Enzimas

UNIVERSIDAD NACIONAL AUTÓNOMA DE MÉXICO

FACULTAD DE CIENCIAS
Laboratorio de Biología Molecular de la Célula l

Reporte No. 3
“Cinética enzimática de la poli fenol oxidasa”

PRESENTA:
Serrano García Estrella
Hernández Ortiz Jorge Eduardo
Utrera Salgado Brenda
Celis Tobón Alma

MÉXICO, D.F. 2012

Objetivos
Ejemplificar cómo se determina la Vo de una enzima y el método paraanalizar la relación entre la (S) y la Vo, lo que nos permitirá calcular la Km y la Vmax.

Hipótesis
La enzima será afín al sustrato y poder calcular la Vmax de la reacción
Variable 1: Afinidad de la enzima al sustrato (Extracto de papa)
Variable 2: Tiempo.
Variable 3: Absorbancia.
Resultados

Tabla 1 Equipo 1 y 2. |
Tiempo/Abs | tubo 1 | tubo 2 | tubo 3 | tubo 4 | tubo 5 | tubo 6 |
0 |------ | 0 | 0 | 0 | 0 | 0 |
15 | ------ | 0.665 | 0.668 | 0.841 | 0.863 | 0.852 |
30 | ------ | 1.024 | 0.969 | 1.023 | 1.031 | 0.962 |
45 | ------ | 1.21 | 0.994 | 1.159 | 1.167 | 1.066 |
60 | ------ | 1.305 | 1.09 | 1.286 | 1.27 | 1.158 |
75 | ------ | 1.383 | 1.171 | 1.378 | 1.36 | 1.253 |
90 | ------ | 1.442 | 1.258 | 1.47 | 1.488 | 1.331 |
105 | ------ | 1.509 | 1.327 | 1.553 |1.604 | 1.388 |
120 | ------ | 1.564 | 1.376 | 1.612 | 1.633 | 1.449 |
135 | ------ | 1.623 | 1.428 | 1.687 | 1.637 | 1.504 |
150 | ------ | 1.665 | 1.48 | 1.745 | 1.675 | 1.548 |
165 | ------ | 1.699 | 1.532 | 1.792 | 1.723 | 1.592 |
180 | ------ | 1.725 | 1.572 | 1.842 | 1.754 | 1.623 |

Tabla dos Equipo 3, 4 y 5.
Tiempo/Abs | tubo 1 | tubo 2 | tubo 3 | tubo 4 | tubo 5 | tubo 6 |0 | | 0 | 0 | 0 | 0 | 0 |
15 | | 0.896 | 0.725 | 0.955 | 0.828 | 0.597 |
30 | | 1.28 | 0.948 | 1.202 | 0.975 | 0.686 |
45 | | 1.482 | 1.121 | 1.33 | 1.086 | 0.806 |
60 | | 1.637 | 1.267 | 1.418 | 1.203 | 0.896 |
75 | | 1.676 | 1.363 | 1.511 | 1.266 | 0.976 |
90 | | 1.725 | 1.447 | 1.577 | 1.346 | 1.053 |
105 | | 1.796 | 1.513 | 1.632 | 1.349 | 1.113 |
120 | | 1.848 |1.562 | 1.6769 | 1.44 | 1.174 |
135 | | 1.883 | 1.612 | 1.715 | 1.48 | 1.228 |
150 | | 1.915 | 1.626 | 1.747 | 1.501 | 1.278 |
165 | | 1.926 | 1.648 | 1.778 | 1.518 | 1.314 |
180 | | 1.948 | 1.671 | 1.805 | 1.579 | 1.38 |
Las gráficas se encuentran en el apéndice elaboradas en papel milimétrico.

Cálculos
----1mL – 6.3 mg
0.1 mL - x
x = 0.63 mg = 0.63x10-3g0.63x10-3g/126g/mol = 5x10-6mol

0.1mL= 0.1x10-3L
M = 5x10-6 Mol/0.1x10-3L = 0.05 M

----1mL – 6.3 mg
0.3mL - x
x = 1.89 mg = 1.89x10-3g

1.89x10-3g/126g/mol = 1.5x10-5mol

0.3mL= 0.3x10-3L
M = 1.5x10-5 Mol/0.3x10-3L = 0.05 M

----1mL – 6.3 mg
0.6 mL - x
x = 3.78 mg = 3.78x10-3g

3.78x10-3g/126g/mol = 3x10-6mol

0.6mL= 0.6x10-3L
M = 3x10-5 Mol/0-6x10-3L = 0.05 M

----1mL – 6.3mg
1.2 mL - x
x = 7.56 mg = 7.56x10-3g

7.56x10-3g/126g/mol = 6x10-5mol

1.2 mL= 1.2x10-3L
M = 6x10-5 Mol/1.2x10-3L = 0.05 M

----1mL – 6.3 mg
1.8 mL - x
x = 11.34 mg = 11.34x10-3g

11.34x10-3g/126g/mol = 9x10-5mol

1.8 mL= 1.8x10-3L
M = 9x10-5 Mol/1.8x10-3L = 0.05 M
----1mL – 6.3 mg
2.4 mL - x
x = 15.12 mg = 15.12x10-3g

15.12x10-3g/126g/mol =1.2x10-4mol

2.4 mL= 2.4x10-3L
M = 1.2x10-4 Mol/2.4x10-3L = 0.05 M

Equipo 1 y 2
Tubo | Operación | Absorbancia/minuto |
2 | 0.0144 x 60 | 0.864 |
3 | 0.0167 x 60 | 1.002 |
4 | 0.012 x 60 | 0.72 |
5 | 0.0109 x 60 | 0.654 |
6 | 0.0119 x 60 | 0.714 |
C = M/Ɛ
Donde C = Concentración sobre minutos
Tubo | C = M/Ɛ | Resultado |
2 | C = (0.864/6.3 mg/mL) | 0.1371 |
3 | C = (1.002/6.3mg/mL) | 0.1590 |
4 | C = (072/6.3 mg/mL) | 0.1142 |
5 | C = (0.654/6.3 mg/mL) | 0.1038 |
6 | C = (0.714/6.3 mg/mL) | 0.1133 |

Equipo 3, 4 y 5
Tubo | Operación | Absorbancia/minuto |
2 | 0.0205 x 60 | 1.23 |
3 | 0.0164 x 60 | 0.984 |
4 | 0.0173 x 60 | 1.038 |
5 | 0.0144 x 60 | 0.864 |
6 | 0.0073 x 60 | 0.438 |

Tubo | C = M/Ɛ | Resultado |
2 | C = (1.23/6.3 mg/mL) |...