and Blood Pressure Control. Curr Opin Nephrol Hypertens. 2014 Mar; 23(2): 106112.
doi: 10.1097/01.mnh.0000441047.13912.56.
BHS (British Hypertension Society). 2008. Angiotensin Converting Enzyme (ACE)
Inhibitors. http://www.bhsoc.org/pdfs/therapeutics/Angiotensin%20Converting%20Enzyme%20%28ACE%29
%20Inhibitors.pdf. Accessed on October 23 2017.
Bhuyan , Bhaskar J and Govindasamy Mugesh. 2011 . Angiotensin Converting Enzyme Inhibitors in the Treatment of
Hypertension . CURRENT SCIENCE, VOL. 101, NO. 7, 10 OCTOBER 2011
Callanan, M., Kaleta, P., OCallaghan, J., OSullivan, O., Jordan, K., McAuliffe, O., et al. (2008). Genome sequence of
Lactobacillus helveticus, an organism distinguished by selective gene loss and insertion sequence element
expansion. J. Bacteriol. 190, 727735. doi: 10.1128/JB.01 295-07
Chopin, A. 1993. Organization and Regulation of Genes for Aminoacid Biosynthesis in Lactic Acid Bacteria. FEMS
Microbiol. Rev. 12, 2137.
Davis, Frank Allston. Kaptopril. https://davisplus.fadavis.com/3976/meddeck/pdf/captopril.pdf. Accesed at 23 October
2017.
Demain, A.L.; Somkuti, G.A. Hunter-Cevera, J.C. Rossmoore, H.W. 1989. Novel Microbial Products for Medicine and
Agriculture. Elsevier Science Publishers: Amsterdam The Netherlands.
Defelice, S.L. 1995. The Nutritional Revolution: Its Impact on Food Industry R&D. Trends Food Sci. Technol. 5961.
Dikmen, Ceren Daskaya, Aysun Yucetepe I, Funda Karbancioglu-Guler, Hayrettin Daskaya and Beraat Ozcelik. 2017.
Angiotensin-I-Converting Enzyme (ACE)-Inhibitory Peptides from Plants. Nutrients 2017, 9, 316;
doi:10.3390/nu9040316.
E. Ryhnen et al. 2001. A New Type of Ripened, Low-Fat Cheese with Bioactive Properties. International Dairy Journal.
11, pp. 441- 447.
Erdmann, K.; Cheung, B.W.Y.; Schroder, H. 2008. The Possible Roles of Food-Derived Bioactive Peptides in Reducing
the Risk of Cardiovascular Disease. J. Nutr. Biochem. 19, 643654.
Fuglsang, A., F. P. Rattray, D. Nilsson, and N. C. B. Nyborg. 2003. Lactic Acid Bacteria: Inhibition of Angiotensin
Converting Enzyme In Vitro and In Vivo. Antonie van Leeuwenhoek 83:2734.
Fujita, H., Yokoyama, K., Yoshikawa, M. Classification and antihypertensive activity of angiotensin I-converting enzyme
inhibitory peptides derived from food proteins. J. Food Sci. 2000, 65, 564569
Gareau M. G., Wine E., Reardon C., Sherman P. M. 2010. Probiotics Prevent Death Caused by Citrobacter
rodentiuminfection in Neonatal Mice. Journal Infect. Dis. 201, 819110.1086/648614.
Giraffa G. 2014. Lactobacillus Helveticus: Importance in Food and Health. Front Microbial 5: 338, 12.
doi:10.3389/fmicb.2014.00338.
Griffiths M. V., Tellez A. M. 2013. Lactobacillus helveticus: The Proteolytic System. Front Microbial. 4:3010.3389/fmicb.
00030.
Hoppe A. 2010. Examination of egg white proteins and effects of high pressure on select physical and functional properties.
Dissertations & Theses in Food Science and Technology. Univ. of Nebraska.
Je, J.Y.; Qian, Z.J.; Lee, S.H.; Byun, H.G.; Kim, S.K. 2008. Purification and antioxidant properties of bigeye tuna
(Thunnus obesus) dark muscle peptide on free radical-mediated oxidation systems. J. Med. Food. 11, 629637.
Je, J.Y.; Park, J.Y.; Jung, W.K.; Park, P.J.; Kim, S.K. 2005. Isolation of Angiotensin I Converting Enzyme (ACE) Inhibitor
from Fermented Oyster Sauce, Crassostrea gigas. Food Chem. 90, 809814
Li, G.H.; Le, G.W.; Shi, Y.H.; Shrestha, S. 2003. Angiotensin IConverting Enzyme Inhibitory Peptides Derived from Food
Proteins and Their Physiological and Pharmacological Effects. Nutr. Res 24, 469486.
Jakala, Pauliina and Heikki Vapaatalo. 2010. Antihypertensive Peptides from Milk Proteins. Pharmaceuticals (Basel). 2010
Jan; 3(1): 251272.
