Microbial population growth

Microbial population growth

A new microorganism has been discovered which at each cell A new microorganism has been discovered which at each cell
division yields three daughters. From the growth rate data below division yields three daughters. From the growth rate data below
calculate the mean time between successive cell divisions? calculate the mean time between successive cell divisions?
t [h] t [h] X[g/dm X[g/dm
3 3
] ]
0.0 0.0 0.10 0.10
0.5 0.5 0.15 0.15
1.0 1.0 0.23 0.23
1.5 1.5 0.34 0.34
2.0 2.0 0.51 0.51
Temperature variation of Growth
Temperature variation of Growth
Johnson, Johnson, Eyring Eyring and and Polisaar Polisaar represent growth of represent growth of E. coli E. coli between between
18 18
o o
C and 46 C and 46
o o
C by the following equation for the specific growth C by the following equation for the specific growth
rate rate : :
Plot this function as log Plot this function as log versus 1/T versus 1/T
Show that this equation can be represented as the product of two Show that this equation can be represented as the product of two function function
whose form is suggested by the plot in above part, and what expl whose form is suggested by the plot in above part, and what explanation anation
rationalizes these two individual functions and the value of abo rationalizes these two individual functions and the value of above ve
parameters? parameters?
In this interpretation of In this interpretation of as function of T, what implicit assumption are as function of T, what implicit assumption are
made with regard to irreversible deactivation? made with regard to irreversible deactivation?
|
.
|

\
|
+
|
.
|

\
|

=
T
T
T
75200
239 exp 1
7520
exp 10 96 . 9
9

Answer Sheet
Answer Sheet
Deactivation line Deactivation line
This decreasing line presented a rapid decrease in growth rate a This decreasing line presented a rapid decrease in growth rate as the temperature s the temperature
approaches the upper limit for survival of the microorganism ten approaches the upper limit for survival of the microorganism tend that the most d that the most
thermally sensitive essential protein denatures and this hypothe thermally sensitive essential protein denatures and this hypothesis also has been sis also has been
confirmed in several instance by genetic studies in which mutati confirmed in several instance by genetic studies in which mutati on of a single gene on of a single gene
has caused a large change in the maximum tolerable temperature f has caused a large change in the maximum tolerable temperature for or
microorganism microorganism
One physical mechanism for this phenomenon is obvious as the tem One physical mechanism for this phenomenon is obvious as the temperature perature
increases, the atoms in the enzyme molecule have greater energie increases, the atoms in the enzyme molecule have greater energies and a greater s and a greater
tendency to move. Eventually, they acquire sufficient energy to tendency to move. Eventually, they acquire sufficient energy to overcome the overcome the
weak interactions holding the globular protein structure togethe weak interactions holding the globular protein structure together, and deactivation r, and deactivation
follows. follows.
Activation line Activation line
This increasing line clarified at low temperature, apparently, t This increasing line clarified at low temperature, apparently, t he metabolic activity he metabolic activity
of cell increase with increasing temperature as the activities o of cell increase with increasing temperature as the activities of its enzyme rises. f its enzyme rises.
Notice in this Notice in this activation line, activation line, that was also commonly called to that was also commonly called to Arrhenius Arrhenius plot, plot,
that that classical classical Arrhenius Arrhenius behavior behavior appears at low temperature, exactly, below of appears at low temperature, exactly, below of
the maximum tolerable temperature for microorganism. the maximum tolerable temperature for microorganism.
The implicit assumption are made regard to irreversible deactiva The implicit assumption are made regard to irreversible deactivation : tion :
irreversibility alteration of whole active forms of enzyme to in irreversibility alteration of whole active forms of enzyme to inactive forms in active forms in
case of a large change in the maximum tolerable temperature for case of a large change in the maximum tolerable temperature for each each
microorganism microorganism
Single and Multiple Substrate Kinetics
A culture grown in a simple medium including 0.3% wt/vol of glucose;
at time t=0 , it is inoculated into a larger sterile volume of the
identical medium that was shown at following result [X(1)]. The same
species cultured in a complex medium is inoculated into a mixture of
0.15% glucose and 0.15% lactose [X(2)]. Evaluate the maximum
specific growth rate
max,
the lag time

t
lag
and the overal yield factors Y
(gram cells per gram substrate), assuming substrate exhausting in
each case. Explain the model kinetic of each case that was tended
with shape of the growth curves. If the model kinetic of the single
substrate is andrew equation, advice the equation of the above
multiple substrate kinetics.

