Laporan Protein BIOKIMIA UNIVERSITAS NEGERI SURABAYA.docx

BIOCHEMISTRY EXPERIMENT REPORT PROTEIN

By: Okta Pr Prisma sma Dya Dyanti nti

1403 14030 02040 20407 73

STATE UNIVERSITY OF SURABAYA MATHEMATIC AND SCIENCE FACULTY BIOLOGY BIOLOG Y DEPARTMENT DEPARTMENT 2015

CHAPTER I INTRODUCTION A. Backg!"#$ !% E&'()*(#+ Protein is the main component in a i!in" ces# $oth pants an% animas& In most

tiss'es o( the $o%y# protein is the ar"est component a(ter )ater& In the $o%y# proteins ha!e a !ery important roe as a $'i%er s'$stance or str'ct're*(ormin" ces# (or e+ampe (or the (ormation o( skin# hair m'sce# ce mem$ranes# heart# i!er# an% ki%neys& Protein is a nat'ra poymer consistin" o( a n'm$er o( 'nits o( amino aci%s that $in% to each other thro'"h an ami%e $on% or pepti%e& Protein aso contain o( a$'min# nitro"en# car$on# hy%ro"en# oksi"en# an% sometimes s'('r or phospate& Proteins can aso $e interprete% as a compe+ or"anic compo'n%s )ith hi"h moec'ar )ei"ht )hich are poymers o( amino aci% monomers inke% to"ether $y pepti%e $on%s& Proteins can $e cassi(ie% $ase% on the shape an%  physica properties o( certain# namey "o$'ar proteins an% protein (i$ers& ,i$er is the str'ct'ra protein materia an% the animas are not )ater so'$e& -o$'ar proteins ten% to $e so'$e an% neary spherica shape& -o$'ar protein pays an important roe in  $ioo"ica acti!ity& Protein contains many eements that compose# so there are se!era tests that can  $e per(orme% to test (or the presence o( a$'min amino aci%# to test its so'$iity# an% there is aso a test to %etermine the pepti%e $on%& There are !ario's )ays to %etermine the  presence o( protein # in this report )i $e %isc'sse% on protein test to pro!in" the eements o( protein# a$'min so'$iity# ninhi%rin test# an% aso pro!in" pepti%e $on% 'sin" $i'ret test& B. S+a+)#g P!,-(* 1& .o) to %etermine/i%enti(y the eements o( protein 2& .o) to %etermine the a$'min so'$iity to !ario's so!ent 3& .o) to %etermine the pepti%e $on% that (orme% protein $i'ret tests 4& .o) to %etermine the presence o( amino aci% 'sin" ninhi%rin test C. A)* !% E&'()*(#+ 1& Determinin"/i%enti(yin" the the eements o( protein& 2& Determinin" the a$'min so'$iity to !ario's so!ent& 3& Determinin" the pepti%e $on% that (orme% protein 'sin" $i'ret test& 4& Determinin" the presence o( amino aci% 'sin" ninhi%rin test& D. B(#(%)+ no)in" the eements insi%e protein# increase% kno)e%"e o( rea%er to check  the (oo% contains protein or not& The coe"e st'%ent kno) the pepti%e $on% that (orme% protein an% ha!e ski to test (oo% 'sin" test ninhi%rin# $i'ret or test the so'$iity o( a$'min&

CHAPTER II THEORITICAL REVIE/ A. P!+()#

Protein the name (rom the -reek protos# )hich means the most important&Proteins are compe+ or"anic compo'n%s o( hi"h moec'ar )ei"ht )hich are poymers o( amino aci% monomers inke% each other $y pepti%e $on%s& 5oec'es containin" protein the a!era"e content o( compost chemica eement that is 60 car$on# 7 hy%ro"en# 23 o+y"en# 28 nitro"en# an% s'('r 0*3 an% 0*3  phosphor's& There are other chemica $on%s in proteins are hy%ro"en $on%s# hy%ropho$ic $on%s# ionic $on%s / $on% eektros Tatik# an% $on%in" !an%er 9aas (orces& Protein can $e 'nsta$e to se!era (actors# namey p.# ra%iation# oy$ean  $arrat're# me%i'm or"anic so!ents# an% %eter"ent& Base% on (orm# so'$iity an% chemica composition# the protein can $e cassi(ie% into three "ro'ps&

