TH62696B - A method for L-Amino acid production using bacteria with the dominant pckA gene expression. - Google Patents

Abstract

DC60 (28/06/47) This invention relates to methods for the production of L-amino acids, such as L-tryptophan, L-phenylalanine and L-. Tyrosine, by means of bacteria of the Escherichia family, and by which the L-Amino acid production of the mentioned bacteria is distinguished by Enhance the function of PEP carboxykinase, which is transcribed by the pckA gene.

Claims (3)

Disclaimer (all) which will not appear on the advertisement page. : ------ 27/10/2017 ------ (OCR) Page 1 of 2 pages Disclaimer 1. Method for producing corrosive L-Amino aromatic acid. Choose from a group that includes L-tryptophan, L-phenylalanine and L-tyrosine by fermentation that included cultivation. Feeds L-amino acid-producing bacteria Escherichia coli, where they are adapted to To increase the function of phosphoenol pyruvate (PEP) carboxykinase, in the cultured feed that Cause the accumulation of L-Amino Acid In culture food And collection of L-amino acids From the cultured feed, the function of PEP carboxy kinase is increased by bending. The sequence that controls the expression of the PEP carboxykinase gene on the bacterial chromosome. So that gene expression is enhanced, or by increasing the number of gene copies, and where bacteria are also modified to allow increased expression of yddG by increasing the number of copies of the read frame. Opens yddG where the yddG gene can be cloned using the primer listed in SEQ ID NO 7 and 8. 2. The claim method 1 where the natural stimulant of the carboxylic PEP gene was found. 3. Claims method 2 where the stronger stimulus is the ppsA stimulant which is stimulated by FruR protein. 1 where the natural sine-dalkano (SD) arrangement of the PEP gene carboxykinase is replaced by a more efficient SD collation. 4 where such a more efficient SD collation is the gene-containing (formula) SD collation 10 from the T7 fajc 1. In the list of collation (B) proteins, which are composed of acid-mediated collation, Amino listed in SEQ ID NO: 2 in collation accounts that include subtraction, substitution, insertion or The addition of one to ten amino acids and which performs the function of PEP carboxykinase. 1. A test that can be prepared from such nucleotide arrangements. Under strict conditions and decoding the protein that performs the function of PEP carboxykinase, under strict conditions it is a condition where the wash is done at 60 ° C, and a salt concentration corresponding to 1 X SSC and 0.1%. SDS 8. Method according to any one of claims 1 through 5, where the PEP carboxyrene gene is transcribed. Protein consisting of amino acid sequences shown in SEQ ID NO: 2 in the collation list 1.1 0. One of the methods of claims 1 through 9 where bacteria are expressed is the protein that is composed of amino acids. Added Of other genes For biosynthesis of amino aromatic acids ------------ Revised August 4, 2017 1. Methods for producing L-Amino aromatic acids selected from Group consisting of L-tryptophan, L-phenylalanine and L-tyrosine by fermentation that included cultivation. Feeds L-amino acid-producing bacteria Escherichia coli, where they are adapted to Provides increased function of phosphoenol pyruvate (PEP) carboxykinase, from cultured food where the function of PEP carboxykinase is enhanced. Up by modifying the order that controls the Expression of the PEP carboxykinase gene on the bacterial chromosome so that gene expression It is either increased, or by increasing the number of copies of the gene, and where the bacteria are also modified. To provide an increased expression of the open read frame (yddG) by increasing the number of copies of the yddG open read frame, where the yddG gene can be cloned using the primer present in SEQ ID NO 7 and 8 - --------- 1. L-amino acid producing bacteria of the Escherichia family, by which the bacteria have been modified. 2. Bacteria according to claim 1, where the function of PEP carboxykinase is induced Distinguished by modifying the arrangement Regulation of the expression of the PEP carboxykinase gene on the bacterial chromosome so that gene expression is dominated or by multiplication. Reproduction of that gene 3. Bacteria according to claim 2, where the natural stimulus The existence of the genes that were mentioned were Replace it with a stronger stimulant. 4. Bacteria according to claim 2, where the natural SD arrangement of the genes mentioned It was replaced by a more efficient SD sequencing. 5. Bacteria according to any of the claims 2 to 4, where the PEP carboxykine gene It originates from bacteria of the Escherichia family 1. In the list of arrangements that include deductions, substitutions, additions or additions of one or more amino acids, and where they are performed. Duty of PEP carboxykinase 1. or test that This can be prepared from that nucleotide stratification under strict conditions and decoding the protein that performs the function of PEP carboxykinase 8. Bacteria according to claim 7, under strict conditions. Is a state Where the rinsing was performed at 60 ํ C, and the salt concentration corresponding to 1x SSC and 0.1% SDS 9. Bacteria according to any of Claims 1 to 8 with the bacteria being modified. In order to have A more prominent expression of the yddG 1 0. Bacteria in any of the claims 1 to 9, where L-amino acids are L-Ami Aromatic Acid No, selected from a group consisting of L-Tryptophan, L-Phenylalanine, and L-Tyrosine 1. 1.Methods for the production of L-Amino aromatic acid Containing Culture Bacteria according to either claim 1 to 10 in the culture medium to cause L-amino acid accumulation in that medium and L-amino acid collection. No from that culture medium 1 2. Method according to claim 11 where L-Amino acid is an amino acid. Romantic L-Amino at Was selected from a group consisting of L-Tryptophan, L-phenylalanine and L-Tyrosine 1 3. Method according to claim 12, whereby bacteria are expressed That is the dominant genes For biological synthesis, amino aromatics