Modification of Proteins

Bimlesh Mann
 Aims
• Blocking the reactions involved in
deterioration of the food e.g. M R
• Improving some physical properties of
proteins e.g. Texture foaming, emulsifying,
gelling
• Improving the nutritional value
1. Increasing the extent of digestibility
2. Inactivation of toxic on other
undesirable constituents
3. introducing essential ingredients such
as amino acids
 Enzymatic Modification
• Limited proteolysis
• Proteolytic modification-improve solubility
e g cereal proteins solubility increase with
increase in the degree of hydrolysis
Improve the moisture sorption and water binding
Moisture uptake related to the Number of ionic
gps-these groups increases on proteolysis due
to release of amino and carboxyl gps
• Improve emulsifying properties-Due to
exposure of buried hydrophobic gps-HLB
• Partial cleavage and unfolding of proteins-
improve foaming properties –Excessive
digestion-loss of globular structure less
stable foam
 Progressive proteolysis
• Bioactive peptides
• Debittering the hydrolysates
 Proteins and Peptides
Nutritional-energy& amino acids
Functional-physicochemical&
sensory
Biological-sp. Biological
properties
potential ingredients for
functional or health
promoting foods
 Bioactive peptides -breakdown
products of proteins
Biologically active peptides or functional
peptides are food-derived peptides that
in addition to their nutritional value
exert a physiological effect in the body.
(Vermeirssen et al., 2004)
 Characterstics of bioactive
peptides
• Perform sp. Biofunction after release
from parent protein
• Activity depend on the a.a. comp.& seq.
• 3-20 a.a.
• Hydrophobic a.a. alongwith
proline,lysine& arginine
• resistant to the action of digestive
peptidases
POTENTIAL APPLICATION AREAS
Physiological Functionality of
Milk-
Derived Bioactive Peptides
ACE INHIBITORY PEPTIDES
• Angiotension converting enzyme(ACE)
hydrolyzes inactive angiotension 1 to the
octapeptide angiotension 2 (vasoconstrictor)-
increase blood pressure .
• ACE also hydrolyze bradykinin –a vasodilator
• ACE inhibitors are antihypertensive peptides.
• Specific inhibitors of ACE proven to be useful
antihypertensive drugs.
• Peptides known as Casokinins.
 OPIOID PEPTIDES

• Having pharmacological
similarities to opium.
• Show contraction of smooth
muscle similar to morphine.
• Casein derived peptides -
Casomorphins
 IMMUNOMODULATING
PEPTIDES
• Stimulates
– macrophage activity against red blood
cells
– phagocytic activity
– proliferation and maturation of T cells
and natural killer cells for defense of new
borne against large number of bacteria
– Obtained from casein and casein by
tryptic and chymotryptic hydrolysis.
 MINERAL BINDING PEPTIDES
• Prevents the precipitation of calcium
phosphate and increase the
concentration of soluble calcium in vitro
and in vivo (lumen of small intestine of
rat).
• CPPs - and caseins - in vivo and in
vitro by gastrointestinal trypsin from
and caseins- can be used as
anticariogenic additive.
• Prevents enamel demineralization.
 ANTIOXIDANT PEPTIDES
• Replenish aging induced loss of defense
system against oxidative stress.
• Inhibitors of lipid peroxidation.
• Direct scavengers of free radicals.
• Agents to chelate transition metal ions that
catalyze generation of radical species.
• Health related functional properties like
antioxidant activity may present a promising
biological benefits of these fermented milks.
ANTIMICROBIAL PEPTIDES
Active against Gram positive & Gram
negative organisms.
Protect against microbial challenges
in intestinal tract.
Support non immune defense in gut.
Increases cell permeability- cell death.
Act as bio-preservative
 Debittering of hydrolysates
• Bitter peptides
1. Nature of protein
2. Protease used
Bitterness is a function of
1. A A Composition
2. A A sequence
Excessive proteolysis with exopeptidases for
debitteringof protein hydrolysates
 Plastein reaction
• Protein hydrolysates at high conc,
-incubate with protease-formation of a ppt
or gel like structure
• Due to condensation and transpeptidation
• The plastein reaction enables peptide
fragments of hydrolysates to join
enzymatically through peptide bonds,
forming a larger Polypeptide
 Reaction
• R-CO-NH-R’ +E-OH↔R-CO-O-E +H2N-R’
↓
R2-NH2 H2O

R-CO-NH-R2 R-COOH
+E-OH +E-OH
 Applications
• Debittering
• Improving nutritional quality
• Improving functional characteristics
• Neutraceutical-Phe free diet
Reactn rate
Nature of AA
Hydrophobic residues preferably linked
together
 Glycosylation
• Introduction of saccharides into proteins
• Make the protein more hydrophilic
• Improve functionality
1. Gluco-CHO+H2N-Protein→glyco-CH=N-Protein
↓NaCNBH3
glyco-CH-NH-Protein
2. Polysaccharides-Dextran+egg albumin,
pectin+WP-60°C→ Conjugate-excellent
emulsifying
3.Lysozyme-heat stability increase by
conjugation with galactomannan
4.Neutral sugars favors the foaming
properties
5.Charged carbohydrates improved the
emulsifying properties
6.Transglutaminase- solubility of the
neoglycoprotein increased markedly
over the native protein in the isoelectric
pt range
 Phosphorylation
• The important proteins of human diet
eg.Milk casein,egg yolk phosvitin
• Introduction of phosphoryl residues
increase the negetive charge and
hydration and changes the functional
properties of the protein.
• Phosphorus oxychloride and sodium
trimeta phosphate used on large scale
 Phosphorylation
• Protein-NH2+POCl2→Protein-NH-PO-23+3HCl
• Protein-OH+POCl2→Protein-OPO-23+3HCl
 Acylation
• Acetic,succinic and maleic anhydride
• Improve the functional properties
• Protein-NH2+Acetic anhydride pH>7
Protein-NH-COCH3+CH3COO-+H+
Change in Isoelectric Pt by blocking NH2
Gp.by acyl residues to lower pH values-
Increase in solubility at lower pH region
 Deamidation
• The deamidation of glutamine and
aspargine in proteins result in the
liberation of the carboxyl gp. and thereby
to an increase in change and hydration.
 Esterification
• The carboxyl gp. Of proteins can be
esterified with methanol containing small
amounts of HCL
• The esterification reduces the No. of
anionic Gps. in the protein and Increases
the Isoelectric Pt.
• Est.of casein increases its thermal stability
• Protein-COOH+CH3OH Protein-
COOCH3 +H2O
 Protein Crosslinking
• The introduction of artificial crosslinks into
food proteins may be useful for improving
the functional properties.
• Transegluminase,peroxidase, polyphenol
oxidase are suitable enzymes for
crosslinking
 Alkylation
• Modification of protein by reductive
methylation of amino Gps with
formaldehyde/NaBH4 retards maillard
reactions.
• The resultant methyl
derivative,depending on the degree of
substitution is less accessible to
proteolysis.
Redox reactions involving cysteine
and cystine
• Disulfide bonds have a strong influence on
the properties of proteins.
• Wheat gluten can be modified by reduction
of its disulfide bonds to sulfahydryl gps
and subsequent reoxidation of these
groups under various conditions