a& Protein (i$ers/ (i$ro's ;n anima protein that is inso'$e in "enera can not $e %estroye% $y en&  $& -o$'ar protein -o$'ar protein is a "o$'ar protein )ith a pepti%e chain )rappe% aro'n% soi%& E+ampes are en# "o$'in =%oes not %isso!e in )ater or  %isso!e itte $y itte# (o'n% in e""s# mik# an% $oo% # 'se(' as a $ack'p an"  protein (o'n% in the see%s o( a pant># an% histones =$asic proteins o( o) moec'ar  )ei"ht# )ater so'$e# (o'n% in the n'ce's o( ces that ?oin )ith %eo+yri$on'ceic aci% * DN;>& c& @on?'"ate protein It is com$ination o( protein an% nonprotein& e+ampes o( )hich are inc'%e% in this "ro'p are (os(oprotein =casein in mik# (os(itin in the yok># "ycoproteins =m'c's secretions># ipoprotein =ce mem$rane># chromoprotein =hemo"o$in# .emocyanin# cytochrome# (a!oproteins> an% n'ceoprotein =a com$ination o(   proteins )ith n'ceic aci%s containe% in the ce n'ce's>& ;mino aci%s are the $'i%in"*$ocks o( proteins& there are 20 amino aci%s are %i!i%e% into t)o "ro'ps# non*enensia amino aci%s an% essentia amino aci%s& 12 kin%s o( amino aci%s in non*enensia pro%'ction $y the $o%y& 9hie the remainin" ei"ht# (orm an essentia amino aci% that m'st $e o$taine% thro'"h (oo%& There are three types o( amino aci%& Non*essentia# essentia# an% semiessentia amino aci%s&

; pepti%e $on% is a chemica $on% (orme% $et)een t)o moec'es )hen the car$o+y "ro'p o( one moec'e reacts )ith the amino "ro'p o( the other moec'e# reeasin" a moec'e o( )ater =.2O>& This is a %ehy%ration synthesis reaction =aso kno)n as a con%ensation reaction># an% 's'ay occ'rs $et)een amino aci%s& The

res'tin" @O*N. $on% is cae% a pepti%e $on%# an% the res'tin" moec'e is an ami%e&The (o'r*atom ('nctiona "ro'p *@=AO>N.* is cae% an ami%e "ro'p or =in the conte+t o( proteins> a pepti%e "ro'p&Poypepti%es an% proteins are chains o( amino aci%s he% to"ether $y pepti%e $on%s# as is the $ack$one o( PN;& 'ch a inka"e is (orme% $y remo!a o( the eements o( )ater =%ehy%ration> (rom the car$o+y "ro'p o(  one amino aci% an% the amino "ro'p o( another&  Pepti%e $on%



the (irst amino aci%s  amino "ro'p (rom other amino aci%

%ipepti%e  .20



 car$o+y "ro'p )ith

Protein str'ct're is !ery compe+ an% pays an important roe in %eterminin" its $ioo"ica acti!ity# protein str'ct're can $e %i!i%e% into the primary str'ct're# secon%ary# tertiary# an% C'aternary& 1& The primary str'ct're e+presse% seC'ence o( amino aci%s in the protein chain an% the ocation o( %is'(i%e $on%s# i( any& Beca'se the protein may contain 100 or  more amino aci% resi%'es so %i((ic't to %escri$e the (orm'a %o)ns& It is there(ore 'se% three*etter a$$re!iation (or each asamamino& ,or e+ampe: ;anin=;a> # -'tamine =-'># ysin=ys>& 2& The secon%ary str'ct're# the reationship space a%?acent amino aci%s in the primary str'ct're# may $e re"'ar an% rec'rrin" perio%icay& Beca'se o( the %ispersion (orces or hy%ro"en $on%s# a ro ike spira poypepti%e chains =apha hei+>& 3& Tertiary str'ct're is the o!era arran"ement an% reationship o( !ario's parts o( a  poypepti%e chain& 4& The C'aternary str'ct're# a protein is sai% to ha!e C'aternary str'ct're )hen the  protein consistin" o( t)o or more poypepti%e chains he% to"ether $y %ispersion (orces =hy%ro"en $on%>& B. S!"c( !% P!+()#

E""s are a (oo% materia (rom catte that ha!e hi"h n'tritiona !a'e# especiay protein# %i"esti$e an% n'tritio's& E"" )hite is cooress iC'i%# containin" )ater )hich is the ar"est component o( an e""& E"" )hites kno)n as the a$'min =a$'s A )hite># $eca'se i( coa"'ate%# e"" )hites )i pro%'ce a )hite precipitate& E"" )hites contain )ater 'p to  o( the tota )ei"ht o( the e"" )hites an% protein more than 0 o( %ry )ei"ht& E"" )hite protein compose% o( protein (i$ers o!om'cin an% "o$'ar proteins& Proteins are )i%ey (o'n% in e"" )hites# amon" others: o!a$'min# cona$'min =o!otrans(errin># o!om'coi%# y (at ="> @ar$ohi%rat ="> @asi'm =m"> Phospor =m"> ,i$er ="> Hinc =m"> itamin B1 =m"> itamin B2 =m"> itamin @ =m">  Niasin =m"> Ener"y =ka>

