Study of Mass Transfer in Viscous Fermentations

8/18/2019 Study of Mass Transfer in Viscous Fermentations

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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


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


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


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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3.2.1 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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3.3.1 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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4.3.1 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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4.4.1 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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5.2.1 Fermenter setup

5.2.2 Viscosity measurement

5.2.3 Fermentation conditions


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~


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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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6.3.1 Microorganism, medium and cultivation procedures

°

°

6.3.2 Analysis


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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


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]


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

Study of Mass Transfer in Viscous Fermentations

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