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Ejercicio Semana 7

The document contains data from experiments measuring initial velocity (Vo) of an enzyme-catalyzed reaction using varying concentrations of substrates AX and B. The data is plotted and analyzed to determine kinetic parameters including Vmax, Km, and K'. Vmax increases with increasing [B] concentration, indicating substrate B acts as an allosteric effector. Lineweaver-Burk plots are used to calculate Km for substrates AX and B at different concentrations. K' is determined and also increases with [B] concentration, demonstrating substrate B is an allosteric activator of the enzyme.

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Carlos Israel Esparza Andrade
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47 views9 pages

Ejercicio Semana 7

The document contains data from experiments measuring initial velocity (Vo) of an enzyme-catalyzed reaction using varying concentrations of substrates AX and B. The data is plotted and analyzed to determine kinetic parameters including Vmax, Km, and K'. Vmax increases with increasing [B] concentration, indicating substrate B acts as an allosteric effector. Lineweaver-Burk plots are used to calculate Km for substrates AX and B at different concentrations. K' is determined and also increases with [B] concentration, demonstrating substrate B is an allosteric activator of the enzyme.

Uploaded by

Carlos Israel Esparza Andrade
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as XLSX, PDF, TXT or read online on Scribd
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Velocidad inicial (mM/min)

INICIAL [AX]
[B]=2.0mM [B]=4.0 mM [B]=6.0 mM
2 250 286 300
2.5 278 323 341
3.3 313 371 395
5 357 435 469
10 417 526 577

Vo versus [AX]
700

600

500 V'máx versus [B]


Vo (mM/min)

400 800 [B]=2.0mM


700 [B]=4.0 mM
300
[B]=6.0 mM
600
200
V'máx (mM/min)
500
100
400
0
1 2 3 4 5 6 7 8 9 10300 11
[AX] (mM) 200
100
1/Vo versus 1/[AX] 0
0 1 2 3 4 5 6
0.004500 [B] mM
0.004000
0.003500
0.003000
0.002500 [B]=2.0mM
1/Vo

[B]=4.0 mM
0.002000
[B]=6.0 mM
0.001500
0.001000
0.000500
0.000000
0.05 0.10 0.15 0.20 0.25 0.30 0.35 0.40 0.45 0.50 0.55
1/[AX]
1/Velocidad inicial (mM/min) [B] V'máx
1/[AX]
[B]=2.0mM 4.0 mM 6.0 mM 2.0 501.0
0.50 0.004000 0.003497 0.003333 4.0 667.1
0.40 0.003597 0.003096 0.002933 6.0 751.9
0.30 0.003195 0.002695 0.002532
0.20 0.002801 0.002299 0.002132
0.10 0.002398 0.001901 0.001733
m 0.00399944 0.00398749 0.0040005
b 0.00199599 0.00149891 0.00132999
Vmax' 0 501.0 667.1 751.9
K' 0 2.00 2.66 3.01
[B] mM 0 2.0 4.0 6.0
sus [B] K' versus [B]
3.5

2.5

2
K'

1.5

0.5

0
4 5 6 7 0 1 2 3 4 5 6 7
] mM [B] mM
K' 1/[B] 1/V'máx 1/V´máx vs 1/[B]
0.00250
2.00 0.500 0.00200
2.66 0.250 0.002000.00150 f(x) = 0.001995743019334 x + 0.000998486776271
3.01 0.167 0.00133 R² = 0.999984934570955
0.00150
1/V'máx

m 0.00199574
b 0.00099849
0.00100
Vmáx 1001.5
Km(AB) KmB 0.00050 2.00
0.00000
0.100 0.150 0.200 0.250 0.300 0.350 0.400 0.450 0.500 0.550
1/[B]
Velocidad inicial (mM/min) 1/Velocidad inicial (mM/min)
INICIAL [B] [AX]=2.0m [AX]=3,3 [AX]=10,0 1/[AX] [AX]=2.0m [AX]=3,3
M mM mM M mM
2 250 313 417 0.50 0.004000 0.003195
4 286 371 526 0.25 0.003497 0.002695
6 300 395 577 0.17 0.003333 0.002532
m 0.0020032 0.0019916
b 0.0029978 0.0021988
Vmax' 0 333.6 454.8
K' 0 0.67 0.91
[AX] mM 0 2.0 3.3

Vo versus [B]
700

600
800
500
700
Vo (mM/min)

400 [AX]=2.0mM
600
[AX]=3,3 mM

V'máx (mM/min)
300 [AX]=10,0 mM 500
200 400

100 300
200
0
1.5 2 2.5 3 3.5 4 4.5 5 5.5 6 6.5
100
[AB] (mM) 0
0
ad inicial (mM/min) [AX] V'máx K' 1/[AX] 1/V'máx
[AX]=10,0
mM 2.0 333.6 0.67 0.500 0.00300 0.00350

0.002398 3.3 454.8 0.91 0.303 0.00220 0.00300


0.001901 10.0 713.4 1.42 0.100 0.00140 0.00250
0.001733 m 0.00399
0.00200

1/V'máx
0.0019933 b 0.0009984
0.00150
0.0014017 Vmáx 1001.6
713.4 Km(BA) alfa KmA 4.00 0.00100

1.42 0.00050
10.0 0.00000
0.050 0.10

V'máx versus [B] K' versus [B]


800 1.6
700 1.4
600 1.2
V'máx (mM/min)

500 1
400 0.8
K'

300 0.6
200 0.4
100 0.2
0 0
0 2 4 6 8 10 12 0 2 4 6 8 10 12
[B] mM [B] mM
1/V´máx vs 1/[AX]
0.00350

0.00300
f(x) = 0.003989998938057 x + 0.000998414857775
0.00250 R² = 0.999910132605519

0.00200
1/V'máx

0.00150

0.00100

0.00050

0.00000
0.050 0.100 0.150 0.200 0.250 0.300 0.350 0.400 0.450 0.500 0.550
1/[B]

[B]

6 8 10 12
mM
V =
TIPO DE MECANISMO

1001.5
Vmáx
Km (AB)
Km(BA)
KA
KB

TIPO DE MECANISMO

Vmáx
KmB
alfa KmA

4.0
4 .0 *  B   2 .0 *  A    A
[A]
4.00 mM
-0.0104
0.0062

* [B] +
Al azar

1001.5 mM/min
2.00 mM

Desplazamiento

1001.5 mM/min
2.0 mM
4.0 mM

[B]
2.0
Tiende a cero

* [A] + [A] [B]


.0 *  A    A  B
𝐾^′=(𝐾_𝑀^𝐵𝐴 (𝐾_𝐵+
[𝐵]))/(𝐾_𝑀^𝐴𝐵+[𝐵])

𝐾^′=(𝐾_𝑀^𝐴𝐵 (𝐾_𝐴+
[𝐴]))/(𝐾_𝑀^𝐵𝐴+[𝐴])

V `max 

K `
V max  A
KM A   A
KM B  A
KM A   A
K'
[B] mM
KB

K'
[AX] mM
KA

V `max 

K `
2.00
2.00
0.0050

0.67
2.00
0.0047

V max  B 
2.66
4.00
-0.0068

0.91
3.30
0.0065

KM B   B 
KM  B 
A

KM   B 
B
3.01
6.00
0.0204

1.42
10.00
-0.0423

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