Master of Science in Chemical Engineering Course in

Bioprocess Technology

Physical Processes in Bioreactors

Physical Processes in Bioreactors

University of San Carlos Department of Chemical Engineering Nasipit, Talamban, Cebu City Telefax: +63323446783 1 Email: chedept@usc.edu.ph

Contents of the Lecture

Fluid flow and mixing Heat transfer Mass transfer

Physical Processes in Bioreactors

Heat Transfer in Bioreactors

In situ sterilization of liquid medium Temperature control during reactor operation Heat-transfer configuration for bioreactors a) Jacketed vessel b) External coil c) Internal helical coil d) Internal baffle-type coil e) External heat exchanger
Physical Processes in Bioreactors 3

Heat-transfer configurations for bioreactors

Physical Processes in Bioreactors

Temperature changes for control of fermentation temperature using cooling water

Fermenter temperature TF

Temperature

Tco

Tci

Distance from cold-fluid inlet

Physical Processes in Bioreactors

Heat Transfer Mechanisms

RECALL, Conduction Convection Radiation

Physical Processes in Bioreactors

Heat Transfer Between Fluids

Th Thw (1) H = hh A (Th Thw ) Tcw Tc Cold fluid

Thw Tcw ) ( (2) H =

(3) H = hc A (Tcw Tc )

Hot fluid (1) Resistances:

(2) (3)

(L) A

1 Rh = hh A 1 Rc = hc A

L Rw = A
Where: h = individual heat transfer coefficient.
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Physical Processes in Bioreactors

Overall Heat Transfer Coefficient

H = UA (Th Tc )
But,

(a)

1 RT = = Rh + Rw + Rc UA 1 1 L 1 = + + UA hh A A hc A
Physical Processes in Bioreactors 8

Fouling Factors

1 1 1 L 1 1 = + + + + U h fh hh hc h fc
Fouling factor of the hot-fluid side Fouling factor of the cold-fluid side

Design equations for heat-transfer systems Equation (a) and energy balance equations
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Estimation of Heat Transfer Coefficients

D b Nu = f Gr Re, Pr, , , L w

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Heat Transfer in Aerobic Fermentations

H rxn
rate of oxygen consumption by cells

Maximum cell concentration supported by heat transfer system

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Heat Transfer in Aerobic Fermentations

9 Heat transfer is severe when: Fluid is viscous Turbulence & high U are difficult to achieve

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Mass Transfer in Bioreactors

9 Conduction (role in bioprocessing) Scale of mixing Solid-phase reaction Interphase transfer 9 Convection

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Diffusion Theory
CA1 JA CA2 x

dC A NA = DAB JA = dx a
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Mass Transfer Across A Phase Boundary

Rate of transfer

N A = kaC A
ics, geometry, hydrodynam k = f fluid properties

Resistance to mass transfer

1 Rm = ka

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Mass Transfer Across A Phase Boundary

Liquid-solid Liquid-liquid Gas-liquid

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Liquid-solid Mass Transfer

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Gas-liquid Mass Transfer

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Oxygen Transfer

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Steps for Oxygen Transfer from gas bubble to cell

i. Transfer from the interior of the bubble to the gas-liquid interface ii. Movement across the gas-liquid interface iii. Diffusion through the relatively stagnant liquid film surrounding the bubble iv. Transport through the bulk liquid v. Diffusion through the relatively stagnant liquid film surrounding the cells vi. Movement across the liquid-cell interface vii. If the cells are in a floc, clump or solid particle, diffusion through the solid to the individual cell viii. Transport through the cytoplasm to the site of reaction
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Analysis of G-L Mass Transfer in Bioreactors

i. ii. iii. iv. Transfer through the bulk gas phase in the bubble is relatively fast The gas-liquid interface itself contributes negligible resistance The liquid film around the bubbles is a major resistance to oxygen transfer In a well-mixed fermenter, concentration gradients in the bulk liquid are minimized & mass-transfer resistance in this region is small. However, rapid mixing can be difficult to achieve in viscous fermentation broths; if this is the case, oxygen-transfer resistance in the bulk liquid may be important. Because single cells are much smaller than gas bubbles, the liquid film surrounding each cell is much thinner than that around the bubbles and its effect on mass transfer can generally be neglected. On the other hand, if the cells form large clumps, liquid-film resistance can be significant. Resistance at the cell-liquid interface is generally neglected. When the cells are in clumps, intraparticle resistance is likely to be significant as oxygen has to diffuse through the solid pellet to reach the interior cells. The magnitude of this resistance depends on the size of the clumps. Intracellular oxygen-transfer resistance is negligible because of the small distances involved.

v.

vi. vii.

viii.

