UNIVERSIDAD NACIONAL EXPERIMENTAL

ROMULO GALLEGOS
AREA DE INGENIERIA AGRONOMICA
DIRECCION DE ESTUDIOS COMUNES
DEPARTAMENTO DE ASIGNATURAS INSTRUMENTALES
CATEDRA DE BIOQUIMICA
PRACTICA N 1:
Identificacin de Aminocidos y Protenas
Introduccin:
Las protenas son sstan!"as #$"!as or%&n"!as 'e a(to peso $o(e!(ar) !onst"t"'a por $!*os
a$"no&!"'os) #e +e%an n pape( $, "$portante en (a estr!tra , '"n&$"!a 'e (a $ater"a -"-a.
So(a$ente /0 a$"no&!"'os '"1erentes se en!entran 1or$an'o parte 'e (as protenas2 s"n
e$3ar%o) no to'as (as protenas !ont"enen (os /0 a$"no&!"'os.
En esta pr&!t"!a -a$os a rea("4ar rea!!"ones !a("tat"-as 'e !o(ora!"5n) #e nos per$"t"r&n
'ete!tar) (a presen!"a 'e %rpos #$"!os en a$"no&!"'os , protenas.
Fundamento de las Reacciones:
1) Reaccin de la ninidrina:
Esta es na rea!!"5n %enera( para !a(#"er a$"no&!"'o , es 'e3"'o a (a presen!"a 'e( %rpo a$"no
6N7/8) 'e $anera 'e #e to'os (os a(1a a$"no&!"'os , por spesto (as protenas 'an pos"t"-a a esta
rea!!"5n. No o3stante) !a(#"er sstan!"a prote"!a #e !onten%a n %rpo a(1a9N7/ ta$3":n 'e
pos"t"-a a esta rea!!"5n) (a rea!!"5n se e1e!t;a en tres etapas.
<.< .9 Re'!!"5n 'e (a n"n*"'r"na o="'a'a por (a a!!"5n 'e( a(1a a$"no&!"'o

<./ .9 7"'r5("s"s 'e( A$"no&!"'o pro'!"'o para 1or$ar n a(1a !eto&!"'o) e( !a( se !ar3o="(a 3a+o
(as !on'"!"ones 'e (a rea!!"5n.



<.> .9 ?"na($ente) e( A$on"a!o 6N7>8 re'!"'o rea!!"ona !on !ant"'a'es e#"$o(ares 'e n"n*"'r"na
o="'a'a , n"n*"'r"na re'!"'a para 1or$ar n !o$pesto 'e !o(or a4( -"o(eta) (a "ntens"'a' 'e(
!o(or es propor!"ona'a a (a !ant"'a' 'e a$"no&!"'os presentes.

