CA2075044C - Production of microorganisms producing l-lysine - Google Patents
Production of microorganisms producing l-lysine Download PDFInfo
- Publication number
- CA2075044C CA2075044C CA002075044A CA2075044A CA2075044C CA 2075044 C CA2075044 C CA 2075044C CA 002075044 A CA002075044 A CA 002075044A CA 2075044 A CA2075044 A CA 2075044A CA 2075044 C CA2075044 C CA 2075044C
- Authority
- CA
- Canada
- Prior art keywords
- microorganisms
- lysine
- strains
- glutamicum
- flavum
- Prior art date
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- Expired - Fee Related
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P13/00—Preparation of nitrogen-containing organic compounds
- C12P13/04—Alpha- or beta- amino acids
- C12P13/08—Lysine; Diaminopimelic acid; Threonine; Valine
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- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Described is a method for the preparation of microorganisms which have an elevated L-lysine production capability by mutation of microorganisms of the genuses Corynebacterium and Brevibacterium with known mutagens in a known manner, characterized in that strains of said genuses are mutated, selecting those wich are resistant to regenerative inhib~~on by 2-azido-.epsilon.-caprolactam.
Description
The production of microorq_anisms producincr L-lysine The present invention relates to a process for producing L-lysine and microorganisms therefor.
L-lysine is an essential amino acid and is widely used as additive to human and animal food. It is also employed in medicine as component of infusion solutions.
L-lysine is obtained by hydrolysis of proteins with acid, by synthesis of D,L-lysine and subsequent resolution of the racemate and by synthesis with.the aid of microorganisms. Microbiological processes for pre-paring.L-lysine are described, for example, in Trends in Biotechnology 1 (1983) 70-74.
We have found an improved process for producing microorganisms which produce L-lysine.
The present invention relates to a process for producing microorganisms which have increased L-lysine productivity comprising:
- mutating strains of mircroorganisms of a genus selected from the group consisting of Corynebacterium and Brevibacterium; and - selecting the strains that are resistant to feedback inhibition by 2-azido-E-caprolactam.
Surprisingly, 2-azido-e-caprolactam has con-siderably higher efficiency in the selection of mutants after mutagenesis than the compounds described in JP 51-19 186 and EP 175 309, such as fluoro- or chloro-caprolactam.
It is thus possible by selection with azidocapro-lactam to increase the lysine productivity of strains by more than 10$.
The mutants according to the invention can be produced by conven°cional mutagenesis, eg. with N-methyl N'-nitro-N-nitrosoguanidine or by U.V. radiation.
la Examples of suitable microorganisms of the genera Corynebacterium (C) and Brevibacterium (B) are the following:
B. amoniagenis, 8. divaricatum, B. flavum, B.
ketoglutamicum, B. lactofermentum, B. linens, B. sp., C.
~o~~o~~
L-lysine is an essential amino acid and is widely used as additive to human and animal food. It is also employed in medicine as component of infusion solutions.
L-lysine is obtained by hydrolysis of proteins with acid, by synthesis of D,L-lysine and subsequent resolution of the racemate and by synthesis with.the aid of microorganisms. Microbiological processes for pre-paring.L-lysine are described, for example, in Trends in Biotechnology 1 (1983) 70-74.
We have found an improved process for producing microorganisms which produce L-lysine.
The present invention relates to a process for producing microorganisms which have increased L-lysine productivity comprising:
- mutating strains of mircroorganisms of a genus selected from the group consisting of Corynebacterium and Brevibacterium; and - selecting the strains that are resistant to feedback inhibition by 2-azido-E-caprolactam.
Surprisingly, 2-azido-e-caprolactam has con-siderably higher efficiency in the selection of mutants after mutagenesis than the compounds described in JP 51-19 186 and EP 175 309, such as fluoro- or chloro-caprolactam.
It is thus possible by selection with azidocapro-lactam to increase the lysine productivity of strains by more than 10$.
