I5CLA1ICN CI CA5LIN IkCM MILk AND

ACID/8A5L nDkCL5I5 AND

NLU1kALI2A1ICN
Alambra
Barrion
Bravo
Cbavz
Escobar
Conong
Yap
IN1kCDUC1ICN
8C1LlnS
lL ls naLurallv occurrlna unbranched
polvmer wlLh monomer unlL called omloo
oclJs
roLelns are chalns of amlno acld
molecules connecLed bv lJ booJs
lL ls a pepLlde ln whlch aL leasL 30 amlno
acld resldues are presenL
lL can be caLeaorlzed ln Lwo Lvpes
llbrous
Clobular
ll88CuS 8C1LlnS
tenu to foim long
anu stiong iepeateu
sets of amino aciu
iesiuues
0nlike globulai
pioteins, it foims
inteimoleculai
inteiactions between
the siue chains of the
iesiuues
CLC8uLA8 8C1LlnS
tenu to folu back
on themselves into
compact units that
appioach neaily
spheioiual shapes
uo not foim
inteimoleculai
inteiactions
between piotein
units
8C1Lln P?u8CL?SlS
iotein that has been hyuiolyzeu oi
bioken uown into its component amino
acius
Peptloe bonos ot protelns are byorolyzeo
by eltber strong aclo or strong base.
,lLk
lL ls a complex bloloalcal fluld wlLh hlah
amounL of proLelns llplds and mlnerals
lL ls a aood source of calclum and
phosphorus buL lL ls deflclenL ln lron and
a poor source of ascorblc acld
,llk proLelns conLaln all 9 essenLlal amlno
aclds requlred bv humans
1he funcLlon of mllk ls Lo supplv nuLrlenLs
such as essenLlal amlno aclds requlred for
Lhe arowLh of Lhe newborn
ASLln
lL ls Lhe maln proLeln ln mllk
lL ls a phosphoproLeln whlch has
phosphaLe aroups aLLached Lo Lhe
hvdroxvl aroups of some of Lhe amlno
aclds sldechalns
aseln exlsLs ln mllk as a calclum salL
calclum caselnaLe
alclum caselnaLe has an lsoelecLrlc polnL
of pP 46
ASLln
aseln conslsts ot a talrly blgb number ot
prollne peptloes, wblcb oo not lnteract.
Tbere are also no olsultloe brloges.
As a result, lt bas relatlvely llttle tertlary
structure. 8ecause ot tbls, lt cannot
oenature
|t ls relatlvely byoropboblc
|t ls touno ln mllk as a suspenslon ot
partlcles calleo caseln mlscelles wblcb
sbow some resemblance wltb surtactant-
type mlscellae
C8!L1lvLS Cl 1PL LxL8l,Ln1
o isolate casein fiom non-fat milk by
isoelectiic piecipitation.
To byorolyze caseln wltb an aclo or a base
I5CLA1ICN CI CA5LIN
IkCM NCNIA1 MILk
(MILk MAGIC)
kCCLDUkL AND kL5UL15
g
powdrd
non-fat dry
milk
--uissolve in mL uistilleu
B

0
--heat the sol'n to
o
C on
hot plate
--take note of initial pB
--auu % CBC00B
uiopwise until pB of
solution ieaches .
case|n
kC1LIN nDkCL5I5 5CnLMA1IC DIAGkAM
case|n
--filtei by giavity
filtiation
wby
{filtrat)
casin
{rsidu)
--uiy between filtei
papeis anu weigh
--calculate % yielu
--uiviue into two
poitions
Ac|d/8ase
hydro|ys|s
Color
ractions
ACID nDkCL5I5 kCCLDUkL
protin isolat
-auu mL 8N B

S0

-label flask, plug with cotton,

anu covei with aluminum foil
-note appeaiance befoie
autoclaving
-autoclave the flask at psi
foi his.
-note appeaiance aftei
autoclaving
-uilute hyuiolyzate with
mL u.B

0 anu tiansfei to
mL beakei
-neutialize the hyuiolyzate by
auuing spatula full of Ba(0B)

-check pB using litmus papei (ieu

puiple): confiim using pB papei
-if not yet neutializeu, auu
satuiateu Ba(0B)

solution
uiopwise
-filtei off piecipitate foimeu anu
wash with mL hot B

0 twice
-If the volume of the filtiate is less
than mL, make up to mL
then pioceeu to coloi ieactions
Acid bydrolyzat
Befoie autoclaving Aftei autoclaving
uioup
Cleai yellow liquiu
Baik biown liquiu with
thin biown iesiuues
uioup
White solution
Black solution with
black piecipitate
uioup
Cleai yellow liquiu
Baik biown liquiu with
thin biown iesiuues
uioup
White solution Black solution
uioup 9
White pieces of casein in
cleai liquiu
Biownish-black
solution with biown
flakes
uioup
Coloilesscleai liquiu
with suspenueu casein
paiticles
Biown to black solution
with light biown
paiticles
8A5L nDkCL5I5 kCCLDUkL
-auu mL boiling B

