Different modes of operation

In biotechnological processes there are three main ways of growing microorganisms in the
bioreactor: batch, fed-batch or continuous

Batch culture :

• Microbes inoculated into a fixed volume of medium and as growth takes place nutrients are
consumed and products of growth (biomass, metabolites) accumulate.
• Continuous change in the nutrient environment
• Induces change in cell metabolism.
• Cell multiplication ceases because of exhaustion or limitation of nutrient(s) and accumulation
of toxic excreted waste products.

Means of prolonging the life of a batch culture and thus increasing the yield by various substrate
feed methods:

(1)by the gradual addition of concentrated components of the nutrient, e.g. carbohydrates, so
increasing the volume of the culture (fed batch) – used for industrial production of baker’s
yeast.

(2) by addition of medium to the culture (continuous) and withdrawal of an equal volume of
used cell-free medium – used in mammalian cell cultivations.
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Paulová, Leona & Brányik, Tomáš. (2013). Advanced Fermentation Processes. 10.1201/b15426-6.
Advantages of Fed-batch culture
• The possibility to prolong product synthesis, the ability to achieve higher cell densities and thus
increase the amount of the product, which is usually proportional to the concentration of the
biomass
• The capacity to enhance yield or productivity by controlled sequential addition of nutrients, and
• The feature of prolonged productive cultivation over the “unprofitable periods” when the
bioreactor would normally be prepared for a new batch.
• Fed-batch is advantageously used in processes
(a)where substrate inhibition or catabolic repression is expected; this problem can be overcome by
using a “safe” concentration of the substrate in batch mode followed by feeding the remaining
substrate within fed-batch operation
(b) where a Crabtree effect (repression of yeast respiratory enzymes by high concentrations of
glucose) is expected; by gradual feeding of the substrate, the production of ethanol by yeasts
can be eliminated under aerobic conditions
(c) where a high cell density is required; a high and constant specific growth rate can be
maintained by exponential feeding of the substrate
(d) where a high production rate should be achieved; cell metabolism can be regulated by precise
sequential feeding of nutrients, and
(e)where a high viscosity of culture broth is expected (e.g., production of dextran or xanthan); a
gradual dilution of the medium can overcome the problems of mixing and oxygen transfer.
Relationship between precalculated profiles of substrate feeding and specific growth rates of cell culture. (a) Constant feeding
rate, (b) linearly increasing feeding rate, and (c) exponential feeding rate.
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How cells grow in continuous culture:
Dilution rate and influent substrate
concentration are the two parameters
controlled in a chemostat to study microbial
growth or to optimize metabolite
production.

Chemostats can hold the exponential phase of growth for extended periods of time.
How cells grow in continuous culture:

Continuous stirred tank reactor

Mass balance for Biomass

- F 𝑋𝑋� + µ 𝑋𝑋� V = 0

𝐹𝐹
From eqn 1 , 𝜇𝜇=D Dilution rate, 𝐷𝐷 =
𝑉𝑉

𝜇𝜇max 𝑠𝑠̅
Critical Dilution Rate 𝐷𝐷𝑐𝑐 =
𝐾𝐾𝑠𝑠 + 𝑠𝑠̅
When a chemostat is operating at Dc, if the dilution rate
is increased further, the growth rate will not be able to Chemostats have been an important tool for the study of
increase (since it is already at μmax) to offset the increase the physiology of microbial growth, as the growth rate and
in dilution rate. the cell number can be controlled independently
The result will be washing out of cells and a decline in
the operating efficiency of the chemostat. Thus, Dc is an
important parameter because if the chemostat is run at
dilution rates less than Dc, operation efficiency is not
optimized; whereas if dilution rates exceed Dc, washout
of cells will occur
Bioreactor

• A bioreactor is any device or vessel that is used to carry out one or more biochemical reactions to convert any
substance (i.e. a substrate) to some product.
• Driven by biocatalyst
• Provides an environment that is conducive to optimal functioning of the biocatalyst
• Submerged culture with the biocatalyst suspended in a nutrient medium in a suitable reactor

• Stirred tank reactors

Fermentation is multiphase

(1) The need to maintain monoseptic operation.

(2) Mixing to ensure suspension of the biocatalyst and attain a relatively homogeneous environment in the bioreactor

(3) Supply of oxygen and removal of carbon dioxide.

(4) Supply of various other nutrients in such a way that the rate of supply does not limit the performance of the biocatalyst.

(5) Heat transfer for temperature control.

(6) Control of the shear stress levels in the bioreactor so that the biocatalyst is not damaged by various hydrodynamic forces.

Fermentation technologists seek to achieve a maximisation of culture

Reference book – Basic Biotechnology by Colin Ratledge & Bjorn Kristiansen

Lab scale bioreactors – 5-10 l

Pilot plant bioreactors - 100–10,000 l

Full-scale industrial bioreactors - 20 000 l and 400 000 l in volume

The management of scale-up requires high capital investment in mixing and aeration, in monitoring and control
devices, and in stringent maintenance of sterility.