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Cheddar cheese is one of the most popular varieties of cheese. The origin of this cheese is the town of Cheddar in Somerset, southeast England. It is available in a ripened form to develop flavor in cheese. Cheddar cheese exhibits various bio-functional properties. These bio-functional properties are attributed due to bioactive peptides. During cheese manufacturing and ripening, those bioactive peptides are produced. Bioactive peptides are short chains of amino acids which exhibit biological effect in humans, such as anti-oxidative, anti-hypertensive, ACE-inhibitory, anti-microbial, opioid peptides, and immunomodulatory properties, etc. These activities fight against free radicals, microbial infections, thrombosis, high pressure, bad cholesterol, diabetes, etc. and simultaneously enhance immunity. Therefore, consumption of cheddar cheese is considered safe and helps in the development of good health.
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Health benets of Cheddar cheese
Supriya Das1, Chandrakanta Sen2, Mahasweta Bhattacharyya2 and Pinaki Ranjan Ray2*
1Dairy Technology Division, ICAR- National Dairy Research Institute, Karnal, Haryana
2Dept. of Dairy Chemistry, West Bengal University of Animal and Fishery Sciences, Kolkata
Email: pinakirray@gmail.com
Received: 13th October 2020 | Accepted: 21st November 2020
Abstract
Cheddar cheese is one of the most
popular varieties of cheese. The origin of this
cheese is the town of Cheddar in Somerset,
southeast England. It is available in a ripened
form to develop avor in cheese. Cheddar
cheese exhibits various bio-functional
properties. These bio-functional properties are
attributed due to bioactive peptides. During
cheese manufacturing and ripening, those
bioactive peptides are produced. Bioactive
peptides are short chains of amino acids
which exhibit biological effect in humans,
such as anti-oxidative, anti-hypertensive, ACE-
inhibitory, anti-microbial, opioid peptides, and
immunomodulatory properties, etc. These
activities ght against free radicals, microbial
infections, thrombosis, high pressure, bad
cholesterol, diabetes, etc. and simultaneously
enhance immunity. Therefore, consumption of
cheddar cheese is considered safe and helps
in the development of good health.
Keywords: Cheddar cheese, Bioactive Peptide,
Antioxidant, ACE-inhibitory, Immunity
Introduction
Cheddar cheese is a fermented dairy
product which was originated in the village
of “Cheddar” in Somerset, England. Cheddar
Gorge on the edge of the town contains various
caverns, which gave the perfect humidity and
consistent temperature for developing the
cheddar. Cheddar cheese is normally ripened
at 6-8°C for 4 to 6 months. It is prepared by
coagulating cow milk with a suitable enzyme
named rennet and acidied by suitable cultures
which produce a wide range of avor, texture.
The processing of cheddar cheese is done by
the stirred-curd process without matting. This
process was discontinued because it produced
many gassy cheeses with off-avors due to
poor sanitary conditions. The quality of the
cheese is mainly improved by the cheddaring
process that results in faster and more amount
of acid production (Ong et al. 2017). The
development of gas-forming organisms such
as coliform is repressed as the pH decreases
below 5.4. When curd pH reaches 5.8 or below
it forms a brous structure that is commercially
acceptable. During the ripening of cheddar
cheese, the indigenous intact caseins are
proteolyzed by proteolytic enzymes and form
smaller peptides, known as bioactive peptides.
They are comparatively short chains of amino
acids that have a biological effect on people
and animals. These are fragments of proteins
that have a positive effect on body functions
and ultimately control the health (Kitts and
Weiler 2003). Bioactive peptides contain
2-20 amino acid residues in length (Korhonen
2009). These are identied in various types of
dairy products such as milk, cheese, yoghurt,
ker. They are inactive within the protein
molecules. Conjugated linoleic acid (CLA)
has been found as a major source in dairy
products, having benecial physiological
effects such as anti-atherosclerotic activities
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and anti-carcinogenic activity (Lock and
Bauman 2004). Proteolysis of cheddar cheese
is governed by proteolytic enzymes mainly
plasmin and chymosin and some starter
cultures. During this proteolysis, bioactive
peptides are formed in cheddar cheese which
manifests various functional properties such as
anti-oxidant, anti-hypertensive, ACE-inhibitory,
and anti-microbial. Bioactive peptides can be
liberated by mainly enzymatic hydrolysis by
digestive enzymes, microbial fermentation
and plant-derived or microorganisms-derived
proteolytic enzymes. Digestive enzymes
mainly pepsin, trypsin, and chymotrypsin help
in enzymatic hydrolysis. On the other hand,
fermentation of milk with a proteolytic starter
culture, and proteolysis by enzymes derived
from microorganisms or plants also help to
release bioactive peptides (Korhonen 2009).
