METHOD FOR THE FERMENTATIVE PRODUCTION OF CADAVERINE

• US 20110039313A1
• Filed: 01/10/2008
• Published: 02/17/2011
• Est. Priority Date: 02/01/2007
• Status: Abandoned Application

• Current Assignee: Evonik Degussa GmbH (Rag-Stiftung)
• Original Assignee: Evonik Degussa GmbH (Rag-Stiftung)
• Inventors: Karau, Andreas, Verseck, Stefan, Eggeling, Lothar, Sahm, Hermann, Haeger, Harald
• Application Number:
  US12/517,923
• Publication Number:
  US 20110039313A1
• Time in Patent Office: Days
• US Class Current: 435/128
• CPC Class Codes: C12N 9/88 Lyases (4.)
• CPC Class Codes: C12P 13/001 Amines; Imines

Abstract

The invention relates to recombinant microorganisms in which polynucleotides which code for lysine decarboxylase are enhanced and, using which, cadaverine (1,5-diaminopentane) is produced fermentatively, with the carbon source used preferably being renewable raw materials such as, for example, glucose, sucrose, molasses and the like.

Claims

1. A cadaverine-producing recombinant microorganism with a high lysine titre, in which polynucleotides which code for a lysine decarboxylase are present in an enhanced dose in comparison to microorganisms which are not modified with regard to this enzyme.

2. The microorganism according to claim 1, in which polynucleotides which code for a protein referred to as lysine/cadaverine antiporter are present in an enhanced dose in comparison to microorganisms which are not modified with regard to this protein.

3. The microorganism according to claim 1, in which the polynucleotides which code for lysine decarboxylase are derived from microorganisms selected from the group consisting of:

Escherichia coli, Bacillus halodurans, Bacillus cereus, Bacillus subtilis, Bacillus thuringensis, Burkholderia ambifaria, Burkholderia vietnamensia, Burkholderia cenocepatia, Chromobacterium violaceum, Selenomonas ruminantium, Vibrio cholerae, Vibrio parahaemolyticus, Streptomyces coelicolor, Streptomyces pilosus, Eikenalla corrodens, Eubacterium acidaminophilum, Francisella tulariensis, Geobacillus kaustophilus, Salmonella typhi, Salmonella typhimurium, Hafnia alvei, Neisseria meningitidis, Thermoplasma acidophilum, Plasmodium falciparum, Kineococcus radiotolerans, Oceanobacillus iheyensis, Pyrococcus abyssi, Porochlorococcus marinus, Proteus vulgaris, Rhodoferax ferrireducens, Saccharophagus degradans, Streptococcus pneumoniae, and Synechococcus sp.

4. The microorganism according to claim 1, in which the polynucleotides which code for the protein referred to as lysine/cadaverine antiporter are derived from microorganisms selected from the group consisting of:

Escherichia coli, Thermoplasma acidophilum and Vibrio cholerae.

5. The microorganism according to claim 1, which takes the form of a recombinant strain of the genera Escherichia or Bacillus or coryneform bacteria.

6. The microorganism according to claim 1, in which the polynucleotides which code for the lysine decarboxylase and the protein referred to as lysine/cadaverine antiporter are derived from microorganisms of the species Escherichia coli.

7. The microorganism according to claim 1, in which overexpression takes place as the result of increasing the copy number of the abovementioned polynucleotides by at least one in comparison with the untransformed microorganism or by combination of the abovementioned polynucleotides with a stronger promoter in comparison with the original strain.

8. The microorganism according to claim 1, which takes the form of a coryneform microorganism and in which one or more of the genes from Corynebacterium selected from the group consisting of:

a) the dapA gene, which codes for dihydrodipicolinate synthase,

b) the gap gene, which codes for glyceraldehyde-3 phosphate dehydrogenase,

c) the zwf gene, which codes for glucose-6 phosphate dehydrogenase, and its alleles,

d) the pyc gene, which codes for pyruvate carboxylase, and its alleles,

e) the mqo gene, which codes for malate-quinone oxidoreductase,

f) the lysC gene, which codes for a feedback-resistant aspartate kinase, and its alleles, and

g) the zwa1 gene, which codes for the Zwa1 protein,

are simultaneously enhanced or overexpressed.

9. The microorganism according to claim 1, in which the lysE gene, which codes for a L lysine export protein, is diminished or switched off.

10. The microorganism according to claim 1, which takes the form of a microorganism of the genus Escherichia, in which one or more of the genes from E. coli selected from the group consisting of

a) the gene which codes for a feedback-resistant aspartate kinase, or alleles,

b) the gene which codes for dihydrodipicolinate synthase,

c) the gene which codes for dihydrodipicolinate reductase,

d) the gene which codes for succinyldiaminopimelate transaminase, and

e) the gene which codes for succinyldiaminopimelate deacylase

are simultaneously enhanced or overexpressed.

11. The microorganism according to claim 1, which is capable of producing L lysine before having been transformed with a lysine decarboxylase gene.

14. A method of producing cadaverine, in which

a) a microorganism according to claim 1 is fermented in a medium under conditions under which cadaverine is formed, and

b) the cadaverine is accumulated in the cells of the microorganism or in the fermentation medium.

15. The method according to claim 11, in which

a) the cadaverine is isolated, and, optionally,

b) further dissolved components of the fermentation liquor and/or the biomass in their entirety or in amounts of ≧

0 to 100% remain in the product which has been isolated.

16. The method according to claim 14, in which coryneform bacteria are employed.

17. The method according to claim 14, in which microorganisms of the genus Escherichia are employed.

12. A vector or plasmid containing a polynucleotide which codes for a lysine decarboxylase and/or a polynucleotide which codes for a protein referred to as lysine/cadaverine antiporter.

13. The vector or plasmid according to claim 12 in which the polynucleotides are derived from Escherichia coli.