La e!a!"5n G(o3a( es A4( V"o(eta
Esta rea!!"5n se 'esarro((a $e+or a p7 netro. To'os (os a$"no&!"'os 'an !o(or a4( -"o(eta)
!on e=!ep!"5n 'e pro("na e *"'ro="pro("na #e 'a !o(or a$ar"((o.
!) Reaccin de "iuret:
Esta rea!!"5n es !ara!terst"!a para a#e((os !o$pestos #e poseen 'os o $as en(a!es pept'"!os.
La 'eno$"na!"5n B"ret se 'e3e a #e e( !o$pesto or%&n"!o B"ret ta$3":n 'a pos"t"-a a esta
rea!!"5n. La rea!!"5n 'e B"ret se 3asa en (as 1or$a!"ones en $e'"o a(!a("no) 'e n !o$p(e+o 'e
!oor'"na!"5n 'e !o(or -"o(eta entre e( "5n !;pr"!o 6C
@/
8 , A &to$os 'e n"tr5%eno) pro-en"entes 'e (as
!a'enas pept'"!as.
#) Reaccin $anto%roteica:
Con esta rea!!"5n po'e$os "'ent"1"!ar (a presen!"a 'e (os a$"no&!"'os aro$&t"!os 6t"ros"na)
tr"pto1ano8.
La rea!!"5n se 3asa en (a n"tra!"5n 'e( an"((o 3en!:n"!o !on a!"'o ntr"!o) pro'!"en'o 'er"-a'os
'e( n"tro3en!eno 'e !o(or a$ar"((o. Estos 'er"-a'os a$ar"((os se tornan anaran+a'os a (a a(!a("n"'a' 'e
(a so(!"5n !on N7AO7 o NaO7. En (as !on'"!"ones 'e (a rea!!"5n (a 1en"(a(an"na es '"1!"( 'e n"trar
por !anto se re#"ere (a presen!"a 'e n !ata("4a'or.
#) Reaccin de &illn:
Esta rea!!"5n es espe!1"!a para e( %rpo *"'ro="1en"(. Por (o #e e( a$"no&!"'o t"ros"na 'a pos"t"-a
a esta rea!!"5n. Se !ree #e e( %rpo *"'ro="1en"( pr"$ero 1or$a n 'er"-a'o n"trato !on e( A!"'o
N"tr"!o 67NO>8) #e (e%o rea!!"ona !on e( $er!r"o , 1or$a n !o$p(e+o 'e !o(or ro+o.
') Reaccin (a)a*uci:
Esta es na rea!!"5n espe!1"!a para e( %rpo Gan"'r"na. Por (o tanto e( a$"no&!"'o ar%"n"na 'a
pos"t"-a a esta rea!!"5n.
La Gana'"na en So(!"5n a(!a("na) 'a n !o(or ro+o en presen!"a 'e a(1a9na1to( e 7"po'ro$"to 'e
So'"o.
PR+,-NTA( .+ PR+ /A"0RAT0RI0:
<.9 De1"na (as s"%"entes pa(a3rasB
A$"no&!"'o Protena Pept"'o
/.9 D"3+e (a estr!tra %enera( 'e (os a$"no&!"'os) "'ent"1"!an'o !a'a no 'e (os %rpos 1n!"ona(es
#e poseen.
>.9 D"3+e a Estr!tra 'e (os s"%"entes a$"no&!"'osB T"ros"na) Le!"na) Met"on"na) L"s"na) Tr"pto1ano
, ar%"n"na
A.9 7a%a (a !(as"1"!a!"5n 'e (os a$"no&!"'os se%;n s %rpo R
PART+ +$P+RI&+NTA/
0"1+TI20(:
a8 Re!ono!er (os Grpos C9a$"nos 'e (os a$"no&!"'os.
38 D"1eren!"ar a$"no&!"'os ("3res 'e !o$pestos !on 'os o $as en(a!es pept'"!os.
!8 I'ent"1"!ar a$"no&!"'os aro$&t"!os.
'8 I'ent"1"!ar e( a$"no&!"'o ar%"n"na.
e8 I'ent"1"!ar na so(!"5n pro3(e$a.
&AT+RIA/+( 3 R+ACTI20(:
T3os 'e ensa,o.
Gra'"((as Met&("!as para t3os 'e ensa,o
BeaDers 'e E00 $(.
P"petas Gra'a'as 'e F $( , 'e <0 $(
P(an!*a 'e Ca(enta$"ento
?ras!os %oteros.
?ras!os 'e -"'r"o
P"n4as 'e Ma'era
So(!"5n 'e n"n*"'r"na a( 0)<G.
So(!"5n 'e 7"'r5="'o 'e So'"o 6NaO78 a( /)F N.
So(!"5n 'e S(1ato 'e Co3re C6SO
A
8 a( <G.
A!"'o Ntr"!o 67NO
>
8 Con!entra'o.
7"'r5="'o 'e A$on"o 6N7
A
O78 !on!entra'o o A$on"a!o 6 N7
>
8
Rea!t"-o 'e M"((5n 6so(!"5n 'e Mer!r"o en A!"'o ntr"!o Con!entra'o8
So(!"5n a(!o*5("!a 'e C9na1to( a( <G.
So(!"5n 'e 7"'r5="'o 'e so'"o 6NaO78 a( FG.
So(!"5n 'e *"po!(or"to 'e So'"o 6NaCLO8 a( <0G.
So(!"5n a( 0)<G 'eB Ar%"n"na) Le!"na) Met"on"na) L"s"na) Tr"pto1ano) T"ros"na.
So(!"ones a( <G 'e A(3;$"na 'e 7e-o.
PR0C+.I&I+NT0
+4%erimento N 1: R+ACCI0N .+ NIN5I.RINA6
a.9 To$e A t3os 'e ensa,o ("$p"os
3.9 Co(o#e /)F $( 'e !a'a na 'e (as so(!"ones 'e a$"no&!"'os o protenas) en '"1erentes t3os 'e
ensa,o) !o$o se $estra en (a 1"%ra