There are four types of enzymes in cheddar
cheese such as coagulant, indigenous milk
proteases, starter and non-starter proteases,
and nally exogenous proteases. Breakdown of
caseins into smaller peptides and amino acids
takes place by different enzymes like plasmin,
pepsin, and chymosin, which helps in the
development of the avor of cheddar cheese
(Roy et al. 2015). The major source of bioactive
peptides is the milk proteins. Our body
systems deal with numerous diseases like the
cardiovascular, digestive, endocrine, immune,
and nervous system which can be treated by
bioactive peptides. Bioactive peptides also
inuence on several biological processes such
as evoking behavioral, nutritional responses,
neurological, hormonal and gastrointestinal.
Several varieties of peptides are formed in
cheese during the ripening process, such as
immunomodulatory peptides, antioxidative
peptides, anti-cholesterimic peptides, ACE
inhibitory peptides, etc. During cheese
ripening, secondary proteolysis also produces
some other bioactive peptides. This present
article enlightens potential bioactive peptides
formed during the ripening of Cheddar cheese
and their health benecial attributes.
Preparation of cheddar cheese
Every variety of cheese has a specic
manufacturing procedure. The manufacturing
procedure of cheddar cheese is expressed in
the below Fig. 1.
Peptides and bioactive peptides
Milk contains two types of proteins
namely casein and whey proteins. Casein
fraction constitutes αS1, αS2, β, and
κ-casein. During cheddar cheese processing
Fig. 1. Flow diagram of Cheddar Cheese
Manufacturing
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casein fraction retains in the curd and the
whey protein portion goes out with whey.
Proteolysis takes place during the ripening
of cheese which leads to the fractionation
of casein. Casein fractions contribute to
specic bioactive peptides from β-casein,
α-casein, and κ-casein. α-casein has opioid
and mineral binding properties (Fernandez-
Tome et al. 2016). β-casein exhibits blood
pressure-lowering activity (Aluko 2015), DPP-
IV inhibitory (Nongonierma and FitzGerald
2016), anti-tumor (Zhao et al. 2016), mineral
binding (Nongonierma et al. 2016), intestinal
stress-reducing effect (Bessette et al. 2016).
κ-casein has blood pressure-lowering (Beltran-
Barrientos et al. 2016) and anti-inammatory
activity (Sawin et al. 2013).
Different cheeses and their bioactivity
There are a variety of cheeses that
contain various bioactive compounds. These
bioactive compounds are health benecial.
The various type of bioactivity exhibited by the
various types of cheeses is shown in Table 1.
Different health-benecial attributes of
cheddar cheese
Bioactive peptides are formed in cheddar
cheese during proteolysis exhibit some bio-
functional properties namely anti-oxidative,
ACE-inhibitory, immunomodulatory, anti-
cholesterimic and antimicrobial. Specic milk
proteins derived bioactive peptides exhibit the
aforesaid bio-functional properties in cheddar
cheese, shown in Table 2.
Antioxidative activity
The antioxidant activity has been
identied in the domains within the milk
proteins (Hernandez et al. 2005). Antioxidative
peptides prevent the formation of free radicals
by free radical scavenging activity. These
free radicals cause coronary-artery disease,
carcinoma, aging, stroke and other heart
Table 1. Variety of cheeses exhibiting bio-functional activity
Cheese Bioactivity found Reference
Cheddar
Anti-oxidant
ACE-inhibitory (Hossain et al. 2018)
Anti-microbial
Anti-oxidant
Anti-hypertensive
(Pritchard et al. 2010)
Antioxidant (Gupta et al. 2009)
Phosphopeptides (Singh et al. 1997)
Angiotensin-Converting Enzyme
(ACE)-inhibitory (Ong et al. 2007)
Gouda Anti-hypertensive (Saito 2000)
Finnish varieties:
Edam, Emmental,
Turunmaa, Cheddar
Angiotensin-converting enzyme
(ACE)-inhibitory
(Korhonen and
Pihlanto 2003)
Italian varieties:
Mozzarella, Crescenza, Italico,
Gorgonzola
Angiotensin-Converting Enzyme
(ACE)-inhibitory (Smacchi and Gobbetti 1998)
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problems. The fermentation of milk is described
as the process for releasing antioxidant
peptides from caseins. Superoxide anion and
hydrogen peroxide both are degraded by lactic
acid bacteria (Kullisaar et al. 2003). Histidine
and proline are the essential residues in the
lipoprotein per-oxidation activity of peptides.
