study of mass transfer in viscous fermentations
TRANSCRIPT
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David Kold
Ph.D. Tesis
April 2010
Study of Mass Transfer in ViscousFermentations- using a Rotating Jet Head Mixing System
Center for Microbial BiotechnologyDepartment of Systems Biology
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Copyright © David Kold 2010
ISBN-9788791494918
Printed in Denmark by
WWW.FRYDENBERG.DK
Baldersgade 12 - 16
DK-2200 Copenhagen N
Frontpage: Broth from two Aspergillus oryzae fermentations.
Left: Fermentation conducted in a Rotating Jet Head fermenter system.
Right: Fermentation conducted in a impeller mixed tank
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Abstract
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η
( ) 8.00.122
t0.4sL
V
Pkuak
−
= appη
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Dansk Resumé
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η
( ) 8.00.122
t0.4sL
V
Pkuak
−
= appη
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Preface
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Nomenclature
Roman Letters
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Greek Letters
a
0
a
0
Subscripts
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Contents
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Chapter 1Introduction
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Chapter 1
Introduction
1.1 Impeller Mixed Fermenters
•
•
•
•
•
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53a DN
P
Po R
=
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a
2RaNDRe
η =a
1.2 Mixing with Jets
1.3 The Rotary Jet Head System
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1.3.1 System Setup
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1.3.2 Power Input Calculation
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2Lkin v½FP =
n
loop
fermg
L
A
p
p
F
F
v
+
=
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0.0
1.0
2.0
3.0
4.0
5.0
0 100 200 300 400 500
Pump Speed [RPM]
M a s s F l o w [ k g s -
1
]
.
( )
FppP Lfermlooppump −=
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,
0.0
0.2
0.4
0.6
0.8
1.0
1.2
0 100 200 300 400 500
Pump Speed [RPM]
P p u m p
[ k W ]
;
;
.
ferm
igg
p
plnRTFP =
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pumpgRJH PPP +=
a
aRe
η
vD RJH =
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1.4 References
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Chapter 2Rheological Characterization ofFermentation Broths
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Chapter 2
Rheological Characterization of FermentationBroths
2.1 Introduction
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2.2 Theory
2.2.1 Viscosity and Rheological Models
=
n
0 ) ( +=
0
0
0
a
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0
n) ( =
n) ( =
0
0 ) ( 0 +=
0 n0 ) ( +=
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2.2.2 Effect of temperature on viscosity
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2.2.3 Conventional Rheometers
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=
3r2
3M =
IO
O
rr
r
−=
Lr 2
M
2
O
=
=n
2
O
I
r
r-1n
2
Lr 2
M
2
I
=
h
r r =
+=3
n1
r2
3M
3
h
r r =
+=3h
nh1
r2
3M 0
3
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2.2.4 The Impeller Rheometer
5
R
3
P DNNP =
Re
cN P =
NDRe
2R=
NM2P =
ND2
cM 3R=
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kNa =
1-na (kN) =
1)(n
1
3RcKND
2M
N
1k
−
=
2.2.5 Online Rheometers
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2.3 Materials and Methods
2.3.1 Rheometers and set up
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3a
2
D
4Q
=
π
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2.3.2 Samples for comparing rheology measurements
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2.4 Results and Discussion
2.4.1 Comparison of Rheometer Setups using Standardized Xanthan
Solutions
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0
5
10
15
20
25
30
0 50 100 150 200
Shear rate [s-1]
S h e a r s t r e s s [ P a ]
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3r
2M
=
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0.0
0.1
0.1
0.2
0.2
0.3
0.3
0.4
0.4
0.25 0.50 0.75 1.00 1.25 1.50
Xanthan Concentration [%]
n [ - ]
REF: CP CP with gap CP with gap- PP CP with gap- CP CP with gap- TA Instruments
0.0
2.0
4.0
6.0
8.0
10.0
12.0
14.0
0.25 0.50 0.75 1.00 1.25 1.50
Xanthan Concentration [%]
K [ N s n m - 2 ]
REF: CP CP with gap CP with gap- PP CP with gap- CP CP with gap- TA Instruments
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0.000
0.050
0.100