T (h) 0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0 7.5 8.0
X(1)(g/l)

0.06 0.08 0.11 0.14 0.20 0.26 0.37 0.49 0.70 0.88 1.04 1.04
X(2)(g/l)

0.06 0.06 0.06 0.07 0.10 0.13 0.18 0.26 0.32 0.43 0.48 0.50 0.52 0.60 0.84 1.0 1.0

Answer Sheet
Specific growth rate
Single substrate Multiple Substrate



Time Lag, single substrate t
lag
0.0h
multiple substrate t
lag
(1) 1.0h t
lag
(2) 1.0h
Yield Factor, Single Substrate Y = (1.04-0.06)/3.0= 0.327
Multiple Substrate Y(1) = (0.48-0.06)/1.5= 0.280
Y(2) = (1.0-0.48)/1.5 = 0.347
Multiple substrate kinetic model:


0
0
max
) / ln(
t t
X X

=
1 1
max
606 . 0
0 0 . 3
) 06 . 0 / 37 . 0 ln(

=

= h h
1 1
max
549 . 0
0 . 1 0 . 3
) 06 . 0 / 18 . 0 ln(
) 1 (

=

= h h
1 1
max
672 . 0
5 . 6 0 . 7
) 6 . 0 / 84 . 0 ln(
) 2 (

=

= h h
2 2 2 2 1 1 1 1
max
/ / 1
1
/ / 1
1
i s i s
K s s K K s s K + +

+ +
=
Microbial Population Growth
A new microorganism has been discovered which at each cell
division yield (CDY) have 3 daughters. From the following result,
calculate the mean time between successive cell division.




T[h] 0.0 0.5 1.0 1.5 2.0
X[g/l] 0.10 0.15 0.23 0.34 0.51
D
t
CDY ) 1 ln(
max
+
=
Answer Sheet
Specific growth rate


Mean Time between successive cell division
1
0
0
max
815 . 0
2
) 10 . 0 / 51 . 0 ln( ) / ln(

= =

= h
t t
X X

h h
CDY
t
D
7 . 1
815 . 0
) 3 1 ln( ) 1 ln(
max
=
+
=
+
=

Temperature effect of Cellular Growth

(I ndonesian Language)

Pertumbuhan Microcystis sp. D102 ditunjukan mengikuti persamaan berikut:
={9.96 . 10
7
. exp [-7520/T]}/{1+ exp [239-75200/T]}, T in K
1. Tunjukan kurva laju pertumbuhan sebagai fungsi suhu
2. Berapa temperature optimumnya?
3. Apabila komposisi membrane sel luar dari mikroba ini seperti yang ditunjukan
pada table berikut dan T
m
adalah 361 K, dengan memperhatikan perolehan
jawaban dari pertanyaan terdahulu, termasuk jenis apakah Microcystis sp. D102
ini?
Membran FA 14:0 14:1 16:0 16:1 16:2
Fraksi Mol 0.2 0.1 0.3 0.2 0.2

Jawab:
1. Kurva Pertumbuhan Microcystis sp. D102

Growth Curve
0
0.01
0.02
0.03
0.04
0.05
0.06
0.07
0.08
25 35 45 55
T [Celcius degree]
S
p
e
c
.