Songisepp and coworkers in 2004, stated
that increasing ripening period enhances the
antioxidant activity of cheese (probiotic cheese).
Numerous peptides are formed in the cheese
during ripening process. These are originated
mainly from the casein as a result of proteolysis.
More peptides are released in the cheeses due
to the change in the proteolytic rate and its
pattern by the addition of adjunct cultures
(Ong et al. 2007). During the cheese ripening
process, major peptides are formed in the
water-soluble extract of cheeses. Antioxidant
peptides derived from milk proteins prevent
the peroxidation of essential fatty acids. Casein
digestion produces phosphorylated peptides.
These peptides exhibit both lypophilic and
hydrophilic antioxidant activity by sequestering
of metal ions and quenching of reactive oxygen
species (Díaz et al. 2003). Proteolysis is the
most tangled biochemical phenomena during
ripening of cheese. Bottesini et al. (2013) also
reported the potential free radical scavenging
activity of water soluble extract from Australian
cheddar cheese which is the result of
proteolysis. Antioxidant activity hikes on 2nd
Table 2. Different milk protein derived bioactive peptides exhibiting
biofunctional activity in cheddar cheese
Milk proteins Bioactive peptides Bio-functional activity Reference
κ-Casein, αs1-Casein
and β-Casein
κ-CN (f 96–102), αs1-CN
(f 1–9), αs1-CN (f 1–7), αs1-CN
(f 1–6), αs1-CN (f 24–32) and
β-CN (f 193–209).
ACE inhibitory Ong L and Shah NP
(2008)
β-casein β-Casomorphins Opioid Muehlenkamp and
Warthesen (1996);
Dionysius et al. (2000)
αS1- casein and
β-Casein
αS1- and β-CN (f1-6) Antioxidative Gupta et al. (2009)
αS1- and αS2- Casein
β-Casein
Several phosphopeptides Sing et al. (1997)
β-casein β-casomorphin-11 Opioid Meisel and
Bockelmann (1999)
κ-Casein Casoplatelin Antimicrobial,
Antithrombotic
Rizzello et al. (2005)
κ-Casein Kappacin Antibacterial Malkoski et al. (2001)
αS1- casein Isracidin Antimicrobial Hill et al. (1974)
αS1- casein αS1casokinin Antimicrobial Rizzello et al. (2005)
αs1-casein, β-caseins Caseinophosphopeptides,
Immunopeptides,
Casomorphins, Casokinins,
Prolactin (Interleukins-1,2,6,
& 10)
Immunomodulatory Dionysius (2000)
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month of ripening and starts decreasing in the
later stage of ripening. This is due to a further
breakdown of bioactive peptides by proteolytic
enzymes like plasmin (Hossain et al. 2018).
Various peptides have been identied in the
highest anti-oxidative fraction that seven of
the eight peptides contain at least one proline
residue and six of these eight peptides contain
more than two proline residues. Reactive
oxygen species is the cause of harmful diseases
namely cardiovascular disease, cancer dialects,
cataracts, neurodegenerative disorders, and
aging (Talegawkar et al. 2009). The body has
its mechanism against these reactive oxygen
species based on antioxidative enzymes such
as superoxide dismutase and catalase and
also endogenous enzyme namely glutathione.
If reactive oxygen species overloads the
antioxidant defense system of body then
oxidative stress occurs. It was concluded
that peptides more than 10Kda have better
antioxidant activity than smaller peptides. Raq
et al. (2017) and Huma et al. (2018) reported
the water-soluble peptides (WSP) extract from
cheddar cheese exhibited radical scavenging
activity and decreases there active oxygen
species (ROS) production in macrophages. It
can be stated that antioxidative properties of
cheddar cheese helps to boost up immunity by
decreasing the chances of free radical diseases.
