Recent Advances in Food Biopeptides:
Production, Biological Functionalities And
Therapeutic Applications
Humaira Hussain
PhD Scholar-BCH
10-arid-1763
Contents
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Introduction
Production of biopeptides
The production of endogenous peptides
Production of biopeptides with enzymatic hydrolysis.
Production of biopeptides with microbial fermentation
Biopeptide generation during in vitro and in vivo proteolytic
digestion
Isolation and purification of bioactive peptides
Extraction of bioactive peptides
Seperation of bioactive peptides
Purification of bioactive peptides
Biopeptide transportation modes and absorption routes
Routes For Peptide’s Transport
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Biopeptide Resistance and Bioavailability
General concerns related to in vitro biological activities
General concerns related to in vivo biological activities
Role of Biopeptides
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In scavenging oxidative stress
As antihypertensive agents
In modulating the immune disorders
Antimicrobial and anti-fungal properties
Opiate-like activity
Anticoagulant activity
• Future prospects
• Conclusion
Introduction
• Many naturally occurring compounds from foods such as rice,
vegetables, fruits, and animal products possess properties that
help to slow disease progression, inhibit patho physiological
mechanisms, or suppress activities of pathogenic molecules.
• Proteins and peptides play significant roles in such activities
• Gaining importance as nutraceuticals that benefit numerous
aspects of health and nutrition.
• Peptides are a kind of health protectant present in nature.
• Recent developments in the field of proteomics offering
promising solutions to solving health problems stimulates the
uses of biopeptides as one of the therapeutic.
• Functional peptides(Bio peptides) are typically produced from
protein via enzymatic hydrolysis.
• The attention that has been given to biopeptides is due to their
therapeutic and medicinal potential.
Production of biopeptides
• The common approaches used in the production of
biopeptides are as follows:
• The production of endogenous peptides
• Production of biopeptides with enzymatic hydrolysis
• Production
of
fermentation
biopeptides
with
microbial
The production of endogenous peptides
• These types of biopeptides usually have an internal origin.
• They are secreted with particular routes either by neurons
granules or from endocrine cells
• Biopeptides are considered as captivating biomarkers in the
living systems (cells, tissue, and body fluids)
• The identification and characterization of endogenous peptides
have proposed many challenges and limitations.
Production of biopeptides with enzymatic
hydrolysis
• The enzymatic hydrolysis is the efficient treatment destined for
the production of biopeptides
• Milk proteins are an important source in generating biopeptides
with different biological activities including antioxidative,
antihypertensive.
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The incorporation of protein hydrolysates into foods has been
increasingly applied in the food industry and is of great interest
for human organisms.
• The benefit of this technology is to enhance the daily diets of
humans.
• The physiological effects of biopeptides in regulating blood
pressure have prompted the scientists to exploit different food
materials such as fish, meat, milk, eggs, beans, and plants.
 For instances, goby is a kind of fish that can be exploited for
deriving biopeptide based hydrolysates.
Production of biopeptides with microbial
fermentation
• The production of biopeptides using microbial fermentation
has drawn much of the dairy industry's attention
• Milk products remain the captivating materials in generating
potent biological peptides by the intervention of particular
microbial proteases.
• Many examples are elaborated on the potentiality of microbial
fermentation in the production of dairy products such as
commercial probiotic bacteria, yogurt bacteria,and cheese
starter bacteria, all these examples being the outcomes of the
process of fermentation .
Biopeptide generation during in vitro
and in vivo proteolytic digestion
• Extraction process is an important step in the production of
high yield of targeted compounds.
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In the current review more emphasis is given to the enzymatic
hydrolysis due to its advantages including
High yield
Efficient process
Time saving treatment
and it can be scaled easily in the laboratory
• It does not leave residual organic solvents and excess of toxic
substances resulting from the secondary metabolites during
long fermentation process of foods.
• Owing to their low molecular weight, peptides (e.g.
polypeptides) behaviors are predictable in unusual fashion in
terms of absorption, digestibility, and solubility affinities
• Modification process of these bio fragments by using
enzymatic methods is preferred in order to improve their
resistance, and bioavailability.
• The enzymatic hydrolysis is the best technique and in most of
the cases it needs to be controlled under specific operational
conditions
Degradation behavior of Proteolytic Enzymes before and after enzymatic hydrolysis.
• The enzymatic hydrolysis has shown a direct influence on the
hydrophobicity.
• Each enzyme is responsible for the generation of different
fragments of peptides ranging between low, middle and high
molecular weight.
• These short peptides are preferred due to their stability and
resistance under gastrointestinal tract conditions, and easy to
be carried by specific precursors, with low transmission
energy.
