Agitation

Agitation is a process by which the cultured microbial cells are maintained in a homogenous suspension along
with nutrients in the fermentation medium

Achieving uniform suspension of microbial cells in homogeneous nutrient medium

Fine aerator without mechanical agitation - lower equipment and power costs (agitation provided only when
broths with low viscosity and low total solids used)

Mechanical agitation required in fungal and actinomycete fermentations

Non-agitated fermentations – aeration sufficient to provide agitation

Herein, vessel with height/diameter ratio of 5:1 used and tall column of liquid requires energy input in production
of compressed air

Structural components of fermenter involved in aeration and agitation :

(a) The agitator (impeller).

(b) Stirrer glands and bearings.

(c) Baffles.

(d) The aeration system (sparger).

The agitator (impeller) :

Required for mixing - bulk fluid and gas-phase mixing

air dispersion

oxygen transfer

heat transfer

suspension of solid particles

Maintaining uniform environment throughout vessel contents.

While building fermenter knowledge of most appropriate agitator, air sparger, baffles, the best positions for
nutrient feeds, acid or alkali for pH control and anti-foam addition required

Specification of agitator size and number, speed and power input needed

Agitators classified as :

Disc turbines

Vaned discs

Open turbines of variable pitch and propellers,

Disc turbine - consists of disc with series of rectangular vanes set in vertical plane around circumference

Vaned disc – consists of discs with series of rectangular vanes attached vertically to underside.

Air from sparger hits underside of disc and is displaced towards the vanes where air bubbles broken into smaller
bubbles.
Vanes of variable pitch open turbine and marine propeller blades attached directly to boss on agitator shaft, air
bubbles do not hit any surface before dispersion by vanes or blades

To mix 3 phases within fermenter :

Liquid phase – containing dissolved nutrients

Gaseous phase – containing oxygen and carbon dioxide

Solid phase – containing cells and solid substrates

Mixing for homogenous conditions and promoting nutrient, gas and heat transfer

Heat transfer is necessary during sterilisation and maintaining temperature during fermentation

Efficient agitation needed in aerobic fermentations – even mixing of oxygen from gaseous phase into liquid phase

Efficient agitation prolongs retention of bubble in suspension, reduces bubble size to increase surface area for
oxygen transfer, prevents bubble coalescence and decreases film thickness at the gas-liquid interface

Suitable shear conditions need to be maintained :

High shear can damage shear-sensitive cells

Low shear leads to cell flocculation or unwanted growth on surfaces (vessel walls, stirrer and electrodes)

Fermenter agitation needs substantial energy input

3 principal mechanisms used :

1. Stirred tank reactors (STRs)

2. Pneumatic systems
3. Hydrodynamic mechanisms

1. Stirred tank reactors (STRs)

Mechanically moving agitators or impellers within a baffled cylindrical vessel

Baffles - Flat vertical plates with width about 1/10 th of vessel diameter

4-6 baffle plates fitted in vessel walls – aids mixing and mass transfer by increasing turbulence, preventing vortex
formation and eliminating dead spaces

Most commonly used and adapted for wide range of fermentation processes

Within each vessel, impeller is connected to external motor that drives the stirrer system

The shaft is passed into fermenter through set of aseptic seals to prevent contamination from agitator assembly
including seal

Specific regulations regarding numbers and types of seals

2 or 3 seals required to minimize risk of fermenter contamination and prevent release of microorganisms into
environment (containment)

Effectiveness of agitation dependent on - design of impeller blades, agitation speed and depth of liquid

Height–diameter aspect ratios of 3:1 or 4:1

STRs must create high turbulence to maintain transfer rates, but must control shear force that is generated
simultaneously to avoid cell disruption (animal and plant cells) of shear-sensitive cells due to excessive stirring

For shear-sensitive cells, modified STRs, airlift or supported biofilm reactors can be used

2. Pneumatic systems :

Include airlift fermenters

Devoid of moving parts; use expansion of compressed gas to bring about mixing

Lower energy requirements and create lesser shear than STRs

Compressed air injected at bottom of internal or external riser column and air bubbles expand in riser causing
upward movement of liquid and its cycling

Large fermenters do not require internal cooling coils – jacket provides sufficient heat transfer due to rapid fluid
movement

3. Hydrodynamic mechanisms :

Use liquid kinetic energy to mix fermenter contents – done by External liquid pump for external circulation and
reinjection, e.g. deep-jet fermenters

Mixing of nutrients and gaseous exchange within fermenter is complex and influenced by :

1. Medium density and rheology

2. Size and geometry of the vessel
3. Amount of power used in system

STRs have agitators with multiple impellers to give well-mixed homogeneous environment

Mon-uniform conditions prevail in over 500L capacity vessels

Flow patterns of fluid as a function of Reynold’s Number (Re):

1. Laminar flow
2. Turbulent flow

High Re – Laminar flow

Low Re – Turbulent flow

Re value marks transition between 2 flow regimes and depends on geometry of impeller and vessel

Re is natural variable or dimensionless number and its magnitude does not require units (numerator and
denominator cancel out)

Dimensionless analysis useful in complex hydrodynamics associated with physical transfer processes operating
within fermenter.

In liquid cultures - rheological behaviour, fluid flow properties – major impact on mixing and mass transfer for
oxygen transfer

Liquids behave as :

1. Newtonian fluids
 2. Non -Newtonian fluids
3. Viscoelasticfluids

Newtonian fluids - obey Newton’s law of viscosity and their viscosity does not vary with shear or agitation rate.

Example - bacterial and yeast fermentations

Equation

Non-Newtonian fluids - viscosity varies with shear or agitation rates, Inhibit high flow dynamics

Example - Mycelial cultures, fluids involving polymeric substrates and products, polysaccharide gels (e.g. xanthan)

Pseudoplastic fluids - decreasing apparent viscosity with increasing shear or agitation rate, in dilatant solutions
opposite occurs.

For Bingham-plastic behaviour, flow does not occur unless stress is first imposed.

Viscoelasticfluids - do not observe normal liquid-state properties when stirred (polymers)

Aeration :

Sparger - device for introducing air into the liquid in a fermenter.

3 types of sparger :

Porous sparger :

Made of sintered glass, ceramics or metal

Primarily used on laboratory scale in non-agitated vessels

Bubble size 10 to 100 times larger than aerator block

Throughput of air is low because – pressure drop across the sparger and fine holes blocked by microbial growth

Orifice Sparger :

Perforated pipes (with or without impellers)

In small stirred fermenters, perforated pipes arranged below the impeller as crosses or rings (ring sparger),
approximately three-quarters of impeller diameter

Air holes drilled under surfaces of tubes making up the cross or ring

Sparger holes must be minimum 6 mm diameter – to avoid blockage and minimise pressure drop

Orifice spargers (without agitation) used in - yeast manufacture, effluent treatment and single-cell protein
production

Nozzle sparger

Used from laboratory to industrial scale in mechanically stirred fermenter designs

Single open or partially closed pipe provides stream of air bubbles

Pipe positioned centrally but away from impeller – avoid impeller flooding by air stream
Lower pressure loss and less blockage

Aeration :

Provide microorganisms with sufficient oxygen in submerged conditions