2&00 0&10 1F&10 11&00 68&00 0&30 0&70 0&0F 0&03 18&00 1&40 3&00

CHAPTER III EXPERIMENTAL METHOD A. T!!- a#$ Ma+()a1& Pro!in" the eements that e+ist in protein

• • • • • • •

Toos : Test T'$e Drop pipette Rack test t'$e @amp test t'$e 5eas'rin" c'p pirit's $'rner   p. 'ni!ersa in%icator 

5aterias: • • • • •

@oncentrate% NaO. E"" )hite oy$ean $ar so'tion Potato so'tion Potato so'tion

2& ;$'min 2 o'$iity in !ario's so!ents • • • • • •

Toos : Test T'$e Drop pipette Rack test t'$e @amp test t'$e 5eas'rin" c'p orte+s

5aterias : • • • • •

o'tion o( a$'min 2  NaO. so'tion 0#2  Na@O3 so'tion 0#2 .@ so'tion 0#2 ;C'a%es

3& Pro!in" the pepti%e $on% constr'cte% protein thro'"h $i'ret test Toos :

•

Test T'$e

• • • • •

Drop pipette Rack test t'$e @amp test t'$e 5eas'rin" c'p orte+s

5aterias : • • • • •

4& Pro!in" the amino aci% thro'"h Ninhi%rin test

o'tion o( protein  NaO. so'tion 10 @'O4 so'tion 0#01 5 oy$ean $ar e+tract Potato e+tractso'tion

• • • • • •

Toos : Test T'$e Drop pipette Rack test t'$e @amp test t'$e 5eas'rin" c'p pirit's $'rner  5aterias:

;r"inin  Ninhi%rin so'tion 0#1 # protein reactant =3 kin%s> • oy$ean $ar so'tion • Potato so'tion • B. P!c($"( P!)#g +( (-(*(#+ +a+ (&)+ )# '!+()# 1& P'ttin" 6 %rops a$'min (rom e"" )hite into %ry test t'$e# then heatin" 'p on •

spirit's $'rner %irecty& o$ser!in" the e!i%ence happenin" =sme# coo'r# an% )ater !apor>& 2& P'ttin" 6 %rops o( a$'min=e"" )hite> into %y test t'$e# a%%e% )ith 10 %rops concentrate% NaO.# then $'rnin" care('y on spirit's $'rner# an% o$ser!in" the sme chan"e% 3& P'ttin" 1 m potato so'tion into %ry test t'$e# then heatin" 'p on spirit's $'rner  %irecty& O$ser!in" the e!i%ence happenin" =sme# coo'r# an% )ater !apor>& 4& P'ttin" 1 m potato so'tion into %y test t'$e# a%%e% )ith 2 m concentrate%  NaO.# then $'rnin" care('y on spirit's $'rner# an% o$ser!in" the sme chan"e% 6& P'ttin" 1 m soy$ean $ar so'tion into %ry test t'$e# then heatin" 'p on spirit's  $'rner %irecty& O$ser!in" the e!i%ence happenin" =sme# coo'r# an% )ater  !apor>& 8& P'ttin" 1 m soy$ean $ar so'tion into %y test t'$e# a%%e% )ith 2 m concentrate%  NaO.# then $'rnin" care('y on spirit's $'rner# an% o$ser!in" the sme chan"e%

CHAPTER IV DATA AND ANALYSIS A. Da+a Ta,-( 1 P!)$)#g T( E-(*(#+ Ta+ E&)+ O# P!+()#

N!

Ma+()a-

T(a+*(#+

1&

;$'min

6 %rops a$'min=e"" )hite> heate% 'p→

6 %rops a$'min=e"" )hite> 10 %rops concentrate% NaO. heate% 'p→ 2&

3&

Potato

oy$ean  $ar

1 m potato so'tion heate% 'p→

1 m potato so'tions 2 m concentrate% NaO. heate% 'p→ 1 m soy$ean $ar so'tions heate% 'p→ 1 m soy$ean $ar so'tions 2 m concentrate% NaO. heate% 'p→

