Industrial Microbiology

INDM 4005

Lecture 6

17/02/04
 Questions for today:

• 1. What is a fermentation system?

• 2. What is the most widely used fermenter?
• 3. What are the other types of fermenter?
• 4. How do you control a fermentation system?
• 5. Why is mass transfer important?
 Lecture Overview

(1) Basic design criteria and limitations

(2) Stirred Tank Reactor (STR)

(3) Modifications and Industrial Examples

 Biotechnological processing

Types of Process
Fermentation Design
Fermenter Design

Performance Optimisation Construction Configuration Control

Stirred Tank Reactor

 What is a Fermenter?
• Vessel or tank in which whole cells or cell-free
enzymes transform raw materials into
biochemical products and/or less undesirable
by-products

• Also termed a Bioreactor

 Fermenter - Basic Function
The basic function of a fermenter is to provide a
suitable environment in which an organism can
efficiently produce a target product that may be
- cell biomass,
- a metabolite,
- or bioconversion product.
 Fermentation System
• In this lecture we will concentrate on
fermenters used in traditional microbial, plant
and animal cell culture

• However with the advent of recombinant DNA

technology alternate systems for producing
specific cell products are now available
Two Types of Fermentation Systems
– closed or open.
– A closed system implies that all the nutrient components are added at
the beginning of the fermentation process and, as a result, the growth
rate of the contained organisms will eventually proceed to zero due
either to diminishing nutrients or accumulation of toxic waste products. A
modification of the batch process is the fed batch system. Here,
volumes of nutrients may be added to augment depletion of nutrients.
Overall, the system, however, remains closed and there is no
continuous flow.
– In contrast to the above types, in the open system, organisms and
nutrients can continuously enter and leave the fermenter.
Fermenter General Functions
What it should be capable of;

• Biomass concentration must remain high

• Maintain sterile conditions
• Efficient power consumption
• Effective agitation
• Heat removal
• Correct shear conditions
• Sampling facilities
• Fermenters range from simple stirred tanks to complex
integrated systems involving varying levels of computer
input.

• Fermenter design involves cooperation in Microbiology,

Biochemistry, Chemical Engineering, Mechanical
Engineering, Economics

• There are 3 groups of bioreactor currently used for

industrial production;
- non-stirred, non-aerated
- non-stirred, aerated (Beer and wine)
- stirred, aerated (Biomass, eg Pruteen)
(Antibiotics)
 Fermenter construction
– All materials must be corrosion resistant to prevent
trace metal contamination of the process
– Materials must be non-toxic so that slight dissolution of
the material or components does not inhibit culture
growth
– Materials of the fermenter must withstand repeated
sterilization with high pressure steam
– Fermenter stirrer system and entry ports be sufficiently
robust not to be deformed under mechanical stress
– Visual inspection of the medium and culture is
advantageous, transparent materials should be used
 Basic fermenter configuration
• A microbial fermentation can be viewed as a three-
phase system, involving liquid-solid, gas-solid, and gas-
liquid reactions.

• The liquid phase contains dissolved nutrients, dissolved

substrates and dissolved metabolites.
• The solid phase consists of individual cells, pellets,
insoluble substrates, or precipitated metabolic products.
• The gaseous phase provides a reservoir for oxygen
supply and for CO2 removal.
Optimisation of the Fermenter System

– Fermenter should be designed to exclude entrance of

contaminating organisms as well as containing the
desired organisms
– Culture volume should remain constant,
– Dissolved oxygen level must be maintained above
critical levels of aeration and culture agitation for aerobic
organisms
– Parameters such as temperature of pH must be
controlled, and the culture volume must be well mixed.
– Therefore a need for control exists
Control of Chemical and Physical Conditions
• Intensive properties (cannot be balanced)
- temperature, concentration, pressure, specific heat

• Extrinsive properties (can be balanced)

- mass, volume, entropy and energy

• Mass and energy levels should balance at the start and finish of
fermentations.

• Combining this with determination of thermodynamic properties and rate

equations we can build computer and mathematical models to control
processes.
 Basic Fermenter Design Criteria
(i). Nature of microbial (or mammalian, plant tissue) cell;
(a) Hydrodynamic characteristics
(b) Mass and Heat Transfer
(c) Kinetics
(d) Genotype and Phenotype

(ii). Environmental Control and Monitoring of the process;

(a) pH, temperature, dissolved oxygen etc.
(b) Asepsis and avoidance of contamination

(iii). Process factors;

(a) Effect on other unit operations
(b) Economics
(c) Potential for scale-up
 Types of Fermenter
• Aerobic fermenters may be classified
depending on how the gas is distributed

• Stirred Tank Reactor

• Airlift
• Loop Reactor
• Immobilised System
 Stirred Tank Reactors
• Most commonly fermenter used
• Made from stainless steel when over 20 Litres
• Height to Diameter ratio 2:1 and 6:1
• Baffles prevent a large central vortex
• Also used to carry coolants in large systems
Stirred Tank Reactor
 STR - Control systems
 An agitator system
 An oxygen delivery system
 A foam control system
 A temperature control system
 A pH control system
 Sampling ports
 A cleaning and sterilization system.
 A sump and dump line for emptying of the reactor.
 Aeration and agitation
• The transfer of energy, nutrients, substrate and metabolite within the
bioreactor must be brought about by a suitable mixing device. The
efficiency of any one nutrient may be crucial to the efficiency of the
whole fermentation.

