Here is brief ppt on industrial production of amino acids - glutamine, lysine, tryptophan. Please share your feedback and queries. Constructive criticism is appreciated. Thank you

1. INDUSTRIAL
PRODUCTION OF
IMPORTANT AMINO
ACIDS
SUBMITTED TO
DR. PAWAN KUMAR
AGARWAL
SUBMITTED BY
PRANSHU CHANDRA
BHUSHAN SINGH NEGI

2. INTRODUCTION
 L-Amino acids find various applications in biotechnology. L-Glutamic acid and its salts
are used as flavour enhancers. Other L-amino acids are used as food or feed additives.
 The amino acid business is a multi-billion dollar enterprise. All twenty amino acids are
sold, albeit each in greatly different quantities.
 The major producers of amino acids are based in Japan, the US, South Korea, China
and Europe.
 Many microbe-based industries have their origins in traditions that go back hundreds or
thousands of years.

3. APPROACHES FOR PRODUCTION
Direct
Chemical
Synthesis
Fermentation
Bioconversion
Using
Enzymes

4. CHOOSING RIGHT METHOD
Available technology
Costs of raw material
Market prices and sizes
Cost of running fermentation versus synthesis reactions
Environmental impact of the process

5. STATISTICS

6. GLUTAMIC ACID
PROPERTY DESCRIPTION
Chemical Name L-glutamic acid
Molecular Formula C5H9NO4
Molecular Weight 147.13 g/mol
Colour Colourless to White
Odour Odourless
Solubility Sparingly soluble in water and insoluble in ethanol or
ether
pH Between 3.0 and 3.5

7. HISTORICAL PERSPECTIVE
 First glutamate production dates back to 1908 when Dr. K. Ikeda, a chemist in Japan,
isolated glutamic acid from kelp and discovered that glutamic acid, after neutralization with
caustic soda, developed an entirely new taste.
 In 1908, Kikunae Ikeda of Tokyo Imperial University isolated the flavour enhancing
principle from the seaweed konbu as crystals of monosodium glutamate (MSG).
 The breakthrough in the production of MSG was the isolation of a bacterium,
Corynebacterium glutamicum. The successful commercialization of monosodium
glutamate (MSG) with this bacterium provided a big boost for amino acid production.

8. PRODUCTION OF GLUTAMATE
 Glutamic acid commercial production by microbial fermentation provides 90%
of world’s total demand, and remaining 10% is met through chemical methods.
 For the actual fermentation the microbial strains are grown in fermenter. The
raw materials used include carbohydrate, peptone, inorganic salts and biotin.
 Biotin concentration in the fermentation medium has a significant influence on
the yield of glutamic acid. Fermentation completes within 2-4 days and, at the
end of the fermentation, the broth contains glutamic acid in the form of its

9.  Glutamate is the most abundant free amino acid in bacterial cytoplasm. In
order to be useful, glutamate producers must do two things well:
o They must overproduce glutamate in excess of their normal metabolic
needs.
o They must excrete it into culture broth
 The most popular Coryneform species include C.glutamicum,
Brevibacterium flavum, Brevibacterium lactofermentum.
 Other glutamic acid-producing organisms include Escherichia coli, Bacillus
megaterium, Bacillus circulans, Bacillus cereus, and Sarcina lutea.
PRODUCTION OF GLUTAMATE
[CONTD.]

10. PRODUCTION OF GLUTAMATE
[CONTD.]

11. FLOW CHART

12. COMPONENT COMPOSITION
Glucose 8%
NH4Cl 0.5%
Corn steep liquor 0.3%
K2HPO4 0.5%
MgSO4·7H2O 0.03%
CaCO3 1%
Deionized water Make up volume
MEDIA COMPOSITION & CONDITIONS
The pH of the medium is normally kept at 7 ± 0.2 and the fermentation is usually initiated
with continuous agitation and aeration for 48 h at 30°C

13. SEPARATION AND PURIFICATION
 Centrifugation is a common method used in industry, can be operate semi-continuous or
continuous basis. Poor centrifugation can be improved by adding flocculation agent. This
agent will neutralize the anionic charges on the surface of microbial cells.
 Filtration is widely used in industry and can be improved by using filter aids. This
separates unnecessary cell debris and impurities.
 Ion exchange is used for the extraction and purification of amino acids form the
fermentation broth on the basis of charge of the solution.
 Electro-dialysis is based on the principle that charged particles move towards the
electrodes in the electric field, a mixture of the required amino acid and contaminant salts
can be separated at isoelectric point. The applications are limited to desalting amino acid
solutions.