0.150
0.200
0.250
0.300
0.350
0.400
n [ - ]
REF:Cone-Plate MV-DIN Wide Gap MV-DIN Narrow Gap FL10
0.00
2.00
4.00
6.00
8.00
10.00
12.00
14.00
K [ N s n m - 2 ]
0.000
0.200
0.400
0.600
0.800
1.000
1.200
0.25 0.50 0.75 1.00 1.25 1.50
Xanthan Concentration [%]
V i s c o s i t y @ 2 0 s - 1 [
P a s ]
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0.000
0.100
0.200
0.300
0.400
0.500
0.600
0.700
0.800
0.900
1.000
0.00 0.25 0.50 0.75 1.00 1.25 1.50
Xanthan Concentration [%]
V i s c o s i t y @ 2
0 s -
1 [
P a s ]
REF: Cone-Plate Marimex FL10
2.4.2 Comparison of Rheometers for Viscosity Measurements in
Fermentation Broths
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0.00
0.01
0.02
0.03
0.04
0.05
280 290 300 310
Temperature [K]
V i s c o s i t y
@ 9
5 s -
1 [
P a s ]
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~
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2.5 Conclusion
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2.6 References
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Chapter 3
Mass Transfer Optimization inFed-batch Aspergillus oryzae Fermentations - using a RotatingJet Head Mixing System
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Chapter 3
Mass Transfer Optimization in Fed-batch
Aspergillus oryzae Fermentations - using a Rotating
Jet Head Mixing System
3.0 Abstract
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3.1 Introduction
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c
3
i tkD
PEDCF =
iD
W
4
k =
ci t
1
V
PEDCF =
i
Lc
Q
Vt =
3
igFl,i NDNQ =
Po
PoNN
g
FlgFl, =
5
i
3
L DNPoP =
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3.2 Materials and Methods
3.2.1 Growth medium
3.2.2 Fermenter setup and cultivation conditions
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°
gemptytotIM PP-PP +=
pumpgRJH PPP +=
pQP LLPump =
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3.3 Results and Discussion
3.3.1 Energy dissipation/circulation function (EDCF)
·
·
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3.3.2 Fermentation with complex soy medium
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3.3.3 Fermentation with defined minimal medium
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3.4 Conclusion
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3.5 References
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Chapter 4Intracellular Shear Stress
Response in Aspergilli
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Chapter 4
Intracellular Shear Stress Response in Aspergilli
4.1 Abstract
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4.2 Introduction
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4.3 Materials and Methods
4.3.1 Sampling for mRNA
4.3.2 Extraction and Purification
4.3.3 cRNA Preparation and Microarray Processing
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4.3.4 Analysis of transcriptome data
4.3.5 Aspergillus niger samples
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4.4 Results and Discussion
4.4.1 Initial statistical survey of effect of mixing system
4.4.2 Gene by gene-examinations
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ORF No. Deduced function Reg.
AO090003000770 Glycosyltransferases group 1 -5.02
AO090003000661 Thiamine pyrophosphate enzyme, central domain -6.12
AO090026000820 Trehalose-phosphatase -1.32
AO090020000603 FGGY family of carbohydrate kinases, C-terminal domain 0.54
AO090003000594 Glycosyltransferase family 2 -0.67
4.4.3 Comparative study
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4.4.4 Pathway-level differences in the 325 genes
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4.4.5 Single gene examinations of the 325 genes
•
•
•
•
•
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4.5 Conclusion
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4.6 References
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Chapter 5Effect of Viscosity on kLa during aFilamentous Fungus Fed-BatchFermentation
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Chapter 5
Effect of Viscosity on kLa during a Filamentous
Fungus Fed-Batch Fermentation
5.1 Introduction
5.1.1 Effect of viscosity on mass transfer in fermenters
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( )-c*cak OTR L=
t
sLV
Pkuak
=
( )ω η app
t
sLV
Pkuak
=
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5.2 Materials and Methods
5.2.1 Fermenter setup
5.2.2 Viscosity measurement
5.2.3 Fermentation conditions
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5.3 Results and Discussion
~
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.
.