R("-+ !% (&'()*(#+ B(%!( A%+( @oor : @ooress  @oor : 9hite  me : (ishy e"" (o'n% $ro)n  @harcoa 9ater !apor : * me : ike $'rne% hair   p. : 10 =Base> 9ater !apor :  @oor : Jeo)ish @oor : Bro)n )hite  (o'n% %ark re%  @harcoa me : * me : ike $'rne% hair  9ater !apor : * 9ater !apor :   p. : 13 =Base> @oor : Bro)nish @oor : Oran"e  yeo)  (o'n% $ro)n  @harcoa me : potato me : B'rne% hair  9ater !apor : * 9ater !apor :   p. : 8 =;ci%> @oor : Jeo)  @oor : Jeo)  me : potato (o'n% $ro)n  @harcoa 9ater !apor : * me : ike $'rne% hair   p. : 13 =Base> 9ater !apor :  @oor : 5iky )hite @oor : Jeo)  me : oy$ean $ar  (o'n% $ro)n  @harcoa 9ater !apor : * me : ike $'rne% hair   p. : 6 =;ci%> 9ater !apor :  @oor : 9hitish @oor : Oran"e  yeo)  (o'n% %ark $ro)n  me : oy$ean $ar  @harcoa 9ater !apor : * me : ike $'rne% hair   p. : 13 =Base> 9ater !apor : 

Ta,-( 2 A-,"*)# 2 S!-",)-)+3 )# Va)!" S!-(#+ N!

P!c($"(

1&

;$'min 2  aC'a%es  !orte+

2&

;$'min 2  NaO. 0&2  !orte+

O,(a+)!# R("-+ B(%!( A%+( @oor : cooress * @oor : cooress * o'$e =homo"eno's so'tion> @oor : cooress * @oor : cooress

* 3&

;$'min 2  .@ 0&2  !orte+

@oor : cooress

* *

4&

;$'min 2  Na@O 3 0&2  !orte+

@oor : cooress

* *

o'$e =homo"eno's so'tion> @oor : cooress o'$e =homo"eno's so'tion> @oor : cooress  co% o'$e =homo"eno's so'tion>

Ta,-( 4 B)"(+ T(+

 No

Proce%'re

O$ser!ations Res't Be(ore

;(ter  

a) a. C"SO (ag(#+ 1a& ;$'min  1 m NaO. 10 → ;$'min: yeo)ish ;$'min  NaO.  !orte+te% K  3 %rops @'O 4 )hite an% transparent @'O4 A t'r$i% )hite =>  NaO.: cooress 0#015 K stir K o$ser!e the an% there are i"ht $'e => @'O4: i"ht $'e coor chan"e K i( coor %oes not co% chan"e# a%% 1*2 %rops @'O 4 2a& E+tract soy$ean $ar  1 m E+tract soy$ean $ar : E+tract soy$ean $ar   NaO. 10 K !orte+te% K  3 )hite =>  NaO.  @'O4: t'r$i%  NaO.: cooress %rops @'O 4 0#015 → stir K i"ht $'e => @'O4: i"ht $'e o$ser!e the coor chan"e K i( coor %oes not chan"e# a%% 1*2 %rops @'O 4 3a& E+tract potato  1 m NaO. E+tract potato : $ro)n E+tract potato NaO.  10 K !orte+te% K  3 %rops => )ith )hite => @'O4 A $ack => )ith @'O4 0#015 K stir K o$ser!e  precipitate at the $ottom )hite => precipitate at the the coor chan"e K i( coor %oes o( react t'$e NaO.:  $ottom o( react t'$e not chan"e# a%% 1*2 %rops @'O 4 cooress @'O4: i"ht $'e ,. B)"(+ (ag(#+ 1$ ;$'min  1 m NaO. 10 → ;$'min: yeo)ish ;$'min  NaO.  Bi'ret & !orte+te% K  3 %rops Bi'ret )hite an% transparent rea"ent A p'rpish )hite rea"ent 0#015 K stir K o$ser!e  NaO.: cooress => the coor chan"e K i( coor %oes Bi'ret rea"ent : i"ht not chan"e# a%% 1*2 %rops Bi'ret  $'e rea"ent 2$ E+tract soy$ean $ar  1 m E+tract soy$ean $ar : E+tract soy$ean $ar  &  NaO. 10 K !orte+te% K  3 )hite =>  NaO.  Bi'ret rea"ent:  NaO.: cooress %rops Bi'ret rea"ent 0#015 → yeo)ish )hite => Bi'ret rea"ent: i"ht stir K o$ser!e the coor chan"e K i( coor %oes not chan"e# a%%  $'e 1*2 %rops Bi'ret rea"ent

3$ &

E+tract potato  1 m NaO. 10 K !orte+te% K  3 %rops Bi'ret rea"ent 0#015 K stir K o$ser!e the coor chan"e K i( coor %oes not chan"e# a%% 1*2 %rops Bi'ret rea"ent

E+tract potato : $ro)n => an% there is  precipitate  NaO.: cooress Bi'ret rea"ent: i"ht  $'e

E+tract potato NaO.  Bi'ret rea"ent A %ark  $ro)n => )ith )hite  precipitate at the $ottom o( react t'$e

Ta,-(  P!)#g T( A*)#! Ac)$ T!"g N)#)$)# T(+ N!

