RU2460779C2 - Method for producing hydroxylated amino acid (versions) and microorganism transformed by dna coding dioxygenase - Google Patents

Method for producing hydroxylated amino acid (versions) and microorganism transformed by dna coding dioxygenase Download PDF

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RU2460779C2
RU2460779C2 RU2010107900/10A RU2010107900A RU2460779C2 RU 2460779 C2 RU2460779 C2 RU 2460779C2 RU 2010107900/10 A RU2010107900/10 A RU 2010107900/10A RU 2010107900 A RU2010107900 A RU 2010107900A RU 2460779 C2 RU2460779 C2 RU 2460779C2
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isoleucine
dioxygenase
hydroxy
bacterial
bacterium
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RU2010107900A (en
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Сергей Васильевич Смирнов (RU)
Сергей Васильевич Смирнов
Наталья Николаевна САМСОНОВА (RU)
Наталья Николаевна Самсонова
Вероника Александровна КОТЛЯРОВА (RU)
Вероника Александровна КОТЛЯРОВА
Юки ИМАБАЯСИ (JP)
Юки ИМАБАЯСИ
Синъичи СУЗУКИ (JP)
Синъичи СУЗУКИ
Масаказу СУГИЯМА (JP)
Масаказу СУГИЯМА
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Закрытое акционерное общество "Научно-исследовательский институт "Аджиномото-Генетика" (ЗАО АГРИ)
Аджиномото Ко., Инк.
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Abstract

FIELD: medicine.
SUBSTANCE: method for producing 4'-hydroxy-L-isoleucine or 4,4'-dihydroxy-L-isoleucine or their salts by: enzymatic conversion in the presence of dioxygenase recovered from the bacterium belonging to a group including Pantoea and Pseudomonas geni, or by cultivation of a first producer bacterium possessing activity of dioxygenase recovered from the second bacterium belonging to a group including Pantoea and Paseudomonas geni, in a nutrient medium containing L-isoleicine and/or 4-hydroxy-L-isoleucine or thier salts and recovery of the end product from the culture fluid.
EFFECT: invention enables high-effective production of said compounds and their salts.
13 cl, 24 dwg, 5 tbl, 11 ex

Description

Текст описания приведен в факсимильном виде.

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Claims (13)

