BALMES, Patricia R.

24 February 2020

ChE-5201 Biochemical Engineering

PROBLEM SET NO. 1

1-2

Histamine, an organic nitrogenous compound which can cause allergic rhinitis, can be
produced in our body through the decarboxylation of the amino acid histidine. Given
that 18 mM of the decarboxylase is used and 5 mM of histidine forms

1. Calculate the Michaelis-Menten constant if the reaction velocity and the TON are
6 mM/s and 37/s respectively.
a. 0.330 M
b. 0.440 M
c. 0.550 M
d. 0.660 M

SOLUTION:

GIVEN: [S] = 5 mM
V = 6 mM/s
TON = 37/s
[E] = 18 mM

𝑉𝑚𝑎𝑥
𝑇𝑂𝑁 = [𝐸 ∗ ]
𝑉𝑚𝑎𝑥 = 𝑇𝑂𝑁[𝐸]
𝑉𝑚𝑎𝑥[𝑆]
𝑉= 𝐾𝑚+[𝑆 ]
𝑇𝑂𝑁[𝐸][𝑆]
𝑉= 𝐾𝑚+[𝑆 ]
37
(18 𝑚𝑀)(5𝑚𝑀)
𝑠
6 𝑚𝑀/𝑠 =
𝐾𝑚+5𝑚𝑀
𝑲𝒎 = 𝟓𝟓𝟎 𝒎𝑴 𝒐𝒓 𝟎. 𝟓𝟓𝟎 𝑴
2. If 7 mM of an anti-histamine competitively inhibits the formation of histamine,
what will be the reaction velocity of the enzyme given that the dissociation
constant of the enzyme-inhibitor complex is 1.4 mM?
a. 1.0128 mM/s
b. 1.1082 mM/s
c. 1.2801 mM/s
d. 1.8210 mM/s

SOLUTION:

GIVEN: [I] = 7 Mm
Ki = 1.4 mM

𝑉𝑚𝑎𝑥[𝑆]
𝑉= [𝐼]
[𝑆]+𝐾𝑚 (1+ )
𝐾𝑠

37
18 𝑚𝑀( )(5𝑚𝑀)
𝑠
𝑉= 7 𝑚𝑀
5 𝑚𝑀+550 𝑚𝑀 (1+ )
1.4 𝑚𝑀

𝑽 = 𝟏. 𝟎𝟎𝟕𝟔 𝒎𝑴/𝒔
3-4

Lactase, also known as B-galactosidase, catalyzes the hydrolysis of lactose to produce

glucose and galactose from milk and whey. Experiments are carried out to determine
the kinetic parameters for enzyme. The initial rate data are as follows.

Lactose Reaction Velocity

Concentration (mol/L-min)
(mol/L)
2.500 1.940
2.270 1.910
1.840 1.850
1.850 1.800
1.250 1.780
0.730 1.460
0.460 1.170
0.204 0.779

3. Calculate Vmax using Lineweaver-Burk Plot.

a. 1.11 mol/L-min
b. 2.22 mol/L-min
c. 3.33 mol/L-min
d. 4.44 mol/L-min

SOLUTION:

Lineweaver- Burk Equation

1 𝐾𝑚 1 1
= ( )+
𝑉 𝑉𝑚𝑎𝑥 [𝑆] 𝑉𝑚𝑎𝑥
1
𝑦 = 𝑚𝑥 + 𝑏 ; 𝑚 = 0.1722 𝑏 = 0.4496 𝑤ℎ𝑒𝑟𝑒 𝑏 =
𝑉𝑚𝑎𝑥
1 𝒎𝒐𝒍
𝑉𝑚𝑎𝑥 = = 𝟐. 𝟐𝟐𝟒𝟑
0.4496 𝑳 − 𝒎𝒊𝒏
4. Calculate Km using Lineweaver-Burk Plot.
a. 0.18 mol/L
b. 0.28 mol/L
c. 0.38 mol/L
d. 0.48 mol/L

