Fish Protein Hydrolysate (FPH)as a source of Bioactiveas a source of BioactivePeptidesPeptides Kashmeera N.A.Kashmeera N.A.IV Sem MSc ZoologyIV Sem MSc ZoologyRoll no: 37Roll no: 37Christ collegeChrist college
Acute protein deficiency in dietof females primary issue inAttappady• Source: Keralanext, May 29 2013, 6:15am• The high infant and neonatal mortality inAttappady is primarily due to acuteprotein malnourishment orKwashiorkor disease while generalmalnutrition or anaemia is only acontributing factor.
1 in 3 of the worldsmalnourished childrenlives in India
• Food proteins have long been recognizedfor their nutritional and functionalproperties.• In recent years, a considerable amount ofresearch has also focused on the liberationof bioactive peptidesbioactive peptides which are encryptedwithin food proteins, with a view to utilizingsuch peptides as functional foodingredients aimed at health maintenance.
• Bioactive peptides have been defined as “food derivedcomponents that, in addition to their nutritional valueexert a physiological effect in the body”.• Within the parent protein sequence, the peptides areinactive and must be released to exert an effect.• These bioactive peptides are usually 2–20 amino acidresidues in length, although, some have been reportedto be >20 amino acid residues.• Bioactive peptides may be absorbed through theintestine where they subsequently enter the circulatorysystem intact to exert various physiological effects, orthey may produce local effects in the digestive tract.
• Food derived bioactive peptides havebeen shown to display a wide range ofphysiological functions including• antihypertensive,• antioxidative,• appetite-suppressive &• Immunity strengthening effects.
PRODUCTION OF BAPPRODUCTION OF BAPImportant source – Fish ProteinImportant source – Fish ProteinHydrolysateHydrolysate
• Enzymatic hydrolysis of proteins producespeptides with specialphysicochemical,functional and biologicalactivities based on the sequence andaminoacid composition.• This is the most common way to producebioactive peptides from whole proteinmolecules/parent proteins.
Fish Protein HydrolysateFish Protein Hydrolysate(FPH)(FPH)• Proteins that are chemically / enzymatically broken downProteins that are chemically / enzymatically broken downinto peptides of varying sizes.into peptides of varying sizes.• Prepared from whole fish/fish muscle/fishery by-productsPrepared from whole fish/fish muscle/fishery by-productsof low value fishes/seafood industry waste.of low value fishes/seafood industry waste.• Good source of bioactive peptides.Good source of bioactive peptides.• Biological activities of FPH may vary based on enzymeBiological activities of FPH may vary based on enzymeused for hydrolysis,degree of hydrolysis, size ofused for hydrolysis,degree of hydrolysis, size ofpeptides,sequence & composition of amino acidpeptides,sequence & composition of amino acidresidues.residues.
• Hydrolysis of the protein substrate.Hydrolysis of the protein substrate.• Hydrolysates are assayed for various bioactivities.Hydrolysates are assayed for various bioactivities.• Crude protein hydrolysates are then fractionated basedCrude protein hydrolysates are then fractionated basedon peptide size (using Ultrafiltration).on peptide size (using Ultrafiltration).• The hydrolysate fraction displaying the highest bioactivityThe hydrolysate fraction displaying the highest bioactivityis then further purified to separate individual peptidesis then further purified to separate individual peptides(using high performance liquid chromatography (HPLC)(using high performance liquid chromatography (HPLC)or gel permeation chromatography).or gel permeation chromatography).• Individual peptide fractions are identified using theIndividual peptide fractions are identified using thecombined techniques of mass spectrometry and proteincombined techniques of mass spectrometry and proteinsequencing.sequencing.• Lastly, a synthetic version of the peptide is synthesizedLastly, a synthetic version of the peptide is synthesizedand the assay is repeated to verify bioactivity.and the assay is repeated to verify bioactivity.
ANTIHYPERTENSIVE / ANGIOTENSIN –IANTIHYPERTENSIVE / ANGIOTENSIN –ICONVERTING ENZYME INHIBITORY PEPTIDESCONVERTING ENZYME INHIBITORY PEPTIDES• Hypertension is one of the major risk factors inthe development of cardio-vasculardiseases,stroke & end stage of renal disease.• Renin – angiotensin system plays a key role inmaintaining blood pressure homeostasis as wellas fluid & salt balance in mammals.• Angiotensin I converting enzyme[ACE] belongsto class zinc protease that needs Zn & Cl- foractivation.
• ACE plays important role in blood pressureACE plays important role in blood pressureregulation by virtue of renin- angiotensin system.regulation by virtue of renin- angiotensin system.• ACE converts inactive form of angiotensin I toACE converts inactive form of angiotensin I topotent angiotensin II.potent angiotensin II.• Therefore inhibition of ACE is importantTherefore inhibition of ACE is importanttherapeutic approach for treatment oftherapeutic approach for treatment ofhypertension.hypertension.• Many ACE inhibitors have been isolated fromMany ACE inhibitors have been isolated fromFPH.FPH.
ANTIOXIDANT ACTIVITY OFANTIOXIDANT ACTIVITY OFBIOACTIVE PEPTIDESBIOACTIVE PEPTIDES• Many diseases like stroke, cancer, diabetes,cataract,arthritis & neuro-degenerative disorders werelinked to damage caused by highly reactive free radicals.• Free radicals are compounds with unpaired electronsthat stabilise themselves by oxidising othermolecules,including proteins, carbohydrates, lipids &DNA.• In this process they create more free radicals, sparkingoff a chain of destruction,leading to oxidative damagesresponsible for most diseases.
• BAPs have capacity to prevent oxidativedamage & to reduce risk of diseases & aging.• Many peptides including those from marinefishes have anti-oxidant activities againstperoxidation of lipids.• Peptides from FPH have free radical scavengingactivity, reducing power & autooxidationinhibition.• Presence of electron donating molecules in thehydrolysate provides the ability to preventoxidation process.
ANTIBACTERIAL PEPTIDESANTIBACTERIAL PEPTIDES• Inhibit growth of gram positive& negativebacteria.• Gram negative more resistant – due to cell wallnature.• Antagonistic activity of bioactive peptides isexerted by different mechanisms.• Most common – pore formation in cell wall.• Pore → lose ions & ATP.• Loss of ions → dissipation of proton motive force→ loss of viability of organisms.
IMMUNOMODULATORYIMMUNOMODULATORYPEPTIDESPEPTIDES• Peptide from salmon hydrolysate showedimmunomodulatory effect on mice.• Enhanced capacity of lymphocyteproliferation,NK cell activity,% of T- helpercells in spleen & secretion of interferon &cytokines.
ANTI-APPETISING PEPTIDESANTI-APPETISING PEPTIDES• Cholecystokinin – produced by intestinalendocrine cells – in response to mealstimulation.• Peptides from crustacean by productshydrolysate stimulate cholecystokinin release.• Such peptides incorporated into formulatedfoods control excess intake of food & therebyobesity.
CONCLUSIONCONCLUSION• Optimal exploitation of BAPs for human nutrition& health possesses an exciting scientific &technological challenge,while at the same timeoffering potential for commercially successfulapplications.• BAPs can be incorporated in the form ofingredients in functional and novel foods, dietarysupplements & even pharmaceuticals with thepurpose of delivering specific health benefits.