P!c($"(

1&

;r"inin  Ninhy%rin 0&1  .eate%

2&

;$'min  Ninhy%rin 0&1  .eate%

3&

oy$ean $ar  Ninhy%rin 0&1  .eate% Potato  Ninhy%rin 0&1  .eate%

4&

O,(a+)!# R("-+ B(%!( A%+( @oor : cooress * @oor : $'e => * Precipitate : * @oor : cear yeo) * @oor : $'eish =>  p'rpe => * Precipitate : )hite @oor : )hite => * @oor : $'e => * Precipitate : )hite

@oor : $ro)n =>

*

@oor : $ackish  $'e => * Precipitate : $'e =>

B. A#a-3) P!)$)#g +( (-(*(#+ +a+ (&)+ )# P!+()# ;$'min (rom e"" )hite has cooress  coor# the sme is (ishy e""& Be(ore •

heatin" 'p# p'ttin" 'ni!ersa p. in%icator an% the p. so'tion is 10# means that a$'min is $ase& ;(ter heate% 'p se!era min'tes# the coor chan"e% into )hite  an% (o'n% $ro)n @harcoa & The sme is ike $'rne% hair  an% aso there are )ater !apors & The coor a(ter heate% 'p chan"e% (rom cooress into )hite  an% (o'n%  $ro)n @harcoas  means that a$'min )hich is the e+ampe o( protein contains o( @ar$on =@> eement& The sme o( a$'min a(ter heate% 'p is ike $'rne% hair# its in%icate% that protein contains nitro"en =N> $eca'se the $'rne% hair sme is the characteristic sme o( Nitro"en& In the e+periment# there are (o'n% )ater !apors   its in%icate% that a$'min contains hy%ro"en=.>& ;nother chan"in" process that happene% is the chan"in" coor o( 'ni!ersa p. in%icator& The a$'min )hen teste% )ith 'ni!ersa p. in%icator# sho)e% that a$'min is $ase p.& Beca'se the a$'min contains @#N# an% . comp'n%# so a$'min contains protein&

•

;$'min 1 m (rom e"" )hite has cooress  coor# a(ter a%%e% )ith NaO. 2 m an% shoke it# the coor is yeo)ish )hite  an% no sme& The p. o( its so'tion is 13 means that a$'minNaO. is $ase so'tion& ;(ter heate% 'p se!era min'tes# the coor chan"e% into $o)n  an% (o'n% %ark re%  @harcoa in the so'tion& The sme is ike $'rne% hair  an% aso there are )ater !apors  ess than a$'min a(ter heate% 'p& ;$'min 1 m a(ter a%%e% )ith NaO. 2 m# the 'ni!ersa p. in%icator is $'e#

means that a$'min an% a$'minNaO. $oth o( them are $ase& 9hen the so'tion o( NaO. an% a$'min# the sme is ike $'rne% hair  means that there are  Nitro"en in its reaction& The coor o( so'tion chan"e into $ro)n  an% (o'n% %ark  re%  @harcoa& The @harcoa means that its so'tion contains car$on& The )ater  !aporation in that reactions means that a$'minNaO. contains .y%ro"en& o# a$'min contains proteins& Potato so'tions has $ro)nish yeo)  coor# the sme is ike potato& Be(ore • heatin" 'p# p'ttin" 'ni!ersa p. in%icator an% the p. is 8# means that potato is aci%ic so'tion& ;(ter heate% 'p se!era min'tes# the coor chan"e% into oran"e  an% (o'n% $ro)n charcoa & The sme is ike $'rne% hair  an% aso there are )ater !apors & Potato so'tion is an aci%ic so'tion# $eca'se it has p.A8 a(ter checke% )ith 'ni!ersa p. in%icator& Be(ore heate% 'p# potato so'tion smet ike potato# $'t a(ter  heate% 'p# the sme is ike $'rne% hair& It is sho)e% that it e+ist Nitro"en& The coor o( so'tion chan"e (rom $ro)nish yeo) ino oran"e an% (o'n% the $ro)n charcoa # the charcoa sho)e% that there are car$on& The )ater !apor o( potato so'tion a(ter heate% 'p is present  & Its so'tion in%icates the characteristics o(   protein# ike Nitro"en# .y%ro"en# an% @ar$on & Potato so'tion 1 m a(ter a%%e% )ith NaO. 2 m an% shoke it# the coor is • yeo)  an% sme ike potato& The p. o( its so'tion is 13 means that  potatoNaO. is $ase so'tion& ;(ter heate% 'p se!era min'tes# the coor  chan"e% into yeo)  an% (o'n% $ro)n  charcoa in the so'tion& The sme is ike $'rne% hair  an% aso there are )ater !apors  hi"her than  potato a(ter heate% 'p& The coor a(ter heate% 'p %oes not (rom yeo)  # $'t (o'n% $ro)n @harcoas  means that a$'min )hich is the e+ampe o( protein contains o(  @ar$on =@> eement& The sme o( a$'min a(ter heate% 'p is ike $'rne% hair # its in%icate% that protein contains nitro"en =N> $eca'se the $'rne% hair sme is the

