JPH0414100B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention provides novel L-threo-2-amino-4-
This invention relates to fluoro-3-hydroxybutyric acid and its production method. Conventionally known fluorine-containing physiologically active substances produced by microorganisms include the antibiotic nucleosidein [Morton et al., Journal.
of American Chemical Society (J.
Am Chem.Soc) Volume 91, Page 1535 (1969)]
However, the production of fluorine-containing amino acids by microorganisms was completely unknown. The present inventors attempted to culture various microorganisms using a medium containing a fluorine compound, and found that Streptomyces cattleya, which belongs to the genus Streptomyces, was found to be a novel L-threo-2 which has bioactive effects.
-Amino-4-fluoro-3-hydroxybutyric acid (hereinafter referred to as γ-fluoro-L-threonine)
This led to the completion of the present invention. It is well known that the development of fluorine compounds as pharmaceuticals has been actively promoted in recent years, and the γ-fluoro-L-threonine of the present invention exhibits antimetabolite and antibacterial effects, and has been widely used in the biochemistry and biochemistry fields. It is not only a useful substance in the field of chemotherapy, but also a substance that is expected to be useful as a starting material in synthetic research for pharmaceuticals containing fluorine. The present invention provides a novel γ-
Regarding fluoro-L-threonine. 1 Appearance: Colorless needle crystals (crystallized from hydroalcohol) 2 Melting point: 181-182℃ (decomposed) 3 Properties: Amphoteric substance (pka'8.25 and 1.83) 4 Elemental analysis value (%): Actual value: C35.19 H5.79 N10.08 Calculated value: C35.04 H5.88 N10.22 (calculated as C ₄ H ₈ NO ₃ F) 5 Molecular weight: 137 (field desorption mass spectrometry) 6 Molecular formula: C ₄ H ₈ NO ₃ F 7 Specific rotation: [α] _D ²⁰ −18° (C = 0.5, H ₂ O) 8 Solubility: Easily soluble in water, dimethylformamide, dimethyl sulfoxide Methanol, ethanol, ethyl acetate, butyl acetate, methyl isobutyl ketone ,benzene,
Insoluble or sparingly soluble in acetone, chloroform, n-hexane, and ether 9 Color reaction: Positive ninhydrin reaction on silica gel thin layer chromatogram (orange-yellow) 10 Stability: Relatively stable in acidic and neutral conditions, slightly unstable in alkali. Stable 11 Silica gel thin layer chromatography Rf
Value: 0.49 Adsorbent: Merck pre-coated silica gel
60F 254 Developing solvent: Acetonitrile: Acetic acid: Water = 3:
1:1 12 Ultraviolet absorption spectrum (H ₂ O): Shows only terminal absorption and no specific ultraviolet absorption maximum. 13 Infrared absorption spectra (main absorption maxima measured on potassium bromide tablets): 3420, 3090, 1630, 1480, 1420, 1350, 1320,
1245, 1165, 1125, 1075, 1055, 1005, 985,
930, 880, 770, 720, 695 (cm ^-1 ) (Figure 1) 14 ¹ H-NMR spectrum: Measured using internal standard DSS in heavy water (90MHz) 4.72ppm (2H, dd, J _HH , 4Hz , J _HF , 46Hz) 4.39ppm (1H, dq, J _HH , 4 and 5Hz, J _HF ,
25Hz) 3.9ppm (1H, d, J _HH , 5Hz) (Figure 2) 15 ¹³ C-NMR spectrum:

[Table] From the above physical and chemical properties, the L-threo of the present invention
The structure of 2-amino-4-fluoro-3-hydroxybutyric acid is as follows. The present invention involves culturing γ-fluoro-L-threonine-producing bacteria belonging to the genus Streptomyces in a fluorine-containing medium, collecting γ-fluoro-L-threonine from the culture solution, and converting it into its salts if necessary. The present invention relates to a method for producing γ-fluoro-L-threonine or a salt thereof. An example of the γ-fluoro-L-threonine-producing bacterium belonging to the genus Streptomyces used in the present invention is Streptomyces Cattleya. Streptomyces cattleya simultaneously produces the novel antibiotics thienamycin and N-acetylenamycin during fermentation. Regarding the manufacturing method, JP-A-51-73191 and JP-A-52-
65294, each is described in detail. The γ-fluoro-L-threonine-producing bacterium isolated from the soil sample was named Streptomyces cattleya after extensive taxonomic research, and its type strain was designated MA-4297, which is part of the Northern Utilization strain. -Research and Development Department, US Department of Agriculture, Peoria, Illinois
61604 USA (Northern Utilization)
Research and Development Division, US
Department of Agriculture, Peoria;
Illinois 61604U.SA), as NRRL8057, and the National Institute of Microbial Technology as Microbiological Research Institute No. 3309, respectively. The mycological properties of the strain are as follows. Morphological Characteristics Sporophytes are compact spirals found as side and terminal branches on aerial hyphae. The spores are oval or cylindrical, 0.9μ x 1.2μ in size, form chains of 10 or more, and the surface of the spores is smooth. Culture Characteristics Tomato Paste - Oatmeal Agar: Vegetative Growth - Underside - Yellow-brown, flat, spreading. Aerial mycelium - pale purple mixed with white (10 gc) Soluble pigment - none. Cyclose nitrate agar: Vegetative growth - colorless, flat, spreading. Aerial mycelium - sparse, white with a pink tinge. Soluble dye - none. Egg Albumin Agar: Vegetative growth - yellow-brown, flat, spreading with a gray to pale purple hue. Aerial Mycelium - Lighter hue of lilac and lilac mixed with some white (10 gc). Soluble dye - none. Glycerol Asparagine Agar: Vegetative growth - underside - tan with a gray to pink