UNIVERSITY INSTITUTE OF

BIOTECHNOLOGY

Bachelor of Sciences (Biotechnology)

Subject Name : Fermentation Technology and Bioprocess
Engineering

Code : BTT-202
TOPIC : Bioreactor design/ Microbial
Fermentation DISCOVER . LEARN . EMPOWER
Dr Vipasha Sharma (E4388)
Microbial Fermentation
(Enzymology, Metabolic pathways and Fermentation aspects)

Course Outcome

CO Title Level
Number

CO1 Ability to understand the basic concept of fermentation Understand

and manipulations to enhance production of bioresource
products.
CO2 Profound grasp of various types of substrates that can be Understand
utilized in the fermentation industry; type of fermentation
that can be efficiently utilized for bioconversion of various
low value substrates into value added products.
CO3 Ability to implement the principles of fermentation Understand
technology for the production of numerous products of https://orbitbiotech.com/fermentorbioreactor-types-body-
huge market value. parts-size-impellers-stirrer-glands-baffles/
CO4 Students approaching the end of their course of study will Understand
be able to make informed choices among opportunities for
work or further post graduate education. 2
 Contents
• Introduction.

• Effect of flow rate on substrate concentration.

 Enzymes.

 Classification of enzymes.

 Fermentor systems.

 Microbial cell breakage systems.

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 Introduction
• The fermentation industry is composed of five major bio-ingredient categories.

• They are:
- Proteins & amino acids.

- Organic acids.

- Antibiotics.

- Enzymes.

- Vitamins & hormones. https://www.livemint.com/Sundayapp/

P2MN64QjTrMG9Kn6cFWu2I/Antibiotics-A-biography.html

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 Introduction (cont.)
 Fermentation industry is driven by:

- The cost and availability of feed-stocks.

- The efficiency of industrial microorganism.

- Fermentation condition and optimization.

- Down stream process and end-product

recovery efficiency.

- Fermentation by-product utilization.

- Utility consumption and labor cost.

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 Fermentation media
 Optimum balance of the media is mandatory for cells propagation and for the maximum production of target metabolite
(end-product).

 Media compositions:

- Carbon source.

- Nitrogen source.

- Minerals.

- Growth factors.

http://www.twipu.com/tag/petriplate
- Precursors (mutants).

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 Industrial microorganisms
• Microbial screening. 93 C

- Wild strains

• Microbial yield improvement

43 C

- Mutation

21 C
- Recombinant DNA
4C

- Genetically engineered

• Microbial selection.

• Industrial microorganism

https://www.slideshare.net/shyrosh/classification-of-bacteria-
49373508 7
 Types of fermentation

 Solid State fermentation (SSF).

 Liquid State fermentation (LSF) Surface culture & submerged culture

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 Solid State Fermentation (SSF)
 SSF process can be defined as microbial growth on particles without presence of free water.

 Particles are a solid culture substrate such as rice or wheat bran saturated with water and
inoculated with (mold, yeast, bacteria) in controlled room temperature.

 It is ideal for growing filamentous fungi.

 It has been used in Asia and developing nations.

 It is more cost effective (smaller vessels lower water consumption, reduced waste water
treatment costs, lower energy consumption, and less contamination problems).

https://www.indiamart.com/proddetail/fermentation-
system-2080691197.html

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 SSF process and applications
Applications:

 Potentially many high value products such as extra-cellular enzymes, primary

metabolites, and antibiotics could be produced in SSF.

 It is estimated that nearly a third of industrial enzyme produced in Japan is

made by SSF process.

 Production of organic and ethanol from starchy substrates.

 Digestibility of fibers and lignocelluloses materials for both human and
animal consumption.

https://en.wikipedia.org/wiki/Solid-state_fermentation

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 Liquid State fermentation (LSF)
[Submerged culture]
- Submerged culture is performed in tanks which can reach in size for over 100,000 gallons.

- It is ideal for the growing unicellular organisms such as bacteria and yeast.

LSF methods:
- Batch fermentation.
- Fed-batch fermentation.
- Continuous fermentation.
- Semi-continuous fermentation.

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 Batch fermentation
 Considered to be a closed system.

 The sterilized media in the fermenter is inoculated with the microorganism.

 Incubation is allowed under the optimum conditions (aeration, agitation, temperature).

 During entire fermentation nothing is added except air, antifoam and acid/base.

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 Fed-Batch fermentation
 It is enhancement of batch fermentation.

 Continue adding the nutrients (feeding) in a small doses during the fermentation.

 The method in controlling nutrients feeding process is by measuring methods.

 The main advantage of fed-batch fermentation is the elimination of catabolite repression (feed-back inhibition).

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14
Microbial growth rate
Secondary
metabolites
Primary metabolites

http://textbookofbacteriology.net/growth_3.html 15
 Batch fermenter system

https://www.easybiologyclass.com/industrial-fermentation-process-batch-fed-batch-and-continuous-fermentation/

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 Continuous fermentation
• It is an open system.

• Continuously sterile nutrient is added and the converted nutrient is taken out from the fermentor.

• In continuous process cell loss as a result of outflow must be balanced by growth of the microorganism.

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 Effect of flow rate on substrate
concentration

https://en.wikipedia.org/wiki/Michaelis%E2%80%93Menten_kinetics

The relationship between biomass (X), the concentration of limiting nutrients (C) ,and the dilution rate (D) are important
factors in continuous
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Continuous fermenter system

https://www.easybiologyclass.com/industrial-fermentation-process-batch-fed-batch-and-continuous-fermentation/

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 Important factors for continuous
fermentation
• The system must be stable for at least 500 hours.

• Maintaining sterile conditions for all period of fermentation time.

• The composition of nutrients must be constant all the time.

• Maintaining the strain stability for constant high production yield (concerning about reverse mutation).

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 Semi-continuous fermentation
• Semi-continuous fermentations, in which a fraction of a fermentation is replaced with fresh media at
regular intervals.

http://www.biologydiscussion.com/microorganisms/growth-of-microorganisms-with-diagram/10138
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 Classification of fermentation
 Classification is according to product formation:
- TYPE I:
A Ex

Substrate P product
E1 E2 E3 E4

Substrate A B C D P product

E1 E2 E3 E4
- Type II:

Es1
Substrate A B C D P-Primary Metabolites
Es2 Es3 Es4

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E F G P-Secondary Metabolites
 Assessment Pattern
Knowledge of the topic will be tested through

• Surprise test

• Multiple choice questions

• General class discussion

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 REFERENCES
• Prescott & Duinn's industrial Microbiology by B. Reed million Publishers Ltd., Connecticut.
• Principles of Fermentation Technology by P.F. Stanbury and A. Whitakergamon Press, NY.
• Principles of Fermentation Technology, Third Edition, PF Stanbery, A. Whitaker and Steve Hall, Elesvier (2016).
• Bioprocess Engineering Basic Concepts, by M Shuler and F. Kargi, Prentice Hall Press (2015)
• Fermentation and Biochemical Engineering Handbook, 2nd Edition., H C. Vogel and C M. Todaro, Elesvier (2014)

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 THANK YOU

For queries
Email: [email protected]