Mtodo

LineweaverBurke
Hanes
EadieHofstee

Eje Y

Eje X

Intercepto
Eje Y

1/v

1/s

1/Vmax

s/v

Km/Vax

v/s

Vmax

La actividad enzimtica de la fosfatasa alcalina se ha medido determ

formado por p-nitrofenol fosfato (pNFF). A continuacin los datos obte

LINEWEAVER-BURK
pNF
sustrato
mMoles

0.1
0.125
0.175
0.25
0.5
1
2
4
8
15
25

pNFF
velocidad
mMoles/L*Min

Y
1/S (L/Mol)

0.405
0.495
0.655
0.885
1.805
3.325
4.905
6.195
7.135
7.795
7.835

1/V (L*Min/mMol)
2.47
2.02
1.53
1.13
0.55
0.30
0.20
0.16
0.14
0.13
0.13

10.00
8.00
5.71
4.00
2.00
1.00
0.50
0.25
0.13
0.07
0.04

HANES
pNF
sustrato
mMoles

0.10
0.13
0.18
0.25
0.50

pNFF
velocidad
mMoles/L*Min

Y
S mM(mMoles/L)

0.41
0.50
0.66
0.89
1.81

S/V (Min)

0.10
0.13
0.18
0.25
0.50

0.25
0.25
0.27
0.28
0.28

1.00
2.00
4.00
8.00
15.00
25.00

3.33
4.91
6.20
7.14
7.80
7.84

1.00
2.00
4.00
8.00
15.00
25.00

0.30
0.41
0.65
1.12
1.92
3.19

EADIE HOFSTEE
pNF
sustrato
mMoles

0.10
0.13
0.18
0.25
0.50
1.00
2.00
4.00
8.00
15.00
25.00

pNFF
velocidad
mMoles/L*Min

Y
V/S (Min-1)

0.41
0.50
0.66
0.89
1.81
3.33
4.91
6.20
7.14
7.80
7.84

V (mMol/L*Min)

4.05
3.96
3.74
3.54
3.61
3.33
2.45
1.55
0.89
0.52
0.31

0.41
0.50
0.66
0.89
1.81
3.33
4.91
6.20
7.14
7.80
7.84

Intercepto
Eje x

Pendiente

-1/Km

Km/Vmax

-Km

1/Vmax

Vmax/Km

-Km

se ha medido determinando la cantidad de p-nitrofenol (pNF) liberado al trat

uacin los datos obtenidos del ensayo a concentracin constante de la enzim

LINEWEAVER-BURK
3.00
2.50
2.00

Vmx
9.60

(1/V)

f(x) = 0.2402667391x + 0.1042411194

R = 0.9984059989

1.50

Column D

Linear (Column D

1.00
0.50

km
0.03

0.00
0.00

2.00

4.00

6.00

8.00

10.00 12.00

(1/S)

Hanes
3.50
3.00
2.50

Vmx

(S/V)

8.53

f(x) = 0.1173035174x + 0.2118228585

R = 0.9985514808

2.00

Column D

1.50

Linear (Column D

1.00
0.50

km
0.02

0.00
0.00

5.00

10.00 15.00 20.00 25.00 30.00

S

1.00
0.50
0.00
0.00

5.00

10.00 15.00 20.00 25.00 30.00

S

EADIE HOFSTEE
10.00
8.00
6.00

Vmx
9.03

(V)

f(x) = - 2.0727678479x + 9.034295781

R = 0.9575112577

Linear (Column

4.00
2.00

km
2.07

Column D

0.00
0.00 0.50 1.00 1.50 2.00 2.50 3.00 3.50 4.00 4.50
(V/S)

pNF) liberado al tratar un sustrato artificial

nstante de la enzima fosfatasa alcalina.

URK

Column D
Linear (Column D)

12.00

5
Column D
Linear (Column D)

30.00

30.00

STEE

95781
Column D
Linear (Column D)

4.00 4.50

La glucovainillina es hidrolizada por accin de la enzima B-glucosidasa en vaini

se midi de acuerdo al mtodo de DNS. A continuacin se muestran los dato
glucosidasa ( 2.5 x10-12 g de enzima purificada en 50 mL de disolucin ).

