Process for producing L-amino acid
View Patent ↗The present invention provides a method of improving efficiency of a fermentative production of an L-amino acid. To be specific, the present invention provides a process for producing a L-amino acid, comprising culturing a microorganism wherein the activity of the protein described in any one of (1) to (3) below is higher than that of the parent strain in a medium to produce the L-amino acid and accumulate the L-amino acid in the medium, and then collecting the L-amino acid from the medium: (1) a protein comprising the amino acid sequence shown by any one of SEQ ID NOS:2, 4, 6 and 8 (2) a protein consisting of the amino acid sequence resulting from deletion, substitution or addition of one or more amino acids in the amino acid sequence shown in any one of SEQ ID NOS:2, 4, 6 and 8, and having L-amino acid transport activity (3) a protein consisting of the amino acid sequence having 80% or more homology to the amino acid sequence shown in any one of SEQ ID NOS:2, 4, 6 and 8, and having L-amino acid transport activity.
1. A process for producing an L-amino acid which comprises:
(a) culturing, in a medium, a modified microorganism having L-amino acid transport activity and having a protein activity,
wherein the protein is (1) a protein comprising the amino acid sequence of SEQ ID NO: 2, 4, or 6 or (2) a protein consisting of an amino acid sequence having 95% or more homology to the amino acid sequence of SEQ ID NO: 2, 4, or 6 and having L-amino acid transport activity, and
wherein the protein activity is higher than that of the parent strain of the modified microorganism in the medium,
(b) producing and accumulating the L-amino acid in the medium, and
(c) collecting the L-amino acid from the medium.
2. The process of claim 1 , wherein the modified microorganism is produced by (i) transformation of the parent strain with a DNA or (ii) modification in the parent strain of an expression regulatory sequence of the DNA, wherein the DNA is
(1) a DNA that encodes (A) a protein comprising the amino acid sequence of SEQ ID NO: 2, 4, or 6 or (B) a protein consisting of an amino acid sequence having 95% or more homology to the amino acid sequence of SEQ ID NO: 2, 4, or 6 and having L-amino acid transport activity,
(2) a DNA comprising the nucleotide sequence of SEQ ID NO: 1, 3, or 5, or
(3) a DNA that hybridizes under stringent conditions with the DNA consisting of the nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 1, 3, or 5 and encodes a protein having L-amino acid transport activity, wherein the stringent conditions are the conditions of incubating a DNA-immobilizing filter and a probe DNA in a solution containing 50% fonnamide, 5×SSC (750 mmol/L sodium chloride, 75 mmol/L sodium citrate), 50 mmol/L sodium phosphate (pH 7.6), 5×Denhardt's solution, 10% dextran sulfate, and 20 μg/L denatured salmon sperm DNA at 42° C. overnight, followed by washing the filter in a 0.2×SSC solution at about 65° C.
3. The process of claim 2 , wherein the microorganism belongs to the genus Escherichia , the genus Corynebacterium , the genus Bacillus , the genus Serratia , the genus Pseudomonas or the genus Streptomyces.
4. The process of claim 3 , wherein the L-amino acid is selected from the group consisting of L-serine, L-glutamine, L-cysteine, L-phenylalanine and L-threonine.
5. The process of claim 1 , wherein the microorganism belongs to the genus Escherichia , the genus Corynebacterium , the genus Bacillus , the genus Serratia , the genus Pseudomonas or the genus Streptomyces.
6. The process of claim 5 , wherein the L-amino acid is selected from the group consisting of L-serine, L-glutamine, L-cysteine, L-phenylalanine and L-threonine.
7. The process of claim 2 , wherein the L-amino acid is selected from the group consisting of L-serine, L-glutamine, L-cysteine, L-phenylalanine and L-threonine.
8. The process of claim 1 , wherein the L-amino acid is selected from the group consisting of L-serine, L-glutamine, L-cysteine, L-phenylalanine and L-threonine.
9. The process of claim 1 , wherein the protein comprises the amino acid sequence of SEQ ID NO: 2, 4, or 6.
10. The process of claim 1 , wherein the protein consists of an amino acid sequence having 95% or more homology to the amino acid sequence of SEQ ID NO: 2, 4, or 6 and having L-amino acid transport activity.
11. The process of claim 2 , wherein the DNA encodes a protein comprising the amino acid sequence of SEQ ID NO: 2, 4, or 6.
12. The process of claim 2 , wherein the DNA encodes a protein consisting of an amino acid sequence having 95% or more homology to the amino acid sequence of SEQ ID NO: 2, 4, or 6 and having L-amino acid transport activity.
13. The process of claim 2 , wherein the DNA comprises the nucleotide sequence of SEQ ID NO: 1, 3, or 5.
14. The process of claim 2 , wherein the DNA that hybridizes under stringent conditions with the DNA consisting of the nucleotide sequence complementary to the nucleotide sequence of SEQ ID NO: 1, 3, or 5, and encodes a protein having L-amino acid transport activity, wherein the stringent conditions are the conditions of incubating a DNA-immobilizing filter and a probe DNA in a solution containing 50% formamide, 5×SSC (750 mmol/L sodium chloride, 75 mmol/L sodium citrate), 50 mmol/L sodium phosphate (pH 7.6), 5×Denhardt's solution, 10% dextran sulfate, and 20 μg/L denatured salmon sperm DNA at 42° C. overnight, followed by washing the filter in a 0.2×SSC solution at about 65° C.