characteristic sme o( Nitro"en& In the e+periment# there are (o'n% )ater !apors   its in%icate% that a$'min contains hy%ro"en=.>& ;nother chan"in" process that happene% is the chan"in" coor o( 'ni!ersa p. in%icator& The a$'min )hen teste% )ith 'ni!ersa p. in%icator# sho)e% that a$'min is $ase p.& Beca'se the a$'min contains @#N# an% . comp'n%# so# potato contains protein& oy$ean $ar so'tion 1 m $e(ore heate% 'p# has miky )hite coo'r# the sme • sti ike soy$ean $ar& The p. o( its so'tion is 8 means that soy$ean $ar is aci% so'tion& ;(ter heate% 'p se!era min'tes# the coor chan"e% into yeo)  an% (o'n% %ark $ro)n  charcoa in the so'tion& The sme is ike $'rne% •

hair  an% aso there are )ater !apors  oy$ean $ar so'tion 1 m a(ter a%%e% )ith NaO. 2 m an% shoke it# the coor  is )hitish yeo)  an% sme ike soy$ean $ar& The p. o( its so'tion is 13 means that soy$ean $arNaO. is $ase so'tion& ;(ter heate% 'p se!era min'tes# the coor chan"e% into oran"e  an% (o'n% $ro)n  charcoa in the so'tion& The sme is ike $'rne% hair  an% aso there are )ater !apors  ess than soy$ean $ar $e(ore heate% 'p& Base% on# the e+periment# soy$ean $ar in aci% or $ase con%ition sti contains

o( protein# $eca'se a(ter heate% 'p# the sme is ike $'rne% hair# then in the so'tion there are $ro)n charcoa means that the so'tion contains @ar$on# an% )hen heate% 'p# the )ater !apor (o'n%& 9ater !apor in%icates the hy%ro"en& o# oy$ean $ar contains protein& A-,"*)# 2 S!-",)-)+3 )# Va)!" S!-(#+ ;t 1 m o( 2 ;$'min p's 1 m o( aC'a%es is cooress& ;(ter !orte+e% (or  one ha(# the so'tion sti cooress& This pro!es that 2 ;$'min p's aC'a%es is so'$e =homo"eno's so'tion>& ;t 1 m o( 2 ;$'min p's 1 m o( NaO. 0&2 is cooress& ;(ter (or one ha( # the so'tion sti cooress& This pro!es that 2 ;$'min p's NaO. 0&2 is so'$e =homo"eno's so'tion>& ;t 1 m o( 2 ;$'min p's 1 m o( .@ 0&2 is cooress& ;(ter (or one ha( # the so'tion sti cooress& This pro!es that 2 ;$'min p's .@ 0&2 is so'$e =homo"eno's so'tion>& ;t 1 m o( 2 ;$'min p's 1 m o( Na@O 3 0&2 is cooress& ;(ter (or one ha( # the so'tion sti cooress& This pro!es that 2 ;$'min p's Na@O3 0&2 is so'$e =homo"eno's so'tion>&