SOLUTION:
𝐾𝑚
𝑚 = 𝑉𝑚𝑎𝑥
𝐾𝑚
0.1722 = 2.2243
𝒎𝒐𝒍
𝑲𝒎 = 𝟎. 𝟑𝟖𝟑𝟎 𝑳
5. In an experiment conducted to evaluate the Michaelis-Menten constant, it was
found out that 1 g of bacteria could decompose the waste at a maximum rate of
35 g/day when the waste concentration was high. It was also found that the same
quantity of bacteria would decompose waste at a rate of 18 g/day when the waste
concentration was 20 mg/L. Calculate the rate of waste decomposition by 2 grams
of bacteria if the waste concentration were maintained at 8 mg/L.
a. 10.4 g/day c. 20.8 g/day
b. 15.6 g/day d. 31.2 g/day

SOLUTION:

GIVEN: V = 18 g/day
Vmax = 35g/day
[S] = 20mg/L

35𝑔 8𝑚𝑔
𝑉𝑚𝑎𝑥[𝑆] 2( )( )
𝑑𝑎𝑦 𝐿
𝑉= [𝑆]+𝐾𝑚
𝑉= 𝑚𝑔 8𝑚𝑔
18.888 +
𝐿 𝐿
𝑔 20𝑚𝑔
𝑔 35 ( ) 𝒈
𝑑𝑎𝑦 𝐿
18 𝑑𝑎𝑦 = 20𝑚𝑔 𝑽 = 𝟐𝟎. 𝟖𝟐𝟔𝟓 𝒅𝒂𝒚
+𝐾𝑚
𝐿
𝑚𝑔
𝐾𝑚 = 18.888 𝐿
 6. The enzymatic hydrolysis of an ester occurs according to the following reaction:
RCOOR + H2O RCOOH + ROH

The following data on the rate of formation of RCOOH at 25 deg C. and pH = 6.5
were obtained.

RCOOR concentration 3.0 4.5 6.0 7.5 9.0 10.5

Reaction velocity 0.051 0.064 0.071 0.079 0.082 0.091

The Michaelis-Menten constant, Km is

a. 4.28 b. 8.08 c. 16.8 d. 34.6

SOLUTION:

Lineweaver-Burke Equation

1 𝐾𝑚 1 1
= ( )+
𝑉 𝑉𝑚𝑎𝑥 [𝑆] 𝑉𝑚𝑎𝑥

1
𝑦 = 𝑚𝑥 + 𝑏 ; 𝑚 = 34.7379 𝑏 = 8.0452 𝑤ℎ𝑒𝑟𝑒 𝑏 =
𝑉𝑚𝑎𝑥
1
8.0452 =
𝑉𝑚𝑎𝑥
𝑉𝑚𝑎𝑥 = 0.1243

𝐾𝑚 = 𝑚𝑉𝑚𝑎𝑥 = 34.7379 (0.1243) = 𝟒. 𝟑𝟏𝟕𝟖

Biochemical Engineering Fundamentals 2e Bailey & Ollis

3.1 Determination of Km and Vmax. Initial rates of an enzyme-catalyzed reaction

for various substrate concentrations are listed in Table 3P1.1.

a. Evaluate Vmax and Km by a lineweaver-Burk Plot

b. Using eadie Hofstee plot, evaluate Vmax and Km
c. Calculate the standard deviation of the slope and intercept for each method.

SOLUTION:

a. Lineweaver-Burk Plot
1 𝐾𝑚 1 1
= ( )+
𝑉 𝑉𝑚𝑎𝑥 [𝑆] 𝑉𝑚𝑎𝑥
1 𝑚𝑜𝑙
= 7.62𝑥10−9 ; 𝑉𝑚𝑎𝑥 = 1.31𝑥108
𝑉𝑚𝑎𝑥 𝐿 − 𝑚𝑖𝑛
𝐾𝑚
𝑚 = 𝑉𝑚𝑎𝑥 = 8.08𝑥10−14 ; 𝐾𝑚 = 1.06𝑥10−5 𝑚𝑜𝑙/𝐿

𝐾𝑚𝑉
b. 𝑣 = − [𝑆]
+ 𝑉𝑚𝑎𝑥
3.2. Batch enzymic reaction. An enzyme with a Km of 1x10-3 M was assayed using
an initial substrate concentration of 3x10-5 M. After 2 min, 5 percent of the substrate
was converted. How much substrate will be converted after 10, 30, and 60 min?