1. Способ получения 4'-гидрокси-L-изолейцина или 4,4'-дигидрокси-L-изолейцина или их солей методом ферментативной конверсии в присутствии диоксигеназы, выделенной из бактерии, принадлежащей к группе, включающей роды Pantoea и Pseumodomonas, и способной конвертировать L-изолейцин в 4'-гидрокси-L-изолейцин и/или 4-гидрокси-L-изолейцин в 4,4'-дигидрокси-L-изолейцин.1. A method of obtaining 4'-hydroxy-L-isoleucine or 4,4'-dihydroxy-L-isoleucine or their salts by enzymatic conversion in the presence of dioxigenase isolated from a bacterium belonging to the group comprising the genera Pantoea and Pseumodomonas, and capable of converting L-isoleucine in 4'-hydroxy-L-isoleucine and / or 4-hydroxy-L-isoleucine in 4,4'-dihydroxy-L-isoleucine. 2. Способ по п.1, отличающийся тем, что указанная диоксигеназа выбрана из группы, состоящей из диоксигеназ с аминокислотной последовательностью SEQ ID No:2, SEQ ID No:4, SEQ ID No:56, SEQ ID No:58 или их вариантов.2. The method according to claim 1, characterized in that said dioxygenase is selected from the group consisting of dioxygenases with the amino acid sequence of SEQ ID No: 2, SEQ ID No: 4, SEQ ID No: 56, SEQ ID No: 58 or their variants . 3. Способ получения 4'-гидрокси-L-изолейцина или 4,4'-дигидрокси-L-изолейцина или их солей в среде в присутствии первой бактерии, обладающей активностью диоксигеназы, выделенной из второй бактерии, принадлежащей к группе, включающей роды Pantoea и Pseumodomonas, и способной конвертировать L-изолейцин в 4'-гидрокси-L-изолейцин и/или 4-гидрокси-L-изолейцин в 4,4'-дигидрокси-L-изолейцин, включающий:
- выращивание указанной первой бактерии в питательной среде, содержащей L-изолейцин и/или 4-гидрокси-L-изолейцин или их соли; и
- выделение 4'-гидрокси-L-изолейцина и/или 4,4'-дигидрокси-L-изолейцина из культуральной жидкости.3. A method of producing 4'-hydroxy-L-isoleucine or 4,4'-dihydroxy-L-isoleucine or their salts in a medium in the presence of a first bacterium having dioxigenase activity isolated from a second bacterium belonging to the Pantoea genus group and Pseumodomonas, and capable of converting L-isoleucine to 4'-hydroxy-L-isoleucine and / or 4-hydroxy-L-isoleucine to 4,4'-dihydroxy-L-isoleucine, including:
- growing said first bacterium in a nutrient medium containing L-isoleucine and / or 4-hydroxy-L-isoleucine or their salts; and
- isolation of 4'-hydroxy-L-isoleucine and / or 4,4'-dihydroxy-L-isoleucine from the culture fluid. 4. Способ по п.3, отличающийся тем, что первая бактерия, обладающая активностью бактериальной диоксигеназы, выбрана из группы бактерий, включающей роды Agrobacterium, Anabaena, Bacillus, Bordetella, Burkholderia, Escherichia, Gloeobacter, Gluconobacter, Mesorhizobium, Methylobacillus, Nostoc, Pantoea, Photorhabdus, Pseudomonas, Psychromonas и Streptomyces.4. The method according to claim 3, characterized in that the first bacterium having bacterial dioxygenase activity is selected from the group of bacteria including the genera Agrobacterium, Anabaena, Bacillus, Bordetella, Burkholderia, Escherichia, Gloeobacter, Gluconobacter, Mesorhizobium, Methylobacillus, Methylobacillus, , Photorhabdus, Pseudomonas, Psychromonas, and Streptomyces. 5. Способ по п.4, отличающийся тем, что первая бактерия, обладающая активностью бактериальной диоксигеназы и выбранная из группы бактерий, включающей роды Pantoea и Pseumodomonas, модифицирована таким образом, что активность бактериальной диоксигеназы повышена.5. The method according to claim 4, characterized in that the first bacterium having bacterial dioxygenase activity and selected from the group of bacteria including the genera Pantoea and Pseumodomonas is modified so that the activity of bacterial dioxygenase is increased. 6. Способ по п.5, отличающийся тем, что активность бактериальной диоксигеназы повышена путем усиления экспрессии гена, кодирующего бактериальную диоксигеназу.6. The method according to claim 5, characterized in that the activity of bacterial dioxygenase is increased by enhancing the expression of a gene encoding a bacterial dioxygenase. 7. Способ по п.6, отличающийся тем, что экспрессия гена, кодирующего бактериальную диоксигеназу, усилена путем трансформации первой бактерии, выбранной из группы, включающей роды Pantoea и Pseumodomonas, молекулой ДНК, кодирующей бактериальную диоксигеназу, способную конвертировать L-изолейцин в 4'-гидрокси-L-изолейцин и/или 4-гидрокси-L-изолейцин в 4,4'-дигидрокси-L-изолейцин.7. The method according to claim 6, characterized in that the expression of the gene encoding the bacterial dioxygenase is enhanced by transforming the first bacterium selected from the group comprising the genera Pantoea and Pseumodomonas, a DNA molecule encoding a bacterial dioxygenase capable of converting L-isoleucine to 4 ' -hydroxy-L-isoleucine and / or 4-hydroxy-L-isoleucine in 4,4'-dihydroxy-L-isoleucine. 