G
r
o
w
t
h

R
a
t
e

[
1
/
d
a
y
]


2. Temperature Optimum: 37
o
C
3. Indeks Ikatan Rangkap Membran Sel (DBI) =0(0.2)+ 1(0.1)+0(0.3)+1(0.2)+2(0.2)=0.7
Persentase Guanin- Sitosin dalam DNA Microcystis sp. D102 (%GC)
%GC= (T
m
(
o
C)-69.3)/41=(88-69.3)/41=0.456
Dari kurva teridentifikasi untuk DBI 0.7 dan %GC 0.456 menunjukan strain adalah
Mesophile, serupa dengan Microcystis aeruginosa IAM 228
Please solve 3 numbers for mark of 90 Please solve 3 numbers for mark of 90
or 4 numbers for mark of 100 or 4 numbers for mark of 100
Number b and e must be done Number b and e must be done
a. a. Calculate Calculate | | of each strain of each strain
at S=0.2 g/dm at S=0.2 g/dm
3 3
? ?
b. Calculate A of each strain!!! b. Calculate A of each strain!!!
Calculate Calculate at 285 K and 310 K at 285 K and 310 K
of each strain! of each strain!
Think deeply and carefully before Think deeply and carefully before
answer these question!!! answer these question!!!
c. Define DBI and GC Content! c. Define DBI and GC Content!
d. Write DBI equation! d. Write DBI equation!
e. For each type of strain e. For each type of strain
Calculate the DBI? Calculate the DBI?
Calculate GC Calculate GC contens contens (GC (GC
DNA DNA
)? )?
What type? Why? What type? Why?
O. O. agardhi agardhi A. A. nidulans nidulans S. S. lividus lividus
Fatty acids contents Fatty acids contents
14:0 14:0 0 0 0.011 0.011 0 0
14:1 14:1 0 0 0.012 0.012 0 0
16:0 16:0 0.292 0.292 0.477 0.477 0.54 0.54
16:1 16:1 0.217 0.217 0.385 0.385 0.1 0.1
16:2 16:2 0.033 0.033 0 0 0 0
18:0 18:0 0.004 0.004 0.037 0.037 0.22 0.22
18:1 18:1 0.073 0.073 0.074 0.074 0.14 0.14
18:2 18:2 0.146 0.146 0 0 0 0
18:3 18:3 0.235 0.235 0 0 0 0
Growth Characteristic Growth Characteristic
T T
m m
355 K 355 K 361 K 361 K 371K 371K
T T 295 K 295 K 301 K 301 K 328 K 328 K
K K
i i
0.001 g/dm 0.001 g/dm
3 3
10 g/dm 10 g/dm
3 3
1 g/dm 1 g/dm
3 3
K K
m m
10 g/dm 10 g/dm
3 3
10 g/dm 10 g/dm
3 3
100 g/dm 100 g/dm
3 3
0.012 0.012 h h
1 1
0.014 0.014 h h
1 1
0.048 0.048 h h
1 1
E E
a a
(T (T- -238.5) 238.5) Kj Kj/mole /mole 128 128 Kj Kj/mole 240 /mole 240 Kj Kj/mole /mole
DNA
GC C
m
T
o
+ = 0 . 41 3 . 69 ) (
K mole
J
R

= 314 . 8 ( )
0 ) (
1 ) (
= =
= ~
=

T f T T
T f T T
T f e A
hab
hab
RT
E
a

i
m
K
s
s
K
+ +
=
1
1
|
Answer Sheet
Answer Sheet
| | | | [h [h
- -1 1
] ] A A [h [h
- -1 1
] ]
Oscillatoria Oscillatoria agardhi agardhi Gomont Gomont 0.003984 0.003984 0.0000478 0.0000478 2.06*10 2.06*10
6 6
Anacystis Anacystis nidulans nidulans 0.01960 0.01960 0.0010584 0.0010584 2.29*10 2.29*10
20 20
Synechococcus Synechococcus lividus lividus 0.001995 0.001995 0.0000958 0.0000958 8.00*10 8.00*10
36 36
[h [h
- -1 1
] ]
(285K) (285K) (310K) (310K) DBI DBI GC GC
DNA DNA
Oscillatoria Oscillatoria agardhi agardhi Gomont Gomont 0.00618 0.00618 1.85 10 1.85 10
- -6 6
1.353 0.310 1.353 0.310 psychrophile psychrophile
Anacystis Anacystis nidulans nidulans 0.00306 0.00306 0.0618 0.0618 0.471 0.471 0.456 0.456 mesophile mesophile
Synechococcus Synechococcus lividus lividus 8.21 10 8.21 10
- -8 8
0.0479 0.0479 0.240 0.240 0.700 0.700 thermophile thermophile
DBI was value that defined content of double bond of cellular me DBI was value that defined content of double bond of cellular membrane fatty acid in the each strains mbrane fatty acid in the each strains
plasma plasma- -membrane. membrane.
Percentage of Percentage of Guanin Guanin & & Cytosin Cytosin in whole of DNA in whole of DNA
( ) { }
{ } FA
n N UFA n
DBI
n