Antimicrobial activity
The activity of antimicrobial peptides is
articulated as a breakdown of the cell membrane,
but the major target is the lipid bi-layer of
the cell membrane. For proper antimicrobial
activity, the most important requirement is the
interaction between antimicrobial peptide and
cell membranes. The antimicrobial peptides
activity is dened as specic membrane-lytic
activity in prokaryotic cell membranes (Floris
et al. 2003). Peptides that acquire antimicrobial
activity are of hydrophobic α-helical peptides.
Antimicrobial peptides can kill target cells
rapidly as compared to antibiotics. A wide
range of activity is shown by antimicrobial
peptides. The chief source of antimicrobial
peptides has been identied in milk, milk
hydrolysates, and fermented milk (Gobbetti et
al. 2004). Cheeses like gorgonzola, Parmigiano
and fossa, etc. do not show any antibacterial
peptides. Besides all cheese Pasta-lata types
of cheeses (Mozzarella, Caciocavallo) proclaim
potential antibacterial peptides. Antimicrobial
Peptide possesses a higher degree of homology
with C-terminal, N-terminal, or whole
fragments. Some examples of antimicrobial
peptides are αS1casokinin, isracidin, kappacin
and casoplatelin, β-casomorphin-11, etc.
Ace-inhibitory peptides
Angiotensin I converting enzyme (ACE)
plays vital role for regulation of blood pressure.
Renin enzyme regulates on angiotensinogen
to form angiotensin I, where decrease
blood volume or reduced blood pressure to
the kidney is perceived. ACE catalyzes the
hydrolysis of angiotensin I (decapeptide) to
angiotensin II (octapeptide). It increases the
blood pressure through vasoconstriction,
which results in higher sodium and water
resorption in the kidneys. Vasodilating peptide
bradykinin and opioid peptide Met-enkephalin
both are inactivated by angiotensin-converting
enzyme (ACE). The activity of ACE increases
with the development of proteolysis. Though,
proteolysis exceeds a particular level then
the ACE inhibition index decrement during
the maturation of cheddar cheeses (Hossain
et al. 2018). ACE-inhibitory activity hikes on
1st month of ripening and starts to decrease
on the later stage of ripening. This is caused
further more breakdown of bioactive peptides
by proteolytic enzymes like plasmin (Hossain
et al. 2018). In the case of enzyme-modied
cheese (EMC), ACE-inhibitory peptides are also
obtained by further hydrolysis of pasteurized
and homogenized cheese with neutrase
enzyme. Stuknytè et al. (2015) reported the
αs1-casein-derived peptides like RYLGY and
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AYFYPEL in cheddar cheese which helps to
lower the blood pressure and also reduces the
chances of cardiovascular diseases.
Opioid peptides
Food proteins are the sources of opioid
peptides. They have an attraction to bind with
opiate receptors and show signicant opiate
activity. Peptides that show opioid activities
are called as ligands of opioid receptors. It can
be reversed by naloxane, which is an opioid
antagonist. Naloxane can block opioid activity
by crossing the blood-brain barrier. It can also
be used as a particular tool for determining the
precise effect of agonist opioid peptides. Some
specic organs like adrenaline gland, gastro-
intestinal tract and the spine show opioid
activity. The various responses reported from
opioid peptides include hypotension, sedation,
inuence on satiation, dullness, stress response,
changes in sexual behavior, changes in body
temperature, respiratory disorder, analgesia,
anxiety etc. The appropriate concentration of
β-casomorphins in the cheese extract was less
than 2 mg/ml which can exhibit sufcient
opioid activity (Pihlanto and Korhonen2015).
At the ripening temperature of cheese some
factors inuencing the enzymatic degradation
of β-casomorphins, pH and salt concentration.
Dionysius et al. (2000) reported several opioid
peptides named β -CM 7 and β -CM 4 from
Australian vintage cheddar cheese. These
peptides provide the analgesia effect against
acute or chronic pain same as morphine drugs,
but does not exhibit any side effects.