• Due to their special structure and high molecular mass,
proteins are more fragile and unstable.
Isolation and purification of bioactive
peptides
• The emergence of robust technologies of purification and
separation science of bioactive systems starts to enforce the
research area of bioactive peptides to be more pronounced and
mechanistic.
• Typically, the variability in molecular weights, charges, affinity
of bioactive peptides during separation and purification caused
many obstacles
• Because of these barriers and challenges related to the purity of
peptides the scientists' proposed robust tools and advance
instrumentation for solving these encountered issues.
Extraction
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Size-exclusion chromatography (SEC)
Solvent extraction
Buffer saline systems.
mobile phase that is used to purify peptides from fermented
protein hydrolysates is a mixture of the three elements
including trifluoroacetic acid (TFA), water, and acetonitrile
• Several steps were followed accordingly starting by
centrifugation
• then the filtration of the supernatant
• finally lyophilization of the sample for further liquid
chromatography purification and separation.
Seperation
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HPLC is the common tool in the separation of bioactive
compounds.
RP-HPLC columns .
In contrast the application of normal HPLC is preferred in case
when the targeted bioactive peptides are hydrophilic peptides.
Other chromatographic techniques including
Capillary electrophoresis (CE),
Capillary isoelectric focusing (CIEF),
Ion-exchange chromatography (IEC) could reach to separate
peptides based on their chargeability affinities either positively
or negatively charged bioactive peptides
Purification
• Other techniques of separation and fractionation that can be
employed in the purification trajectory are
• Ultrafiltration,
• Crystallization,
• Partition chromatography .
• Many of the national and international health centers used these
materials to enrich and supplement certain products in order for
them to maintain and serve many of the biological activities
including antioxidative stress, antihypertensive.
Biopeptide transportation modes and
absorption routes
• Their molecular weight is very low in comparison to the molecular
mass of protein.
• Short peptides comprising one or two amine bonds are penetrated
through the brush border membrane by either
• Physical transmission phenomenon
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By the intervention of specific proteins transporter agents .
• The short peptides have been found to be more active and their
rapidity of absorption is favored, thus, in certain cases their
penetration routes are expected to be much vital than amino acids.
• For instance, some specific carrier of peptide named as PepT1
which highly necessitates the driving force resulting from the
electrochemical proton gradient.
• Moreover, various intracellular peptidases could be involved
• Other amino acids residues will be oriented to the portal
circulation via specific transporter agents.
Routes For Peptide’s Transport
• Basically, there are 5 principal routes for peptides:
1. The paracellular route, which is based on the diffusion
mechanism, depending on water-soluble peptides, but
independent on energy sources
2. The passive diffusion which is an auto-mechanism
independent on energy source ,for hydrophobic peptides
3. By the intervention of specific transporter, this mode of
transport usually acts more in the presence of short peptides
having hydrolysis-resistance .
4. Endocytosis,, which is usually adaptable with the large polar
peptides .
5. Lymphatic system, this mode of transmission, in fact more
suitable to the highly lipophilic peptides, because they can be
retained easily into the portal circulation
Biopeptide Resistance and Bioavailability
• Low molecular mass and other properties like hydrophobicity are
expected to be one of the dominating factors responsible for their
biological activities
• Peptide chain comprising two to six amino acids have been found to
be transmitted into portal circulation readily than those having long
amino acid residues.
• Hepatic metabolism can be escaped when the transmission of this
active systems occur via the gastrointestinal lymphatic route.
• Presence of proline or double proline on the lateral chain of
peptides, because they contribute to maintain good resistance and
rigidity for peptide.
• Excess amount of biopeptides lead to subsaturation of peptide
transporters.
• Once these active substances remain for a long time within the
stomach and under the action of gastric enzyme secretion and
later by pancreatic enzymes, the biopeptides in this case will
become more fragile and their transportation will be retarded.
• For this reason, the microencapsulation is needed to maintain
this stability
General concerns related to in vitro
biological activities
• Cell model culture is considered the first choice for fast
screening of the biological activity of such vital substances,
• The manipulation of this model is easier and more reproducible
than animal studies.
• The role of these chemical elements have involved in
maintaining vital mechanisms such as, antibacterial activity,
anticancer activity.
General concerns related to in vivo
biological activities
• Basically, once the biological activities such as antioxidative,
are more pronounced during in vitro manipulations screening,
in vivo study should be carried out.
• An accepted animal model, which is called “Spontaneously
Hypertensive Rats”, is one of the animal strains that is getting
much acceptance by all researchers.
