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CURVA ESTÁNDAR

Concentració
Tubo Glucosa (L) Agua (L) DNS (L) n glucosa
(g/L)
1 (Blanco) 0 0.003 0.003 0
2 0.0005 0.0025 0.003 0.5217
3 0.001 0.002 0.003 1.0433
4 0.0015 0.0015 0.003 1.565
5 0.002 0.001 0.003 2.0867
6 0.0025 0.0005 0.003 2.6083
7 0.003 0 0.003 3.13

Volumen 0.003 L
[Gluc]inicial 3.13 g/L

SOLUCIÓN SACAROS

Tubo Sacarosa (L) Buffer/Enzima (L) Agua (L) DNS(L)

1 (Blanco) 0 0.003 0.003 0.006


2 0.0005 0.003 0.0025 0.006
3 0.001 0.003 0.002 0.006
4 0.0015 0.003 0.0015 0.006
5 0.002 0.003 0.001 0.006
6 0.0025 0.003 0.0005 0.006
7 0.003 0.003 0 0.006

Volumen 0.006 L
Concentración 50
Sacarosa g/L

CONCENTRACIÓN GLUC
[𝐶𝑔𝑙𝑢]=5.5875[𝐴𝑏𝑠]+0.03

Absorbancia
Tubo [Glu] (g/L)
(dilución 1:40)
1 (Blanco) 0 0
2 0.088 0.522
3 0.159 0.918
4 0.202 1.159
5 0.248 1.416
6 0.234 1.337
7 0.253 1.444

ACTIVIDAD ENZIMÁT

𝑈𝐴/𝐿=(((𝐶_𝑖−𝐶_1))/ 〖��〗 _𝑔𝑙𝑢𝑐𝑜𝑠𝑎 )⁄�

0.0009

Tubo [Glu] (g/L) Tiempo (min) UA/L 0.0008


1 (Blanco) 0 10 0
0.0007
2 0.522 10 0.0002895824
3 0.918 10 0.0005097874 0.0006
4 1.159 10 0.0006431509
5 1.416 10 0.000785819 0.0005

6 1.337 10 0.0007423983 0.0004


7 1.444 10 0.0008013264
0.0003

PM glucosa 180.156 g/mol 0.0002

0.0001

0
0 0.2

CONCENTRACIÓN DE SUSTRAT
[𝐶_𝑠𝑠𝑎𝑐 ]
=𝐶_(𝑠𝑎𝑐,0)− 〖 2𝐶 〗 _𝑔𝑙𝑢𝑐

Tubo [Glu] (g/L) [Sac] inicial (g/L) [Sac residual] (g/L)

1 (Blanco) 0 0 0
2 0.522 4.167 3.123
3 0.918 8.333 6.497
4 1.159 12.5 10.183
5 1.416 16.667 13.835
6 1.337 20.833 18.158
7 1.444 25 22.113
PARÁMETROS CINÉTIC

Velocidad
Tubo [Glu] (g/L) [Sac residual] (g/L) Tiempo (min) (g/L min)
1 (Blanco) 0 0 10 0
2 0.522 3.123 10 0.052
3 0.918 6.497 10 0.092
4 1.159 10.183 10 0.116
5 1.416 13.835 10 0.142
6 1.337 18.158 10 0.134
7 1.444 22.113 10 0.144

1/𝑉=𝐾𝑚/𝑉𝑚𝑎�∙1/([𝑆])+1/𝑉𝑚𝑎�
C
25

1/[Sac] 1/[Velocidad]
20

0.320 19.168 f(x) = 45.2385493344x +


R² = 0.9883720243
0.154 10.888 15
0.098 8.631
0.072 7.064 10
0.055 7.477
0.045 6.927 5

0
0 0.05 0.1 0

Ch
([𝑆])/𝑉=1/𝑉𝑚𝑎�∙[𝑆]
+𝐾𝑚/𝑉𝑚𝑎� 180

160
[Sac] [Sac]/[Velocidad]
140 f(x) = 5.0579610159x + 38.5
R² = 0.9718997134
3.123 59.8671011437 120

6.497 70.7362795403 100


10.183 87.8818478003 80
13.835 97.7273904547
60
18.158 135.7661513922
22.113 153.1736672122 40

20

0
0.000 5.000 10.000
40

20

0
0.000 5.000 10.000
CURVA ESTÁNDAR

Absorbancia
(dilución 1:20) Curva Estándar
3.5
0
0.085 3 f(x) = 5.5875165739x + 0.0300293755
0.181 R² = 0.985019752
2.5
0.309
0.321 2
0.476
1.5
0.551
1

