- Suraj Lal Gupta

Dissolved O2 is required for aerobic fermentation. O2 is highly insoluble in water. Solubility of O2 in water at 30 oC, 1 atmospheric pressure is nearly 8 mg/L. Obligate aerobes need dissolved O2 continuously for growth. Oxygen starvation might hinder growth. Mass transfer of O2 has serious implications in the growth regime of aerobic micro-organisms.

Usually there are three types of mass transfer (solutes) in fermentation.

- Liquid

liquid (downstream processing) - Liquid solid (pellets, flocs, immobilized cells etc.) - Liquid gas (gaseous solutes) Oxygen s transfer is liquid gas type. O2 transfer occurs through gaseous film and liquid film (two-film theory).

At steady-state, CAGi and CALi are in equilibrium. CAGi / CALi = m (distribution Factor)

Since solubility of O2 in water is very less, liquid-side resistance is large. Hence k a is much smaller than k a.
G L

Consequently, K a is roughly equal to k a.

L L

C*AL is equal to solubility of the gas A in the given solvent.

The rate at which oxygen is consumed by cells in fermenters determines the rate at which it must be transferred from gas to liquid. Factors influencing COD: - Cell species - Growth phase - Nature of carbon source (degree of reduction of carbon)

Qo = oxygen uptake rate per unit volume qo = specific oxygen uptake rate x = cell concentration When dissolved oxygen concentration decreases below certain value (Ccrit), qo becomes linearly dependent on CL.

Above Ccrit, qo becomes constant. Dissolved oxygen conc. must be greater than Ccrit At all time during fermen-tation.

qo depends on biochemical nature of the cell and its nutritional environment. Choice of substrate also affects oxygen demand. Glucose is consumed rapidly.

8 steps are involved in transfer of O2 from gas bubbles to the cell. a) Transfer through gas bulk phase. The resistance offered in this step is neglected. (Oxygen being sparingly soluble in water, liquid-side resistance dominates) The transfer rate is relatively fast in this step.

b) Transfer through gas-liquid interface. The resistance at gas-liquid interface in most cases is negligible. c) Transfer through liquid film surrounding the gas bubble. The resistance in this step is significantly high.

d) Transfer through bulk liquid. In well-mixed fermenter, resistance in this step is not significant. In viscous broth (mycelia), proper mixing being difficult to achieve, resistance in this step becomes significant.

e) Transfer through liquid film surrounding the cells. This film is much thinner than the liquid film surrounding the gas bubbles. Resistance in this step is neglected. For large clumps, this resistance may be significant.

f) Transfer through liquid-cell interface. Resistance in this step is neglected. g) Transfer through solid to the cells (in case of floc, clump etc.). Resistance is likely to be significant, depending on size of the clamps.

h) Transfer through the cytoplasm to the site of reaction. Resistance is negligible due to small distance covered. For suspended, well-mixed culture, significant resistance is offered in step 3. At steady-state:

kLa is used to characterize the mass-transfer capability of a fermenter. = driving force for oxygen transfer. As a thumb rule, kL in fermentation liquids is about 3-4 x 10^-4 m/s for bubbles greater than 2-3 mm diameter. Once size of bubbles exceed 2-3mm, kL becomes relatively constant.

Focus should be on improving the interfacial area rather than kL. Bubble behavior strongly affects the value of kLa. For a given volume of gas, more interfacial area is provided if the gas is dispersed into many small bubbles rather than a few large ones. Besides the above, small bubbles create high gas holdup.

Relatively large bubbles should be employed in viscous cultures. kL decreases with decreasing bubble diameter below 23 mm (due to surface effects). surface tension dominates, bubble surface behaves as rigid spheres with immobility hindering mass transfer.

Coalescence is undesirable as it decreases interfacial area and gas hold-up. Salts act to suppress coalescence, so, it is not a major problem in fermentation broth. Under typical fermentation conditions, increasing the stirrer speed improves the value of kLa. Increasing gas flow rate doesn t affect kLa much, except at very low gas flow rate.

Effect of antifoam agents: Most antifoam agents tend to decrease surface tension leading to increase of interfacial area. At the same time, value of kL lowers down. In silicone oil-based antifoams, kLa overall decreases necessitating use of mechanical foam breakers (rotating disk, centrifugal foam breaker etc.).

Effect of temperature and pressure: Solubility of oxygen in water decreases as temperature increases while value of kL increases. Between 10 and 40 degree Celsius, mass transfer increases. Above 40, it decreases. Mass transfer increases with increase in partial pressure of oxygen.

Effect of cells: Cells with complex morphology generally leads to lower transfer rate. Cells interfere with bubble break-up and coalescence. Cells, proteins and other molecules which adsorb at gas-liquid interface cause interfacial blanketing which reduces the contact area between gas and liquid.

Effect of agitation: In an aerated, non-viscous system, kLa depends on power dissipated in the vessel. For most of the fermentation broth being in turbulent flow, power dissipated is directly proportional to density of the broth. P = c*d*(N^3)*(D^5) So, kLa is affected by impeller rotational speed and its diameter.

Agitation affects kLa by: 1) increasing area available for oxygen transfer by dispersing the air in the culture fluid in the form of small bubbles. 2) delaying escape of air bubbles from the liquid broth.

contd 3) preventing coalescence of the air bubbles resulting in formation of bubbles of larger size. 4) decreasing the thickness of the liquid film at the gas-liquid interface by creating turbulence in the culture broth.

y Doran, P.M. (1995) Bioprocess Engineering Principles,

Elsevier Science and Technology books. y Treybal, R.E. (1968) Mass- Transfer Operations, 2nd edn., McGraw-Hill, Tokyo. y Whitaker A., Stanbury P.F. and Hall S. J. (1995) Principles of Fermentation Technology, Pergamon Press, Oxford

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