Bioprocessing - Upstream and Downstream Processing of Fermentation.pptx

1. BIO-PROCESSING
SUBMITTED BY: 21SGM06, M.SC. MICROBIOLOGY
SUBMITTED TO: DR. K. KAVITHA, ASSISTANT
PROFESSOR, PG AND RESEARCH DEPARTMENT
OF MICROBIOLOGY, SADAKATHULLAH APPA
COLLEGE (AUTONOMOUS), TIRUNELVELI
(AFFILIATED TO MANONMANIAM SUNDARANAR
UNIVERSITY, ABISHEKAPATTI)

2.  Bio-processing refers to the use of biological systems, such as
microbes, cells and enzymes, to produce biologics or other
products.
 Bio-processing can involve both upstream and downstream
processing steps.
 Bio-processing is defined as a production of value added
material from the living source.
BIO-PROCESSING

3. Inoculum
development
Preparation
of culture
media
Sterilization
UPSTREAM PROCESS

4. FERMENTATION
Fermentation
Fermentation
organism
Fermentation
media
Fermenter
Sterilisation
Very important
Not
sterilised
Media
contamination
Lower yield of
product
Inhibit product
synthesis

5. Product
recivery Formulation
Formulation
and effluent
treatment
DOWNSTREAM PROCESSING

6.  Upstream process encompasses any technology that leads to
the synthesis of a product.
 Refers to the initial stages of industrial bio processing,
including the selection and optimization of micro organisms,
designing and controlling fermentation processes, and
harvesting microbial biomass or metabolites. It is a critical
step in determining the quality and quantity of the final
product.
 The upstream process mainly includes:
 Culture isolation and screening,
 Inoculum preparation,
 Media formulation and sterilization,
 Equipment preparation and sterilization.
I. UPSTREAM PROCESS

7.  Culture isolation refers to the process of isolating and
purifying a specific micro organism from a mixed culture or
sample. As a result we can get a pure culture of the desired
organism.
A) CULTURE ISOLATION AND SCREENING

8.  There are some techniques employed for pure culture
isolation:
MICRO MANIPULATOR
ISOLATION TECHNIQUES

9.  The screening process is a crucial in upstream processing as
it determines the quality and yield of the final product. The
selection of right micro organism can significantly enhance
the production efficiency and reduce the cost of
manufacturing.
SCREENING OF MICRO ORGANISMS
screening
Primary
secondary

10.  Primary screening involves the identification and selection of
micro organisms with desirable properties for the production
of a specific product.
 This can include screening for high yield, productivity, and
stability under various environmental conditions.
 Primary screening may also involve genetic modification or
engineering of micro organisms to improve their performance
in the fermentation process.
 The goal of primary screening is to identify the most
promising micro organisms for further optimization and scale-
up in downstream processing.
PRIMARY SCREENING

11. Primary
screening
Organic acid
producing
microbes
(using CaCo₃)
Enrichment
culture
technique
(defined
media)
Extracellular
metabolites
producing
microbes
(auxanography)
Antibiotic
producing
microbes
(crowded plate)
TYPES OF PRIMARY SCREENING

13.  Secondary screening is a follow-up process to the initial
screening process in upstream processing.
 It involves further testing of the selected micro organisms to
confirm their ability to produce the desired product
consistently and efficiently.
 This process helps to identify any potential issues or
limitations that may arise during large-scale production.
 It is useful in sorting of micro organisms that have real
commercial value.
 Physical, chemical and biological properties are also
determined during secondary screening.
 The result of secondary screening process are used to
optimize the fermentation conditions and develop a robust
production process. This ensures that the final product meets
the required quality standards and is produced in cost-
effective manner.
SECONDARY SCREENING

14.  Inoculum preparation is a crucial step that involves growing a
small amount of selected micro organism to be used in the
main fermentation process.
 The purpose of inoculum preparation is to provide a sufficient
number of viable and active cells that can be used to initiate
the main fermentation process.
 The chemical composition of the inoculum media used in
upstream processing varies depending on the specific micro
organism and the product being produced.
 The media typically contains a carbon source, nitrogen source,
minerals, vitamins, and other nutrients required for the cell
growth.
B) PREPARATION OF INOCULUM

16. Composition of media Constituent of major
source
Supplied as
Carbon source Molasses, malted barley,
starch, dextrins etc.
Glucose, sucrose, maltose,
lactose etc.
Nitrogen source Corn steep liquor, yeast
extract, peptones, soya
bean meal.
Ammonium salts, amino
acids, peptides.
Minerals Added with magnesium,
phosphorus, potassium,
sulfur, calcium, chlorine.
Water has Cobalt, copper,
iron, manganese,
molybdenum, zinc.
C) MEDIA FORMULATION AND
STERILIZATION

17.  Sterilization of the media is a crucial step to ensure the
sterility of the media containing nutrients and sterility of
bioreactor.
 Media must be free from contamination before entering in
fermentor.
 Sterilization of media is commonly achieved by heat, and
other physical methods like filtration, radiation and
biochemical treatments.
 It is important to note that the sterilization process must be
carefully monitored and validated to ensure that it is effective
and consistent. This includes regular testing of the media for
microbial contamination and ensuring that all equipment and
materials used in the sterilization process are properly
maintained and calibrated.
STERILIZATION OF MEDIA

18. D) STERILIZATION OF EQUIPMENTS

20.  Fermentation process is a key component of upstream
processing, where micro organisms such as bacteria, yeast or
fungi are grown in a controlled environment to produce a desired
product.
 Process involves several steps such as, growth and harvest.
 Inoculation involves adding the micro organisms to the sterilized
media in the bioreactor, where they begin to multiply.
 The growth phase involves optimizing the conditions within the
bioreactor, such as temperature, pH and oxygen levels, to
promote the growth and metabolism of the micro organisms.
 During this process, the organisms consume nutrients in the
media and produce the desired product. The product can be
harvested by separating the microbes from the media through
methods such as centrifugation or filtration.
II. FERMENTATION

21. BATCH, FED BATCH, CONTINUOUS
FERMENTATION

22.  Downstream processing is used to purify and isolate the
product for use in various industries. It involves several steps,
including separation, purification and formulation.
 Separation involves separating the product from any
remaining organisms or impurities in the media. This can be
done through methods such as filtration, centrifugation or
chromatography.
 Purification involves further refining the product to remove
any remaining impurities and ensure its quality and purity.
This can involve additional chromatography steps or other
purification techniques.
 Formulation involves preparing the purified product for its
intended use, such as adding stabilizers or preservatives to
increase the shelf-life or adjusting the concentration for
specific applications.
III. DOWN STREAM PROCESSING

23. STEPS IN DOWNSTREAM PROCESSING

24.  Bio-processing has become an essential tool in the production
of a wide range of products, from life-saving medicines to
sustainable fuels.
 Bio-processing offers a more efficient and environment
friendly alternative to traditional manufacturing methods.
 As technology continues to advance, the potential fro bio-
processing to transform industries and improve our world is
truly limitless.
CONCLUSION