·
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5.4 Conclusion
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5.5 References
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Chapter 6
Characterization of Mass Transferin a Rotary Jet Head Fermenterduring Xanthomonas campestris fermentations
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Chapter 6
Characterization of Mass Transfer in a Rotary Jet
Head Fermenter during Xanthomonas campestris
Fermentations
6.1 Abstract
η
( ) 8.00.122
t0.4sL
V
Pkuak
−
= appη
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6.2 Introduction
6.2.1 Xanthan producing bacteria
6.2.2 Composition and properties
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°
6.2.3 Applications for xanthan gum
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6.2.4 Industrial xanthan production:
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6.3 Materials and Methods
6.3.1 Microorganism, medium and cultivation procedures
°
°
6.3.2 Analysis
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6.4 Results and Discussion
6.4.1 Suitability of the RJH system for Xanthomonas campestris
growth
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6.4.2 Xanthan production
6.4.3 Broth rheology
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0.00
0.10
0.20
0.30
0.40
0.50
0.60
0.70
0 20 40 60 80Time [h]
n
0.0
5.0
10.0
15.0
20.0
25.0
K
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0.00
0.01
0.02
0.03
0.04
0.05
0.06
V i s c o s i t y @ 1
2 5 0 s -
1 [ P a s ]
0.00
0.01
0.02
0.03
0.04
0.05
0.06
0 10 20 30 40 50 60 70 80
Time [h]
V i s c o s i t y @ 1
2 5 0 s - 1 [
P a s ]
)
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6.4.4 Suitability of the RJH system for aeration of Xanthomonas
campestris fermentation broths
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0
20
40
60
80
100
120
O 2 s a t u r a t i o n [ % ]
A i r f l o w [ l m i n - 1 ]
0
20
40
60
80
100
120
0 10 20 30 40 50 60 70 80
Time [h]
O 2 s a t u r a t i o n [ % ]
A i r f l o w [ l m i n - 1 ]
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0.000
0.005
0.010
0.015
0.020
0.025
0.030
0.035
0.040
q ( C O 2 ) q ( O 2 ) [ m o l l - 1 h
- 1 ]
0.000
0.005
0.010
0.015
0.020
0.025
0.030
0.035
0.040
0 10 20 30 40 50 60 70 80
Time [h]
q ( C O 2 ) q ( O 2 ) [ m o l l - 1 h
- 1 ]
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~
0.00
0.10
0.20
0.30
0.40
0.50
0 10 20 30 40 50 60 70 80
Time [h]
k L a [ s - 1 ]
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0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
1.8
2.0
P / V [ k w m - 3 ]
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
1.8
2.0
0 10 20 30 40 50 60 70 80
Time [h]
P / V [ k w m - 3 ]
6.4.5 Effect of viscosity on kLa
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6.4.6 Effect of power input on kLa
0.00
0.50
1.00
1.50
2.00
2.50
3.00
3.50
150 200 250 300 350
Pump speed [RPM]
P r e s s u r e [ b a r ]
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0
5
10
15
20
25
30
35
40
0 1 2 3 4 5
Ppump /V [kW m
-3
]
D O T [ % ]
-4.00
-3.00
-2.00
-1.00
0.00
-2.0 -1.0 0.0 1.0 2.0ln(Ppump /V)
l n ( k L a )
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6.4.7 Effect of superficial gas velocity
•
•
•
•
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( ) ( )appk
η ω β α lnV
Pln
ak lnuln tLs −
−
=
-3.0
-2.0
-1.0
0.0
-8.0 -7.0 -6.0 -5.0
ln(us)
l n ( k L a / k ) - l n
( P / V ) - l n ( a p p
)
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6.5 Conclusion
( ) 8.00.122
t0.4sL
V
Pkuak
−
= appη
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6.6 References
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Chapter 7
Preliminary Results, Further Workand Conclusion
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Chapter 7
Preliminary Results, Further Work and Conclusion
7.1 Preliminary Results
7.1.1 Feed strategy optimization during Aspergillus oryzae fed-batch
fermentations
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-14
-12
-10
-8
-6
-4
0.00 20.00 40.00 60.00
Time [h]
l n ( q ( C O 2 ) )
0
50
100
150
200
250
A i r f l ow
[ l mi n-1
]
1
23
7.1.2 Cellulase production with Trichoderma reesei
°
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7.2 Further Work
7.2.1 Scale up of the Rotary Jet Head system
7.2.2 Computational Fluid Dynamics
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7.3 General Conclusions
( ) 8.00.122
t0.4sL
V
Pkuak
−
= appη
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7.4 References
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Appendix I
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LOCUS Coef F.p.value PFAM_NAME PFAM_DESCRIPT IO N
AO090003000770 -5.0178 0.00001 Glycos_transf_1 Glycosyl transferases group 1 1
AO090102000482 -4.1403 0.00003 GST_N Glutathione S-transferase, N-terminal domain 2
AO090103000165 -3.6399 0.00004 MFS_1 Major Fac il it ator Superfami ly 3
AO090010000018 -4.371 0 .00005 p450 Cytochrome P450 4
AO090020000500 -5.1678 0.00008 KR KR domain 5
AO090020000501 -3.8496 0.00009 KR KR domain 6
AO090103000167 -3.863 0 .00011 PP-binding Phosphopantetheine attachment site 7
AO090103000166 -2.6965 0.00012 ABC_membrane ABC transporter transmembrane region 8
AO090012000174 -2.6367 0.00012 Patat in Patat in -l ike phospho lipase 9
AO090003000392 -3.3587 0.00014 UCH Ubiqu it in carboxyl -t erminal hydro lase 10
AO090206000078 -2.3036 0.00015 DUF1774 Fungal protein of unknown function (DUF1774) 11
AO090009000634 -5.0503 0.00015 ADH_zinc_N Zinc-binding dehydrogenase 12AO090005000652 -2.8658 0.00015 AA_permease Amino acid permease 13
AO090003001320 -3.7142 0.00017 CoaE Dephospho-CoA kinase 14
AO090701000780 -2.2457 0.00017 FHA FHA dom