=
:
0
Chemical Reaction Kinetics
Anondho Wijanarko, Ir. M.Eng.

Multisite Enzyme Kinetics
Question
Suppose that an enzyme has two active sites so that substrate is converted to
product via the reaction sequence:

P E ES
P ES ESS
ESS S ES
ES S E
k
k
k
k
k
k
+
+
+
+

4
3
2
2
1
1
) (
) ( ) (
) ( ) (
) (


Derive a rate expression for P formation by assumming quasy steady state for
(ES) and for (ESS).

Answer:
quasy steady state for (ES) and for (ESS).
0 ) (
0 ) (
=
=
ESS
ES
dt
d
dt
d

and then,
( ) ( ) ( ) 0
3 2 2
=

ESS k ESS k ES k

( ) | | ( ) 0
3 2 2
= +

ESS k k ES k

( ) | | ( ) ESS k k ES k + =
3 2 2

( ) ( ) ES
k k
S k
ESS
+
=
3 2
2
(1) (25%)
( ) ( ) ( ) ( ) 0
4 2 2 1 1
= +

ES k ESS k S ES k ES k ES k

( )( ) ( ) ( )( ) 0
3 2 2 4 1 1
= + + +

ESS k k S ES k ES k k ES k

( )( ) ( ) ( )
( )
0
3 2
2
3 2 2 4 1 1
=
+
+ + +

k k
S ES k
k k S ES k ES k k ES k

( )( ) 0
4 1 1
= +

ES k k ES k

( )
4 1
1
k k
ES k
ES
+
=

(2) (50%)
Constant enzyme concentration
( ) ( ) ( ) ( ) ES
k k
S k
ES E ESS ES E E
o
3 2
2
+
+ + = + + =

( )
|
|
.
|

\
|
+

|
|
.
|

\
|
+
+ + =
|
|
.
|

\
|
+
+ + =
4 1
1
3 2
2
3 2
2
1 1
k k
ES k
k k
S k
E ES
k k
S k
E E
o

|
|
.
|

\
|
|
|
.
|

\
|
+

|
|
.
|

\
|
+
+ + =
4 1
1
3 2
2
1 1
k k
S k
k k
S k
E E
o

|
|
.
|

\
|
|
|
.
|

\
|
+

|
|
.
|

\
|
+
+ +
=
4 1
1
3 2
2
1 1
k k
S k
k k
S k
E
E
o
(3) (70%)
( )
|
|
.
|

\
|
|
|
.
|

\
|
+

|
|
.
|

\
|
+
+ +

+
=

4 1
1
3 2
2
4 1
1
1 1
k k
S k
k k
S k
E
k k
S k
ES
o
(75%)
( ) ( ) ES
k k
S k
ESS
+
=
3 2
2

( )
(
(
(
(
(

|
|
.
|

\
|
|
|
.
|

\
|
+

|
|
.
|

\
|
+
+ +

+
|
|
.
|

\
|
+
=


4 1
1
3 2
2
4 1
1
3 2
2
1 1
k k
S k
k k
S k
E
k k
S k
k k
S k
ESS
o
(80%)
derivative rate expression for P formation:
( ) ( ) ES k ESS k P
dt
d
4 3
+ =

|
|
.
|

\
|
+
+
|
|
.
|

\
|
|
|
.
|

\
|
+

|
|
.
|

\
|
+
+ +

+
=

3 2
2
3 4
4 1
1
3 2
2
4 1
1
1 1
k k
S k
k k
k k
S k
k k
S k
E
k k
S k
P
o
dt
d
(85%)
if:
4
3
2
3 2
2
1
4 1
1
, ,
k
k
k
k k
K
k
k k
K =
+
=
+
=