• On the other hand,, due to its similar biological behaviors to
the human organisms, particularly in case of an essential
hypertensive human.
• Researches on antihypertensive activity of biopeptides used
SHR
Role of biopeptides in scavenging
oxidative stress
• Many scientists verified the antioxidative properties of milkenriched with fish oil and during the assessment they noted that
the lower molecular fractions (3–10 kDa andb3 kDa) are found to
have the most effect on iron-chelating activity rather than other
fractions having high molecular weight.
• Strong oxygen radical- scavenging effect exhibited in a sample of
egg white protein.
Role of biopeptides as antihypertensive
agents
• Biopeptides can be defined as authentic components with monobi-and multi-functional biological activities exhibited from their
amino acid residues
• Capable to serve different mechanism of action like intonation,
stimulation, depression, and inhibition once they exist with
adequate dosages in the living organism.
• The antihypertensive activity of bovine milk-derived biopeptides
and other food proteins such as legumes/beans gelatin, and yeast
has been reviewed extensively .
Role of biopeptides in modulating the
immune disorders
• However, the immunomodulatory properties of biopeptides are
seemed to be not appreciated.
• This is perhaps due to the many challenges occurred during the
practical application, due to the complexity of chemical reaction
and mechanism effect resulting by the end of primary and
secondary metabolites within the living system.
• Fraction F6 that is obtained from the fermented skim milk
displayed a potent immuno stimulating function on the
proliferation of murine spleen lymphocytes.
• Lactoferricin can be categorized among the potent peptides
Biopeptides with antimicrobial and
anti-fungal properties
• Antimicrobial biopeptides comprised of less than 50 amino acid
residues with a molecular mass of less than 10 kDa.
• The majority of amino acid residues are hydrophobic in the
nature.
• They can exhibit an efficient role in the host defense against the
most frequent pathogenic bacteria
• Agar diffusion assay, which is named as inhibition zone assay is
used to identify them
Biopeptides with anticancer and antitumor activities
• Protein hydrolysates exhibit a pivotal biological role against
cancer cells
• A recent study has been successfully identified an anti-cancer
food from shellfish Mytilus-coruscus by using pepsin
hydrolysates.
• Biopeptides has become an alternative molecules compared to
other efficient drugs because of safety point of view.
• Peptides that have the ability of inducing the apoptosis in tumor
cells are implicated to be as effective anticancer agents .
Biopeptides with opiate-like activity
• kind of peptide that can be found in bovine milk.
• Reported to play a central role in the nervous network within
human body.
• Exhibit similarity mode of action of morphin.
• This type of peptide acts by adhering with targeted cell receptor.
• For example µ-receptor, which can induce the emotional behaviors,
and it found to suppress as well the motility of intestine.
Biopeptides with anticoagulant activity
• Used to decline the bleeding in the same time to regenerate and
reconstitute the damaged regions at the wall of the blood vessel.
• 13 of plasma serine proteases are encrypted in the mechanism of
blood coagulation .
• Many of the marine organisms such as Tegillarca granosa (Blood
cockle),,Mytilus edulis(blue mussel) used derive biopeptides as an
anticoagulant agent.
• A potent anticoagulant biopeptides have been isolated from marine
macro and microalgae strains.
Future prospects
• Food biopeptides are versatile materials for regulating human
bodily functions.
• Still potential exists to verify their bio-resistance to ensure their
intact absorption into portal circulation.
• Best way to maintain this bio-resistance is by microencapsulation
technique.
• There is need to emphasize on the resulting metabolites (primary
and secondary) effect and their communication at cellular level by
coupling both of proteomic and of genomic sciences in
combination with chemical engineering.
Conclusion
• Due to biophysical and biochemical properties biopeptides are
valued among the potential ingredients having the capability to
regulating food intake, scavenging oxidative stress, reducing
the risk of cardiovascular diseases and modulating the immune
disorders once they exist with an adequate dosage in the living
organisms.
• Insufficient findings on immunomodulating properties
compared to that done for antioxidative and antihypertensive
activities. This is because of the lack of expertise in the area of
immunology and due to the absence of a real linkage between
major fields such as pharmacy, medicine, biotechnology, and
chemical engineering.
References
• Sami Saadi, Nazamid Saari, Farooq Anwar, Azizah
Abdul Hamid, Hasanah Mohd Ghazali. Recent
Advances in Food Biopeptides: Production,
Biological
Functionalities
And
Therapeutic
Applications, Department of Chemistry, Universiti of
Sargodha, Sargodha 40100, Pakistan. Biotechnology
advances.2014.12.003