TH73889A - Microorganisms that produce L-amino acids, and methods for L-amino acid production? - Google Patents

Abstract

DC60 (14/06/56). L-amino acids are produced by culturing microorganisms which have the ability to produce L-amino acids but have been modified so that the expression of the yibE gene is increased. L-amino acids are collected from culture media or from microorganisms. L-amino acids are produced by culturing microorganisms which have the ability to produce L-amino acids but have been modified so that the expression of the yibE gene is increased. L-amino acids are collected from culture media or from microorganisms.

Claims (16)

Revised 27/05/2015 1. Microorganisms capable of producing L-amino acids, selected from a group comprising Enterobacteriaceae, Coryneform bacteria, and methanol-dependent bacteria, exhibiting 90% or greater similarity to the amino acid sequence 17-315 of sequence number 2, and modified so that the expression of the ybjE gene, which encodes a protein with L/amino acid excretion activity, is increased. 2. Microorganisms according to claim 1, where the expression of the ybjE gene is increased by adding... 3. A microorganism under Reputation 1 or 2 in which the ybjE gene is a DNA identified in (c) or (d) as follows: (c) DNA containing the base sequences 49-948 of sequence number 1; (d) DNA in which the base sequences 49-948 of sequence number 1 are hybridized under strict conditions and encoding a protein with L-amino acid excretion activity. 4. One microorganism under Reputation 1 to 3 in which the L-amino acid excretion activity is enhanced by the increased expression of the ybjE gene. 5. One microorganism under Reputation 1 to 4 in which the L-amino acid excretion activity is enhanced by the increased expression of the ybjE gene. L-amino acids or their acid analogs. L-amino acids are enhanced by altered expression of the ybjE gene, resulting in increased levels. 6. One microorganism according to one of the claims 1 through 5, in which the L-amino acid is one or two or more selected L-amino acids from a group consisting of L-lysine, L-arginine, L-ornithine, L-histidine, L-isoleucine, L-threonine, L-proline, L-phenylalanine, L-cysteine, and L-glutamic acid. 7. One microorganism according to one of the claims 1 through 6, in which the microorganism belongs to the genus Escherichia. 8. One microorganism according to one of the claims 1 through 6, in which the microorganism belongs to the genus Methylophilus or Methylobacillus. 9. A method for the production of L-amino acids, specifically selected from one of the following microbial claims (1 to 8), using a culture medium, resulting in biosynthesis and ----------------------------------------------------------------- Revised 17/04/58 1. Microorganisms capable of producing L-amino acids, selected from a group consisting of Enterobacteriaceae, Coryneform bacteria, and methanol-fed bacteria, exhibiting 90% or greater similarity to the amino acid sequence of amino acid numbers 17-31 of sequence number 2, and modified so that the expression of the ybjE gene, which encoding a protein with L-amino acid-deficient activity, is increased. 2. Microorganisms according to claim 1, where the expression of the ybjE gene is increased by adding... 3. A microorganism under Reputation 1 or 2 in which the ybjE gene is a DNA specified in (c) or (d) as follows: (c) DNA containing the base sequences 49-948 of sequence number 1; (d) DNA hybridized with the base sequences 49-948 of sequence number 1 under strict conditions and encoding a protein with L-amino acid excretion activity. 4. One microorganism under Reputation 1 to 3 in which the specific characteristic is enhanced L-amino acid excretion activity by the expression of the ybjE gene modified to be increased. 5. One microorganism under Reputation 1 to 4 in which the specific characteristic is acid tolerance. L-amino acids or their acid analogs. L-amino acids are enhanced by the expression of a modified ybjE gene, which is increased by 6. One microorganism according to one of the claims 1 through 5, in which the L-amino acid is one or two selected L-amino acids from a group consisting of L-lysine, L-arginine, L-ornithine, L-histidine, L-isoleucine, L-threonine, L-proline, L-phenylalanine, L-cysteine, and L-glutamic acid. 7. One microorganism according to one of the claims 1 through 6, in which the microorganism belongs to the genus Escherichia. 8. One microorganism according to one of the claims 1 through 6, in which the microorganism belongs to the genus Methylophilus or Methylobacillus. 9. A method for the production of L-amino acids. 