FOOD AND BIOPROCESS TECHNOLOGY

UNIT I
Introduction to Bioprocess Technology: History and Scope- Bioreactor: Design, parts and
accessories, functions- Modes of Operation of fermenter – Batch & continuous - Types of
reactors - Bubble column, Fluidized bed reactor, plug flow reactor.
Introduction
Bioprocess engineering encompasses the design, operation, control, and optimization
of biochemical processes involving various biological pathways or reactions mediated by
living cells of animals, plants and microorganisms or enzymes under controlled conditions
for the efficient biotransformation of raw material into a range of products at requisite scales.
The product may be directly useful as food, medicine or the industrial compounds or
indirectly in the way of bioprocess without any direct product formation as in the
detoxification of industrial wastes or treatment of factory effluents. For production of
fermented foods, micro-organisms have been used from the very past.
Products of Industrial Microbiology
 Pharmaceutical chemicals : antibiotics and steroid drugs
 Commercially valuable chemicals: solvents, enzymes
 Food supplements: yeast, bacteria, algae, Single cell Proteins, amino acids, yougurt,
cheese, butter, fermented foods
 Alcoholic beverages: beer, wine
 Vaccines
Fermentation:
The conventional definition of fermentation is the breakdown (metabolism) of the
larger molecules, for example carbohydrates, into simpler ones under the influence of
microorganisms for their enzymes.
Industrial fermentation is comprised of two main stages
• Upstream Processing (USP)
• Downstream Processing (DSP)
Upstream Processing
Involves all factors and processes leading to and including the fermentation. It
consists of three main areas:
1. Producer microorganism- This include processes for
a. obtaining a suitable microorganism
 b. strain improvement to increase the productivity and yield
c. maintenance of strain purity
d. preparation of suitable inoculum
2. Fermentation media
3. Fermentation Process
Downstream Processing
It is the collective term for the processes that follows fermentation i.e.
a. cell harvesting
b. cell disruption
c. product purification from cell extracts or the growth medium
Scales of fermentation:
Bench top scale: This scale is up to 3 liters.
Research scale: This scale is usually 5–50 liters
Lab scale: This scale can range from 0.5–100 liters.
Pilot scale: This scale can have a working volume of 10–350 liters.
Industrial scale: Working volume ranges from 300 L to 150,000 L.
Bioreactors/ fermenter:
The heart of fermentation (or bioprocessing) technology is the fermenter (or
bioreactor). A bioreactor is basically a device in which the organisms cells are cultivated and
motivated to form the desired products. It is a system designated to give right environment
for optimal growth and metabolic activity of the organism. Traditional fermenters are made
up of wood or slate.
In recent years, stainless steel bioreactors are in use. A high quality stainless steel
does not corrode or leak toxic metals into the growth medium. The size of the bioreactor is
highly variable ranging from 20 liters to 250 million liters or even more.
General requirements of the bioreactor are as follows:
1) The vessel should be robust and strong enough to withstand the various treatments
required such as exposure to high heat, pressure, and strong chemicals and washings and
cleanings.
2) The vessel should be able to be sterilized and maintain stringent aseptic conditions
over long periods of the actual fermentation process.
3) The vessel should be equipped with stirrers or mixers to ensure mass transfer
processes occur efficiently.
4) It should have sensors to monitor and control the fermentation process.
5) It should be provided with an inoculation point for aseptic transfer in the inoculum.
6) Sampling valve for withdrawing a sample for different tests.
7) Baffles should be provided in case of stirred fermentor to prevent vertex formation.
8) It should be provided with a facility for intermittent addition of an antifoam agent.
 9) In case of aerobic submerged fermentation, the tank should be equipped with an
aerating device.
10) Provision for controlling temperature and pH of fermentation medium.
11) Manhole should be provided at the top for access inside the fermentor for different
purposes.
Design and Major Parts of fermentor
1. Impellers/ Agitator
2. Baffles
3. Inoculation port
4. Sparger/ Aerator
5. Sampling point
6. pH control device
7. Temperature control system
8. Foam control device
9. Bottom drainage system
Impeller/ Agitator
 Impellers are an agitation device. They are mounted on the shaft and introduced in the
fermentor through its lid.
 They are made up of impeller blades and the position may vary according to its need.
 These impellers or blades are attached to a motor on lid.
 The important function of an impeller is to mix micro-organisms, media and oxygen
uniformly.
 Impeller blades reduce the size of air bubbles and distribute these air bubbles
uniformly into the fermentation media.
 Impeller also helps in breaking foam bubbles in the head space of fermentor. This
foam formed during fermentation process can cause contamination problem and this
problem is avoided by the use of impellers.
Baffles
 Baffles are mounted on the walls of a fermentor.
 The important function of baffles is to break the vortex formed during agitation
process by the impellers. If this vortex is not broken, the fermentation media may spill
out of fermentor and this may result in contamination as well as can lead to different
problems. So it is important to break the vortex formed by using a barrier.
 Baffles acts as a barrier which break the vortex
Inoculation Port
 Inoculation port is a device from which fermentation media, inoculum and substrate
are added in the fermentation tank.
 Care should be taken that the port provides aseptic transfer.
 The inoculation port should be easy to sterilize.
Spargers/ Aerator
 A Sparger is an aeration system through which sterile air is introduced in the
fermentation tank.
 Spargers are located at the bottom of the fermentation tank.
 Glass wool filters are used in a sparger for sterilization of air and other gases.
 The sparger pipes contain small holes of about 5-10 mm. Through these small holes
pressurized air is released in the aqueous fermentation media.
 The air released is in the form of tiny air bubbles. These air bubbles help in mixing of
media.
Sampling point
 Sampling point is used for time to time withdrawal of samples to monitor
fermentation process and quality control.
 This sampling point should provide aseptic withdrawal of sample.
pH Control device
 The pH controlling device checks the pH of media at specific intervals of time and
adjusts the pH to its optimum level by addition of acids or alkalis.
 Maintaining pH to its optimum level is very important for growth of micro-organism
to obtain a desired product.
Temperature control
 Temperature control device generally contains a thermometer and cooling coils or
jackets around fermentor.
 During the fermentation process, various reactions take place in the fermentor. Heat is
generated and released in the fermentation media. This increase in temperature is
detrimental to the growth of micro-organisms, which may slow down the
fermentation process. So, it is necessary to control the rise in temperature. This is
done by passing cool water through the coils or jackets present around fermentor.
Parts of fermentor and its function
Foam controlling device
 A Foam controlling device is placed on the top of fermentor with an inlet into
fermentor. This device contains a small tank containing anti-foaming agent.
 Foam is generated during fermentation. It is necessary to remove or neutralize this
foam with the help of anti-foaming agents, or the media may spill out of fermentor
and lead into contamination and a mess.
Bottom drainage system
 It is an aseptic outlet present at the bottom of fermentor for removal of fermented
media and products formed
Modes Fermentation Process
Fermentation process is divided depending on the feeding strategy of the
culture and medium as follows.
i. Batch Fermentation
ii. Continuous Fermentation
iii. Fed batch Fermentation
i. Batch Fermentation
It is a fermentation process that involves adding all nutrients and
substrate at the beginning, and then allowing the process to proceed in a
controlled environment until the desired end product concentration is achieved
and it is then closed system. Microbes consume the provided raw material as the
reaction progresses, depleting nutrient volume. Batch fermentation is used to
produce biomass, secondary metabolites, and various food products.
When cells are grown in a batch reactor, they go through a series of stages
known as
1. Lag phase
2. Exponential phase
3. Stationary phase
4. Death phase

