TH100672A - L-glutamic acid producing bacteria and methods for producing L-glutamic acid - Google Patents

Abstract

DC60 Acid-producing bacteria L-Amino, of the genus Pantoea, Enterobacter, Klebsiella or Erwinia, where bacteria are modified by gene reproduction to prevent the rpoS gene from functioning properly by cultivating in the medium to cause Has accumulated acid L-glutamic acid in the medium and L-glutamic acid is collected from that medium. Bacteria that produce acid L-Amino, of the genus Pantoea, Klebsiella or Erwinia, where bacteria are modified by gene reconstruction to make the rpos gene inactive because it is cultured in the medium to cause the substance. Accumulation of acid L-glutamic acid in the medium and L-glutamic acid is collected from that medium.

Claims (9)

Updated 05/01/2015 1. Methods for producing L-glutamic acid involved A) culturing L-glutamic acid-producing bacteria in a medium, which included bacteria of the genera Pantoea, Enterorbacter, Serratia, Klebsiella, and Erwinia, and B) harvesting L-glutamic acid from the medium, where the bacteria were genetically modified to inactivate the rpoS gene. The rpoS gene encoded a selected protein from a group consisting of: (A) a protein containing the amino acid sequence SEQ ID NO:2, and (B) a protein containing the amino acid sequence SEQ ID NO:2 but including substitutions, deletions, insertions, or additions of one to five amino acid segments, and which functioned as the S sigma factor of RNA polymerase. 2. Method according to claim 1, where bacteria are cultured at pH 3 to 5. 3. Method according to claim 1, where L-glutamic acid is accumulated along with precipitation in the medium during culture. 4. Method according to claim 1, where the rpoS gene is inactivated by a method selected from the group. It consists of reducing the expression of the rpoS gene or by interfering with the rpoS gene and its components. 5. Method according to claim 1, where the rpoS gene is selected from a group consisting of: (a) DNA containing the nucleotide sequence of SEQ ID NO: 1 and (b) DNA which can be recombined with a sequence matching the nucleotide sequence of SEQ ID NO: 1 under strict conditions containing a salt concentration corresponding to 60°C, 1xSSC, 0.1% SDS and where such DNA transcribes the protein with the S sigma factor function of RNA polymerase. 6. Method according to claim 1, where the bacterium is of the genus Pantaea agglomerans or Pantoea anantis. ---------------------------------------- 1. Acid production method. L-glutamic acid was collected using two methods: A) Culturing L-glutamic acid-producing bacteria in a medium, belonging to the genera Pantoea, Enterobacter, Serratia, Klebsiella, and Erwinia; and B) Collecting L-glutamic acid from the medium, where the bacteria were genetically modified to inactivate the rpoS gene, and the rpoS gene encoded a selected protein from a group consisting of: (A) a protein containing the amino acid sequence SEQ ID NO: 2; and (B) a protein containing the amino acid sequence SEQ ID NO: 2 but including substitutions, deletions, insertions, or additions of 1 to 5 amino acid segments, and which functions as the S factor of RNA polymerase. 2. Method according to claim 1, where the bacteria were cultured at pH 3 to 5. 3. Method according to claim 1, where L-glutamic acid accumulates and precipitates in the medium during culture. 4. Method according to claim 1, where the rpoS gene is rendered inactive by a method selected from a group consisting of either reduction of rpoS gene expression or by interference with the rpoS gene and its components. 5. Method according to claim 1, where the rpoS gene is selected from a group consisting of: (a) DNA containing the nucleotide sequence of SEQ ID NO: 1 and (b) DNA capable of matching a sequence matching the nucleotide sequence of SEQ ID NO: 1 under strict conditions containing a salt concentration corresponding to 60°C, 1xSSC, 0.1% SDS, and where such DNA transcribes the functional factor S sigma protein of RNA polymerase. 6. Method according to claim 1, where the bacterium is of the genus Pantoea agglomerans or Pantoea ananatis. 4. Bacteria pursuant to claim 1, whereby the rpoS gene is selected from a pool consisting of: (a) DNA containing the nucleotide sequence of SEQ ID NO: 1 and (b) DNA which can be recombined with a sequence matching the nucleotide sequence of SEQ ID NO: 1 or a tracer which can be prepared from the nucleotide sequence under strict conditions, and where such DNA transcribes the factor S sigma protein of RNA polymerase. 5. The method for producing L-glutamic acid involves culturing bacteria as described in point one of claims 1 through 4 in the medium and harvesting L-glutamic acid from that medium. Which of claims 1 through 4 pertains to the medium and the harvesting of L-glutamic acid from the medium? 6. Method under claim 5, where bacteria are cultured at pH 3 to 5. 7. Methods under claim 5 or 6 where L-glutamic acid accumulates and precipitates in the medium during cultivation. 8. A method for improving the growth of L-glutamic acid-producing bacteria under acidic conditions involving the non-functioning of the rpoS gene through gene editing. 9. Method according to claim 8, where the microorganism belongs to a selected genus consisting of Pantoea, Enterobacter, Klebsiella, or Erwinia.