Amino Acids Production

1. Technical aspects of
production of Amino
acids
Munna Iype Joy- 315009
M-Tech( FSQM) - NIFTEM

2. Amino acids
Amino acids are used as flavor enhancers
(monosodium glutamic, serine, aspartic acid)
and as nutritional enhancers
Glutamic acid, lysine and methionine account
for the majority, by weight, of amino acids sold.
MSG is the most predominant form of amino
acid used in the food sector.
All twenty amino acids are sold, albeit each in
greatly different quantities

4. Strategies used for aa’
over - expression
•Deletion of genes coding enzymes for
aa’ degradation and over expression of
aa’ gene.
(Glutamate by C. Glutamicum)
•Inactivation of amino acid degradation
pathways that are interfering with high
product yield
( threonine production by E. Coli)

5. Transport reactions can be
used for optimization in
three ways
1.Highly active nutrient uptake is a
prerequisite
for efficient synthesis of amino
acids.
2.Have to be actively secreted in order
to be accumulated in the surrounding
medium
3.The above process may be
counteracted by the activity of amino
acid uptake systems

6. •amino acid excretion seems to be a
common event in the prokaryotic world
•amino acid uptake systems may be of
significance for amino acid production
in two different ways
•Lipid compostiton & modification of
mycolic acids in cell wall in C.
glutamicum leads to increase in aa’
excretion

8. Technical aspects
aa’
productio
n
Fermantati
on
Extracti
on
Chemic
al
Synthes
is
Enzyma
tic
Synthes
is

9. 1. Fermentation
1.Batch
2.Fedbatch
3.Continuous

10. Shaking
flasks
First seed
tank(1-2k L)
Second seed
tank(10-20k
L)
Production
tank(50-
500k L)
1. Fermentation

11. Filtration (Rotary vacuum filter)
Crystallization Ion Exchange
1. Fermentation
Downstream Processing
Solubility is minimum at
isoelectric point
Lowering temperature
Precipitation with salts
( ammonium) , metals (zinc)
Followed by treatment with
treatment with alkali or acid to
get free/ acid from of aa’
Using ion exchange resins aa’
adsorbed , pH is important
Cation exchange resins &
anion exchange resins
pH less than IEP – Cationic
pH greater than IEP – Anionic
Aa’ is eluted by treating with
counterion
Used when other methods fails

12. Glutamate
• Non essential amino acid
•Most important amino acid in food
biotechnology
•First amino acid to be produced by
fermentation
•More than 1,000,000 tons of glutamate
are produced annually
• For production of L- glutamate C.
Glutamicum is used
•Fed – batch type is used

13. Process
• Carbon source - glucose, potassium phosphate,
magnesium sulfate, yeast extract
• Nitroen source - urea
• Carbon source - cane or beet molasses and starch
hydrolyzates from corn or cassava
• Nitroen source - Ammonia and ammonium sulfate
For seed cultures
For large cultures
• Oleic acid (0.65 ml/l) might be added at the beginning of the fermentation to
encourage glutamate excretion
• After 14 hours of growth, temperature increased to 38 from 32ºC
• Broth is transferred into recovery tanks.
• Yield 100g/l broth.

14. l-lysine production
process with
C. glutamicum
•Starch hydrolsate, sucrose – C
Source
•Ammonium sulfate – nitrogen
source
•170g/l lysine accumulation

15. • Pure enzymes are used.
2. Enzymatic Synthesis

16. (S)- aspartic acid is produced by adding ammonia to fumaric acid

17. 3. Chemical Synthesis
1. Can be used for large scale synthesis
2. Typical disadvantage is; gives racemic mixture of
enantiomeric forms
3. Chemical synthesis is predominant in methionine, alanine
and glycine.
4. Process involves resolution into D and L forms and then
recycling the unwanted enantiomer.
5. For this purpose we use an enzyme called aminoacylase
produced by Aspergillus niger.
6. This step makes the process expensive and hence
chemical synthesis is limitted to few amino acids

18. β-Cyanopropionaldehyde then converted
to glutamic acid by the Strecker process in
which the aldehyde is converted to the
amino analog of a cyanohydrin, which is
then hydrolyzed to glutamic acid.
Chemical Synthesis Of Glutamic
acid

19. Chemical Synthesis Of Methionine
• The addition of methyl mercaptan to
acrolein takes place by a
nucleophilic mechanism. Attack of
the conjugate base of methyl
mercaptan (pK a = 10.7) gives a
resonance-stabilized anion, which
then accepts a proton on carbon to
give the addition product, β-
methylthiopropionaldehyde.
• β-Methylthiopropionaldehyde is
then converted to methionine by the
Bucherer method, a modification of
the Strecker method in which
ammonium carbonate takes the
place of ammonia.

20. 4. Extraction From Natural
Sources
• Standard procedure is hydrolysis with aqueous
acid, followed by capture of the amino acids by
passage of the hydrolysate over a strongly
acidic ion exchange resin.
• After the resin is washed with water, elution with
aqueous ammonia frees the amino acids, which
are collected in fractions.
• Extraction is the most economical process for
the production of both (S )-tyrosine and (R,R)-
cystine. Reduction of (R,R)-cystine gives (R )-
cysteine.

21. Isolation of Glutamic Acid from
Protein Hydrolysates
 Predominant method before fermentation.
 Gluten is the main raw material ( contains
upto 95% glutamic acid by weight).
Gluten
Aq. HCl
Hydrolysis
Hydrolysate
Acidification
by conc. HCl
Cooling &
glutamic
acid hydrochloride
crystals
Filtration
(insoluble
humic
materials
removed)
Filtrate
Conc. Under
high pressure
pH adjustment to
3.2
Glutamic acid crystals

22. Thank You…