TH95082A - A method for producing L-amino acids using bacteria of the family. Enterobacteriaceae - Google Patents

Abstract

DC60 (15/10/50) This invention discloses methods for producing L-Amino acids are described, for example L-threonine, L-lysine, L- Histidine, L-phenylalanine, L-arginine, L-tryptophan, L-glutamic acid using bacteria of the family Enterobacteriaceae, whereby the bacteria have been modified To achieve the predominance of the function of the original alcohol dehydrogenase transcribed by the adhE gene or the alcohol dehydrogenase mutant resistant to the inactivity of oxygen.

Claims (11)

------23/03/2018------(OCR) Page 1 of 2 pages. Patents: 1. Method for L-amino acid production by fermentation comprising: A) Culturing in ethanol-containing culture medium of L-amino acid-producing Enterobacteriaceae bacteria with alcohol dehydrogenase, and B) Isolation of L-amino acids from the culture medium, where the gene encoding the said alcohol dehydrogenase is expressed under the control of an unnatural activator which functions under aerobic culture conditions. 2. Method according to Patent 11, where the said unnatural activator is selected from a group consisting of ptac, Plac, Ptrp, ptr0, PR, and PL. 3. Method according to Patents 1 or 2, where the said alcohol dehydrogenase is resistant to... 4. Any of the methods under claims 1 to 3, where the alcohol dehydrogenase is of bacterial origin selected from a group comprising Escherichia coli, Erwinia carotovora, Salmonella typhlmurium, Shigella flexneri, Yersenia pestis, Pantoea ananatis, Lactobacillus plantarum, and Lactococcus lactis. 5. Any of the methods under claims 1 to 4, where the alcohol dehydrogenase consists of the amino acid sequence provided in sequence 2, except that the glutamic acid sequence at position 568 is replaced by another amino acid sequence other than the aspartic acid sequence. 6. Any of the methods under claims 1 to 4, where the alcohol dehydrogenase... This refers to the amino acid sequence provided in sequence:2, except that the glutamic acid sequence at position 568 is replaced by the lysine sequence 7. According to claim 5 or 6, the said dehydrogenel alcohol has at least one additional mutation which can lead to improved growth of the said bacteria. 8. Liquid medium in which ethanol is the sole carbon source. Method according to claim 7, where the said additional mutations were selected from the group consisting of: A) Substitution of the glutamic acid sequence at position 560 in sequence 2 with another amino acid sequence; B) Substitution of the phenylalanine sequence at position 566 in sequence 2 with another amino acid sequence; C) Substitution of the glutamic acid sequence, methionine sequence, tyrosine sequence, isosine sequence, and alanine sequence at positions 22, 236, 461, 554, and 786, respectively, in sequence 2. with another set of amino acid compounds, and a) their combination. 9. Method according to claim 7, where such additional mutations are selected from groups consisting of: A) the replacement of the glutamic acid compound at position 560 in sequence:2 with the lysine compound; B) the replacement of the phenylalanine compound at position 566 in sequence:2 with the valine compound; C) the replacement of the glutamic acid compound, methionine compound, tyrosine compound, isoleucine compound, and alanine compound at positions 22, 236, 461, 554, and 786, respectively, in sequence:2 with the glycine compound, valine compound, cysteine compound, sherine compound, and valine compound, respectively, and a) their combination. 10. Any one of the methods according to claims 1 through 9, where the bacteria produce L-amino acids. Such as, the genus selected from the group comprising Escherichia, Enterobacter, Erwinia, Klebsiella, Pantoea, Providencia, Salmonella, Serratia, Shigella, and Morganella 11. Any one of the methods of claims 1 through 10, where such L-amino acids are selected from the group comprising L-threonine, L-lysine, L-histidine, L-phenylalanine, L-arginine, L-tryptophan, L-glutamic acid, and L-leucine ------------ 1. Methods for the production of L-amino acids were: A) Culturing in an ethanol-containing culture medium of bacteria of the family Enetrrobacteriaceae containing alcohol dehydrogenase, and B) Isolation of L-amino acids from that culture medium, where the gene transcribing the alcohol dehydrogenase was expressed under the control of an unnatural activator in which oxygen was present during the culture process. 2. The method pursuant to claim 1, where the aforementioned unnatural stimulus is selected from a group comprising Ptac, Ptac, Ptrp, Ptrc, PR, and PL. 3. A method under claim 1 or 2, whereby the alcohol dehydrogenase is resistant to inactivation in the presence of oxygen. 4. Any of the methods under Claim 1 or 3, where the alcohol dehydrogenase is of a selected bacterial origin from a group comprising Escherichia coli, Erwinia carotovora, Salmonella typhimurium, Shigella flexneri, Yersenia pestis, Pantoea ananatis, Lactobacillus plantarum, and Lactococcus lactis. 5. Any of the methods under Claims 1 through 4, where the alcohol dehydrogenase consists of the amino acid sequence provided in sequence 2, except that the glutamic acid sequence at position 568 is replaced by a sequence of amino acids E other than the aspartic acid sequence. 6. Any of the methods under claims 1 through 4, where the alcohol dehydrogenase consists of the amino acid sequence provided in sequence 2, except that the glutamic acid sequence at position 568 is replaced by the lysine sequence. 7. A method pursuant to claim 5 or 6, whereby the said alcohol dehydrogenase has at least one additional mutation which can result in improved growth of such bacteria in a liquid medium in which ethanol is the sole carbon source. 8. Methods under claim 7, where such additional mutations are selected from a group consisting of: A) the substitution of the glutamic acid sequence at position 560 in sequence 2 with another amino acid sequence; B) the substitution of the phenylalanine sequence at position 566 in sequence 2 with another amino acid sequence; C) the substitution of the glutamic acid sequence, the methionine sequence, the tyrosine sequence, the isoleucine sequence, and the alanine sequence at positions 22, 236, 461, 554, and 786, respectively, in sequence 2 with another amino acid sequence; and D) their combination. 9. The method pursuant to claim 7, where such additional mutations are selected from a group consisting of: A) the substitution of the glutamic acid series at position 560 in sequence:2 with the lysine series; B) the substitution of the phenylalanine series at position 566 in sequence:2 with the valine series; C) the substitution of the glutamic acid series, methionine series, tyrosine series, isoleucine series, and alanine series at positions 22, 236, 461, 554, and 786, in sequence:2 with the glycine series, valine series, cysteine series, serine series, and valine series, respectively; and D) their combination. 10. Any of the methods under Claims 1 through 9, where the bacteria belong to a selected genus from the group comprising Escherichia, Enetrrobacter, Erwinia, Klebsiella, Pantoea, Providencia, Salmonella, Serratia, Shigella, and Morganella. 11. Any of the methods under Claims 1 through 10, where such L-amino acids are selected from a group consisting of L-threonine, L-lysine, L-histidine, L-phenylalanine, L-arginine, L-tryptophan, L-glutamic acid, and L-leucine.