Different phases of microbes in batch reactor

1. Lag Phase
In lag phase the microbial population remains constant as there is no
growth. However it is the period of intense metabolic activity. Factors
Influencing the Lag Phase
 Chemical composition of the fermentation media influences the
length of the lag phase. Longer lag phase is observed if the inoculum
is transferred into a fresh medium of different carbon source as that of
the medium in which the cells are grown. This is because the cells
 need to induce the enzymes required for the metabolism of the new
substrate.
 Age of the inoculum. If the inoculum is in exponential growth phase,
it will exhibit shorter lag in the fresh medium.
 Concentration of the inoculum
 Viability and morphology of the inoculum
2. Exponential Phase / log phase
 Cell divides with increasing frequency till it reaches the maximum
growth rate. At this point logarithmic growth begins and cell numbers or
cell biomass increase at a constant rate.
3. Stationary Phase
 The specific growth rate of the microorganism continues decelerating
until the substrate is completely depleted.
 Overall growth rate has declined to zero and there is no net change in cell
numbers/ biomass i.e. rate of cell division equals rate of cell death.
 Microorganisms are still metabolically active, metabolizing intracellular
storage compounds, utilizing nutrients released from lysed cells and in
certain cases produce secondary metabolites.
4. Death Phase
Cells die at constant rate and often undergo lysis.
ii. Continuous fermentation
This is an open system. It involves the removal of culture medium
continuously and replacement of them with a fresh sterile medium in a
bioreactor. Reactors which include chemo stat and turbido stat bioreactors are
used. Examples: production of antibiotics, organic solvents, beer, ethanol and
SCP
Types of Continuous Culture
There are two types of continuous culture techniques:
a) Turbidostat: In this, the turbidity or the optical density is measured as per
the cell count. The turbidity level is used to track the amount of nutrients
added and products removed. It necessitates the integration of a real-time
feedback sensor.
b) Chemostat: This method is used to grow the microbial cells continually in
any specific phase of growth. A portion of the product containing actively
diving cells in a specific phase is always left in the fermenter during
product recovery. This serves as an inoculum for the nutritional supply to
come. The microorganisms’ desired stage is constantly maintained in this
sort of fermentation.
 Batch Vs Continuous fermentation
Basis of Batch Continuous
Comparison Fermentation Fermentation