Jo, H.Y.; Jung, W.K.; Kim, S.K. 2008. Purification and characterization of a novel anticoagulant peptide from marine
echiuroid worm, Urechis unicinctus. Process Biochem. 2008, 43, 179184.
Kim, S.Y.; Je, J.Y.; Kim, S.K. Purification and characterization of antioxidant peptide from hoki (Johnius balengerii)
frame protein by gastrointestinal digestion. J. Nutr. Biochem. 2007, 18, 3138
Korhonen H and H. Philanto. 2003. Food Derived Food-derived Bioactive Peptides Opportunities for Designing Future
Foods. Current Pharmaceutical Design, 91, pp. 297-1308.
Lee, D.H., Kim, J.H. Park, J.S., Choi, Y.J., Lee, J.S. Isolation and characterization of a novel angiotensin-I-converting
enzyme inhibitory peptide derived from the edible mushroom Tricholoma giganteum. Peptides 2004, 25, 621627.
Leppl, Anne P. 2000. Bioactive peptides derived from bovine whey proteins:opioid and ace-inhibitory peptides. J. Food
Science and Tech.
Leo, De Leo F1, Panarese S, Gallerani R, Ceci LR. 2009. Angiotensin converting enzyme (ACE) inhibitory peptides:
production and implementation of functional food.
Lpez-Fandio, R., J. Otte, and J. Van Camp. 2006. Physiological, Chemical and Technological Aspects of Milk-Protein-
Derived Peptides with Antihypertensive and ACE-inhibitory Activity. Int. Dairy J. 11:12771293.
Maruyama, S.; Miyoshi, S.; Tanaka, H. Angiotensin-I-converting enzyme inhibitor derived from Ficus carica. Agric. Biol.
Chem. 1989, 53, 27632769.
Nakamura et al., 1955. Antihypertensive Effect of Sour Milk and Peptides Isolated from it that are Inhibitors to Angiotensin
Converting Enzyme. Journal of Dairy Science, 78, pp. 1253- 1257 (1995a).
_______________1955. Purification and Characterization of Angiotensin I-converting Enzyme Inhibitors from Sour Milk.
Journal of Dairy Science, 78, pp. 777-783 (1995b).
Ondetti, M.A. 1977. Design of Specific Inhibitors of Angiotensin-Converting Enzyme: New class of orally active
antihypertensive agents. Science 1977, 196, 441444. 7.
Pihlanto-Leppl et al., 1998. Angiotensin I Converting Enzyme Inhibitory Peptides Derived from Bovine Milk Proteins.
International Dairy Journal, 8, pp. 325-331.
Skeggs, L.T.; Kahn, J.R.; Shumway, N.P. 1956. The Preparation and Function of the Hypertension Converting Enzyme.
J. Exp. Med. 1956, 103, 295299.
Skrzypczak, K., W. Gustaw, A. Wasko. 2015. Health-Promoting Properties exhibited by Lactobacillus helveticus strains.
http://www.actabp.pl/pdf/4_2015/2015_1116.pdf. Accessed on October 22 2017.
Slattery, L., OCallaghan, J., Fitzgerald, G. F., Beresford, T., and Ross, R. P. 2010. Invited Review: Lactobacillus helveticus
a Thermophilic Dairy Starter Related to Gut Bacteria. J. Dairy Sci. 93, 44354454. doi: 10.3168/jds.2010-3327.
Spiller, H.A. 2014. Angiotengsin Converting Enzyme (ACE) Inhibitors. Encyclopedia of Toxicology (Third Edition).
Taverniti V, Guglielmetti S. 2012. Health-Promoting Properties of Lactobacillus helveticus. Front Microbiol 3: 392, 1
13. doi: 10.3389/ fmicb.2012.00392.
Unal, Gulfem and A. Sibel Alkalin. 2006. Milk Proteins and Hypertensions. Anno 17 - No. 6 AgroFOOD Industry.
Wu Q, Jia J, Yan H, Du J, Gui Z. 2015. A Novel Angiotensin- Converting Enzyme (ACE) Inhibitory Peptide from
Gastrointestinal Protease Hydrolysate of Silkworm Pupa (Bombyx mori) protein. Biochemical Characterization
and Molecular Docking Study.
Yamamoto, N., A. Akino, and T. Takano. 1994. Antihypertensive Effect of the Peptides Derived from Casein by An
Extracellular Proteinase from Lactobacillus helveticus Cp790. J. Dairy Sci. 77:917922.
Yamamoto, T. Takano. 1999. Antihypertensive Peptides Derived from Milk Proteins. Nahrung, 43(3) , pp. 159-164.
Yokoyama, K.H.; Chiba, H.; Yoshikawa, M. 1992. Peptide Inhibitors for Angiotensin-I-Converting Enzyme from
Thermolysin Digest of Ried bonito. Biosci. Biotechnol. Biochem. 1992, 56, 15411545.