NAME: Gabayno, Blessy A.

DATE:February 20,2020

Section: ChE-5201

Biochemical Engineering

For numbers 1 and 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

For numbers 3 and 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

PROBLEMS:

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 0.95(3𝑥10−5 ) ) + (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

PROBLEMS:

SOLUTION:
 𝐾_1+𝐾2 4.4𝑥104 +(103 )
a. 𝐾𝑚 = = = 4.5𝑥10−5 𝑀
𝐾1 109
𝑉𝑚+𝑆
b. 𝑉 = 𝐾𝑚+𝑆 ; 𝑟2 = 𝑘2 + 𝑘1 − 𝑘−1 = 999.957 𝑥106 𝑉𝑚 = 𝑟2 𝐸𝑜
(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. = 𝑉𝑚𝑎𝑥 ([𝑆]) + 𝑉𝑚𝑎𝑥
𝑉
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