SOLUTION:

Given: Km=1x10-3 M
[S]initial = 3x10-5 M
[𝑆]𝑖
𝐾𝑚 (𝑙𝑛 [𝑆]𝑓 ) + ([𝑆]𝑖 − [𝑆]𝑓)
𝑉𝑚𝑎𝑥 =
𝑡𝑓 − 𝑡𝑖

3𝑥10−5
1𝑥10−3 (𝑙𝑛 ) + (3𝑥10−5 − 0.95(3𝑥10−5 ))
0.95(3𝑥10−5 )
𝑉𝑚𝑎𝑥 = ; [𝑆]𝑓 = 0.95[𝑆]𝑖
2

𝑉𝑚𝑎𝑥 = 2.64 𝑥10−5 𝑀/𝑚𝑖𝑛

3𝑥10−5
1𝑥10−3 (𝑙𝑛 [𝑆]𝑓
) + (3𝑥10−5 − [𝑆]𝑓)
−5
2.64 𝑥10 = ; 𝐴𝑓𝑡𝑒𝑟 10 𝑚𝑖𝑛𝑠
10 − 2
[𝑆]𝑓 = 𝟐. 𝟒𝟒 𝒙𝟏𝟎−𝟓 𝑴

3𝑥10−5
1𝑥10−3 (𝑙𝑛 [𝑆]𝑓
) + (3𝑥10−5 − [𝑆]𝑓)
−5
2.64 𝑥10 = ; 𝐴𝑓𝑡𝑒𝑟 30 𝑚𝑖𝑛𝑠
30 − 2
[𝑆]𝑓 = 𝟏. 𝟒𝟓 𝒙 𝟏𝟎−𝟓 𝑴

3𝑥10−5
1𝑥10−3 (𝑙𝑛 [𝑆]𝑓
) + (3𝑥10−5 − [𝑆]𝑓)
−5
2.64 𝑥10 = ; 𝐴𝑓𝑡𝑒𝑟 60 𝑚𝑖𝑛𝑠
60 − 2
[𝑆]𝑓 = 𝟔. 𝟒𝟔 𝒙 𝟏𝟎−𝟔 𝑴
Bioprocess Engineering Basic Concepts by Shuler and Kargi

SOLUTION:
𝐾_1+𝐾2 4.4𝑥104 +(103 )
a. 𝐾𝑚 = = = 4.5𝑥10−5 𝑀
𝐾1 109
𝑉𝑚+𝑆
b. 𝑉 = ; 𝑟2 = 𝑘2 + 𝑘1 − 𝑘−1 = 999.957 𝑥106 𝑉𝑚 = 𝑟2 𝐸𝑜
𝐾𝑚+𝑆
(999.957 𝑥106 )(10−6 ) + 10−3
𝑉= = 9.58𝑥10−4 𝑀
4.5𝑋10−5 + 10−3
SOLUTION:

a. For I=0, No inhibitor

b. For 1/Vm = 0.2 and Vm=5 mM/h
𝑉𝑚[𝑆]
𝑉= 𝐼
𝐾𝑚 (1 + 𝐾1) + [𝑆]
1.3𝑚𝑀 5(0.5)
= 1.3
ℎ 0.83 (1 + ) + 0.5
𝐾1
𝐾1 = 𝟏. 𝟖𝟐 𝒎𝑴
SOLUTION:

1 𝐾𝑚 1 1
a. 𝑉
= 𝑉𝑚𝑎𝑥 ([𝑆]) + 𝑉𝑚𝑎𝑥
𝑉𝑚
𝑆𝑙𝑜𝑝𝑒 = 9.2𝑥10−3 = 𝑦 − 𝑖𝑛𝑡𝑒𝑟𝑐𝑒𝑝𝑡 = 𝑉𝑚 = 0.04 𝑚𝑙/ℎ
𝐾𝑎
0.04 𝒖𝒈
𝐾𝑎 = −3
= 𝟒. 𝟑𝟓
9.2𝑥10 𝒍
60
b. 𝑉 = 𝑉𝑚(1 + 𝐾𝑎) = 0.04 (1 + 4.35) = 𝟎. 𝟓𝟗 𝒎𝒍/𝒉