0.5

0
0 0.1 0.2 0.3 0.4 0.5 0.6

OLUCIÓN SACAROSA

Absorbancia
[Sac] inicial (dilución 1:40)
0 0
4.17 0.088
8.33 0.159
12.5 0.202
16.67 0.248
20.83 0.234
25 0.253

CENTRACIÓN GLUCOSA
TIVIDAD ENZIMÁTICA

Chart Title
0.0009

0.0008

0.0007

0.0006

0.0005

0.0004

0.0003

0.0002

0.0001

0
0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6

ACIÓN DE SUSTRATO RESIDUAL


RÁMETROS CINÉTICOS

Chart Title
1/Vmax 4.4095
Km/Vmax 45.239
Vmax 0.22678308
f(x) = 45.2385493344x + 4.4094714602
R² = 0.9883720243 Km 10.2594398

0.05 0.1 0.15 0.2 0.25 0.3 0.35

Chart Title
1/Vmax 5.058
Km/Vmax 38.554
f(x) = 5.0579610159x + 38.5541014019
Vmax 0.1977066
R² = 0.9718997134 Km 7.62238039

5.000 10.000 15.000 20.000 25.000


5.000 10.000 15.000 20.000 25.000
CURVA ESTÁNDAR

Concentración
Tubo Glucosa (L) Agua (L) DNS (L) glucosa (g/L)
1 (Blanco) 0 0.003 0.003 0
2 0.0005 0.0025 0.003 0.522
3 0.001 0.002 0.003 1.043
4 0.0015 0.0015 0.003 1.565
5 0.002 0.001 0.003 2.087
6 0.0025 0.0005 0.003 2.608
7 0.003 0 0.003 3.13

Volumen 0.003 L
[Gluc]inicial 3.13 g/L

SOLUCIÓN SACAROSA

Tubo Sacarosa (L) Buffer/Enzima (L) Agua (L) DNS(L)

1 (Blanco) 0 0.003 0.003 0.006


2 0.0005 0.003 0.0025 0.006
3 0.001 0.003 0.002 0.006
4 0.0015 0.003 0.0015 0.006
5 0.002 0.003 0.001 0.006
6 0.0025 0.003 0.0005 0.006
7 0.003 0.003 0 0.006

Volumen 0.006 L
Concentración 50
Sacarosa g/L

CONCENTRACIÓN GLUCOSA

[𝐶𝑔𝑙𝑢]=0.2794[𝐴𝑏𝑠]+0.03
Absorbancia
Tubo [Glu] (g/L)
(x40)
1 (Blanco) 0 0
2 3.52 1.013
3 6.36 1.807
4 8.08 2.288
5 9.92 2.802
6 9.36 2.645
7 10.12 2.858

ACTIVIDAD ENZIMÁTICA

𝑈𝐴/𝐿=(((𝐶_𝑖−𝐶_1))/ 〖��〗 _𝑔𝑙𝑢𝑐𝑜𝑠𝑎 )⁄�

0.0009

Tubo [Glu] (g/L) Tiempo (min) UA/L 0.0008


1 (Blanco) 0 10 0
0.0007
2 1.013 10 0.00056256
3 1.807 10 0.00100301 0.0006
4 2.288 10 0.00126976
5 2.802 10 0.00155512 0.0005

6 2.645 10 0.00146827 0.0004


7 2.858 10 0.00158614
0.0003

PM glucosa 180.156 g/mol 0.0002

0.0001

0
0 0.2 0.4

CONCENTRACIÓN DE SUSTRATO RE

[𝐶_𝑠𝑠𝑎𝑐 ]
=𝐶_(𝑠𝑎𝑐,0)− 〖 2𝐶 〗 _𝑔𝑙𝑢𝑐

[Sac
Tubo [Glu] (g/L) [Sac] inicial (g/L) residual]
(g/L)
1 (Blanco) 0 0 0
2 1.013 4.167 2.140
3 1.807 8.333 4.719
4 2.288 12.5 7.925
5 2.802 16.667 11.063
6 2.645 20.833 15.543
7 2.858 25 19.285

PARÁMETROS CINÉTICOS

Tiempo Velocidad (g/L


Tubo [Glu] (g/L) [Sac residual] (g/L) (min) min)
1 (Blanco) 0 0 10 0
2 1.013 2.140 10 0.101
3 1.807 4.719 10 0.181
4 2.288 7.925 10 0.229
5 2.802 11.063 10 0.280
6 2.645 15.543 10 0.265
7 2.858 19.285 10 0.286

