This document discusses the production of the essential amino acid L-lysine through fermentation. It begins by defining amino acids and classifying them as essential and non-essential. It then introduces L-lysine, describing its history of commercial production starting in 1956. The document outlines the two pathways for lysine biosynthesis and describes the industrial production process using Corynebacterium glutamicum. This involves upstream processes like culture optimization and media formulation and downstream processes such as separation, extraction and purification to produce purified L-lysine. Finally, applications of L-lysine in food, beverages, pharmaceuticals and other industries are listed.

1. SEMESTER – IV
FERMENTATION & INDUSTRIAL MICROBIOLOGY
SEMINAR TOPIC : L-LYSINE AMINO ACID
SUBMITTED BY,
G. PONSUTHA
II M.SC MICROBIOLOGY
REG NO : 20201232516113
SUBBMITED TO,
Dr. S. VISWANATHAN
HEAD DEPARTMENT OF
MICROBIOLOGY

2. What is amino acid?
 Amino acid are building blocks of proteins.
 They are the group of organic components containing two functional groups
amino acid and carboxyl.
 The amino group(-NH2 ) is basic while the carboxyl group (-COOH) is acidic in
nature.
 And a side chain (R group) specific to each amino acid is also present.

3. classification
 All the 20 aminoacid are not needed to be taken in the diet.
 Based on the nutritional requirements amino acid can be classified into two
classes:
 Essential amino acid
 Non essential amino acid
Essential amino acid:
 The amino acid which cannot be synthesized by the body and need to be supplied
through the diet are called essential amino acid.
 (Eg) 10 amino acids are essential amino acid
 Arginine, Valine, Histidine, Isoleucine, Leucine, Lysine, Methionine, Phenylalanine,
Threonine, Tryptophan.

4.  Non essential amino acid :
 There are synthesized be the body, and do not need not be supplied in the diet.
 There are – Glycine, Alanine, Serine, Cysteine, Aspartate, Asparagine, Glutamate,
Glutamine, Tyrosine, and Proline.
 Some microorganisms are capable of production certain amino acid such as
glutamic acid and tryptophan.

5. Introduction of lysine
 L-lysine is an essential amino acid for the nutrition of human and other animals. It is used for
supplementing cereal proteins lacking this amino acid. Molecular formula(C6H14N2O2)
 Thus, protein quality of certain foods (e.g. wheat based food) is improved growth and tissue synthesis. This
amino acid has also bee used medically as a nutrient.
 Kinoshita et al(1958) first reported the fermentative production of L-lysine using homoserine auxotroph's
of Corynebacterium glutamicum.
 Food source of lysine are high protein foods such as eggs, meat, etc.
 Solubility: sparingly soluble in water.
 Covering more than 90% of total world amino acid production.
 80% fermentation
 20% chemical synthesis

6. HISTORY
1889
• Lysine first isolate from casein
1956
• Lysine was commercially introduced as feed
1978
• The first production of fermented l-lysine was produced
by a Japanese company kyowa Hakko Kogyo.

7. PRODUCTIONOF LYSINE
Lysine biosynthesis
 Production of lysine naturally takes place by two biosynthesis pathway
1. The DAP (Diaminopimelic Acid)pathway found in Bacteria (e.g.
Corynebacterium glutamicum), Algae, Higher plant, Oomycota
2. The AAA (alpha-amino-adipate) Pathway found in Fungi and
Euglenoids.

8. DAP pathway

9. AAA PATHWAY

10. INDUSTRIALLYSINE PRODUCTION

11. I
Inbound
logistics
Raw materials
Microbial
culture
Media
U
Upstrea
m
processe
s
Culture
optimization
Media
formulation
F
FERMEN
TAION
Media ultrazation
and lysine
production
D
Down stream
process
separation
Extraction
Purification
o
Out
bound
logistics
Packaging of l-
lysine

12. Raw materials
Fermentation media
 Carbon source cane molasses
 Nitrogen source corn steep liquor / soybean meal
 Minerals and salt KH2PO4 / K2HPO4 / CaCO3
 Trace elements Corn steep liquor
 Anti foaming agent PEG-2000, silicone base oil.

13. Microbial culture
 Corynebacterium glutamicum
 Gram positive, rod shaped bacterium
 They live in chain or in clusters , usually present in soil.
 Non motile
 Non sporing
 Non pathogenic bacterium

14. Inoculum development
Pre culture slant
100
ml
500
ml
1000
ml
L.F
5L
SEED
FERMENTER
50L
PILOT
FERMENTER
100L
TO PRODUCTION FERMENTOR

15. Media formulation

16. FERMENTAION
FERMENTAION
PROCESS
• Submerged
fermentation
• Aerobic
fermentation
Mode of
operation
• Batch process
• Fed-batch
process
Fermenter type
• Stirred tank
fermenter
• Airlift
fermenter
Process
parameter
• Optimum pH
7.2
• Temperature –
35 -370C
• Time 100 hrs.
production
cycle

17. Down stream processing
Separation
• Cell removal
• L-lysine
recovery
Extraction
• Further removal
of impurities
• concentration
Purification
• Purified l-lysine
ready
•.

18. SEPERATION OF L-LYSINE

19. EXTRACTION OF L-LYSINE
L-LYSINE CONCENTRATED
(ACIDIFIED L-LYSINE)
L-LYSINE
MOTHER
LIQUOR
HCL
MOTHER
LIQUOR
(CANE
MOLLASES
LIQUID
L-LYSINE HCL

20. PURIFICATION OF LYSINE
SPRAY DRYER PLANT

22. PRODECT RECORY
 The fermentation broth is sent to an ultra filtration system for the removal of cell
debris and other.
 Subsequently, the liquor form ultra filtration is fed to ion exchange columns,
where l-lysine is selectively absorbed.
 The absorbed l-lysine is eluted from the ion exchange resin by washing with an
aqueous ammonia solution.

23. PRODUCT CONCENTRATION AND DRYING
 The L-Lysine eluted from the ion-exchange column is mixed with mother liquor
from the product filtration step and concentrated by evaporation.
 The concentrated lysine solution is acidified with hydrochloric acid and free is
converted to lysine HCL.

24. CRYSTALLIZER
 Crystallization is also a chemical solid-liquid separation techniques.
 In which mass transfer of solute from the liquid solution to a pure solid
crystallization phase occur.
 Chemical engineering crystallization occurs In a crystallizer.
 TYPES
 Cooling
 Concentration
 Reaction (or) equilibrium displacement

25. APPLICATION AND USES
In food - nutritional supplements, food decorants
In beverages – nutritional sport drink
In pharmaceutical – preventing and treating cold sores causes by simplex labialis virus
In health and personal
care – cosmetics, personal care products, baby products, bath products, eye
In agricultural/animal feed/
Poultry – animal feed and used In fertilizer
In other industries – as intermediate to produce other organic chemicals.

26. References
 INDUSTRIAL MICROBIOLOGY-A.H.PATEL(Page no.160-161)
 https://www.slideshare.net/MdMominulIslam18/industrial-production-of-amino-
acid-llysine
 https://www.slideshare.net/robmondal/lysine-production-123963762

27. BENEFIT OF TAKING SEMINAR
 I can improve self confident.
 Learn about new industrial technologies.
 It can improve teaching skill.

28. Thank you