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Oxygen demand during cultivation of yeast cells

In the culture medium the O2 solubility is CO2* = 1.1 mmol/l = 35.2 mg / 1 (ppm), Which in case of air saturation reduces to CO2* = 35.2 x 0.209 = 7.36 ppm The typical O2 demand of the yeast fermentation is in the following range Q O2 = 0.3 g O2 / (g Dm h) Hence, for biomass concentration of 20 g Dm/l the oxygen demand is (d) This implies that the number of complete saturations of the fermentation broth amounts to (e) This is not an easy task, particularly, if higher biomass concentrations are desired which are usual in practice (for instance 60 g DM/l). Physical Processes in Bioreactors 22 n-Sat = 6 / 0.00736 = 816 h-1 Q O2 = 0.3 x 20 = 6 g O2 / h (c) (b) (a)

Critical oxygen concentration To avoid limitations of cell growth and product formation by lack of oxygen it is necessary that a certain oxygen concentration is maintained in the culture media. At next, an estimation is done to find out the conditions under which the biochemical reactions involved in cell growth are the limiting steps. Usually, the highest oxygen consumption rates are observed during cell growth. The largest mass transfer rates for oxygen (mol/cm3.s) are obtained for CL = 0, hence max = kL a CL On the other hand, the maximum oxygen consumption during cell growth is given by rmax = max X . 1/YO2 with the yield coefficient YO2 = generated biomass / consumed moled O2 It is understood that the main resistance for oxygen utilization is predominantly limited by microbial metabolism if or max >> rmax kLa CL* >> 1/YO2 max X , respectively.
Physical Processes in Bioreactors 23

(1

(2)

(3)

(4) (5)

From the biochemical viewpoint this implies that always sufficient oxygen is available to except the electron pairs which pass through the respiratory chain. To estimate the critical oxygen concentration which leads to growth limitation by lack of oxygen a kinetic law of Monod type is assumed to apply for oxygen, i.e., r = max x
CL 1 K O 2 + C L YO 2

(6) OUR oxygen uptake rate

CL 1 K O 2 + C L YO 2

Then the stationary oxygen balance is given by OTR = Oxygen transfer rate kL a (CL* - CL) = max x With the assumption CL* >> CL it follows

(7)

CL = CL* YO K O k a ' / max X

2 2 L

1 YO 2CL * kLa ' / max X

(8)

It can be anticipated as a guideline that no mass transfer limitation for oxygen exists at the gas/liquid interface if eq. (38) applies (9) ( = C , crit ) C >3 K
L

O2

In this case there is no mass transfer resistance for oxygen and the entire growth process is controlled by the biochemical reactions in the cell machinery. However, if CL < CL, crit ( = Ko2 ), oxygen can be the limiting substrate and hence the oxygen input into the culture media is the limiting step of the process (Fig. 9). Physical Processes in Bioreactors 24

Fig.9

Typical values of the critical O2 concentration (at sufficient supply with other substrates) CL* (at air saturation) = 0.23 mmol/l Microorganisms T, C Azetobacter vinlandii 30 E. coli 37.8 15 Pseudomonas denitroficans 30 Yeast 34.8 20 P. chrysogenum 24 30 as a rule of thumb that 0.003 < CL, crit < 0.05 mmol/l Physical which corresponds to an air saturation of 1 Processes to 25% in Bioreactors

CL, Crit mmol/l 0.03 0.008 0.003 0.009 0.0046 0.0037 0.022 0.009It can be assumed
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Scale-up
The performance of a bioreactor depends on process specific data (stoichiometric coefficients, thermodynamic and kinetic data) and transport parameters (hydrodynamics, heat and mass transfer properties). Specific data are scale independent and the transport parameters may be drastically dependent on the reactor size. For instance, the oxygen solubility (CL*) is a thermodynamic quantity which depends only on the medium composition and the temperature while the actual oxygen concentration in the culture medium is a complex function of the biochemical kinetics and the scale dependent transport parameters (kLa, , etc.). Phenotypical appearance of an organism is determined by both its genotype and the living conditions in the culture. Kossen has proposed to select production strains in laboratory scale at conditions which prevail in large reactors. Instead of looking for a baseball player in a soccer field, one should do scale down of reactors for proper screening of microbes.

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The different methods for scale-up can be summarized as follows: (1) Fundamental methods (modeling of the process on the basis of the balances for mass, heat and momentum which requires detailed knowledge of all transport effects and their interaction) (2) Semifundamental methods (solution of simplified balance equations) (3) Dimensional analysis (is only of limited value as not always applicable) (4) Rules of thumbs, know how (5) Trial and error In todays fermentation industry only methods (4) and (5) are established. The often used rules of thumbs imply: Do the scale-up in such a way that the culture conditions (environment) and some typical engineering parameters (P/V, , kLa, Re, ) are kept constant. Scale-up criteria used in industry % of industry 30% 30% 20% 20% *utip = stirrer tip velocity
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Criterium used P/V constant kLa constant utip constant* Po2 constant

Examples for a successful scale-up on the basis of different criteria are given in Fig. 10. The intrinsic know how is the choice of the correct criteria for the specific process. Different scale-up criteria and their consequences Scale-up from a 10 litre reactor to 10 m3 which is a linear scale factor of 10.

Criterium: Constancy of P/V N(-1) utip Re

P P/V 103 105 102 0.1 1 102 0.1 10-4

Value in 10 m3 scale N(-1) ND 0.22 1 0.1 10-2 2.15 10 1 0.1

Re 21.5 102 10 1

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Some criteria cannot be fulfilled in practice. An example is the requirement for equal mixing times in small and large scale. For many stirrer systems we have the empirical relation
p3 = 300 D5

(10)

Considering this, the equality of mixing times ( = idem) implies that

( P/V P/V )S = ( 2 ) l 2 D D

(11)

(P/V)l = (P/V)S ( D l ) 2 DS For a linear scale factor of 10 (0.01 to 10) and (P/V)S = 2 kW/m3 it follows (P/V)l = 200 kW/m3

(12)

Such a high energy input in a bioreactor is nearly impossible and would be extremely costly.

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