Microorganism and method for the fermentative production of an organic-chemical compound
The invention relates to a microorganism which produces and/or secretes an organic-chemical compound, wherein the microorganism has increased expression, compared to the particular starting strain, of one or more protein subunits of the ABC transporter having the activity of a trehalose importer, said microorganism being capable of taking up trehalose from the medium; and to a method for the production of an organic-chemical compound, using the microorganism according to the invention, wherein accumulation of trehalose in the fermentation broth is reduced or avoided.
1. An isolated Corynebacterium glutamicum bacterium which produces and/or secretes an L-amino acid during fermentation, wherein:
a) compared to other bacteria of the same species and strain, said isolated bacterium comprises increased expression of polypeptides comprising the amino acid sequence of SEQ ID NO:8 and SEQ ID NO:10, wherein increased expression of said polypeptides is due to the presence of a larger number of DNA sequences encoding said polypeptides and/or due to DNA sequences encoding said polypeptides being functionally linked to a promoter which is stronger than a promoter of a gene encoding said polypeptides in wild type bacteria of the same species and strain;
b) compared to other bacteria of the same species and strain, said isolated bacterium comprises increased trehalose importer activity, said activity being sufficient to reduce the amount of trehalose accumulated in fermentation broth by 70% or more when said isolated bacterium is cultured in a fermentation broth.
2. The isolated Corynebacterium glutamicum bacterium of claim 1 , wherein compared to other bacteria of the same species and strain, said isolated bacterium comprises increased trehalose importer activity, said activity being sufficient to reduce the amount of trehalose accumulated in fermentation broth by 80% or more when said isolated bacterium is cultured in a fermentation broth.
3. The isolated Corynebacterium glutamicum bacterium of claim 1 , wherein compared to other bacteria of the same species and strain, said isolated bacterium comprises increased trehalose importer activity, said activity being sufficient to reduce the amount of trehalose accumulated in fermentation broth by 90% or more when said isolated bacterium is cultured in a fermentation broth.
4. The isolated Corynebacterium glutamicum bacterium of claim 1 , wherein compared to other bacteria of the same species and strain, said isolated bacterium comprises increased trehalose importer activity, said activity being sufficient to reduce the amount of trehalose accumulated in fermentation broth by 95% or more when said isolated bacterium is cultured in a fermentation broth.
5. The isolated Corynebacterium glutamicum bacterium of claim 1 , wherein said L-amino acid is L-lysine.
6. The isolated Corynebacterium glutamicum bacterium of claim 1 , wherein compared to other bacteria of the same species and strain, said isolated bacterium comprises increased trehalose importer activity, said activity being sufficient to reduce the amount of trehalose accumulated in fermentation broth by 90% or more when said isolated bacterium is cultured in a fermentation broth.
7. The isolated Corynebacterium glutamicum bacterium of claim 1 , wherein, compared to other bacteria of the same species and strain, said isolated bacterium comprises increased expression of an additional polypeptide, wherein said additional polypeptide comprises the amino acid sequence of SEQ ID NO:4, and wherein increased expression of said additional polypeptide is due to the presence of a larger number of DNA sequences encoding said additional polypeptide and/or due to DNA sequences encoding said additional polypeptide being functionally linked to a promoter which is stronger than a promoter of a gene encoding said additional polypeptide in wild type bacteria of the same species and strain.
8. The isolated Corynebacterium glutamicum bacterium of claim 7 , wherein compared to other bacteria of the same species and strain, said isolated bacterium comprises increased trehalose importer activity, said activity being sufficient to reduce the amount of trehalose accumulated in fermentation broth by 80% or more when said isolated bacterium is cultured in a fermentation broth.
9. The isolated Corynebacterium glutamicum bacterium of claim 7 , wherein compared to other bacteria of the same species and strain, said isolated bacterium comprises increased trehalose importer activity, said activity being sufficient to reduce the amount of trehalose accumulated in fermentation broth by 90% or more when said isolated bacterium is cultured in a fermentation broth.
10. The isolated Corynebacterium glutamicum bacterium of claim 7 , wherein said L-amino acid is L-lysine.
11. The isolated Corynebacterium glutamicum bacterium of claim 1 , wherein, compared to other bacteria of the same species and strain, said isolated bacterium comprises increased expression of an additional polypeptide, wherein said additional polypeptide comprises the amino acid sequence of SEQ ID NO:6, and wherein increased expression of said additional polypeptide is due to the presence of a larger number of DNA sequences encoding said additional polypeptide and/or due to DNA sequences encoding said additional polypeptide being functionally linked to a promoter which is stronger than a promoter of a gene encoding said additional polypeptide in wild type bacteria of the same species and strain.
12. The isolated Corynebacterium glutamicum bacterium of claim 11 , wherein compared to other bacteria of the same species and strain, said isolated bacterium comprises increased trehalose importer activity, said activity being sufficient to reduce the amount of trehalose accumulated in fermentation broth by 80% or more when said isolated bacterium is cultured in a fermentation broth.
13. The isolated Corynebacterium glutamicum bacterium of claim 11 , wherein compared to other bacteria of the same species and strain, said isolated bacterium comprises increased trehalose importer activity, said activity being sufficient to reduce the amount of trehalose accumulated in fermentation broth by 90% or more when said isolated bacterium is cultured in a fermentation broth.
14. The isolated Corynebacterium glutamicum bacterium of claim 11 , wherein said L-amino acid is L-lysine.
15. The isolated Corynebacterium glutamicum bacterium of claim 1 , wherein, compared to other bacteria of the same species and strain, said isolated bacterium comprises increased expression of additional polypeptides, wherein said additional polypeptides comprise the amino acid sequence of SEQ ID NO:4 and the amino acid sequence of SEQ ID NO:6, and wherein increased expression of said additional polypeptides is due to the presence of a larger number of DNA sequences encoding said additional polypeptides and/or due to DNA sequences encoding said additional polypeptides being functionally linked to a promoter which is stronger than a promoter of a gene encoding said additional polypeptide in wild type bacteria of the same species and strain.
16. The isolated Corynebacterium glutamicum bacterium of claim 15 , wherein, compared to other bacteria of the same species and strain, said isolated bacterium comprises increased trehalose importer activity, said activity being sufficient to reduce the amount of trehalose accumulated in fermentation broth by 80% or more when said isolated bacterium is cultured in a fermentation broth.
17. The isolated Corynebacterium glutamicum bacterium of claim 15 , wherein, compared to other bacteria of the same species and strain, said isolated bacterium comprises increased trehalose importer activity, said activity being sufficient to reduce the amount of trehalose accumulated in fermentation broth by 90% or more when said isolated bacterium is cultured in a fermentation broth.
18. The isolated Corynebacterium glutamicum bacterium of claim 15 , wherein said L-amino acid is L-lysine.
19. A method for the fermentative production of an L-amino acid, comprising the steps of:
a) culturing the isolated bacterium of claim 1 in a medium to produce a fermentation broth, wherein said isolated bacterium produces and/or secretes said L-amino acid;
b) accumulating said L-amino acid in the fermentation broth of a); and
c) recovering said L-amino acid.
20. The method of claim 19 , wherein said L-amino acid is L-lysine.