The mutants according to the invention can be produced by conven°cional mutagenesis, eg. with N-methyl N'-nitro-N-nitrosoguanidine or by U.V. radiation.
la Examples of suitable microorganisms of the genera Corynebacterium (C) and Brevibacterium (B) are the following:
B. amoniagenis, 8. divaricatum, B. flavum, B.
ketoglutamicum, B. lactofermentum, B. linens, B. sp., C.
~o~~o~~
- 2 - O.Z. 0050/41780 acetoacidophilum, C. acetoglutamicum, C. glutamicum, C.
lilium and C. sp. Preference is given to B, flavum and C.
glutamicum, especially B. flavum ATCC 21.474 and C.
glutamicum ATCC 21526. The latter has the special advan-Cage that it is homoserine-dependent and, moreover, is resistant to S-(2-aminoethyl)-L-cysteine.
As already indicated, it is beneficial for the strains to be homoserine-dependent. It is also useful for the strains to be resistant to S-(2-aminoethyl)-L-cys-teine. If the strains do not possess this resistance it can be introduced as described in US 3,707,441 by treating the strains with N-methyl-N'-vitro-N-nitroso-guanidine and subsequent selection.
Example Corynebacterium glutamicum ATCC 21 526 was treated with 250 ~g/ml N-methyl-N'-vitro-N-nitrosoguani-dine in tris/maleic acid buffer, pH 6.0, at 30°C for 30 min. The cells were then washed with 0.1 M tris buffer, pH 7.2, plated on minimal agar plates and then incubated at 28°C for from 4 to 14 days.
The minimal agar had the following composition:
20 g/1 agar 0.1 g/1 MnS04-H20 2 g/1 ( NH4 ) ZSO4 100 ~gf 1 biotin 0.5 g/1 RHZPO4 30 mg/1 each Met, Thr, Leu 0.5 g/1,K2HpO4 4 g/1 lactate 0 .4 g/1 MgS04 ~ 7H20 0. O1 g/1 FeSO, ~ 7H20 pH = 7 . 0 The colonies- producing lysine amongst those growing on the agar plates after incubation were identi-fied. The strains which produced 10% more lysine than the initial strain were isolated.
lilium and C. sp. Preference is given to B, flavum and C.
glutamicum, especially B. flavum ATCC 21.474 and C.
glutamicum ATCC 21526. The latter has the special advan-Cage that it is homoserine-dependent and, moreover, is resistant to S-(2-aminoethyl)-L-cysteine.
As already indicated, it is beneficial for the strains to be homoserine-dependent. It is also useful for the strains to be resistant to S-(2-aminoethyl)-L-cys-teine. If the strains do not possess this resistance it can be introduced as described in US 3,707,441 by treating the strains with N-methyl-N'-vitro-N-nitroso-guanidine and subsequent selection.
Example Corynebacterium glutamicum ATCC 21 526 was treated with 250 ~g/ml N-methyl-N'-vitro-N-nitrosoguani-dine in tris/maleic acid buffer, pH 6.0, at 30°C for 30 min. The cells were then washed with 0.1 M tris buffer, pH 7.2, plated on minimal agar plates and then incubated at 28°C for from 4 to 14 days.
The minimal agar had the following composition:
20 g/1 agar 0.1 g/1 MnS04-H20 2 g/1 ( NH4 ) ZSO4 100 ~gf 1 biotin 0.5 g/1 RHZPO4 30 mg/1 each Met, Thr, Leu 0.5 g/1,K2HpO4 4 g/1 lactate 0 .4 g/1 MgS04 ~ 7H20 0. O1 g/1 FeSO, ~ 7H20 pH = 7 . 0 The colonies- producing lysine amongst those growing on the agar plates after incubation were identi-fied. The strains which produced 10% more lysine than the initial strain were isolated.
Claims (5)
1. A process for producing microorganisms which have increased L-lysine productivity comprising:
- mutating strains of mircroorganisms of a genus selected from the group consisting of Corynebacterium and Brevibacterium;
- and selecting the strains that are resistant to feedback inhibition by 2-azido-.epsilon.-caprolactam.
- mutating strains of mircroorganisms of a genus selected from the group consisting of Corynebacterium and Brevibacterium;
- and selecting the strains that are resistant to feedback inhibition by 2-azido-.epsilon.-caprolactam.
