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EP-3778901-B1 - A NOVEL PROMOTER, AND A METHOD FOR PRODUCING L-AMINO ACID BY USING SAME

EP3778901B1EP 3778901 B1EP3778901 B1EP 3778901B1EP-3778901-B1

Inventors

  • LEE, JI YEON
  • CHANG, JIN SOOK
  • KIM, HYUNG JOON
  • YOON, BYOUNG HOON
  • CHOI, SUN HYOUNG
  • CHOI, YUNJUNG

Dates

Publication Date
20260506
Application Date
20190327

Claims (13)

  1. A polynucleotide having promoter activity, comprising a nucleotide sequence of SEQ ID NO: 1, wherein the nucleotides corresponding to the 53rd and 55th nucleotides of SEQ ID NO: 1 are substituted with T; or the nucleotides corresponding to the 53rd and 55th nucleotides of SEQ ID NO: 1 are substituted with T and the nucleotide corresponding to the 60th nucleotide of SEQ ID NO: 1 is substituted with G.
  2. The polynucleotide according to claim 1, wherein the polynucleotide consists of the nucleotide sequence of SEQ ID NO: 2 or 3.
  3. The polynucleotide according to claim 1 or 2, wherein the polynucleotide is operably linked to a gene encoding a target protein.
  4. A vector comprising the polynucleotide of claim 1 or 2; and a gene encoding a target protein operably linked to the polynucleotide.
  5. The vector according to claim 4, wherein the target protein is phosphoribosyl-ATP pyrophosphatase (HisE), ATP phosphoribosyltransferase (HisG), or a combination thereof.
  6. A microorganism of the genus Corynebacterium, comprising the polynucleotide of claim 1; and a gene encoding a target protein operably linked to the polynucleotide.
  7. The microorganism according to claim 6, wherein the polynucleotide consists of the nucleotide sequence of SEQ ID NO: 2 or 3.
  8. The microorganism according to claim 7, wherein the target protein is phosphoribosyl-ATP pyrophosphatase: (HisE), ATP phosphoribosyltransferase (HisG), or a combination thereof.
  9. The microorganism according to claim 8, wherein, in the ATP phosphoribosyltransferase (HisG), the 233rd and 235th amino acids of the HisG amino acid sequence of SEQ ID NO: 16 are substituted with histidine (H) and glutamine (Q), respectively.
  10. The microorganism according to any one of claims 6 to 9, wherein the microorganism of the genus Corynebacterium is Corynebacterium glutamicum.
  11. A method for producing a target substance, comprising: culturing the microorganism of the genus Corynebacterium of any one of claims 8 to 9 in a medium; and recovering the target substance from the medium, wherein the target substance is an amino acid.
  12. A method for preparing a fermented composition, comprising fermenting by culturing the microorganism of the genus Corynebacterium of any one of claims 6 to 9 in a medium.
  13. A fermented composition comprising the microorganism of the genus Corynebacterium of any one of claims 6 to 9.

Description

Technical Field The present disclosure relates to a novel polynucleotide having promoter activity and a method for producing a target substance using said polynucleotide having promoter activity and more specifically, , a vector and a microorganism of the genus Corynebacterium comprising the polynucleotide, a method for producing a fermented composition using the microorganism, and the fermented composition. Background Art L-Amino acids are the basic building blocks of proteins and are used as important materials such as pharmaceutical raw materials, food additives, animal feeds, nutritional supplements, pesticides, bactericides, etc. Among these, L-glutamic acid is a representative amino acid produced by fermentation and has a unique, distinctive taste (umami taste), and thus is an important amino acid widely used in the food field as well as in the medical field and other animal feed fields. Further, glycine is mainly used as a flavor enhancer in the food industry because of its sweet taste, and is used with natural flavor enhancers to enhance taste. Furthermore, glycine is also used for its antioxidant activity, buffering action, etc., and in terms of medicine, it is used in infusion solutions, antacids, multi-amino acid preparations, and nutritional supplements. A typical method for producing amino acids includes a fermentation method using a microorganism of the genus Brevibacterium or Corynebacterium (Amino Acid Fermentation, Gakkai Shuppan Center: 195-215, 1986) or using Escherichia coli or microorganisms of the genera Bacillus, Streptomyces, Penicillum, Klebsiella, Erwinia, Pantoea, etc. (US Patent Nos. 3,220,929 and 6,682,912). In addition, such amino acids are also produced by an industrial method using a synthetic process such as the monochloroacetic acid method, the Strecker method, or the like. Additionally, various studies have been conducted for efficiently producing amino acids; for example, efforts have been made to develop microorganisms or fermentation process technologies for producing amino acids with high efficiency. Particularly, approaching methods for specific to target materials have been developed, such as enhancement of expression of genes encoding enzymes involved in the biosynthesis of the amino acids in the strain of the genus Corynebacterium or deletion of genes unnecessary for the biosynthesis of amino acids (Korean Patent Nos. 10-0924065 and 1208480). In addition to these methods, a method for removing genes that are not involved in the production of amino acids and a method for removing genes whose functions for producing amino acids are not specifically known have also been utilized. However, there is still a growing need to study methods for efficiently producing amino acids with high yield. Technical Problem The present inventors have made efforts to develop a method capable of simultaneously producing several amino acids, and as a result, they have developed a novel polynucleotide having the promoter activity of the present disclosure, and have found that the novel polynucleotide can improve glycine productivity while maintaining the glutamic acid productivity of the strain, thereby completing the present disclosure. Technical Solution The invention is set out in the appended set of claims. An objective of the present disclosure is to provide a polynucleotide having promoter activity, wherein, in the nucleotide sequence of SEQ ID NO: 1, the 53rd and 55th nucleotides are substituted with T; or the 53rd and 55th nucleotides are substituted with T and the 60th nucleotide is substituted with G. Another objective of the present disclosure is to provide a vector comprising the polynucleotide; and a gene encoding a target protein operably linked to the polynucleotide. Still another objective of the present disclosure is to provide a microorganism of the genus Corynebacterium comprising the polynucleotide; and a gene encoding a target protein operably linked to the polynucleotide. Still another objective of the present disclosure is to provide a method for producing a target substance, comprising: culturing the microorganism of the genus Corynebacterium in a medium; and recovering a target substance from the medium. Still another objective of the present disclosure is to provide a method for preparing a fermented composition, comprising fermenting by culturing the microorganism of the genus Corynebacterium in a medium. Still another objective of the present disclosure is to provide a fermented composition prepared by the above method. Advantageous Effects The novel promoter of the present disclosure is introduced into a microorganism producing amino acids to increase the production amounts of the amino acids in the microorganisms. In particular, in the case of producing the amino acids by using the novel promoter, glycine, which has been prepared by an existing synthetic method, can be produced by a fermentation method, and further, glutamic acid and glycine can be produced si