# Presentation: 417364

## Amino Acids Production 

**Amino Acids Production**** **

## Rexim is 100%  Degussa since 20 years

**Rexim is 100%  Degussa since 20 years**

**Headquarter :**	Courbevoie , France,  (Paris)
**Production Plants for Pharmaceutical Grade L-amino acids** :
- 		**Ham** ,  France
- 		**Konstanz** , Germany
- 		**Nanning** , China (2001)
**Production Plants for Feed Grade Aminoacids**
- 		**Blair ,Mobile** , USA (L-lys ,DL-met)
- 		**Antwerp** , Belgium (DL-met )
- 		**Wesseling** , Germany ( DL-met )
- 		**Slovakia** ( L-lys ,L-thr )
**   **

## Processes

**Processes**

**Production Plants for Pharmaceutical Grade L-Amino Acids** :
- 		Ham ,  France : Extraction from Proteins hydrolyzates
- 			           Purification from lower grades ( feed,...)
- 		Konstanz , Germany :      Enzymatic resolution
- 		Nanning , China  : Biocatalysis and fermentation
**Production Plants for Feed Grade Amino acids**
**	****	**Synthesis
- 		Fermentation
**   **

## L-Amino Acids Extraction from Proteins Hydrolysates1/8

**L-Amino Acids Extraction from Proteins Hydrolysates****1/8**

**Serine ,  Threonine ,**
**Proline ,  Hydroxyproline,**
**Alanine,  Valine,**
**Isoleucine ,  Leucine**
**Phenylalanine,**
**Histidine,  Arginine**
- ( asp, glu, gly, lys, tyr ,met)
**   **

## Extraction from Proteins Hydrolysates2/8 :Process

**Extraction from Proteins Hydrolysates****2/8 :Process**

- Selection Purification of natural Proteins
- Acidic Hydrolysis
- Filtration
- Ion exchange 	chromatographic steps
- Charcoal treatment
- (Ultrafiltration)
- Crystallization 1
- Charcoal treatment
- Ultrafiltration
- Crystallization 2 ,Drying
**   **

## Extraction from Proteins Hydrolysates3/8

**Extraction from Proteins Hydrolysates****3/8**

**Selection Purification of natural Proteins and**
**pure Chemicals:**
-    Keratins from feathers ( chicken ,ducks ,turkeys )
- 	Gelatin from pigs
- 	Bovine free materials
- 	Chemicals like NH3, HCl, NaOH, H2SO4,alcohols
- 	Regular Suppliers Audits
**   **

## Extraction from Proteins Hydrolysates4/8

**Extraction from Proteins Hydrolysates****4/8**

**Acidic Hydrolysis of Proteins:**
- Conditions choosen for complete disappearance of proteinic
- materials : only short peptides (2-4 units ) remain.
- Use of a large excess of concentrated HCl.
- Keratin is heated 6 hrs at 105°C
- Gelatin is heated 4 hrs at 120°C
**   **

## Extraction from Proteins Hydrolysates5/8

**Extraction from Proteins Hydrolysates****5/8**

**Ion exchange chromatography**** :**
- Main step achieving separation of single amino acids.
- Separation based on ionic properties but also on differences
- on sizes and hydrophobicities (  peptides separated aswell)

## Extraction from Proteins Hydrolysates6/8

**Extraction from Proteins Hydrolysates****6/8**

**Activated mineral ****Charcoal**** treatment :**
- Purification based on surface adsorption :
- Charcoal fixes coloured entities and macromolecules ( endotoxines... )

## Extraction from Proteins Hydrolysates7/8

**Extraction from Proteins Hydrolysates****7/8**

**Ultrafiltration ****:**
- Main Purpose is  biological purification : removal of endotoxines
- for making infusion grade amino acids .
- Done on  membranes with cut off of 5000 to 10 000 daltons;
- Efficiency measured by limulus test.
**Crystallization ****:**
- A recognized method for purification as demonstrated by HPLC

## Extraction from Proteins Hydrolysates8/8

**Extraction from Proteins Hydrolysates****8/8**

**Biological Safety**
**TSE** Expert reports from Pasteur Institute since 1992
**Viral Safety** Validation report (Pasteur Institute /1998 ) using  :
- 		Encephalomyocarditis virus
- 		Canine Parvovirus
- classified as « resistant » and « highly resistant » by the CPMP
- (Committee for Proprietary Medicinal Products )

## Purification from lower Grade ( feed )

**Purification from lower Grade ( feed )**

**Typical examples : asp ,glu , gly ,lys**
**Process uses activated charcoal,ultrafiltration and **
**Recrystallization**
**Origins:**
**	****	****asp ****: bioconversion of fumaric acid**
**	****	****glu , lys**** : fermentation **
**	****	****gly ****: synthesis**

## 	Enzymatic resolution at Konstanz		L- Methionine1.  DL- Methionine ->  N-acetyl-DL-Methionine2. acylase -> L-met + N-acetyl-D-Met3. ion exchange separation-> L- Methionine-> N-acetyl-D-Met   4. -> N-acetyl-DL-Methionine

**	****Enzymatic resolution at Konstanz****	****	****L- Methionine****1.****  ****DL- Methionine ->  N-acetyl-DL-Methionine****2.**** **acylase** ****-> L-met + N-acetyl-D-Met****3.**** **ion exchange separation**-> L- Methionine****-> ****N-acetyl-D-Met   4. -> N-acetyl-DL-Methionine**

## Biocatalysis in China 

**Biocatalysis in China **

- Fumaric acid + NH**3** + fumarase  **L-aspartic acid**
- Bioreaction, filtration, precipitation , filtration, drying
- L-aspartic + aspartate decarboxylase       **L-alanine**
- Bioreaction, filtration, precipitation , filtration, drying

## Fermentation in China 

**Fermentation in China **

- Sugar + Cornsteep + NH**3**  **L- Valine**
- Biomass removal, ion exchange ,crystallisation ,filtration, drying
- Sugar + Cornsteep + NH**3**  **L- Isoleucine**
- Biomass removal, ion exchange ,crystallisation ,filtration, drying