2. A process according to claim 1, wherein the step of mutating is performed with N-methyl-N'-nitro-N-nitrosoguanidine or by U.V. radiation.
3. A process according to claim 1 or 2, wherein the microorganisms are selected from the group consisting of B. amoniagenis, B. divaricatum, B. flavum, B. keto-glutamicum, B. lactofermentum, B. linens, B. sp., C. acetoacidophilum, C. acetoglutamicum, C. glutamicum, C. lilium and C. sp.
4. A process according to claim 3, wherein the microorganisms are B. flavum or C. glutamicum.
5. A process according to claim 4, wherein the microorganisms are B. flavum having ATCC deposit number 21.474 or C. glutamicum having ATCC deposit number 21526.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DEP4023576.9 | 1990-07-25 | ||
| DE4023576A DE4023576A1 (en) | 1990-07-25 | 1990-07-25 | METHOD FOR PRODUCING L-LYSINE PRODUCING MICROORGANISMS |
| PCT/EP1991/001316 WO1992001785A1 (en) | 1990-07-25 | 1991-07-13 | Method of preparation of l-lysine-producing microorganismes |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2075044A1 CA2075044A1 (en) | 1992-01-26 |
| CA2075044C true CA2075044C (en) | 2001-07-10 |
Family
ID=6410961
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002075044A Expired - Fee Related CA2075044C (en) | 1990-07-25 | 1991-07-13 | Production of microorganisms producing l-lysine |
Country Status (7)
| Country | Link |
|---|---|
| EP (1) | EP0540556B1 (en) |
| JP (1) | JPH05508543A (en) |
| CA (1) | CA2075044C (en) |
| DE (2) | DE4023576A1 (en) |
| FI (1) | FI106043B (en) |
| HU (1) | HU211056B (en) |
| WO (1) | WO1992001785A1 (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE4204361A1 (en) * | 1992-02-14 | 1993-08-19 | Degussa | METHOD FOR PRODUCING L-LYSINE BY FERMENTATION OF CORYNEFORM BACTERIA |
| KR100688575B1 (en) * | 2004-10-08 | 2007-03-02 | 삼성전자주식회사 | Non volatile semiconductor memory device |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0175309A3 (en) * | 1984-09-14 | 1987-11-11 | Toray Industries, Inc. | A method for producing l-lysine |
-
1990
- 1990-07-25 DE DE4023576A patent/DE4023576A1/en not_active Withdrawn
-
1991
- 1991-07-13 CA CA002075044A patent/CA2075044C/en not_active Expired - Fee Related
- 1991-07-13 EP EP91912793A patent/EP0540556B1/en not_active Expired - Lifetime
- 1991-07-13 HU HU9202242A patent/HU211056B/en not_active IP Right Cessation
- 1991-07-13 JP JP91511992A patent/JPH05508543A/en active Pending
- 1991-07-13 DE DE59103718T patent/DE59103718D1/en not_active Expired - Lifetime
- 1991-07-13 WO PCT/EP1991/001316 patent/WO1992001785A1/en active IP Right Grant
-
1992
- 1992-12-28 FI FI925887A patent/FI106043B/en active
Also Published As
| Publication number | Publication date |
|---|---|
| CA2075044A1 (en) | 1992-01-26 |
| HU211056B (en) | 1995-10-30 |
| EP0540556B1 (en) | 1994-11-30 |
| EP0540556A1 (en) | 1993-05-12 |
| DE4023576A1 (en) | 1992-01-30 |
| DE59103718D1 (en) | 1995-01-12 |
| FI925887A0 (en) | 1992-12-28 |
| HUT68739A (en) | 1995-05-17 |
| FI106043B (en) | 2000-11-15 |
| WO1992001785A1 (en) | 1992-02-06 |
| FI925887A (en) | 1992-12-28 |
| HU9202242D0 (en) | 1992-10-28 |
| JPH05508543A (en) | 1993-12-02 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request | ||
| MKLA | Lapsed |