TH55146A - Methods for cultivating amino acid-producing bacteria and methods for preparing amino acids by fermentation with the created amino acid-producing bacteria. - Google Patents

Abstract

DC60 (17/12/42) A method for producing Coryneform bacteria with improved amino acid or nucleic acid production results involves steps of inducing mutations in the activator sequence of the amino acid or nucleic acid biosynthesis gene on the Coryneform bacteria chromosome to make it closer to the corresponding sequence, or introducing changes in the activator sequence of the amino acid or nucleic acid biosynthesis gene on the Coryneform bacteria chromosome by recombination of the gene to make it closer to the corresponding sequence to obtain amino acid or nucleic acid producing mutants of Coryneform bacteria, culturing the mutants, and producing the desired amino acid or nucleic acid in large quantities. This method can create mutants that can appropriately increase or control the expression of the intended gene without the use of plasmids, and similarly can produce amino acids in high yields through recombination or mutation. The method for producing Coryneform bacteria with improved amino acid or nucleic acid production results consists of steps... This involves inducing mutations in the activation sequence of amino acid or nucleic acid biosynthesis genes on the chromosomes of Coryneform bacteria to approximate their alignment, or introducing changes in the activation sequence of amino acid or nucleic acid biosynthesis genes on the Coryneform bacteria chromosome. By recombining the genes to approximate their alignment, the goal is to obtain amino acid or nucleic acid-producing mutants of Coryneform bacteria. The mutants are then cultured to produce the desired amino acid or nucleic acid in appropriate quantities. This method can create mutants that can enhance or appropriately control the expression of the intended gene without the use of plasmids, and similarly, can produce amino acids in high yields through recombination or mutation.

Claims (17)

1. The method for producing Coryneform bacteria with improved amino acid or nucleic acid production involves steps such as inducing mutations in the activation sequence of the amino acid or nucleic acid biosynthesis gene on the Coryneform bacteria chromosome to bring it closer to the corresponding sequence, or importing changes in the activation sequence of the amino acid or nucleic acid biosynthesis gene on the Coryneform bacteria chromosome by recombination of the gene to bring it closer to the corresponding sequence. To obtain amino acid or nucleic acid-producing Coryneform bacteria mutations, the mutants are cultured and selected to produce the desired amino acid or nucleic acid in large quantities. 2. Method of Claim 1, where amino acids are selected from a group consisting of glutamic acid, lysine, arginine, serine, phenylalanine, proline, and nucleic acids are selected from a group consisting of iodine, guanosine, akenosine, adenosine, and nucleotides. 3. Method of Claim 1, where the amino acid is glutamic acid and the activator for the biosynthetic gene is selected from a group comprising activators of genes producing glutamate dehydrogenase (GDH), citrate synthase (CS), isocitrate synthase (JCDH), pyruvate dehydrogenase (PDH), and adinote (ACO). 4. Method of claim 3, where the glutamate dehydrogenase (GHD) gene activator has at least one DNA sequence selected from a group consisting of (i) CGGTCA, TTGTCA, TTGACA and TTGCCA in domain-35, (ii) the TATAAT sequence or a sequence similar to TATAAT but where the ATAAT metabolite is replaced by another metabolite and where it does not inhibit activator function in domain-10, and (iii) a combination of (i) and (ii). 5. Method of Claim 4, where the triggers for GDH are TGGTCA in domain-35, and TATAAT in domain-10, or TTGTCA in domain-35, and TATAAT in domain-10. 6. The method described in Claim 3, where the actuator for CS has (i) a TTGACA alignment in domain-35, (ii) a TATAAT alignment in domain-10, or (iii) a combination of (i) and (iii) a combination of (i) and (iii), and this alignment does not interfere with the function of the actuator. 7. Method of Claim 3, where at least one of the first and second actuators for ICDH has (i) alignment of TTGCCA or TTGACA in domain-35, (ii) alignment of TATAAT in domain-10, or (iii) a combination of (i) and (ii), and the alignment does not interfere with the function of the actuator. 8. Method of claim 3, where the activator for PDH has (i) TTGCCA alignment in domain-35, (ii) TATAAT alignment in domain-10, or (iii) a combination of (i) and (ii), and the alignment does not inhibit the function of the activator. 9. Method of claim 1, where the amino acid is arginine, and the activator for the biosynthesis gene for arginanosucciase synthase. 10. The method of claim 9, where the activator for argininosuccinate synthase has at least one DNA sequence selected from the group comprising (I) TGCCA, TTGCTA and TTGTCA in domain-35, (ii) the TATAAT sequence or the TATAAT sequence but where the ATAAC conjugate is replaced by another conjugate that does not interfere with the activator function in domain-10, and (iii) the combination of (I) and (ii). 11. Method of claim 10, where the activator for argininosuccinate synthase has at least one DNA sequence selected from the group comprising (I) TTGTCA in domain-35, (iii) the TATTAAT sequence in domain-10, and (iii) a combination of (I) and (iii). 12. A gene that synthesizes glutamic acid with the activators mentioned in claims 4 through 8. 13. The gene that synthesizes arginine with the activator mentioned in claim 10. 14. A glutamic acid-producing choreform bacterium containing the glutamic acid synthesis gene listed in claim 12. 15. Arginine-producing coryneform bacteria containing the arginine synthesis gene mentioned in claim 13. 16. A method of producing amino acids or amino acid supernatants, which consists of steps of culturing coryneform bacteria created by the methods in Reservations 1 through 11 and resulting in the production of enhanced amino acids or nucleic acids, or coryneform bacteria of Reservations 14 or 15 in the culture medium, and by accumulating the intended amino acids or nucleic acids in the medium, and collecting them from the culture medium. 17. A method for producing L-glutamic acid by fermentation, which consists of the steps of culturing coryneform bacteria that produce L-glutamic acid resistant to 4-fluoroglutamic acid in a liquid medium to create and by accumulating L-glutamic acid in the medium and collecting it from the culture medium.