1/𝑉=𝐾𝑚/𝑉𝑚𝑎�∙1/([𝑆])
+1/𝑉𝑚𝑎� Chart T
25

1/[Sac] 1/[Velocidad]
20

0.467 9.867 f(x) = 45.2385493344x + 4.4094714


R² = 0.9883720243
0.212 5.534 15
0.126 4.371
0.090 3.569 10
0.064 3.780
0.052 3.500 5

0
0 0.05 0.1 0.15

Chart Titl
([𝑆])/𝑉=1/𝑉𝑚𝑎�∙[𝑆]
+𝐾𝑚/𝑉𝑚𝑎� 80

70
f(x) = 2.7867725319x + 13.0869154628
60 R² = 0.9820089548

50
Chart Titl
80

70
[Sac] [Sac]/[Velocidad] f(x) = 2.7867725319x + 13.0869154628
60 R² = 0.9820089548
2.140 21.1121460409 50
4.719 26.1173609359
40
7.925 34.6435665725
11.063 39.4887961181 30
15.543 58.7594864226
20
19.285 67.4882065898
10

0
0 5 10
CURVA ESTÁNDAR

Absorbancia Absorbancia
(dilución 1:20) (x20)
0 0
0.085 1.7 Curva Estándar
0.181 3.62 3.5
0.309 6.18
3
0.321 6.42 f(x) = 0.2793758287x + 0.0300293755
R² = 0.985019752
0.476 9.52 2.5
0.551 11.02
2

1.5

0.5

0
0 2 4 6 8 10 1

SOLUCIÓN SACAROSA

Absorbancia Absorbancia
[Sac] inicial (dilución 1:40) (x40)
0 0 0
4.167 0.088 3.52
8.333 0.159 6.36
12.5 0.202 8.08
16.667 0.248 9.92
20.833 0.234 9.36
25 0.253 10.12

CONCENTRACIÓN GLUCOSA
ACTIVIDAD ENZIMÁTICA

Chart Title
0.0009

0.0008

0.0007

0.0006

0.0005

0.0004

0.0003

0.0002

0.0001

0
0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6

NTRACIÓN DE SUSTRATO RESIDUAL


PARÁMETROS CINÉTICOS
0.35

0.3

0.25

0.2

0.15

0.1

0.05

0
0 5 10 15 20 25

Chart Title
1/Vmax 4.4095
Km/Vmax 45.239
Vmax 0.22678308
f(x) = 45.2385493344x + 4.4094714602
R² = 0.9883720243 Km 10.2594398

0.05 0.1 0.15 0.2 0.25 0.3 0.35

Chart Title
1/Vmax 2.7868
Km/Vmax 13.087
f(x) = 2.7867725319x + 13.0869154628
R² = 0.9820089548
Chart Title

f(x) = 2.7867725319x + 13.0869154628 Vmax 0.35883451


R² = 0.9820089548 Km 4.69606717

5 10 15 20 25
ándar

8 10 12
CURVA ESTÁNDAR

Concentración Absorbancia
Tubo Glucosa (L) Agua (L) DNS (L) glucosa (g/L) (dilución 1:20)
1 (Blanco) 0 0.003 0.003 0 0
2 0.0005 0.0025 0.003 0.522 0.085
3 0.001 0.002 0.003 1.043 0.181
4 0.0015 0.0015 0.003 1.565 0.309
5 0.002 0.001 0.003 2.087 0.321
6 0.0025 0.0005 0.003 2.608 0.476
7 0.003 0 0.003 3.13 0.551

Volumen 0.003 L
[Gluc]inicial 3.13 g/L

SOLUCIÓN SACAROSA

Tubo Sacarosa (L) Buffer/Enzima (L) Agua (L) DNS(L) [Sac] inicial

1 (Blanco) 0 0.003 0.003 0.006 0


2 0.0005 0.003 0.0025 0.006 4.167
3 0.001 0.003 0.002 0.006 8.333
4 0.0015 0.003 0.0015 0.006 12.5
5 0.002 0.003 0.001 0.006 16.667
6 0.0025 0.003 0.0005 0.006 20.833
7 0.003 0.003 0 0.006 25

Volumen 0.006 L
Concentración 50
Sacarosa g/L

CONCENTRACIÓN GLUCOSA

[𝐶𝑔𝑙𝑢]=0.2794[𝐴𝑏𝑠]+0.0015
Absorbancia
Tubo [Glu] (g/L)
(x40)
1 (Blanco) 0 0
2 0.088 0.026
3 0.159 0.046
4 0.202 0.058
5 0.248 0.071
6 0.234 0.067
7 0.253 0.072