;$'min is one o( the e+ampe o( protein& In this e+periment )e 'se ;$'min 2 to pro!e the ;$'min so'$iity in !ario's so!ents& ;n% in the a treatment# the %ata sho)e% that in !ario's so!ents# 2 o( a$'min can $e so'$e& This  pro!es that the !ario's so!ents =aC'a%es# NaO. 0&2 # .@ 0&2 # Na@O 3 0&2> are s'ita$e so!ent& Be(ore !orte+e% $et)een so'te an% so!ent is can not mi+e%  per(ecty# in this treatments the ('nction o( !orte+ is to make the so'tions mi+e% competey so a(ter !orte+e% the so'tion can mi+e% per(ecty& Besi%e that so'$e  proteins (orm cooi%a or rea so'tions the sta$iity o( protein so'tions mainy %epen%s on the eectric char"e o( the partices# their %e"ree o( hy%ration an% temperat're& Protein# )hich as a res't o( the %enat'ration a"ent action ost its cooi%a character# 's'ay precipitates (rom the so'tion& B)"(+ T(+ a. C"SO (ag(#+ ;t this e+periment a$'min a%%e% )ith 1 m NaO. 10 then it • !orte+te% in 1&16 min'tes& ;(ter !orte+te% the coor chan"es to yeo)ish )hite an% transparent& Then it a%%e% $y 3 %rops 0&01 5 @'O4 rea"ent an% the coor chan"es to t'r$i% )hite => an% there are •

i"ht $'e => cots in the react t'$e& ;t this e+periment e+tract soy$ean $ar a%%e% )ith 1 m NaO. 10 then it !orte+te% in 1&16 min'tes& ;(ter !orte+te% the coor chan"es to )hite =>& Then it a%%e% $y 3 %rops 0&01 5 @'O 4 rea"ent an% the

•

coor chan"es to t'r$i% i"ht $'e =>& ;t this e+periment e+tract potato a%%e% )ith 1 m NaO. 10 then it !orte+te% in 1&16 min'tes& ;(ter !orte+te% the coor chan"es to $ro)n => )ith )hite => precipitate at the $ottom o( react t'$e& Then it a%%e% $y 3 %rops 0&01 5 @'O 4 rea"ent an% the coor chan"es to

 $ack => )ith )hite => precipitate at the $ottom o( react t'$e& ,. B)"(+ (ag(#+ ;t this e+periment a$'min a%%e% )ith 1 m NaO. 10 then it • !orte+te% in 1&16 min'tes& ;(ter !orte+te% the coor chan"es to yeo)ish )hite an% transparent& Then it a%%e% $y 3 %rops 0&01 5 •

Bi'ret rea"ent an% the coor chan"es to p'rpish )hite =>& ;t this e+periment e+tract soy$ean $ar a%%e% )ith 1 m NaO. 10 then it !orte+te% in 1&16 min'tes& ;(ter !orte+te% the coor chan"es to )hite =>& Then it a%%e% $y 3 %rops 0&01 5 Bi'ret rea"ent an% the coor chan"es to yeo)ish )hite =>&

•

;t this e+periment e+tract potato a%%e% )ith 1 m NaO. 10 then it !orte+te% in 1&16 min'tes& ;(ter !orte+te% the coor chan"es to $ro)n => an% there is precipitate at the $ottom o( react t'$e& Then it a%%e%  $y 3 %rops 0&01 5 Bi'ret rea"ent an% the coor chan"es to %ark $ro)n

=> )ith )hite precipitate at the $ottom o( react t'$e& P!)#g +( a*)#! ac)$ +!"g #)#)$)# +(+ ;t 6 %rops o( ;r"inin in the reaction t'$e an% then a%%e% 6 %rops o(  0&1 Ninhy%rin# the so'tion is cooress& ;(ter heate% in the )ater  'nti $oi# the coor o( so'tion is $'e => $'t there is not  precipitate& The $'e coor is in%icate that the so'tion containin" amino aci%& ;t 6 %rops o( ;$'min in the reaction t'$e an% then a%%e% 6 %rops o(  0&1 Ninhy%rin#

the coor o( so'tion is cear yeo) => & ;(ter 

heate% in the )ater 'nti $oi# the coor o( so'tion is $'eish p'rpe => an% there is precipitate )ith )hite coor& The $'eish p'rpe is in%icate that the so'tion containin" amino aci%& ;t 6 %rops o( e+tract soy$ean $ar in the reaction t'$e an% then a%%e% 6 %rops o( 0&1 Ninhy%rin# the coor o( so'tion is )hite => & ;(ter  heate% in the )ater 'nti $oi# the coor o( so'tion is $'e => an% there is precipitate )ith )hite coor& The $'e coor is in%icate that the so'tion containin" amino aci%& ;t 6 %rops o( e+tract potato in the reaction t'$e an% then a%%e% 6 %rops o( 0&1 Ninhy%rin# the coor o( so'tion is $ro)n => & ;(ter  heate% in the )ater 'nti $oi# the coor o( so'tion is $ackish $'e = > an% there is precipitate )ith $'e => coor& The $ackish $'e coor  is in%icate that the so'tion containin" amino aci%& C. D)c")!# Pro!in" the eements o( protein • 1& In those e+periment# %i% not 'se itm's paper# $'t 'sin" 'ni!ersa p.

