Proteolytic enzymes

For B.Pharm, 4th sem

1. Proteolytic Enzymes

2. Proteolytic Enzymes
Enzymesinvolvedindegradationof
proteinsarecalledasproteasesor
proteolyticenzymes
Industriallyavailableproteolyticenzymes
producedbymicroorganismsareusually
mixturesofendopeptidases and
exopeptidases.
Proteins Polypeptides
Polypeptides amino acids
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Endopeptidases
Exopeptidases

3. Cont …
Becauseofthecomplexstructuresandhigh
molecularweightsofproteinsenzymicproteolysis
isextremelycomplicated.
Mostproteasesarequitespecificwithregardto
whichpeptidelinkagestheycansplit
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4. List of proteolytic enzymes
a.Papain
b.Ficain
c.Bromelain
d.Pepsin
e.Rennin
f.Cathepsin
g.Trypsin
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5. A) Papain
◦Foundinpapaya
◦papain,alsoknownaspapayaproteinaseI
◦BroadpH(3-11)andtemperaturestability
Forthisreasonverypopularforavarietyoffood
applications.
1.Usedasameattenderizeroninferiormeatcuts(canalsouse
sliceofpineappleonmeat)
Theenzymemakesitswayintothemuscleand
hydrolyzesprimarilyconnectivetissueproteins(collagen
etc.)andsoftensmuscle
Havetouselowamounttopreventliquefactionof
muscle
IfyoumixrawpapayaintoJell-Oitwillnotformagel

6. Application of papain
Meat Tenderizer-Papain( a protease
found in papaya) is used as meat
tenderizer to soften meat for cooking

7. Cont…
Papain can be used to dissociate cells
in the first step ofcell
culturepreparations.
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8. Ficain
Ficainalsoknownas
ficinisanenzymethatisderived
fromfigslatex
Itisofafamilyofproteasesknownas
thecysteineendopeptidases
Itisoneofthemostcommonlyusedfor
differentiatingmanybloodgroupantigens
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9. Bromelain
Bromelainisanenzymegenerallyfoundinthe
stemsandfruitsofpineapples.
Accordingtheirsourcesenzymesaredividedinto
A.Stembromelain.
B.Fruitbromelain.
Theproteolyticenzymesaresulfhydrylproteases,
sinceafreesulfhydrylgroupofacysteineside
chainisrequiredforfunction
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10. Bromelain is present in all parts of the
pineapple plant but the stem is the
most common commercial source
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11. Applications of bromelain
Alongwithpapain,bromelainisoneofthemost
popularproteasestouseformeattenderizing
Cookedpineappledoesnothaveatenderizing
effect,astheenzymesareheat-labile
Preparationofmeatballs
Systemicenzymetherapy(combinationsof
proteolyticenzymessuchasbromelain,trypsinand
papain)hasbeenusedinthetreatmentofbreast
cancer
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12. Pepsin
Pepsinisanenzymewhosezymogen(pepsinogen)is
releasedbythechiefcellsinthestomachandthatdegrades
foodproteinsintopeptides
ItwasdiscoveredbyTheodorSchwannwhoalsocoinedits
namefromtheGreekwordpepsis,meaning"digestion
Pepsinisoneofthreeprincipalprotein-degradingenzymein
the digestive system, the other two
beingchymotrypsinandtrypsin
Pepsinismostefficientincleavingpeptidebondsand
preferablybreaksataromaticaminoacidssuch
asphenylalanine,tryptophan,andtyrosine
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13. Cont…
PepsinismostactiveinacidicenvironmentsPepsinexhibits
maximalactivityatpH2.0andisinactiveatpH6.5
InmostcasesPepsinogenisactivatedbyHCl
Pepsinsshouldbestoredatverylowtemperatures(between
−80°Cand−20°C)topreventautolysis(self-digestion
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14. Applications of pepsin
Pepsiniscommonlyusedinthe
preparationofF(ab')2fragmentsfrom
antibodies
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15. Cont…
Powderofpepsinisoftenusedasa
dietarysupplementtoaiddigestion
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16. Rennin
RenninorChymosin isaproteaseproducedby
newbornruminantinthefourthstomachtocurdlethemilk
theyingest
ThenativesubstrateofChymosinisK-caseincleavaesat
aminoacidslikephenylalanineandmethionine
BovinechymosinisnowproducedrecombinantlyinE.
coli,Aspergillusnigeralternativeresource.
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17. Application of rennin
It is widely used in the production
ofcheese
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18. Cathepsin
Cathepsinsareproteasesthatdegradeproteins)
foundinallanimalsaswellasotherorganisms.
Enzymecathepsinsfoundonlyinthelysosome
Mostofthemembersbecomeactivatedatthelow
pHfoundinlysosomes
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19. Cont….
Thereareapproximatelyadozenmembers
ofthisfamily,whicharedistinguishedby
theirstructure
CathepsinA(serineprotease)
CathepsinB(cysteineprotease)
CathepsinC(cysteineprotease)
CathepsinD(aspartylprotease)
CathepsinE(aspartylprotease)
CathepsinF(cysteineproteinase)
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20. Trypsin
Trypsinisaproteasefoundinthedigestive
system ofmany vertebrates,where
ithydrolysesproteins.
Trypsinisproducedinthepancreasastheinactive
proteasetrypsinogen.
Trypsincleavespeptidechainsmainlyat
the carboxylside of the amino
acidslysineorarginine
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21. Application of trypsin
InaAnimaltissueculturelab,trypsinisusedtore-
suspendcellsadherenttothecellculturedishwall
duringtheprocessofharvestingcells
Usedinproteomicsexperimentstodigestproteins
intopeptidesformassspectrometryanalysis
As a baking enzyme to improve the workability of
dough;
in the extraction of seasonings and flavourings
from vegetable or animal proteins and in the
manufacture of sauces
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22. Cont…
Toimprovethetextureoffishproducts
Totenderizemeatalongwithpapain
Forcoldstabilizationofbeer
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23. Cont…
Intheproductionofhypoallergenicfoodwhere
trypsinbreakdownspecificallergenicproteinsinto
nonallergenicpeptides
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