ACTIVIDAD ENZIMÁTICA

𝑈𝐴/𝐿=(((𝐶_𝑖−𝐶_1))/ 〖��〗 _𝑔𝑙𝑢𝑐𝑜𝑠𝑎 )⁄�

0.0009

Tubo [Glu] (g/L) Tiempo (min) UA/L 0.0008


1 (Blanco) 0 10 0
0.0007
2 0.026 10 1.448E-05
3 0.046 10 2.5492E-05 0.0006
4 0.058 10 3.216E-05
5 0.071 10 3.9294E-05 0.0005

6 0.067 10 3.7123E-05 0.0004


7 0.072 10 4.007E-05
0.0003

PM glucosa 180.156 g/mol 0.0002

0.0001

0
0 0.2 0.4

CONCENTRACIÓN DE SUSTRATO RES

[𝐶_𝑠𝑠𝑎𝑐 ]=𝐶_(𝑠𝑎𝑐,0)− 〖 2𝐶 〗
_𝑔𝑙𝑢𝑐

[Sac
Tubo [Glu] (g/L) [Sac] inicial (g/L) residual]
(g/L)
1 (Blanco) 0 0 0
2 0.026 4.167 4.114
3 0.046 8.333 8.241
4 0.058 12.5 12.384
5 0.071 16.667 16.525
6 0.067 20.833 20.700
7 0.072 25 24.856

PARÁMETROS CINÉTICOS
0.008

0.007

0.006
Tiempo Velocidad (g/L
Tubo [Glu] (g/L) [Sac residual] (g/L) 0.005
(min) min)
0.004
1 (Blanco) 0 0 10 0
0.003
2 0.026 4.114 10 0.003
3 0.046 8.241 10 0.005 0.002
4 0.058 12.384 10 0.006 0.001
5 0.071 16.525 10 0.007
0
6 0.067 20.700 10 0.007 0 5
7 0.072 24.856 10 0.007

1/𝑉=𝐾𝑚/𝑉𝑚𝑎�∙1/([𝑆])
+1/𝑉𝑚𝑎� Chart Title
25

1/[Sac] 1/[Velocidad]
#DIV/0! #DIV/0! 20

0.243 383.330 f(x) = 45.2385493344x + 4.4094714602


R² = 0.9883720243
0.121 217.748 15
0.081 172.596
0.061 141.260 10
0.048 149.522
0.040 138.527 5

0
0 0.05 0.1 0.15 0.2

Chart Title
([𝑆])/𝑉=1/𝑉𝑚𝑎�∙[𝑆]
+𝐾𝑚/𝑉𝑚𝑎� 4000

3500
f(x) = 92.5002927602x + 1058.4793481576
3000 R² = 0.9589918813
2500
Chart Title
4000

3500
[Sac] [Sac]/[Velocidad] f(x) = 92.5002927602x + 1058.4793481576
3000 R² = 0.9589918813
0.000 #DIV/0!
4.114 1577.207 2500
8.241 1794.569 2000
12.384 2137.449
1500
16.525 2334.342
20.700 3095.051 1000
24.856 3443.170 500

0
0.000 5.000 10.000 15.000
ESTÁNDAR

Concentración
glucosa real
0
0.0261 Curva Estándar
0.0522 0.18
0.0783
0.16
0.1043 f(x) = 0.2793758287x + 0.0015014688
0.14
0.1304 R² = 0.985019752
0.12
0.1565
0.1
0.08
0.06
0.04
0.02
0
0 0.1 0.2 0.3 0.4 0.5 0.6

N SACAROSA

Absorbancia
(dilución 1:40)
0
0.088
0.159
0.202
0.248
0.234
0.253

ACIÓN GLUCOSA
D ENZIMÁTICA

Chart Title

0.2 0.4 0.6 0.8 1 1.2 1.4 1.6

DE SUSTRATO RESIDUAL
ROS CINÉTICOS
0.008

0.007

0.006

0.005

0.004

0.003

0.002

0.001

0
0 5 10 15 20 25 30

Chart Title
1/Vmax 3.0331
Km/Vmax 52.253
Vmax 0.32969569
2385493344x + 4.4094714602
83720243 Km 17.2275889

0.1 0.15 0.2 0.25 0.3 0.35

Chart Title
1/Vmax 119.81
Km/Vmax 568.57
x + 1058.4793481576
Chart Title

x + 1058.4793481576 Vmax 0.00834655


Km 4.7455972

10.000 15.000 20.000 25.000 30.000


0.6