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Derive the rate equation for the following enzyme reaction using the BriggsHaldane approach.
k1
k3
S+E
ES
ES
P+|E
k2
Steady-state
Cell Conc.
Cx ,g/l
2.00
1.20
2.40
2.33
1.25
3. Prokaryotic cells
Cell wall
..
.
.
.. .
..
..
.
..
..
Typical.. prokaryotic cells
.
Cell membrane
Ribosome
Nuclear region
cytoplasm
.. prokaryotic cells
Typical
The prokaryotic cell is unit of structure, two group microbial, bacteria and blue
green algae. Prokaryotic cell is simple and small. Prokaryotic cell is not
compartmentalized by unit membrane. Prokaryotic cell have two structurally
distinguished two internal region: cytoplasm and nuclear region. Cytoplasm contains
graing dark spot due to the present of ribosome.
Ribosome is the site of important biochemical reaction for protein synthesis. The
nucleur contain deoxyribose nucleuic acid (DNA) ,which contain genetic information
that determine the production of protein and structure .
The prokaryotic cell is surrounded by cell wall and a cell membrane. The cell wall
is considerable thicker than cell membrane, and it protect cells from external
influence. The cell membrane serve as the surface which other cells components
attack and important cell function take place.
4. The basic procedures of recombinant DNA techniques have two factor. They are
(1) to join the DNA segment to the molecule and they can be replicated .
(2) to provide environment that allow the reproduction of join DNA molecules .
The produce for the flow chart of recombinant DNA techniques are
DNA
Foreign
Stricky end
DNA Ilagse
Tranaformation
Transformation cell
Fig : The procedure of recombinant DNA .
Figure show the procedure of recombinant DNA techniques. The plasmid cut in
definite size with restriction enzyme. The DNA of the foreign cleaved with restriction
enzyme. Some of fragment have interest gene. The plasmid and genome fragment mixed
and joined by the DNA Ligases. The recombinant plasmids are introduced into the
bacteria and coculativation of plasmid and bacteria.
s
1 .solution
k1
S+E
k3
ES
ES
k2
The intermediate of reaction with respect to time is neglected.
dCES = 0
dt
rCES = k1CsCE- k2 CES- k3 CES = 0
k1CsCE = k3 CES + k2CES
k1CsCE = (k2 +k3 ) CES
CE = (k2 + k3 ) CES
k1Cs
rp = k3 CES
P+|E
(1)
(2)
By enzyme balance,
CEO = CE + CES
= (k2 + k3 ) CES
k1Cs
= ( k2 + k3 +1) CES
k1Cs
CEO = (k2 + k3 + k1Cs) CES
k1Cs
By dividing by k1
3
CES
CEOCS
k2 + k3 +Cs
k1
k2
k1
rmax = k3CEO
kM = k2+k3 k2
k1
k1
2. Solution
CS0
Cs
kM = 0.03 mol/L
rmax =13mol 60 min = 780 mol
L min 1 hr
L hr
(a) v =?
xs =95%
Cso =10 mol / L
F = 10 L / hr
CS= CSO (1_ xs)
= 10 (1 _ 0.95) = 0.5 mol /L
At the CSTR,
Input _ Output + general = 0
FCSO _ FCS +rsv
=0
F (CSO _CS)
= _ rsv
F (CSO _Cs)
= rpv
F (CSO _CS)
= rmax Cs V
KM +CS
F
V
= rmax Cs
(KM+CS)(CSO_CS)
F
V
= 780* 0.5
(0.03+0.5) (10-0.5)
F
V
= 390
0.53 * 9.5
F
V
V
= 77.46 hr-1
V
V
=F
77.46hr-1
= 10 L/hr
77.46
= 0.129 liter.
2 .(b)
CS0
Plug Flow
Cs
t
rmax
0
Cso
t
KM +CS dCs = rmax t
Cs Cs
0
Cso
Cso
kM dCs + dCs = rmax t
Cs
Cs
kMlnCso + ( Cso _Cs ) = rmaxt ( plus = t batch )
kMlnCso + ( Cso _Cs ) = rmax
0.03 ln 10 + (10-0.5) = 780
0.5
0.0899 + 9.5 = 780
9.95899 = 780
= 0.0123hr
= V / F = 0.0123hr
V = 0.0123 10 L/hr
V = 0.123 Liter
6. Soln:
(a) rx = ?
(b) rp = ?
=
hr
max Cs
ks Cs
=
ks 1
1
+
max Cs max
y= mx+c
D = hr-1
CS g/l
0.084
0.1
0.16
0.198
0.242
0.054
0.16
0.13
0186
0.226
1
Cs
11.9
10
6.25
5.05
4.13
18.5
12.6
7.25
5.38
4.42
From graph,
C=
1
= 1.6
max
1
= 0.625
1.6
ks
0.8
m=
=
= 0.667
max
1.2
max =
ks = 0.42 g/l
6. (a) rx = ?
max Cs
rx =
ks Cs
0.625Cs
0.42 Cs
rx =
6. (b) rp = ?