Glucovainillina
Glucosa
SUSTRATO
VELOCIDAD
X
Y
(mMoles)
(mMoles/L*Min)
1/S (L/Mol)
1/V(L*Min/mMol)
1.68E-03
1.16E-06
5.95E+02
8.62E+05
3.25E-03
2.75E-06
3.08E+02
3.64E+05
5.25E-03
3.89E-06
1.90E+02
2.57E+05
1.13E-02
4.78E-06
8.89E+01
2.09E+05
3.29E-02
5.95E-06
3.04E+01
1.68E+05
5.38E-02
6.98E-06
1.86E+01
1.43E+05

Glucovainillina
Glucosa
SUSTRATO
VELOCIDAD
x
y
(mMoles)
(mMoles/L*Min)
mM(mMoles/L)
min
1.68E-03
1.16E-06
1.68E-03
3.25E-03
2.75E-06
3.25E-03
5.25E-03
3.89E-06
5.25E-03
1.13E-02
4.78E-06
1.13E-02
3.29E-02
5.95E-06
3.29E-02
5.38E-02
6.98E-06
5.38E-02

Glucovainillina
Glucosa
SUSTRATO
VELOCIDAD
x
(mMoles)
(mMoles/L*Min)
MIN-1
1.68E-03
1.16E-06

1.45E+03
1.18E+03
1.35E+03
2.35E+03
5.52E+03
7.70E+03

y
mMOL/L*Min
6.90E-04

1.16E-06

3.25E-03
5.25E-03
1.13E-02
3.29E-02
5.38E-02

2.75E-06
3.89E-06
4.78E-06
5.95E-06
6.98E-06

8.46E-04
7.41E-04
4.25E-04
1.81E-04
1.30E-04

2.75E-06
3.89E-06
4.78E-06
5.95E-06
6.98E-06

glucosidasa en vainillina y glucosa; la determinacin de glucosa como producto de hidr

muestran los datos obtenidos de los ensayos a concentracin constante de la enzim
de disolucin ).

LINEWEAVER-BURK
Vmx
1.13202E-005

1.00E+06

km
0.0135445675

6.00E+05

8.00E+05

(1/V)

f(x) = 1196.4949646636x + 88338.4334522297

R = 0.9477911339
Column D
Linear (Column

4.00E+05
2.00E+05
0.00E+00
0.00E+00

5.00E+02

1.00E+03

(1/S)

HANES

Vmx
7.73982E-006

1.00E+04
8.00E+03

km
120535129.84

6.00E+03
(S/V)

f(x) = 129202.147006445x + 932.9211840144

R = 0.9908555419
Column D

Linear (Colu

4.00E+03
2.00E+03
0.00E+00
0.00E+00

2.00E-02

4.00E-02

6.00E-02

EADIE
8.00E-06
6.00E-06

f(x) = - 0.0060786978x + 7.30459436097289E-00

EADIE

Vmx
7.00E-06

8.00E-06

km

6.00E-06

0.0061
Axis Title

4.00E-06

f(x) = - 0.0060786978x + 7.30459436097289E-00

R = 0.7512388591
Column D

Linear (Co

2.00E-06
0.00E+00
0.00E+00

5.00E-04
Axis Title

1.00E-03

como producto de hidrlisis

constante de la enzima B-

R-BURK

8338.4334522297
Column D
Linear (Column D)

1.00E+03

ES

5x + 932.9211840144

02

Column D
Linear (Column D)

6.00E-02

IE

86978x + 7.30459436097289E-006

IE

86978x + 7.30459436097289E-006
91
Column D
Linear (Column D)

1.00E-03

La penicilina es hidrolizada y con ello inactivada por la penilasa, una enzima presente el algu
La cantidad de penicilina hidrolizada en un minuto en disolucin de 10 mL que contiene 10e
concentracin de penicilina no cambia apreciblemente durante el ensayo, representar 1/v fre

