CINÉTICA ENZIMÁTICA

Extracción – Actividad Enzimática Total

AT ( minμmol∙ mL )= ε ∙ L ∙ PM
∆ A∙ϕ
caseína
∙ 10 6

AT ( minμmol∙ mL )= 10,1 g
0,0426 ∙10
−1
1 g
∙10 6
( 100 mL )( )
cm
∙1 cm∙ 23600
mol

AT ( minμmol∙ mL )=178,7212 minμmol∙ ml

AT ( minμmol∙ mL )=178,7212 mlU

Actividad Enzimática Específica

AE ( min∙molmg )= [ prot
AT
]
p
( % proteína ) ( )∙ M enzima
[ prot ] mg =
[ ] v
∙10 3
ml V muestra

10
[ prot ] =
( 100
mg/ml ) ∙5,5 ml
∙ 10 3
10 7,8 ml

mg
[ prot ] =5,102
ml

μmol
178,7212
min ∙ml
AE=
mg
5,102
ml

U
AE=35,029
mg
PURIFICACIÓN

1º. Purificación

AT ( minμmol∙ mL )= ε ∙ L ∙ PM
∆ A∙ϕ
caseína
∙ 10 6

AT ( minμmol∙ mL )= 10,1 g
0,047 6 ∙10
−1
1 g
∙10 6
( 100 mL )( )
cm
∙1 cm∙ 23600
mol

AT ( minμmol∙ mL )=1 99,6979 minμmol∙ ml

AT ( minμmol∙ mL )=199,6979 mlU

AE ( min∙molmg )= [ prot
AT
]
p
( %proteína ) ( )∙ M enzima
[ prot ] mg =
[ ] v
∙ 103
ml V muestra

25
[ prot ] =
( 100
mg/ml ) ∙5,5 ml
∙ 10 3
107,8 ml

mg
[ prot ] =12 ,755
ml

μmol
199,6979
min ∙ ml
AE=
mg
3 , 57 1
ml

U
AE=55 , 9 22
mg

2º. Purificación
AT ( minμmol∙ mL )= ε ∙ L ∙ PM
∆ A∙ϕ
caseína
∙ 10 6

AT ( minμmol∙ mL )= 10,1 g
0 , 012∙ 10
−1
1 g
∙10 6
( 100 mL )( )cm
∙1 cm∙ 23600
mol

AT ( minμmol∙ mL )=50,344 minμmol∙ml

AT ( minμmol∙ mL )=50,344 mlU

AE ( min∙molmg )= [ prot
AT
]
p
( %proteína ) ( )∙ M enzima
[ prot ] mg =
[ ] v
∙ 103
ml V muestra

5
[ prot ] =
( 100
mg/ml ) ∙5,5 ml
∙ 10 3
107,8 ml

[ prot ] =2 ,55 1 mg
ml

μmol
50,344
min∙ ml
AE=
mg
2 , 55 1
ml

U
AE=19 , 735
mg