T3o NH< T3o NH / T3o NH > T3o NH A
Le!"na a( 0.<G A(3$"na 'e *e-o a( <G So(!"5n Pro3(e$a A%a Dest"(a'a 6Test"%o8
!.9 Con na P"peta %ra'a'a ("$p"a) a%re%e < $( 'e So(!"5n 'e N"n*"'r"na a( 0)<G a !a'a t3o 'e
ensa,o.
'.9 Co(o#e (os t3os 'e ensa,o en n 3aIo 'e a%a *"r-"ente por nos <0 $"ntos.
e.9 Ret"re (os t3os 'e( 3aIo 'e $ar"a , 'e+e en1r"ar en (a %ra'"((a.
1.9 Anote ss O3ser-a!"ones , res(ta'os
T3o NH <B
JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
JJJJJJJJJJJJJJJJJJJJJJJJJJJJJ Res(ta'oB JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
T3o NH /B JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
JJJJJJJJJJJJJJJJJJJJJJJJJJJJ Res(ta'oB JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
T3o NH >B
JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
JJJJJJJJJJJJJJJJJJJJJJJJJRes(ta'oB JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
T3o NH AB
JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
JJJJJJJJJJJJJJJJJJJJJJJJJRes(ta'oB JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
E=per"$ento NH /B R+ACCI0N .+ "I-R+T6
a.9 To$e > t3os 'e ensa,o ("$p"os
3.9 Con na p"peta %ra'a'a ("$p"a) !o(o#e /)F $( 'e !a'a na 'e (as so(!"ones 'e a$"no&!"'os o
protenas) en '"1erentes t3os 'e ensa,o) !o$o se $estra en (a 1"%ra


T3o NH< T3o NH / T3o NH >
Le!"na o L"s"na a( 0.<G A(3$"na 'e 7e-o a( <G So(!"5n Pro3(e$a
!.9 Con na p"peta %ra'a'a ("$p"a) a%re%e 0)F $( 'e NaO7 /)F N a !a'a t3o 'e ensa,o.
'.9 A%re%e > %otas 'e so(!"5n 'e s(1ato !;pr"!o a( <G a !a'a t3o 'e ensa,o.
E.9 A%"te !a'a t3o 'e ensa,o
e.9 O3ser-e s" se 'esarro((a a(%;n Co(or.
1.9 Anote ss O3ser-a!"ones , res(ta'os
T3o NH <B
JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
JJJJJJJJJJJJJJJJJJJJJJJJJJJJJ Res(ta'oB JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
T3o NH /B JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
JJJJJJJJJJJJJJJJJJJJJJJJJJJJ Res(ta'oB JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
T3o NH >B
JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
JJJJJJJJJJJJJJJJJJJJJJJJJRes(ta'oB JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
+$P+RI&+NT0 N #: R+ACCI0N $ANT0PR0T+ICA6
a.9 To$e A t3os 'e ensa,o ("$p"os
3.9 Con na P"peta Gra'a'a ("$p"a) !o(o#e < $( 'e !a'a na 'e (as so(!"ones 'e a$"no&!"'os o
protenas) en '"1erentes t3os 'e ensa,o) !o$o se $estra en (a 1"%ra

T3o NH< T3o NH / T3o NH > T3o NH A
Tr"pto1ano a( 0.<G Met"on"na a( 0.<G A(3$"na 'e *e-o a( <G So(!"5n Pro3(e$a
!.9A%re%e !on na p"peta %ra'a'a) 0)F $( 'e 7NO> !on!entra'o en !a'a t3o) en (a !a$pana 'e
e=tra!!"5n.
'.9 Ca("ente (os t3os en n 3aIo 'e a%a *"r-"ente por <0 $"ntos.
'.9Ret"re (os t3os 'e( 3aIo , ':+e(os en1r"ar.
e.9A%re%e (enta$ente < $( 'e A$on"a!o 6N7AO78 !on!entra'o en (a !a$pana 'e e=tra!!"5n ,
o3ser-e e( !a$3"o 'e !o(or a$ar"((o *a!"a anaran+a'o en (a "nter1ase.
1.9 Anote ss o3ser-a!"ones , res(ta'os
T3o NH <B
JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
JJJJJJJJJJJJJJJJJJJJJJJJJJJJJ Res(ta'oB JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
T3o NH /B
JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
JJJJJJJJJJJJJJJJJJJJJJJJJJJJ Res(ta'oB JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
T3o NH >B
JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
JJJJJJJJJJJJJJJJJJJJJJJJJRes(ta'oBJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
T3o NH AB
JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
JJJJJJJJJJJJJJJJJJJJJJJJJRes(ta'oB JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
+$P+RI&+NT0 N ': R+ACCI0N .+ &I//0N6
a.9 To$e A t3os 'e ensa,o ("$p"os
3.9 Con na P"peta Gra'a'a ("$p"a) !o(o#e <)F $( 'e !a'a na 'e (as so(!"ones 'e a$"no&!"'os o
protenas) en '"1erentes t3os 'e ensa,o) !o$o se $estra en (a 1"%ra