Here, the fermentation process is

In this type of fermentation is performed in
continuously ongoing with persistent
batches where the raw materials and microbial
Meaning nutrient intake and the product is
inoculum are introduced at the beginning and
simultaneously recovered during the
product is recovered at last.
ongoing reaction.

Type of system Closed system Open system

Suitable of generation of secondary Used for production of primary

Application
metabolites. metabolites.
• Less labor
• Highly efficient
• Less automation
Advantages • Less time consuming
• Only one batch is wasted in case of
• High product yield.
contamination.
• More labor demand
• Less efficient
• High level automation
Disadvantages • More time consuming
• Huge volume of the raw material and
• Low product yield
product is wasted if contaminated.
Microbial Microbes are cultivated in the limited supply of Microbes are grown under the constant
Growth nutrients. supply of nutrients.
Nutrients are supplied continuously
Nutrient supply Nutrients are added at the initial stage only.
throughout the process.

Microbes keep on growing at optimum rate Microbes grow exponentially with

Growth rate
until the nutrients are exhausted. constant growth rate.

Comprise of Lag, log stationary and decline

Growth Phases Constitutes lag and log phases.
phases.
Product is continuously discharged
Product is recovered after the completion of the
Product harvest during the ongoing fermentation
fermentation reaction.
process.
 Yield Product yield is low. Product yield is high.
Level of Can be performed manually and requires less
High range of automation is necessary.
Automation automation.
Chances of
Low High
contamination
Turnover Rate Low High
Done in 1-2 shifts of 5-7 days based on the
Time Duration Requires 24x7 shifts.
desired product.
Internal Doesn't change; Internal equilibrium is
Changes as the reaction proceeds.
Changes maintained.
Fermenters Large fermenters Small fermenters
Initial
Low High
investment
Product Quality May vary from batch to batch. Remains consistent.

iii. Fed batch system

It is a combination of both batch and continuous systems. In this,
additional nutrients are added to the fermentors as the fermentation is in
progress. This extends the time of operation, but the products are harvested at
the end of the production cycle as in batch fermenter.
Types of Bioreactors.
Based on design of the bioreactors they can be grouped into the following types:
1. Continuous stirred tank bioreactor
2. Bubble column bioreactor
3. Airlift bioreactors
4. Packed bed bioreactors
5. Photobioreactors
6. Plug flow reactor
7. Trickling bed reactor
Continuous stirred tank bioreactor:
A continuous stirred tank bioreactor consists of a cylindrical vessel with motor driver
central shaft that supports one or more agitator(impeller).
 The shaft is fitted at the bottom of the bioreactor. The no. of impeller is variable and
depends on the size of the bioreactor. Different types of impellers (concave bladed, marine
propeller etc.,) are in use.
In stirred tank bioreactor, the air is added to the culture medium under pressure
through a device called sparger. The sparger may be the ring with many holes or a tube with
single orifice. The sparger along with impeller (agitators) enables better gas distribution
system throughout the vessel. The bubbles generated by sparger are broken down to small
ones by impellers and dispersed throughout the medium. This enables the creation of a
uniform and homogenous environment throughout the bioreactor.
Bubble column bioreactor:
In the bubble column bioreactor the air or gas is introduced at the base of the column through
perforated pipes or plates, or metal micro porous spargers. The flow rate of the air/gas
influences the performance factors-O2 transfer; mixing .The bubble column bioreactor may
be fitted with perforated plates to improve performance. The vessel used for bubble column
bioreactors is usually cylindrical with an aspect ratio of 4-6 (i.e., high to diameter ratio)
Bubble column bioreactor:

Airlift bioreactors:
Also known as tower reactor, an airlift bioreactor can be described as a bubble
column containing a drought tube .Many types of airlift bioreactors are currently in use today.
Air is typically fed through a sparger ring into the bottom of a central drought tube that
controls the circulation of air and medium. Air flows up the tube forming bubbles and
exhaust gas leaves at the top of the column. The degassed liquid then flows downward and
the product is drained from the tank. There are two types
a.) Internal loop airlift bioreactor: Has a single container with a central draft tube that
creates interior liquid circulation channels.
b.) External-loop airlift bioreactor: This system consists of a riser and an external down-
comer, which are connected at the bottom and the top respectively. As the injected air at the
bottom of the riser creates gas bubbles that begin to rise through the main tank, exhausted gas
leaves at the top and the resulting heavier solution descends through the down comer.
Airlift bioreactors:

Packed bed bioreactors:

A bed of solid particles with biocatalysts on or within the matrix of solids, packed in a
column constitute a packed bed bioreactor. The solid may be porous or non porous gels, and
may be compressible or rigid in nature. A nutrient broth flows continuously over the
immobilized biocatalyst. The product obtained in the packet bed bioreactor are released into
the fluid and removed .While the flow of the fluid can be upward or downwards, downflow
under gravity is preferred.
Packed bed bioreactors:
Photobioreactors:
These are the reactor specialized for fermentation and can be carried out either by
exposing to sunlight or artificial illumination. Since artificial illumination is expensive, only
the outdoor photo-bioreactor are preferred. Certain important compounds are produced by
employing photo bioreactors e.g., B carotene, asthaxanthin. The photoreactors are made up of
glass or more commonly transparent plastic. The array of tubes or flat panels constitute light
receiving system (solar receiver).The culture can be circulated through the solar receiver by
methods such as using centrifugal pumps or airlift pump. It is essential that the cells are
continuous circulation without forming sediments. Further adequate penetration of sunlight
should be maintained. The tubes should also be cooled to prevent rise in temp.
Photobioreactors are usually operated in a continuous mode at a temp in the range 25-40oC.
Microalgae and cynobacteria are normally used.

Photobioreactors

A plug flow reactor (PFR) :

A plug flow reactor (PFR) is a chemical reactor that uses a tube or pipe to
continuously process reactants into products. The reactants enter the reactor at one end and
flow through in a "plug-like" fashion, exiting at the other end.
 Primary Metabolites Vs. Secondary Metabolites

Basis for Comparison Primary Metabolites Secondary Metabolites

Primary metabolites are the Secondary metabolites are the organic

compounds that are directly compounds that are produced by
involved in the metabolic various organisms that are not directly
Definition pathways of an organism involved in the growth, development,
necessary for its growth, or reproduction of the organism but
development, and are essential in the ecological and
reproduction. other activities.

Primary metabolites are also Secondary metabolites are also termed

Also termed
termed as central metabolites. as specialized metabolites.

Primary metabolites are

Secondary metabolites are produced
produced during the growth
during the stationary phase of the
Growth phase phase of the organism. This
organism. This phase of growth is also
phase of growth is also
termed as ‘idiophase’.
termed as ‘trophophase’.

Primary metabolites are Secondary metabolites are produced

Quantity
produced in large quantities. in small quantities.

It is easier to extract primary It is difficult to extract secondary

Extraction
metabolites. metabolites.

Primary metabolites are not

Secondary metabolites are species-
species-specific and thus
Specificity specific and thus are different in
might be identical in some
different organisms.
organisms.

Primary metabolites are

Secondary metabolites are involved in
involved in the growth,
Involved in ecological functions and species
development, and
interactions.
reproduction of organisms.

Primary metabolites might Secondary metabolites are not a part

Structural component form the molecular structure of the molecular structure of the
in organisms. organism

Secondary metabolites are used in

Primary metabolites are used
various biotechnological procedures
Importance in various industries for
for the formation of drugs and other
different purposes.
compounds.

Primary metabolites are not Secondary metabolites are active

Defensive action active in the defense against foreign invaders and might be
mechanism. involved as a defense mechanism.
 Examples of primary
Some examples of secondary
metabolites include proteins,
metabolites include steroids, essential
Examples enzymes, carbohydrates,
oils, phenolics, alkaloids, pigments,
lipids, vitamins, ethanol,
antibiotics, etc.
lactic acid, butanol, etc.