The culture medium used to grow E. kaisocheenum is given in table below.

The culture was conducted in a bioreactor

(nominal volume = 80m3, working volume = 40 m3) operated at 32C and dilution rate of 0.4 h-1. Oxygen-enriched air
(31% oxygen) at 1.05 atm was provided and delivered at the bottom of the reactor at a rate of 0.75 VVM. The power
required for liquid agitation at this aeration rate was 1,100 W/m3 with impeller speed of 2 rps. Under these conditions,
the cell yield on oxygen is 1.3 g dcw/g O2 while cell yield on glucose is 0.6 gdcw/g glucose. Clearly state your assumptions.
a. Determine the maximum steady-state cell concentration under these conditions.
b. If a large-scale bioreactor is used and the homogeneity of temperature and dissolved oxygen is
difficult to achieve, calculate the new oxygen transfer rate when temperature at inlet and outlet
stream is 30C and 34C respectively. (Note: Use log-mean concentration difference for driving force)
c. Calculate the fraction of gas present in the culture medium.
Component
Glucose
NH4NO3
Na2SO4
KH2PO4
Trace metals

g/L
40
4
1.5
5.2
-

Cell composition by mass is C:50%, N:15%, P:3% and S:1%.

P
Kla = 0.022 g
VL

0.95

.vs

0.67

P 2 .N .D 3
Pg = 1.11 o 0.56 i
Q

0.45

Richards correlation:
0.4

Po , [ Hp]

.vs 0.5 = 7.63.H + 2.37

V
L

KLa = hr-1
VL = liquid volume, m3
vs = superficial velocity of air based on bioreactor cross-section, m/hr
Pg = power consumed for agitation in gassed system, W
Po = power consumed for agitation in ungassed system, W
N = impeller speed, rps
Q = volumetric of air working volume per minute (VVM)
Di = impeller diameter, m

GIVEN:
VR = 80 m3
VL = 40 m3
T = 32oC
D = 0.40 hr-1
Oxygen-enriched air (31% oxygen) at 1.05 atm
Q = 0.75 VVM
Pg/VL = 1,100 W/m3
YX/O2 = 1.30 g dcw/g O2
YX/S = 0.60 g dcw/g glucose
REQUIRED:
a) Xmax at steady-state
b) OTR if Tin = 30oC and Tout = 34oC
c) !
SOLUTION:
a) The growth of the cell can be limited by (1) oxygen-transfer limitation or (2) availability of necessary nutrients.
Thus, the approach is to evaluate the cell densities, X, that can be sustained by the two aforementioned factors,
respectively, and by comparing the results, we select the maximum possible cell concentration for the given
conditions.

"#$ = "&$
12

'() (+,(
+,( ) =

34/67

Assume standard reactor:

Dt = HL
40 : ; =

<
=

>? 7 (>? )

>? = 3.7067 :

'() = 0.022 1,100

G.HI

0.75

: KLM
40 : ; N. O.
;
: N. O. :LP
R
3.7067 7 : 7
4

60

:LP
M

G.ST

= 525.56 M UV

W=

X67
1 KY:
KY: [
= 0.02761

=
:Z "
+,( 36.22
:Z "7
7
(@ 30] +, ^_M` "7 )
[

+,(
=

X67
0.31 1.05 KY:
:Z "7
=
= 11.7892

KY: [
W
[
0.02761
:Z "7

525. 56 M UV

11.7892

:Z"7
0 =
[

0.40 2

:Z cdN
1.30
:Z "7

e = fg. hi j klm/n maximum cell concentration achievable based on the given aeration condition.


40
2=

Z
[

6 12
1 180
0.50

= if

j klm
pqrsk tu lqvptu qwqxyqpxyxz{
n

4
2=

Z
[

2 14
1 80
0.15

5.2
2=

Z
[

1 31
1 136
0.03

= |. ii

= i|. }h

j klm
pqrsk tu uxzvtjsu qwqxyqpxyxz{
n
j klm
pqrsk tu ~tr~tvtr qwqxyqpxyxz{
n

1.5
2=

Z
[

1 32
1 142
0.01

= ii. g

j klm
pqrsk tu ryv qwqxyqpxyxz{
n

By comparing the results, the maximum cell concentration that can be achieved is limited by the availability of
the nitrogen source. ANSWER: 9.33 g dcw/L.


b)

at 30oC:

X67
1 KY:
KY: [
W= =
= 0.02695

+,( 37.1 :Z "7
:Z "7
[

X67
0.31 1.05 KY:
:Z "7

+,(
=
=
= 12.0779

KY: [
W
[
0.02695
:Z "7

at 34oC:
X67
1 KY:
KY: [
W= =
= 0.02830

+,( 35.34 :Z "7
:Z "7
[

X67
0.31 1.05 KY:
:Z "7

+,( ]?
=
=
= 11.5018

KY: [
W
[
0.02830
:Z "7

+,(
+,(

+,(
+,(

ln

]?? +,( +,( ?

+,(
+,( ]??

+,(
+,( ?

"#$ = 525.56 M UV

11.7875

11.5018 0 (12.0779 0)
= 11.7875
11.5018 0
ln
12.0779 0

:Z"7
jf
= h|}. g

[
n v

c)
1100

:;

40 : ; = 1.11

XG

3.7067
3
0.75 G.IS
2

; G.=I

XG = 61,057.06 = 81.88 X

81.88
40

G.=G

G.IG

0.75 40 60
R
3.7067 7
4

= 7.63 W + 2.37

W = 1.9438 :

! =

O_:` KLM
W
1.9438 :
=
=
= g. ig
O_:` KLM + O_:` L_Lc
W + W( 1.9438 : + 3.7067 :