1
dCx
YX / P dt
max C s C x
1
=
YX / P k s C s
rp =
YX
P =
YX
P =
=
C x
Cx
=
C p
Cp
YX
P = 0.27
max C s C x
1
YX / P k s C s
0.625C S
rp =
1
0.27 0.42 C S
rp =
2.31C S
0.42 C S
= k3CES
dC ES
= k1CSCE k2 CES k3 CES 0 (2)
dt
Substituting Eq (1) into Eq (2) confirms that the rate of product, formation and that of the
substrate consumption are the same, that is ,
dC p
dC S
r=
== k3 CES
dt
dt
If we assume that the total enzyme contents are conserved,
CEO = CE + CES (3)
Substituting Eq (3) into Eq (2) for CE, and rearranging for CES
C Eo C S
CES =
k 2 k 3 k1C S
k 3C EO C S
rmax C S
dC S
r=
== k 2 k3
=
CS
K M CS
dt
dt
k1
While in the Briggs- Haldane approach, it is equal to ( k2 + k3 ) / k1 .
dC p
CS0
Cs
2. KM = 0.03 mol/lit
Umax = 13 mol/lit
CSO = 10 mol/lit
F = 10 L / hr
1hr
1
= Lit / min
60 min 6
= 0.129 Lit.
(b) V =?
FC SO
CS
dC S
C
max S
dt
K M CS
CCSSO
CCSOS
K M CS
dC S max t0 dt
CS
KM
dC S CCSOS dC S max t
CS
CS
(C SO C S ) max t
CS
10
V
0.03 ln
(10 0.5) 13
0.5
10 / 60
V =0.123lit
K M ln
..
.
Cell wall
Cell membrane
Ribosome
Nuclear region
cytoplasm
...
.
..
.
..
..
.
Plasmld
..
..
..
.
Restrictio
n enzyme
..
Foreign
DNA
'Stricky
end'
DNA Ilagse
Tranaformation
Transformed cell
Fig, Method for the production of recombinant DNA
Fig shows the overall procedure for the production of recombinant DNA .The
plasmid is cut at a number of defined sites with a restriction enzyme. The DNA of a
foreign genome is cleaved with a restriction enzyme. Some of the resulting fragments may
have the gene of interest. Plasmids and genome fragments are mixed and joined by DNA
ligase. The recombinant plasmids are introduced into bacteria by cocultivation of plasmids
and bacteria.
5. Growth Cycle for Batch Cultivation
If you inoculate unicellular microorganisms into a fresh sterilized medium and
measure the cell number density with respect to time and plot it, you may find that there
are six phases of growth and death,as shown in Fig. They are:
1. Lag phase: A period of time when the change of cell number is zero.
2. Accelerated growth phase: The cell number starts to increase and the division
rate increases to reach a maximum.
3. Exponential growth phase: The cell number increases exponentially as the cells
start to divide. The growth rate is increasing during this phase, but the division rate which
is proportional to d ln Cno / dt, is constant at its maximum value, as illustrated in fig .
10
12
10
8
t
Fig Typical growth curve of unicellular organisms: (A) lag phase; (B) accelerated
growth phase; (C) exponential phase; (D) decelerated growth phase; (E) stationary phase;
(F) death phase.
At the end of the lag phase, when growth begins, the division rate increases
gradually and reaches a maximum value in the exponential growth period, as shown by the
rising inflection at B in fig. This transitional period is commonly called the accelerated
growth phase and is often included as a part of the lat phase.
In unicellular organisms, the progressive doubling of cell number results in a
continually increasing rate of grown in the population. A bacterial culture undergoing
balanced growth mimics a first-order autocatalytic chemical reaction. Therefore, the rate of
the number density (Cn) of bacteria present at that time:
dC n
rn
C n where the constant is known as the specific growth rate
dt
hr 1 .The specific growth rate should not be confused with the growth rate, which has
11
different units and meaning. The growth rate is the change of the cell number density with
time, while the specific growth rate is
1 dC n d ln C n
C n dt
dt
The growth of microbial populations is normally limited either by the exhaustion of
available nutrients or by the accumulation of toxic products of metabolism. As a
consequence, the rate of growth declines and growth eventually stops. At this point a
culture is said to be in the stationary phase. The transition between the exponential phase
and the stationary phase involves a period of unbalanced growth during which the various
cellular components are synthesized at unequal rates. Consequently, cells in the stationary
phase have a chemical composition different from that of cells in the exponential phase.
The stationary phase is usually followed by a death phase in which the organisms
in the population die. Death occurs either because of the depletion of the cellular reserves
of energy, or the accumulation of toxic products. Like growth, death is an exponential
function. In some cases, the organisms not only die but also disintegrate, a process called
lysis.
6. (a) Cxi= 0
, Csi = 0
C
max S
K S CS
KS
1
1
max C S max
K 1
1
1
S
D max C S maz
K
1
1
, slope S
Plot Vs
D CS
max
Intercept
D = hr-1
0.084
0.1
0.16
0.198
0.242
max
CS g/l
1
Cs
0.054
11.9
0.16
10
0.13
6.25
0186
5.05
0.226
4.13
KS
1
0.69 , Intercept
Slope =
= 1.2
max
max
18.5
12.6
7.25
5.38
4.42
max 0.8
x C x
0.833C S
.C x
0.575 C S
F (C Pi C p ) rPV 0
12
F (C Pi C p ) rpV
C P C Pi
V C Pi C P
F
rP
rP
V CP
F
rP
CP
CP
For S.S CSTF with sterile feed, rP
m
D
1
,
m
rP C P
max C S
K S Cs
max C P
.C B
=
K S CS
From Manod, rP C P .
plot
1
1
Vs
D
CB
straight line
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