Process for the fermentative preparation of organic chemical compounds using coryneform bacteria in which the SugR gene is present in attenuated form
AssigneesEvonik Degussa GmbH, Forschungszentrum Julich GmbH
InventorsBrigitte Bathe, Bastian Blombach, Bernhard Eikmanns, Verena Engels, Georg Thierbach, Volker Wendisch
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Abstract
The invention relates to a recombinant coryneform bacterium which secretes an organic chemical compound and in which the sugR gene which codes for a polypeptide having the activity of an SugR regulator has been attenuated. The invention further relates to a processes for using this bacterium for the fermentative preparation of organic chemical compounds.
Claims18
A recombinant coryneform bacterium comprising a SugR regulator, wherein:
- a) said recombinant coryneform bacterium:
- i) secretes an organic chemical compound;
- ii) has the ability to utilize as a carbon source one or more of the compounds selected from the group consisting of:
- glucose, fructose, sucrose, and acetic acid;
- and
- iii) comprises a feedback resistant aspartate kinase;
- and
- b) said SugR regulator comprises all of the sequence of SEQ ID NO:
- 2 except for one or more differences selected from the group consisting of:
- i) the amino acid at position 37 is selected from the group consisting of:
- L-alanine, glycine, L-isoleucine and L-proline;
- ii) the amino acid at position 38 is selected from the group consisting of:
- L-alanine, glycine, L-isoleucine and L-proline;
- iii) the amino acid at position 39 is selected from the group consisting of L-alanine, glycine, L-isoleucine and L-proline;
- iv) the amino acid at position 40 is L-proline;
- v) the amino acid at position 72 is selected from the group consisting of:
- L-alanine, glycine, L-glutamic acid and L-aspartic acid;
- vi) the amino acid at position 101 is selected from the group consisting of:
- L-arginine, L-lysine, L-phenylalanine, L-methionine, L-glutamine, L-tryptophan, L-tyrosine and L-glutamic acid;
- vii) the amino acid at position 105 is selected from the group consisting of L-proline, L-phenylalanine, L-isoleucine, L-methionine, L-glutamine, L-tryptophan and L-tyrosine;
- viii) the amino acid at position 210 is selected from the group consisting of:
- L-alanine, L-arginine, and L-proline;
- and
- ix) the amino acid at position 216 is selected from the group consisting of:
- L-alanine, L-glutamic acid, L-isoleucine and L-tryptophan.
A recombinant coryneform bacterium comprising a gene encoding the SugR regulator of SEQ ID NO:
- 2, wherein:
- a) said recombinant coryneform bacterium:
- i) secretes an organic chemical compound;
- ii) has the ability to utilize as a carbon source one or more of the compounds selected from the group consisting of:
- glucose, fructose, sucrose, and acetic acid;
- and
- iii) comprises a feedback resistant aspartate kinase;
- and
- b) said gene also comprises in its promoter region, the nucleotide sequence of all of SEQ ID NO:
- 13 except for one or more mutations selected from the group consisting of:
- i) replacement of the nucleobase thymine at position 7 of SEQ ID NO:
- 13 by guanine;
- ii) deletion of one or more of the nucleobases from position 2 to 7 of SEQ ID NO:
- 13;
- iii) deletion of one or more of the nucleobases between position 52 and 57 of SEQ ID NO:
- 13;
- and
- iv) replacement of one or more of the nucleobases adenine or guanine between position 52 and 57 of SEQ ID NO:
- 13 by thymine or cytosine.
Dependencies are inferred from the claim text.