1. AN INTRODUCTION TO FERMENTATION PROCESS

2. What is fermentation Conversion of sugar to alcohol using yeast.
Chemical conversion of carbohydrates into alcohols or acids.
The process is often used to produce wine and beer
Also employed in preservation to create lactic acid in sour foods such as pickled cucumbers, kimchi and yogurt.
The science of fermentation is known as zymology.
With the goal of producing a specific chemical product.

3. The chemistry of fermentation
Aerobic & anaerobic cellular respiration
Glycolysis
Alcoholic fermentation
Lactic acid fermentation

4. Aerobic Cellular Respiration
Aerobic means ‘with air”. This type of respiration needs oxygen for it to occur so it is called aerobic respiration
Glucose + oxygen  Carbon dioxide + Water + Energy
The chemical equation is:
𝑪 𝟔 𝑯 𝟏𝟐 𝑶 𝟔 + 𝟔𝑶 𝟐 → 𝟔𝑪𝑶 𝟐 +𝟔 𝑯 𝟐 𝑶+𝟐𝟗𝟎𝟎 𝐤𝐣
3 stages – glycolysis
– citric acid cycle
– electron transport chain

5. Stages of Aerobic Celllular Respiration
In glycolysis, a net of 2 molecules of ATP, or chemical energy, are produced.
The citric acid cycle produces another 2 molecules of ATP
The electron transport chain produces 28 molecules of ATP
Oxygen is used in aerobic cellular respiration as the final electron acceptor in the electron transport chain, which is part of why it’s able to create so much ATP

7. Anaerobic Cellular Respiration
In anaerobic cellular respiration, the only step of this process that occurs is glycolysis

8. What is fermentation
It is a process by which the living cell is able to obtain energy through the breakdown of glucose and other simple sugar molecules without requiring oxygen
Fermentation results in the production of energy in the form of two ATP molecules, and produces less energy than aerobic process of cellular respiration
Louis Pasteur in the 19th century used the term fermentation in a narrow sense to describe the changes brought about by yeast and other microorganism growing in the absence of air (anaerobically)
He also recognized that ethyl alcohol and carbon dioxide are not the only product of fermentation
Fermentation is also used more broadly to refer to the bulk growth of microorganisms on a growth medium often with the goal of producing a specific chemical product

10. Lactic Acid Fermentation
In lactic acid fermentation, the pyruvic acid from glycolysis is reduced to lactic acid by NADH, which oxidized to NAD+.
This commonly occurs in muscle cells.
Lactic acid fermentation allows glycolysis to continue by ensuring that NADH is returned to its oxidized state (NAD+)

11. Products of Fermentation
The end products of fermentation differ depending on the organism
Lactic acid and lactate, carbon dioxide, and water – produced from many bacteria, fungi, protists and animals cells (notably muscle cells in the body)
Ethyl alcohol, carbon dioxide and water – produced from yeast and most plant cells

12. Products of Fermentation
Fermentation products include:
Food products :
Milk - yogurt, kefir, fresh and ripened cheeses
Fruits - wine, vinegar
Vegetable – pickles, soy sauce
Meat – fermented sausage, salami
Industrial chemical:
Solvent – acetone, butanol, ethanol, enzymes, amino acids
Specialty chemicals – vitamins, pharmaceuticals

13. Products of fermentation

15. The range of fermentation process
There are five major groups of commercially important fermentations:
Those that produce microbial cells (or biomass) as the product.
Those that produce microbial enzymes
Those that produce microbial metabolites.
Those that produce recombinant products.
Those that modify a compound which is added to the fermentation - the transformation

16. The component parts of a fermentation process
The formulation of media to be used in culturing the process organism during the development of the inoculums and in the production fermenter.
The sterilization of the medium, fermenters and ancillary equipment.
The production of an active, pure culture in sufficient quantity to inoculate the production vessel.
The growth of the organism in the production fermenter under optimum conditions for product formation.
The extraction of the product and its purification.
The disposal of effluents produced by the process.

17. Interrelationships between the six component parts

18. Industrial penicillin manufacturing
Case Study
Industrial penicillin manufacturing

19. What is penicillin?
Group of antibiotics produced by the Penicillium fungi
Effective against actively growing Gram positive bacteria
Some penicillin like amoxicillin are also effective against Gram negative bacteria, except Pseudomonas aeruginosa.
It is a group of closely related compounds, not a single compound
Examples: Amoxicillin, ampicillin, phenooxymethlpenicillin
Around 50 drugs that are penicillin

20. Discovery and Production of Penicillin

22. How to produce penicillin
By fermentation
Fermenters
Purpose of fermenter – provide contained, controlled and homogeneous environment in which the fermentation can proceed in a manner that is both safe and practical and which optimizes the particular objective of the fermentation.
Other primary factors include cost, reliability and safety.
For reactor being designed for specific purpose, there are a number of important parameters that will greatly affect performance.

23. Fermenter consideration
Reactor size: optimum rates of production
Reactor configuration: mechanical agitation or will a bubble column
Mode of operation: will it be fed or continuously fed?
Conditions inside the reactor: how will conditions (pH, temperature, …) be controlled?
Economic requirements:
Easy to operate aseptically
Reasonably flexible regarding process requirements
Low power consumption
Stable under fluctuating conditions
Cheap, robust, simple and well understood for scale- up

24. Specific conditions for penicillin production
Most penicillin form filamentous broths. This means they can be difficult to mix due to their high viscosity. Also increasing viscosity of the broth can hinder oxygen transfer
Solution: bubble column (air lift reactors) – would distribute the oxygen equally and also to agitate the medium
Penicillin is an aerobic organism; oxygen supply is critical
Optimum pH for penicillin growth is 6.5: maintain pH efficiently
Strain stability problems (mutations): careful strain maintenance is required
Biomass doubling is about 6h: provisions must be made

25. Media consideration
Provide all the elements required for the synthesis of cell materials and the formation of desired product
Provide favorable environment for the culture in question
Be cost effective

26. Media Formulation

27. Primary and Secondary Metabolismcc

28. Production method Primary or secondary production?
Secondary metabolites are only produced in times of stress when resources are low and the organism must produce these compounds to kill off its competitors to allow t to survive
It is these conditions that we wish to duplicate in order to achieve the maximum amount of product from our fermentation

29. Overall process Seed culture Addition of solvent Medium
Heat sterilization
Fermentation
Biomass removal
Centrifugation extraction
Extraction
Fluid bed drying
Final product (Penicillin G)
Addition of solvent

30. Overall process

44. Quiz 6

45. Question 1
Identify at least 10 types of food, beverages and/or snack that were produced (at any level) with the aid of microorganisms

46. Question 2
Provide an overview of the production of one of the product you answered in Question 1