T3o NH< T3o NH / T3o NH > T3o NH A
T"ros"na a( 0.<G Le!"na a( 0.<G A(3$"na 'e *e-o a( <G So(!"5n Pro3(e$a
!.9 Con na p"peta o !on n %otero) a%re%e <F %otas 'e( Rea!t"-o 'e M"((5n a !a'a t3o 'e ensa,o.
'.9 Ca("ente (os t3os en n 3aIo 'e a%a *"r-"ente por nos <0 $"n.
e.9 Ret"re !on !"'a'o (os t3os 'e ensa,o 'e( 3aIo 'e Mara , !o(5#e(os en (a %ra'"((a
1.9 Anote ss O3ser-a!"ones , res(ta'os.
T3o NH <B
JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
JJJJJJJJJJJJJJJJJJJJJJJJJJJJJ Res(ta'oB JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
T3o NH /B
JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
JJJJJJJJJJJJJJJJJJJJJJJJJJJJ Res(ta'oB JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
T3o NH >B
JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
JJJJJJJJJJJJJJJJJJJJJJJJJRes(ta'oBJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
T3o NH AB
JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
JJJJJJJJJJJJJJJJJJJJJJJJJRes(ta'oB JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
+$P+RI&+NT0 N 7: R+ACCI0N .+ (A8A,-C5I6
a.9 To$e A t3os 'e ensa,o ("$p"os
3.9 Con na P"peta Gra'a'a ("$p"a) !o(o#e <)F $( 'e !a'a na 'e (as so(!"ones 'e a$"no&!"'os o
protenas) en '"1erentes t3os 'e ensa,o) !o$o se $estra en (a 1"%ra

T3o NH< T3o NH / T3o NH > T3o NH A
Ar%"n"na a( 0.<G Met"on"na a( 0.<G A(3$"na 'e *e-o a( <G So(!"5n Pro3(e$a
!.9 A%re%e !on na p"peta ("$p"a) 0)F $( 'e 7"'r5="'o 'e So'"o 6NaO78 a( FG a !a'a t3o 'e ensa,o.
'.9 A%re%e !on n %otero) / %otas 'e a(1a9na1to( a( <G a !a'a t3o 'e ensa,o.
e.9 A%re%e / !on n %otero) / %otas 'e *"po!(or"to 'e So'"o a( <0G a !a'a t3o.
1.9 Anote ss O3ser-a!"ones , Res(ta'os.
T3o NH <B
JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
JJJJJJJJJJJJJJJJJJJJJJJJJJJJJ Res(ta'oB JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
T3o NH /B
JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
JJJJJJJJJJJJJJJJJJJJJJJJJJJJ Res(ta'oB JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
T3o NH >B
JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
JJJJJJJJJJJJJJJJJJJJJJJJJRes(ta'oBJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
T3o NH AB
JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
JJJJJJJJJJJJJJJJJJJJJJJJJRes(ta'oB JJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ
PR+,-NTA( .+ P0(T /A"0RAT0RI0
<.9 KQ: %rpos 1n!"ona(es se re-e(an en (a rea!!"5n 'e n"n*"'r"naL
/.9 KEs a'e!a'a (a rea!!"5n 'e n"n*"'r"na para '"st"n%"r a$"no&!"'os 'e protenasL E=p"#e
>.9 E=p("#e e( so , (a "$portan!"a 'e (a rea!!"5n 'e B"retL
A.9 KCon #e rea!!"5n '"st"n%"ra ste') n a$"no&!"'o 'e na protenaL
F.9 Es!r"3a (a 1or$(a estr!tra( 'e (a t"ros"na e "'ent"1"#e e( %rpo responsa3(e 'e (a rea!!"5n
'e M"((5n.
E.9 KQ: otros a$"no&!"'os 'an pos"t"-a a (a rea!!"5n 'e M"((5nL
M.9 KLa L"s"na po'ra 'ar pos"t"-a a (a rea!!"5n Xantoprote"!aL
N.9 Des!r"3a (a 1or$(a estr!tra( 'e (a Ar%"n"na , seIa(e e( %rpo %an"'"na.
"I"/I0,RAFIA:
CONN. EE , STUMP?. B"o#$"!a ?n'a$enta(. Se%n'a E'"!"5n. E'"tor"a( L"$sa9
O"(e, S.A. <PP/.
DANIEL) LQ. An' L. NEAL. La3orator, "n 3"o!*e$"str,. V. O? CORNELL) NeR CorD.
LITOACS C. 3"o#$"!a E=per"$enta(. Un $ana( 'e La3orator"o. E'"!"ones O$e%a. S.A.
Bar!e(ona) EspaIa) <PP0.