• For the three phases, the stirring of a bioreactor brings about the
following:

 Dispersion of air in the nutrient solution

 Homogenisation to equalise the temperature and the
concentration of nutrients throughout the fermenter
 Suspension of microorganisms and solid nutrients
 Dispersion of immiscible liquids
 Basic features of a stirred tank bioreactor

Agitation system
The function of the agitation system is to
 provide good mixing and thus increase mass
transfer rates through the bulk liquid and bubble
boundary layers.
 provide the appropriate shear conditions required
for the breaking up of bubbles.
• The agitation system consists of the agitator and the baffles.
• The baffles are used to break the liquid flow to increase
turbulence and mixing efficiency.
 Agitator design and operation

Radial flow impellers - Rushton turbine

The most commonly used agitator in microbial fermentations

Like all radial flow impellers, the Rushton turbine is designed to provide the high
shear conditions required for breaking bubbles and thus increasing the oxygen
transfer rate.
 Mass Transfer
• One of the most critical factors in the operation of a
fermenter is the provision of adequate gas exchange.

• Oxygen is the most important gaseous substrate for

microbial metabolism, and carbon dioxide is the most
important gaseous metabolic product.

• For oxygen to be transferred from a air bubble to an

individual microbe, several independent partial resistance’s
must be overcome
 Oxygen Mass Transfer Steps

Gas bubble
Liquid film

1 Microbial cell
2

4
6
1) The bulk gas phase in the bubble
7
2) The gas-liquid interphase 5
3) The liquid film around the bubble
4) The bulk liquid culture medium
5) The liquid film around the microbial cells
6) The cell-liquid interphase
7) The intracellular oxygen transfer resistance
 Air lift reactors
• In such reactors, circulation is caused by the
motion of injected gas through a central tube with
fluid re-circulating through the head space where
excess air and the by-product CO2 disengage.

• The degassed liquid then flows down the annular

space outside the draught tube
Airlift reactors

Effluent gas

Draught tube

Inlet air
 Airlift reactors
Advantages
• Low shear
• Easier to maintain sterility
• Increased oxygen solubility (KLa)
• Can allow large vessels

Disadvantages
• High capital cost
• High energy costs
• Hard to control conditions
• Foaming hinders gas -liquid separation
 SOME MODIFICATIONS
•(i) Important in tank reactor design:
•1. Continuous flow (activated sludge waste treatment)
•· Suitable when substrate at low conc.
•· Allows greater control on growth rate\ cell physiology

•2. Immobilised cells - may be membrane (e.g. hollow fibre reactor),

immobilised onto support such as ceramic (e.g packed-bed) or in polymers
(e.g alginate beads)
•· Increases rate of reaction
•· Microenvironment created protects cells e.g. from shear damage

•3. Low energy aeration\ mixing Air-lift, draft-tubes, loop reactors etc.
•· Increase height to diameter ratio. Increased path length of bubble,
improves mass transfer
•· Results in decreased shear levels, important in floc systems.
 SOME MODIFICATIONS
• (ii) Industrial examples of modified STR / bioreactors
• (i) Waste treatment. - Activated sludge system.
• Characterised by: Low substrate conc. Therefore require (a) recycle of
biomass, (b) continuous operation, (c) Low cost aeration / mixing.

• (ii) Brewing - Cylindro-conical fermenter;

• Note no aeration but gas produced by yeast cells contributes to mixing,
closed to capture carbon dioxide produced, cone helps sedimentation of
yeast, Low shear environment promotes flocculation.

• (iii) Tissue culture - low shear, anchored and immobilised systems.

• (iv) Solid-state fermentations e.g. silage, mushroom production etc.

 In Summary
Major considerations include
1. Bioreactor size - to provide required production capacity
2. Mass transfer - to provide nutrients to cells, well dispersed,
adequate oxygen etc
3. Control systems
(a) temperature, pH, etc.
(b) sterilisation/ aseptic operation
(c) representative sampling
(d) heat transfer - example sterilisation of media
4. Requirement for asepsis / containment
 Critical Concepts or Questions

• What are the objectives in fermenter design?

• Draw a diagram of a STR
• How does a STR relate to structure and function?
• How can fermentation systems be controlled?
 Conclusion
• This lecture introduced the various parameters
involved in design of an industrial fermenter.

• Using a STR it illustrated the optimisation and

control of a fermentation system.