14. ION EXCHANGE
CHROMATOGRAP
HY
ELECTRODIALYSI
S

15.  Solvent extraction has only limited applications. The distribution coefficients of amino acids
between organic solvent and water phases are generally small.
 Decolorisation is performed to get rid of the colored impurities in the fermentation broth. It is
based on the fact that amino acids (especially the non-aromatic amino acids) do not adsorb onto
activated charcoal. Although the treatment is very effective, some of the amino acid is lost during
this step.
 After the glutamic acid crystal cake is separated from the acidified fermentation broth it is added
to the NaOH solution and converted into monosodium glutamate (MSG), which has strong umami
taste.
 The MSG is cleaned by using active carbon and the clean MSG solution is concentrated by
heating and the monosodium glutamate crystal is formed.
 The crystal produce are dried with a hot air in a closed system. Then, the crystal is packed in the
SEPARATION AND PURIFICATION
[CONTD.]

16. Nutritional supplement in food production
MSG is used as flavor enhancer
Agriculture/Animal Feed
As an intermediate in manufacturing of various
organic chemicals
APPLICATIONS OF GLUTAMIC ACID

17. LYSINE
PROPERTY DESCRIPTION
Chemical Name L-lysine
Molecular Formula C6H14N2O2
Molecular Weight 146.19 g/mol
Colour Colourless
Odour Odourless
Solubility Highly soluble in water and insoluble in organic solvents
pH Between 3.0 and 3.5

18. PRODUCTION OF LYSINE
 The pathway for the synthesis of L-lysine is complex, and an outline of it is depicted in given
figure. This metabolic pathway is also involved in the formation of 3 other amino acids, namely
methionine, threonine and isoleucine.
 Based on the biosynthetic pathway and the regulatory steps certain improvements have been
made in the strains of C. glutamicum and B. flavum for overproduction of lysine.
 The export or release of L-lysine from the cells into the surrounding medium occurs through a
lysine-export (LysE) carrier protein. It participates in lysine transport. The exporter system is
very efficient active process to export large quantities of intracellular lysine.

19. PRODUCTION OF LYSINE [CONTD.]

20. PRODUCTION OF LYSINE [CONTD.]
PARAMETER DESCRIPTION
Carbon Source Cane Molasses
Nitrogen source Corn steep liquor
Minerals and Salts K2HPO4 , CaCO3
Antifoam PEG-2000
pH 7.2
Temprature 35-370C
Time 100 hours

21. FLOW CHART

22. Nutritional supplement in sports drinks
Lysine is used for preventing and treating cold sores
Agriculture/Animal Feed
As an intermediate in manufacturing of various organic chemicals
Food deodorant in potatoes, rice and flour, canned food etc
APPLICATIONS OF LYSINE

23. TRYPTOPHAN
PROPERTY DESCRIPTION
Chemical Name L-tryptophan
Molecular Formula C11H12N2O2
Molecular Weight 204.229 g/mol
Colour White to slightly yellowish-white
Odour Odourless
Solubility Slightly soluble in acetic acid, ethanol; insoluble in ethyl
ether; Soluble in water
pH Between 3.0 and 3.5

24.  The industrial production of tryptophan is biosynthetic and is based on the
fermentation of serine and indole using either wild-type or genetically modified
bacteria such as B. amyloliquefaciens, B. subtilis, C. glutamicum or E. coli.
 These strains carry either mutations that prevent the reuptake of aromatic
amino acids or multiple/overexpressed trp operons. The conversion is
catalyzed by the enzyme tryptophan synthase.
PRODUCTION OF TRYPTOPHAN

25. PRODUCTION OF TRYPTOPHAN [CONTD.]

26. PRODUCTION OF TRYPTOPHAN
[CONTD.]
PARAMETER DESCRIPTION
Carbon Source Cane Molasses
Nitrogen source Corn steep liquor
Minerals and Salts K2HPO4 , CaCO3
Antifoam Silicon RD
pH 6.8
Temprature 35-370C
Time 72 hours

27. L-Tryptophan is widely used as Leavening agent, Oxidant &
antioxidant in food production
L-Tryptophan is widely used as nutritional supplement in
beverages.
L-Tryptophan is used as sedatives and psychotropic drug
L-Tryptophan is widely used as Amino acid whitening agents in
Cosmetics.
L-Tryptophan is widely used as nutritional supplement in
Agriculture/Animal Feed.
APPLICATIONS OF TRYPTOPHAN

28. THANK YOU