Immunomodulating peptides
Immune cell functions can be enhanced
by bioactive peptides which are derived from
cheeses and fermented foods. In the case of
atopic humans, it may also reduce allergic
reactions. In the gastrointestinal tract, it increases
mucosal immunity (Korhonen and Pihlanto
2003). The proliferation of human lymphocytes
can be modulated by the peptides which are
liberate during the fermentation of milk with
lactic acid bacteria.91 peptides have been
identied in Emmental cheese out of them 28
peptides exposed several bioactive properties
such as mineral carrying, antimicrobial,
antihypertensive, immunomodulatory
activities (Gagnaire and Langlais 2005).
Various immunomodulatory peptides named
immunopeptides, casomorphins, casokinins,
prolactin (interleukins-1,2,6, & 10) have been
found in cheese. Dionysius et al. (2000)
reported several immunomodulatory peptides
along with ACE inhibitory peptides in cheddar
cheese. Caseinophospho peptides are one
of the most important immunomodulatory
peptides which have been found in numerous
cheeses like gouda, cheddar etc.
Anti-diabetic peptides
There is a worldwide increase in type 2
diabetes which is undesirable. Type 1 and type
2 diabetes are the major forms of diabetes and
it also contributes 10% and 90% of the total
world populatin (El-Sayed and Awad 2019).
Insulin deciency results various symptoms
in the body such as an increase in blood
lipid, total cholesterol, and free fatty acids
at the liver in large amounts. Whey protein
ingestion causes more secretion of insulin
than micellar casein. As per WHO, there are
about 3.2 million deaths per year due to this
diabetes. Insulin secretion can be increased
by whey and casein ingestion. Whey protein
ingestion helps to secret insulin more rapidly
that casein ingestion (Nilsson et al. 2007). A
greater in vivo insulinotropic effect in cheese is
observed due to hydrolysed milk as compared
to un-hydrolysed milk proteins (Power et al.
2009).Whey protein derived bioactive peptide
named tripeptide Ile-Pro-Ala, released from
β-lactoglobulin hydrolysis, reduces glucose
levels and stimulates insulin (Tulipano et
al. 2011). Han et al. (2013) conducted an
experiment and reported that β CM-7 peptide
reduces the blood glucose level in rat. Garcıa-
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Nebot et al. (2014) reported that antioxidative
peptides reduce chances of diabetes by
suppressing the ROS. No specic research work
has been found regarding antidiabetic property
of cheddar cheese. This article encourages
the further research regarding this property in
cheese, especially in cheddar cheese.
Anti-thrombotic activity
Thrombus formation in blood circulatory
system leads to enhance the chances of several
deadly diseases like cardiovascular diseases,
cerebral venous sinus thrombosis (CVST),
etc. Fermented milk products like cheddar
cheese is a surplus sources of anti-thromobotic
peptides which helps to prevent this diseases.
Casoplatelins is the major anti-thrombotic
peptides released in fermented product by
hydrolysing κ–casein (Raq et al. 2020). This
casoplatelins inhibits the brinogen binding
process with platelets. Raq et al. (2017)
concluded that the anti-thrombotic property
of cheddar cheese increased with progression
of ripening period.
Anti-cholesteremic activity
Fermented dairy products exhibit
cholesterol lowering activity due to having
several bioactive peptides. Low density
lipoprotein is known as bad cholesterol as it
enhances the chances of heart attack and stroke.
This bad cholesterol is found in the wall of
blood vessels. On the other side, high density
lipoprotein is known as good cholesterol as it
carries low density lipoprotein to liver from
where it is further ushed from the body. It
also helps to protect nerves and make health
cells and hormones. Mainly β -lactoglobulin
hydrolysed bioactive peptide named lactostatin
exhibits anti-cholesteremic activity. Nagaoka et
al. (2001) reported that peptide with the IIAEK
sequence, isolated from β -lactoglobulin tryptic
hydrolysate, exhibit cholesterol lowering
activity. Hussain et al. (2020) revealed that
phytosterol esters enriched cheddar cheese
exhibited greater anti-cholesteremic activity
than normal cheese.