|

then:
|
|
.
|

\
|
+
|
|
.
|

\
|
|
|
.
|

\
|

|
|
.
|

\
|
+ +
=
2
1 2
4
1
1
1 1
K
S
K
S
K
S
E k
K
S
P
o
dt
d
|
(90%)
if: o
E k
4 max
=

|
|
.
|

\
|
+
|
|
.
|

\
|
+ +
=
|
|
.
|

\
|
+
|
|
.
|

\
|
+ +
=
2
2
1
max
2
2 1 1
4
1
1
1
1
1
K
S
K
S
S
K K
S
K
S
K
S
K
S
E k
K
S
P
o
dt
d
|

|

(100%)

derivative rate expresion for P formation:
+ +
+
=
2
1
2
max
1
1
K
S
S
K
K
S
P
dt
d
|

in case
1 = |
,then
+ +
+
=
2
1
2
max
1
1
K
S
S
K
K
S
P
dt
d

(Webb Equation) (125%)

Catatan: kebenaran jawaban hingga baris penurunan persmaan kinetik dinilai
berdasarkan persentase nilai dari soal ini adalah persentase yang ada di sebelah
kanan dari baris penurunan persamaan kinetik yang dilakukan oleh mahasiswa.
Cellulose Hydrolysis Kinetics
Cellulose Hydrolysis Kinetics
The enzymes which degrade cellulose, producing the The enzymes which degrade cellulose, producing the dimer dimer cellobiose cellobiose, a , a
simplified reaction network can be written : simplified reaction network can be written :
Where G Where G
1 1
, G , G
2 2
are insoluble cellulose and soluble are insoluble cellulose and soluble cellubiose cellubiose and E and E
1 1
is is
indicative of the enzyme involved in the slowest step leading to indicative of the enzyme involved in the slowest step leading to cellobiose cellobiose. .
Derive a reaction rate of this hydrolysis studies? Derive a reaction rate of this hydrolysis studies?
1 2 1 2
1 2 1 1
1 1 1 1
3
3
2
1
E G E G
E G E G
E G E G
k
k
k
k

+
+
+
Answer Sheet
Answer Sheet
Fast reaction Fast reaction
Slow reaction Slow reaction
Inhibition step Inhibition step
Intermediate species Intermediate species
1 2 1 2
1 2 1 1
1 1 1 1
3
3
2
1
E G E G
E G E G
E G E G
k
k
k
k

+
+
+
| | | |
1 1 1 2
& E G E G
| | | | | | | |
1 2 1 1 1 0
E G E G E E + + =
| |
| |
0
0
1 2
1 1
=
=
dt
E G d
dt
E G d
| | | | | |
| | | | | | 0
0
1 2 3 1 2 3
1 1 2 1 1 1
=
=

E G k E G k
E G k E G k
| | | | | |
| | | | | |
1 2
3
3
1 2
1 1
2
1
1 1
E G
k
k
E G
E G
k
k
E G
=
=

| | | |
2 1
0
1
3
3
2
1
1 G G
E
E
k
k
k
k

+ +
=
Rate of cellulose hydrolysis reaction
Rate of cellulose hydrolysis reaction
| |
| | | | | | | |
| |
| | | | | | | | | | | |
| |
| | | |
| |
| |
| | | |
2 1
0
1 1
2
1 1 1
2
1 2
3
3
3 1 2 3 1 1
2
1
2
2
1 2 3 1 2 3 1 1 2
2
3
3
2
1
1 G G
E
G k
dt
G d
E G k
dt
G d
E G
k
k
k E G k E G
k
k
k
dt
G d
E G k E G k E G k
dt
G d
k
k
k
k

+ +
=
=
+ =
+ =

| |
| | | |
2 1
0
2
2
3
3
2
1
1 G G
E
k
dt
G d
k
k
k
k

+ +
=