LINEWEAVER BURK
[penicilina]
moles hidr/Mn
x
y
sustrato M(Mol/L)
velocidad (Mol/L*Min)
1/s (Mol/L) 1/v (L*Min/Mol)
1.00E-06
1.10E-10
1.00E+06
9.09E+09
3.00E-06
2.50E-10
3.33E+05
4.00E+09
5.00E-06
3.40E-10
2.00E+05
2.94E+09
1.00E-05
4.50E-10
1.00E+05
2.22E+09
3.00E-05
5.80E-10
3.33E+04
1.72E+09
5.00E-05
6.10E-10
2.00E+04
1.64E+09

HANES
[penicilina]
moles hidr/Mn
X
Y
sustrato M(Mol/L)
velocidad (Mol/L*Min)
S (Mol/L)
S/V (Min)
1.00E-06
1.10E-10
1.00E-06
3.00E-06
2.50E-10
3.00E-06
5.00E-06
3.40E-10
5.00E-06
1.00E-05
4.50E-10
1.00E-05
3.00E-05
5.80E-10
3.00E-05
5.00E-05
6.10E-10
5.00E-05

9.09E+03
1.20E+04
1.47E+04
2.22E+04
5.17E+04
8.20E+04

EADIE-HOFSTEE
[penicilina]
moles hidr/Mn
X
Y
sustrato M(Mol/L)
velocidad (Mol/L*Min)
V/S (Min-1) v (Mol/L*Min)
1.00E-06
1.10E-10
1.10E-04
1.10E-10
3.00E-06
2.50E-10
8.33E-05
2.50E-10
5.00E-06
3.40E-10
6.80E-05
3.40E-10
1.00E-05
4.50E-10
4.50E-05
4.50E-10
3.00E-05
5.80E-10
1.93E-05
5.80E-10

5.00E-05

6.10E-10

1.22E-05

6.10E-10

nzima presente el algunas bacterias resistentes a la penicilina. El peso de esta enzima en Staphylococ
0 mL que contiene 10e-9 g de penicilasa purificada se midi como funcin de la concentracin de pe
yo, representar 1/v frente a 1/[s] a partir de los siguientes datos:

LINEWEAVER-BURK
1.00E+10

VMX
0.000000001

Axis Title

KM

5.00E+09

f(x) = 7622.510221609x + 1460192678.32255

Column D
R = 0.9999306967
Linear (Column D)

0.00E+00
0.00E+001.00E+062.00E+06

7622500000000

Axis Title

HANES
1.00E+05
8.00E+04

Vmx
0.000000001

6.00E+04

f(x) = 1487092463.31889x + 7414.7018072152

R = 0.9999507388
Column D

S/V 4.00E+04

km
7414700000000

Linear (Column D)

2.00E+04
0.00E+00
0.00E+00

5.00E-05

1.00E-04

EADIE
7.00E-10
6.00E-10

Vmx

f(x) = - 5.11922250403315E-006x + 6.78269107227111E-010

5.00E-10 R = 0.999126021

7E-010
km

4.00E-10

Column D

3.00E-10

Linear (Column D)

2.00E-10
1.00E-10
0.00E+00
0.00E+00 5.00E-05 1.00E-04 1.50E-04

6.00E-10

f(x) = - 5.11922250403315E-006x + 6.78269107227111E-010

5.00E-10 R = 0.999126021
V

5.00E-06

4.00E-10

Column D

3.00E-10

Linear (Column D)

2.00E-10
1.00E-10
0.00E+00
0.00E+00 5.00E-05 1.00E-04 1.50E-04
V/S

ma en Staphylococcus aureus es de 29.6 kDa.

ncentracin de penicilina. Suponiendo que la

60192678.32255
Column D
Linear (Column D)

018072152
Column D
Linear (Column D)

69107227111E-010

Column D

Linear (Column D)

69107227111E-010

Column D

Linear (Column D)