8. Способ по п.4, отличающийся тем, что первая бактерия, обладающая активностью бактериальной диоксигеназы и выбранная из группы бактерий, включающей роды Agrobacterium, Anabaena, Bacillus, Bordetella, Burkholderia, Escherichia, Gloeobacter, Gluconobacter, Mesorhizobium, Methylobacillus, Nostoc, Photorhabdus, Psychromonas и Streptomyces, модифицирована таким образом, что активность бактериальной диоксигеназы сообщена.8. The method according to claim 4, characterized in that the first bacterium having bacterial dioxygenase activity and selected from the group of bacteria including the genera Agrobacterium, Anabaena, Bacillus, Bordetella, Burkholderia, Escherichia, Gloeobacter, Gluconobacter, Mesorhizobium, Methylobacushab, Ny Psychromonas and Streptomyces are modified so that bacterial dioxygenase activity is reported. 9. Способ по п.8, отличающийся тем, что активность бактериальной диоксигеназы сообщена путем трансформации первой бактерии по п.8 молекулой ДНК, кодирующей бактериальную диоксигеназу, способную конвертировать L-изолейцин в 4'-гидрокси-L-изолейцин и/или 4-гидрокси-L-изолейцин в 4,4'-дигидрокси-L-изолейцин.9. The method according to claim 8, characterized in that the activity of the bacterial dioxygenase is reported by transforming the first bacterium according to claim 8 by a DNA molecule encoding a bacterial dioxygenase capable of converting L-isoleucine to 4'-hydroxy-L-isoleucine and / or 4- hydroxy-L-isoleucine to 4,4'-dihydroxy-L-isoleucine. 10. Способ по пп.3, 7, 9, отличающийся тем, что указанная бактериальная диоксигеназа выбрана из группы, состоящей из бактериальных диоксигеназ с аминокислотной последовательностью SEQ ID No:2, SEQ ID No:4, SEQ ID No:56, SEQ ID No:58 или их вариантов.10. The method according to claims 3, 7, 9, characterized in that said bacterial dioxygenase is selected from the group consisting of bacterial dioxygenases with the amino acid sequence of SEQ ID No: 2, SEQ ID No: 4, SEQ ID No: 56, SEQ ID No: 58 or their variants. 11. Бактерия-продуцент 4'-гидрокси-L-изолейцина или 4,4'-дигидрокси-L-изолейцина или их солей, трансформированная молекулой ДНК, кодирующей диоксигеназу, выделенную из бактерии, принадлежащей к группе, включающей роды Pantoea и Pseudomonas, и способной конвертировать L-изолейцин в 4'-гидрокси-L-изолейцин и/или 4-гидрокси-L-изолейцин в 4,4'-дигидрокси-L-изолейцин.11. A bacterium producing 4'-hydroxy-L-isoleucine or 4,4'-dihydroxy-L-isoleucine or their salts, transformed with a DNA molecule encoding a dioxygenase isolated from a bacterium belonging to the group comprising the genera Pantoea and Pseudomonas, and capable of converting L-isoleucine to 4'-hydroxy-L-isoleucine and / or 4-hydroxy-L-isoleucine to 4,4'-dihydroxy-L-isoleucine. 12. Бактерия-продуцент по п.11, отличающаяся тем, что указанная бактерия принадлежит к группе, включающей роды Agrobacterium, Anabaena, Bacillus, Bordetella, Burkholderia, Escherichia, Gloeobacter, Gluconobacter, Mesorhizobium, Methylobacillus, Nostoc, Pantoea, Photorhabdus, Pseudomonas, Psychromonas и Streptomyces.12. The producing bacterium according to claim 11, characterized in that said bacterium belongs to the group including the genera Agrobacterium, Anabaena, Bacillus, Bordetella, Burkholderia, Escherichia, Gloeobacter, Gluconobacter, Mesorhizobium, Methylobacillus, Nostoc, Pantoeus, Porodom, Photorud, Photorhab Psychromonas and Streptomyces. 13. Бактерия-продуцент по пп.11 и 12, отличающаяся тем, что указанная бактериальная диоксигеназа, кодируемая трансформирующей молекулой ДНК, выбрана из группы, состоящей из бактериальных диоксигеназ с аминокислотной последовательностью SEQ ID No:2, SEQ ID No:4, SEQ ID No:56, SEQ ID No:58 или их вариантов. 13. A producer bacterium according to claims 11 and 12, characterized in that said bacterial dioxygenase encoded by a transforming DNA molecule is selected from the group consisting of bacterial dioxygenases with the amino acid sequence of SEQ ID No: 2, SEQ ID No: 4, SEQ ID No: 56, SEQ ID No: 58 or their variants.
RU2010107900/10A 2009-08-21 2010-03-04 Method for producing hydroxylated amino acid (versions) and microorganism transformed by dna coding dioxygenase RU2460779C2 (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2822892C1 (en) * 2019-09-17 2024-07-15 Бэйджин Дабэйнун Биотекнолоджи Ко., Лтд. Mutant polypeptide hydroxyphenylpyruvate dioxygenase, coding gene thereof and use thereof

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2006106311A (en) * 2006-03-01 2007-09-20 Закрытое акционерное общество "Научно-исследовательский институт Аджиномото-Генетика" (ЗАО АГРИ) (RU) METHOD FOR PRODUCING 4-HYDROXY-L-ISOLEUCIN OR ITS SALTS
WO2008044614A1 (en) * 2006-09-28 2008-04-17 Ajinomoto Co., Inc. Method for producing 4-hydroxy-l-isoleucine

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2006106311A (en) * 2006-03-01 2007-09-20 Закрытое акционерное общество "Научно-исследовательский институт Аджиномото-Генетика" (ЗАО АГРИ) (RU) METHOD FOR PRODUCING 4-HYDROXY-L-ISOLEUCIN OR ITS SALTS
WO2008044614A1 (en) * 2006-09-28 2008-04-17 Ajinomoto Co., Inc. Method for producing 4-hydroxy-l-isoleucine

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2822892C1 (en) * 2019-09-17 2024-07-15 Бэйджин Дабэйнун Биотекнолоджи Ко., Лтд. Mutant polypeptide hydroxyphenylpyruvate dioxygenase, coding gene thereof and use thereof

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