Conclusion
Cheddar cheese is the most popular
cheese worldwide and also a plethora of
biologically active peptides. It helps in the
maintenance of different body functions. By
consuming cheddar cheese different diseases
can be prevented due to the presence of
bioactive peptides. Anti-oxidant activity
scavenges the free-radicals formed in our body
and prevents us from various ROS assisted
diseases. The anti-microbial activity of cheddar
cheese protects us from microbial infections
of internal organs and keeps us healthy. ACE-
inhibitory activity helps us to control our blood
pressure level. Immunomodulatory activity
boosts our immunity. Opioid activity keeps our
nervous system healthy. Anti-diabetic activity
keeps the insulin secretion stable and keeps us
away from diabetes. Overall, regular cheddar
cheese consumption makes us healthy and safe
from many deadly diseases.
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Chapter
This chapter describes the manufacture of Cheddar cheese and related dry-salted cheese varieties. Curd formation, whey separation, cheddaring, milling, salting, and pressing are described in detail, including an illustration of how the microstructure of this cheese develops. The chapter also explains the main factors that determine Cheddar cheese quality including the chemical composition, texture, and flavour of the cheese. It summarizes the role of lipolysis, proteolysis, starter, nonstarter lactic acid bacteria, or adjunct cultures in the development of Cheddar flavor during ripening. Although there is no universal standard for measuring Cheddar quality, different grading and assessments of Cheddar cheese are also presented including sensory evaluation and the use of instrumental analysis to measure key aspects of Cheddar cheese ripening.
Article
The generation of bioactive peptides (BAPs) from dietary proteins has been widely studied. One of the main limitations of a broader application of BAPs in functional foods may arise from their low potency. Therefore, the search for more potent structures is crucial. Quantitative structure–activity relationship (QSAR) has been widely applied in drug discovery and some examples may also be found in the study of BAPs. The aim of this review was to assess the efficiency of QSAR for the discovery of novel and potent BAPs, derived from food protein sources. A wide range of bioactive properties including antioxidant, antimicrobial, angiotensin converting enzyme (ACE), renin and dipeptidyl peptidase IV (DPP-IV) inhibition as well as bitter peptides has been investigated with QSAR. Some studies have identified structural requirements for specific bioactivities, which generally confirmed findings from earlier studies carried out on those BAPs. However, discrepancies are found across analyses, possibly due to the quality of the peptide datasets as well as the descriptors used to build QSAR models. It appears to date that only a limited number of QSAR studies conducted with BAPs have subsequently carried out confirmatory studies and evaluated promising peptide sequences in vivo. This suggests that more research is needed in order to advance knowledge in the area of BAP discovery using QSAR.
Chapter
Milk proteins and milk protein-derived peptides have been widely studied for their health enhancing properties. This chapter presents the updated scientific knowledge on the bioactive properties of milk protein-derived peptides. The different bioactive properties which have been attributed to milk protein-derived peptides are discussed. These include mineral binding, cardioprotective, antidiabetic, satiating, opioid, antimicrobial, immunomodulatory, anticancer and antioxidant activities. The structure-function relationship is presented for the aforementioned bioactive properties based on current scientific knowledge. For each bioactive property, the data obtained in vitro is discussed, followed by an analysis of the information obtained from animal and human intervention studies. To date, most studies have been conducted in vitro. However, an increasing number of in vivo studies testing the efficacy of milk protein-derived peptides are being conducted. In certain instances, the in vivo studies have confirmed the bioactivity of specific milk protein-derived peptides or milk protein hydrolysates. However, conflicting data still exist in the scientific literature, which demonstrates that the bioactive properties observed in vitro do not always translate in vivo. Detailed knowledge of the peptide sequences responsible for the bioactive properties, together with a better understanding of the bioavailability and stability of these peptides in vivo may help to enhance the development of milk protein hydrolysates with health promoting capabilities in humans. Ultimately, this may lead to the approval of health claims by the relevant regulatory agencies.
Chapter
Fermented milk and soybean products naturally have high nutritional value and many health-promoting effects, which may be attributed to the release of bioactive peptide sequences. Peptides and peptide fractions with bioactive properties (e.g. immunomodulatory, hypocholesterolemic, anti-hypertensive) have been isolated from fermented dairy products and in many traditional fermented soy foods.Among bioactive food peptides, those with anti-hypertensive activity have received special attention owing to the high prevalence of hypertension in Western countries. Peptide fractions have been reported to decrease systolic blood pressure in spontaneously hypertensive rats and in mild hypertensive human volunteers.This chapter reviews the current literature on bioactive peptides released during fermentation of milk and soybean, with a special focus on peptides relevant to cardiovascular health.
