Calculos para Inhibicin enzimticaTubo123456

C piruvato (mM)0.060.090.150.250.360.5

Tiempo (min)Absorbancia

0.51.351.1171.0821.1560.9741.372

11.321.0721.0561.10.9091.288

1.51.2961.0241.0241.0310.8391.202

21.2720.9720.990.980.7711.122

2.51.2490.9290.9570.9240.7031.047

31.2290.8850.920.8690.6330.972

3.51.2110.8450.8920.8120.570.904

41.1920.8030.8670.7650.510.831

4.51.1760.7620.840.70.4510.761

51.160.7220.8170.6470.3960.694

Actividad0.0017354850.0036582530.0025303960.0046779030.0054103860.006242753

Volumen de reaccion (mL)111111

Se conocen los datos de la C de piruvato (mM) Graficar Abs vs tiempo para obtener la pendiente (Abs/min)

Cantidad de enzima Se calcula tomando en cuenta la concentracin de la protena en la fraccin, en este caso F3. Concentracin F3= 38.63 ug/uL= 38.63 mg/mL Dilucin= (1:100) Se agregaron 10 uL de F3 diluida

Unidades

Clculo para 0.06 mM Pendiente= 0.0417 Usar pendientes positivas Volumen de la reaccin= Volumen final de la celda= 1.0 mL

Una vez que se obtienen todas las actividades:

Concentracin PiruvatoActividad especifica1/ [S]1/Actividad[S][S]/v

0.060.00173548516.66576.200.0634.57

0.150.0025303966.66395.190.1559.27

0.250.0046779034213.770.2553.44

0.360.0054103862.77184.820.3666.53

0.50.0062427532160.180.580.09

Grfica de Lineweaver-Burk ( Actividad especfica vs Concentracin)

Grfica Hanes

Ecuacin de Lineweaver-Burk

Y= 1/Vo X= 1/[C] m= (Km/Vmax) b= 1/Vmax

Ecuacin de Hanes

Y= C/v X= [C] m= (1/Vmax) b= (Km/Vmax)

Inhibidor

Tubo123456

C piruvato (mM)0.060.090.150.250.360.5

Tiempo (min)Absorbancia

0.50.9771.0981.0421.0291.0311.002

10.9711.0921.02511.0020.967

1.50.9481.0751.0060.9690.9750.933

20.9351.0530.9880.9410.9460.903

2.50.9191.0350.9690.9110.9180.865

30.9071.0140.950.8810.8910.828

3.50.8940.9990.9330.8520.8630.789

40.880.9830.9140.8230.8360.752

4.50.8680.9680.8950.7940.810.713

50.8540.9490.8770.7650.7820.673

Actividad0.0011611520.001440.0015315550.0024430.0022973330.00303814

Volumen de reaccin (mL)111111

ConcentracinActividad1/ [S]1/Actividad[S][S]/v

0.060.00116115216.6666667861.21362010.0651.6728172

0.150.0015315556.66666667652.93097830.1597.93964674

0.250.0024429974409.33321980.25102.3333049

0.360.0022973332.77777778435.28731880.36156.7034348

0.50.003038142329.14876710.5164.5743836

Para conocer el tipo de inhibicin, se comparan todos los grficos