10. A method for producing L-amino acids that are specific for the cultivation of microorganisms according to one of the claims 1 to 8, in a quantity of 1, using a culture medium with methanol as the primary carbon source, resulting in the biosynthesis and accumulation of L-amino acids in the culture medium or in the microorganism, and the collection of such L-amino acids from the culture medium or in the microorganism. From the culture medium or microorganism: 11. Methods according to claims 9 or 10, where the said L-amino acid is one or two or more basic amino acids selected from a group consisting of L-lysine, L-arginine, L-ornithine, L-histidine, L-isoleucine, L-threonine, L-proline, L-phenylalanine, L-cysteine, and L-glutamic acid. 1. Microorganisms capable of producing L-amino acids were modified so that the expression of the ybjE gene was increased. The ybiE gene was selected from a group consisting of: (a) DNA containing the nucleotide sequence of SEQ ID NO: 1; (b) DNA hybridized under strict conditions containing the nucleotide sequence of SEQ ID NO:1, and this DNA encodes an L-amino acid exporting protein; (c) DNA containing the nucleotide sequence of nucleotide numbers 49 to 948 in SEQ ID NO:1; and (d) DNA hybridized under strict conditions containing the nucleotide sequence of nucleotide numbers 49 to 948 in SEQ ID NO:1, and this DNA encodes an L-amino acid exporting protein. 2. Microorganisms under claim 1 where the expression of the ybjE gene is increased by increasing the number of copies of the ybjE gene or by modifying the sequence that controls the expression of the ybjE gene. 3. A microorganism pursuant to claim 1 where the amino acid sequence of the protein encoded by the ybjE gene is selected from a pool consisting of SEQ ID NO: 2, 9, and 10 nucleotide sequences, in which such protein is capable of exporting L-amino acids. 4. A microorganism pursuant to claim 1 in which the ybjE gene is selected from a group consisting of: (a) DNA containing the nucleotide sequence of SEQ ID NO: 1; and (b) DNA recombinable under strict conditions with the nucleotide sequence of SEQ ID NO: 1, or a probe that can be prepared from the nucleotide sequence of SEQ ID NO: 1; and in which the DNA encoding an L-amino acid export protein is selected. 5. A microorganism pursuant to claim 1 in which the ybjE gene is selected from a group consisting of: (a) DNA with a nucleotide sequence of 49 to 948 nucleotides in SEQ ID NO: 1; and (b) DNA recombinable under strict conditions with a nucleotide sequence of 49 to 948 nucleotides in SEQ ID NO: 1, or a probe which can be prepared from a nucleotide sequence of 49 to 948 nucleotides in SEQ ID NO: 1; and in which the DNA encodes an L-amino acid export protein. 6. In any one of the microorganisms described in claims 1 through 5, where the L-amino acid export capacity of such microorganism is increased, the expression of the ybjE gene is increased. 7. Any microorganism matching claims 1 through 6 whose tolerance to amino acids or amino acid-like organisms is increased by increasing the expression of the ybjE gene. 8. Microorganisms according to claims 1 through 7 in which L-amino acids are selected from a group consisting of L-lysine, L-arginine, L-ornithine, L-histidine, L-isoleucine, L-threonine, L-proline, L-phenylalanine, L-cysteine, and L-glutamic acid. 9. Any microorganism listed in claims 1 through 8 that belongs to the family Enterobacteriaciae. 10. The microorganisms under claim 9, which belong to the family Enterobacteriaciae, of the genus Escherichia. 11. A microorganism under any of the claims in Claims 1 through 8, which is a coryneform bacterium. 12. Any of the microorganisms listed in Claims 1 through 8, which are bacteria capable of digesting and absorbing methanol. 13. The microorganisms under claim 12, which are bacteria capable of digesting and absorbing methanol, are of the genus Methylophilus or Methylobacillus. 14. The method for the production of L-amino acids involves the cultivation of microorganisms according to one of the claims 1 through 13 in a medium that creates and causes the accumulation of L-amino acids, and the collection of L-amino acids from the medium or microorganisms. 15. A method for the production of L-amino acids consisting of the cultivation of microorganisms according to patents 12 to 13 in a medium with methanol as the primary carbon source for the creation and accumulation of such L-amino acids, and the collection of L-amino acids from the medium or microorganisms. 16. The method pursuant to claims 14 to 15 in which L-amino acids are selected from a group consisting of L-lysine, L-arginine, L-ornithine, L-histidine, L-isoleucine, L-isoleucine, L-threonine, L-proline, L-phenylalanine, L-cysteine, and L-glutamic acid.