in%icator an% sho) that the so'tion is $ase so'tion $eca'se a(ter checke% )ith coo'r scae# the coor sho)s the p. is $ase aro'n% 13& 2& The 'ni!ersa p. in%icator sho)e% the aci%ity o( so'tion& The 'ni!ersa p. in%icator sho)e% that the so'tion is $ase& Base content on protein means that the protein contains o( amino aci%# $eca'se amino aci% is the main str'ct're% (orme% protein& I( there is an amino aci%s# the protein sho)e% that contains o+y"en# hy%ro"en# an% nitro"en too&

CHAPTER V CLOSURE A. C!#c-")!# 1. M!-)c T(+ 5atose has $ackish p'rpe rin" in the mi%%e o( so'tion# thickess are =

># means that the matose containe% the most car$ohy%rates an% the s)eetest (a!o'r# the res't is %i((erent (rom theory $eca'se the e+perimenter %roppe% the .2O4 not in the same porpotion& -'cose

an% s'ccrose a itte more

car$ohy%rates than actose# se'ose# an% (r'ctose& I( the coo'r o( rin" that (orme% in the mi%%e o( so'tion more %arker or t'r$i%# it means that the so'tion containe% hi"her car$ohy%rates# an% the taste is more s)eet& 2. B(#($)c+ T(+ -'cose# (r'ctose# actose# an% matose (orme% the $rick re% coo'r o(   precipitation in the $ase o( test t'$e# means that those materia can re%'ce& In s'crose so'tion# the precipitation is )hite => $eca'se s'crose containin" t)o monosakri%a = (r'ctose an% "'cose > are @harcoa atoms n'm$er 1 at "'cose )ith @harcoaLs atom n'm$er 2 in (r'ctose $y atoms o( o+y"en& 4. S(-)6a#!%% T(+ ei)ano(( test is to i%enti(y there are or not ketose "ro'p =(r'ctose>& Base% on this e+periments sho)e% that (r'ctose an% s'crose pro%'ces $rick*re% coor  that i%enti(ies contain ketose in the monosacchari%e car$ohy%rate& ;my'm# "'cose# (r'ctose# actose# an% ce'ose %o not sho)e% the re% $rick coo'r   $eca'se it %oes not ha!e keton "ro'p# these car$ohy%rates contain o( a%ose "ro'p& . I!$)#( T(+

Io%ine test 'se% to %etect any poysacchari%e& ;my'm# @e'ose an% -yco"en )hich a%%e% )ith )ater an% NaO. 8N then heate% 'p# the so'tion sti t'r$i% $eca'se

there are no coor%ination $on% $et)een Io%i%e ion in

hei+s& amy'm# ce'ose# an%

"yco"en )hich react )ith .@ or in aci%

atmosphere thereLs the chan"in" coor (rom coo're% into cooress& In aci% con%ition NaI an% NaIO3 chan"e% into I 2 $y chori%e aci% 5. Ca,!3$a+( +(+ !# Ma#g! In moisch test# 'nripe# ha( rippene%# an% ripe man"o contains car$ohy%rates# an% the ripe man"o contains the hi"hest car$ohy%rate& In Bene%ict test# 'nripe# ha(rippene%# an% ripe man"o (orme% the $rick re%  precipitation# means that man"o can re%'ce the ion @'2 into @'& In ei)ano(( test# Mnripe man"o has o)er cate"ory o( $rick*re% coo'r# (oo)e% $y ha(*ripe man"o# an% the hi"hest e!e o( $rick*re% coo'r is ripe man"o& ; o( the man"o )as $rick*re% coo'r# so the conc'sion in 'nripe man"o# ha(*rippene% man"o# an% ripe man"o contain ketose "ro'p# especiay (r'ctose an% s'crose& Io%ine test res'te% that 'nripe man"o an% ha(rippene% man"o contains the amy'm $eca'se the coo'r a(ter reaction is p'rpe& The ripe man"o contains "'cose $eca'se compete% hy%roysis amy'm into "'cose& B. S"gg(+)!# '""estion (rom the e+perimeneter o( Biochemistry sho'% $etter 'n%erstan% the proce%'re o( e+periment# (in% the iterat're $e(ore %o the e+periment& The e+perimenter sho'% thro'"h )hen %roppe% the so'tion or e+tract# an% minimi