Article
Water-soluble peptides from Mozzarella, Italico, Crescenza, and Gorgonzola cheeses were fractionated by reverse-phase fast protein liquid chromatography. Peptide fractions with inhibitory activity to amino- and endo-peptidases from Lactobacillus delbrueckii ssp. bulgaricus B397, Streptococcus thermophilus 305, and Lactococcus lactis ssp. cremoris Wg2 were found. Enzymes from Lactobacillus casei ssp. casei 2752 were less sensitive. Endopeptidase from Lactobacillus casei ssp. casei 2752 also had a different response to the effect of some inhibitors. It probably showed limited differences in catalysis and substrate positioning. Most of these inhibitory peptides were also effective in reducing the activity of the Pseudomonas fluorescens ATCC 948 endopeptidase and the angiotensin I-converting enzyme. Inhibitory peptide fractions from Mozzarella, Italico, and Crescenza cheeses had a certain degree of hydrophobicity while the peptide fraction from Gorgonzola cheese eluted in the initial part of the acetonitrile gradient. One of the inhibitory peptides contained in the water-soluble extract of Crescenza cheese was further purified and sequenced. It corresponded to the β-casein fragment 58-72.
Article
This study examined the presence of antimicrobial, antioxidant and antihypertensive peptides in three commercially available Australian Cheddar cheeses. Peptide extracts as well as fractionated peptide extracts were examined. Commercial cheese A peptides exhibited the greatest inhibition against Bacillus cereus and also commercial cheese A fractionated peptides greater than 10 kDa showed the highest inhibition against B. cereus. Commercial cheese A peptides also showed the highest inhibition of 2,2-diphenyl-1-picrylhydrazyl (DPPH), a free radical used to measure antioxidant activity. All cheese fractionated peptides greater than 10 kDa demonstrated higher inhibition of DPPH after fractionation. Antihypertensive peptides were determined by inhibition of the angiotensin-converting enzyme (ACE). Overall, commercial cheese A had the lowest concentration required to inhibit ACE and commercial cheese A fractionated peptides lower than 5 kDa had the lowest inhibition after fractionation. These preliminary findings suggest that peptide extracts of three commercial Australian Cheddar cheeses exhibit antimicrobial, antihypertensive and antioxidant properties.
Article
The aim of the study was to examine the release of angiotensin-converting enzyme (ACE)-inhibitory peptides in Cheddar cheeses made with starter lactococci and Bifidobacterium longum 1941, B. animalis subsp. lactis LAFTI® B94, Lactobacillus casei 279, Lb. casei LAFTI® L26, Lb. acidophilus 4962 or Lb. acidophilus LAFTI® L10 during ripening at 4 and 8 °C for 24 weeks. ACE-inhibitory activity of the cheeses was maximum at 24 weeks. Cheeses made with the addition of Lb. casei 279, Lb. casei LAFTI® L26 or Lb. acidophilus LAFTI® L10 had significantly higher (P < 0.05) ACE-inhibitory activity than those without any probiotic adjunct after 24 weeks at 4 and 8 °C. The IC50 of cheeses ripened at 4 °C was not significantly different (P > 0.05) to that ripened at 8 °C. The lowest value of the IC50 (0.13 mg mL−1) and therefore the highest ACE-inhibitory activity corresponded to the cheese with the addition of Lb. acidophilus LAFTI® L10. Several ACE-inhibitory peptides were identified as κ-CN (f 96–102), αs1-CN (f 1–9), αs1-CN (f 1–7), αs1-CN (f 1–6), αs1-CN (f 24–32) and β-CN (f 193–209). Most of the ACE-inhibitory peptides accumulated at the early stage of ripening, and as proteolysis proceeded, some of the peptides were hydrolyzed into smaller peptides.
Article
Preclinical and clinical studies suggest that whey proteins can reduce postprandial glucose levels and stimulate insulin release in healthy subjects and in subjects with type 2 diabetes by reducing dipeptidyl peptidase-4 (DPP-4) activity in the proximal bowel and hence increasing intact incretin levels. Our aim was to identify DPP-4 inhibitors among short peptides occurring in hydrolysates of β-lactoglobulin, the major whey protein found in the milk of ruminants. We proved that the bioactive peptide Ile-Pro-Ala can be regarded as a moderate DPP-4 inhibitor.