0 anu
.g Ba(0B)

-label flask, plug with cotton,

anu covei with aluminum foil
-note appeaiance befoie
autoclaving
-autoclave the flask at psi
foi his.
-note appeaiance aftei
autoclaving
-uilute hyuiolyzate with
mL u.B

0 anu tiansfei to
mL beakei
protin isolat
-neutialize the hyuiolyzate by
auuing .mL N B

S0

-check pB using litmus papei (ieu

puiple): confiim using pB papei
-if not yet neutializeu, auu 8N
B

S0

solution uiopwise
-filtei off piecipitate foimeu anu
wash with mL hot B

0 twice
-If the volume of the filtiate is less
than mL, make up to mL
then pioceeu to coloi ieactions
Bas bydrolyzat
Befoie autoclaving Aftei autoclaving
uioup Clouuy solution Yellow clouuy solution
uioup
Yellowish foim, smooth
textuie
uibiu, cuiu foims,
yellow coloi
uioup
Soliu light yellow
piouuct
Yellowish liquiu with
small amounts of white
piecipitate
uioup 8
uioup Clouuy white solution
Yellowish soulition with
lumps of unuissolveu
casein
uioup
Soliu light yellow
piouuct
Yellowish liquiu with
small amounts of white
piecipitate
eicent Yielu
uioup .%
uioup
8.%
uioup
.%
uioup
.9%
uioup .%
uioup .%
uA1A Anu 8LSuL1S
eicent Yielu
uioup 9%
uioup 8
uioup 9
9.9%
uioup
uioup 8%
uioup .%
L8Ln1 ?lLLu
DI5CU55ICN
8lnlLLS lnvCLvLu ln lSCLA1lCn Cl ASLln
Isoelectiic iecipitation
Aciu Byuiolysis
Alkaline Byuiolysis
Neutialization
lSCLLL18l 8Lll1A1lCn
iecipitation- foimation of a soliu in a solution oi
insiue anothei soliu uuiing a chemical ieaction. When
the ieaction occuis in a liquiu, the soliu foimeu is
calleu the piecipitate.
he piecipitation fiom suspension of a piotein when
the pB is at the isoelectiic point
lSCLLL18l P
iotein is uncbargd
ositive chaige = Negative chaige
Ninimizeu inteimoleculai iepulsions
Bisplays minimum watei solubility
lSCLLL18l P
pH of milk = .
Isolctric pH of casin= .
Actic Acid- phosphate gioups piesent in
casein is piotonateu anu the neutial piotein
piecipitates.
causes the casein micelles to uestabilizeaggiegate by
uecieasing the pB.
aggiegation occuis as a iesult of entiopically uiiven
hyuiophobic inteiactions
piecipitates casein by coagulation
lncreases solublllLv of oraanlc calclum and phosphorous
ln Lhe mlcelle
pH= 4.6 pH< 4.6
soIubIe insoIubIe
lSCLLL18l 8Lll1A1lCn
pH= 4.6 pH> 4.6
soIubIe
AS 1PL P lALLS 1PL PA8CL Cn ASLln lALLS Anu l1
8Lll1A1LS PLnL ,lLk u8uLLS AS l1 SCu8S C8
1PL ASLln 8Lll1A1LS ,C8L C,LL1LL? A1 LCW
P
ASLln ,lLLLL
A: a submicll: B: protruding cbain: C: Calcium pbospbat: D: -casin: E:
pbospbat groups
Alu P?u8CL?SlS
Acld hvdrolvsls proceeds wlLhouL
racemlzaLlon and wlLh less desLrucLlon of
cerLaln amlno aclds (S 1 8) Lhan alkallne
LreaLmenL
lL ls mosL llkelv Lhe meLhod of cholce ln Lhe
analvsls of proLelns and polvpepLldes
1rvpLophan ls desLroved bv acld hvdrolvsls
ln acld hvdrolvsls Lhe acld lLself also acLs as a
caLalvsL
?lelds burrvbrown soluLlon
Au1lLAvlnC
Beating with stiong B

S0

heie is complete uestiuction of tiyptophan, a seveie

loss of cysteine anu minoi losses of seiine anu
thieonine.
8ASL P?u8CL?SlS
$erlne, Tbreonlne, ysteln, ano Arglnlne
are oestroyeo ln base byorolysls.
|t ls not useo mucb because lt oestroys
more amlno aclos compareo wltb aclo
byorolysls.
8a (OH)
2
Tbe base ltselt also acts as a catalyst.
nLu18ALlZA1lCn
Ba(0B)
-
neutializing agent
ioceuuie foi isolating mixtuie of fiee
amino acius
CnLuSlCn
Casein was isolateu thiough isoelectiic
piecipitation with the use of a weak
aciu, acetic aciu. he isoelectiic pB of
casein is .
Casein isolateu was hyuiolyzeu using a
stion aciu (B

S0

) anu a stiong base

(Ba(0B)

).
W is uestioyeu in aciu hyuiolysis
S,,R,C aie uestioyeu in base hyuiolysis.