CHE142

IMMOBILIZED
ENZYME SYSTEMS
Ababat
Abolencia
Alcantara
Atienza
Ayson
PROBLEM 3-10
 REQUIRED
mg mg
A. Vmax
mlmin
unit of enzyme ; KM
ml
cal
B. Activation Energy ( )
gmole
C. Determine if the immobilized enzyme is diffusion limited.
 SOLUTION
A. Plotting the product concentration versus time
350

300

250
Poduct Concentration
(mg dextrose/ml)

200
Soluble data

150 Immobilized data

100

50

0
0 50 100 150 200 250 300 350 400 450
Time (min.)
Working Reaction Equation:
glucoamylase
Water + Starch Dextrose
P-dextrose
Molecular Weight of Dextrose: 180 g
Molecular Weight of Water: 18 g

Amount of water used = Amount of Dextrose Produced

18 g of Water used = 18 g of Dextrose Produced
18
1 mole of Water = = 0.1 moles of Dextrose Produced
180
Starch Remaining ( [ S] ):
S = S 0 Dextrose formed Water used
S = S 0 P 0.1 P
S = S 0 0.9P

Mass Balance Equation : S = S 0 0.9P

dP d Dextrose 21
Rate Equation : = = =
dt dt 21
Setting up the data and tabulating the graph parameters
Time (min) Product Concetration (mg Soluble Immobilized
dextrose/ml) x ( 1/[S] ) y (1/v) x ( 1/[S] ) y (1/v)

Soluble Immobilized
0.006361323 0.535714286 0.003130478 0.12864494
0 12 18.4
15 40 135 0.007575758 0.410958904 0.004662005 0.230769231

30 76.5 200 0.010085729 0.842696629 0.006410256 0.416666667

45 94.3 236
0.012029352 0.583657588 0.008090615 0.625
60 120 260
75 135.5 258 0.016666667 0.967741935 0.009803922 0.725

90 151.2 262 0.021715527 0.955414013 0.009633911 3.75

105 150.4 266
120 155.7 278 0.031328321 1.875 0.00998004 3.75

135 160.1 300 0.030637255 2.830188679 0.010351967 1.25

150 164.9 310

0.035880875 3.409090909 0.011655012 0.681818182
165 170 306
225 316 0.041823505 3.125 0.015151515 1.5

415 320 0.051046452 2.941176471 0.01754386 3.75

 B) Plotting ln Vm versus 1/T for determining the activation
energy for soluble and immobilized enzyme reactions gives

T Vmsol Vmimmo lnVmsol (y) lnVmimo (y) 1/T (x)

0.00335
298 0.62 0.8 -0.478035801 -0.223143551 5705
0.00324
308 1.42 1.4 0.350656872 0.336472237 6753
0.00314
318 3.6 3 1.280933845 1.098612289 4654
0.00304
328 8 6.2 2.079441542 1.824549292 878
3 0.00295
338 16 11 2.772588722 2.397895273 858
2.5

1.5

1
y = -8298.6x + 41.18
0.5 y = -6582.4x + 35.65

0
0.0029 0.00295 0.003 0.00305 0.0031 0.00315 0.0032 0.00325 0.0033 0.00335 0.0034

-0.5

-1
Soluble Immobilization Linear (Soluble) Linear (Immobilization)
 C) Since Km (soluble) = Km (apparent) there is no diffusion
limitation.