hue, flat, spreading. Aerial mycelium - light purple with some white (10 gc) Soluble pigment - none. Yeast Extract - Glucose + Salt Agar: Vegetative Growth - Yellow-brown color with a grayish pink hue. Aerial mycelium - pale purple mixed with pinkish white (10 gc). Soluble dye - none. Yeast extract - malt extract agar: Vegetative growth - yellowish brown. Aerial mycelium - pale purple mixed with pinkish white (10 gc). Soluble dye - none. Peptone-iron-yeast extract agar: Vegetative growth - yellowish brown. Aerial mycelium - none. Soluble dye - slight browning of the medium. Melanin - negative. _H2S production - negative. Nutrient Agar: Vegetative growth - light yellow-brown color. Aerial mycelium - none. Soluble dye - none. Nutrient starch agar: Vegetative growth - cream to tan color. Aerial mycelium - none. Soluble dye - none. Starch hydrolysis - moderate. Nutritional gelatin agar: Vegetative growth - cream colored. Aerial mycelium - none. Soluble dye - none. Liquefaction of gelatin - moderate. Gelatin stub: Vegetative growth - tan color. Aerial mycelium - none. Soluble dye - none. Liquefaction of gelatin - moderate. Potato plug: Vegetative growth - moderate, yellowish brown. Aerial mycelium - sparse, gray to pinkish [-] white. Soluble dye - none. Leffler's Serum Medium: Vegetative growth - cream colored. Aerial mycelium - none. Soluble dye - none. Liquefaction - none. Skimmed milk agar: Vegetative growth - tan color. Aerial mycelia - sparse, once white. Soluble dye - slight browning of the medium. Hydrolysis of casein - positive. Litmus Milk: Vegetative growth - tan to brown. Aerial mycelium - none. Color - No soluble dye, litmus indicator turns blue. Coagulation and/or peptonization - partial peptization, becoming alkaline. Skimmed milk: Vegetative growth - tan color. Aerial mycelium - none. Soluble dye - none. Coagulation and/or peptonization - partial peptonization, becoming alkaline. Tyrosine Agar: Vegetative growth - yellowish brown. Aerial mycelium - mixture of light purple (10 gc) and white. Soluble dye - none. Tyrosine degradation - positive. The above judgments were obtained after 3 weeks of culturing at 28°C unless otherwise stated. The PH of the media used in these studies was approximately neutral, i.e. PH
It is 6.8-7.2. The color representations used in the description are as defined in the Color Harmony Manual, 4th Edition (1958), Container Corporation of America, Chicago, Illinois. The ability of Streptomyces Cattleya to assimilate or utilize various carbohydrates was also tested. For this purpose, the microorganisms were grown for 3 weeks at 28° C. on basal synthetic medium (Pridham and Gottlieb) containing 1% carbohydrate. The pH of the culture medium used in the study is generally neutral (6.8-7.2).
Table 1 shows the utilization of these carbohydrate sources by Streptomyces Cattleya. + means good growth ±
indicates poor growth and - indicates no growth on the particular carbohydrate.

[Table] The effects of growth amount, oxygen demand, and nitrate on microorganisms due to temperature changes are as follows. Temperature range (yeast extract - glucose + salt agar): 28°C - good 37°C - moderate 50°C - no growth Oxygen requirement (yeast extract - glucose + puncture culture in salt agar): aerobic nitrate reduction - positive more than γ - Although we have explained about fluoro-L-threonine-producing bacteria, the properties of actinomycetes are not constant, and the properties of actinomycetes are not constant. It is well known that γ-fluoro-L, which belongs to the genus Streptomyces, can be mutated by artificial mutation methods such as nitrosoguanidine, ethyl methanesulfonate, etc.) and cell fusion methods. - Includes all strains that produce threonine. Fluorine compounds contained in the production medium used in the present invention include, for example, sodium fluoride, potassium fluoride, calcium fluoride, magnesium fluoride, ammonium fluoride, and barium fluoride, which dissolve in the medium and release fluorine ions. or inorganic fluorine compounds such as monofluoroacetic acid, m-fluoro-DL-phenylalanine, p-fluoro-DL
- Organic fluorine compounds such as phenylalanine and 4-fluoroglutamic acid may be mentioned, and any fluorine-containing compound that can be used by γ-fluoro-L-threonine-producing bacteria belonging to the genus Streptomyces may be used. The elementary compounds may be used alone or in combination. Furthermore, commercially available organic nitrogen sources containing various fluorine compounds (for example, soybean casein manufactured by Kishida Chemical Co., Ltd.) can also be used.The concentration of fluorine compounds in the culture medium depends on the microorganism used, culture conditions [-], or It should be determined considering the type of elementary compound, but usually 0.001 to 0.5%
(W/V) concentration is appropriate. The addition time is γ-
It is effective as long as fluoro-L-threonine production continues, and may be carried out from the beginning of the culture or at an appropriate time after the start of the culture. In addition to the above-mentioned fluorine compounds, the culture medium used in the present invention utilizes known nutrients for actinomycetes, such as glucose, glycerin, sucrose, succinic acid, acetic acid, etc. as carbon sources, and nitrogen sources such as glucose, glycerin, sucrose, and acetic acid. contains peptone, meat extract, yeast, cottonseed meal, corn steep liquor, casein, soybean meal, amino acids (e.g. glutamic acid, methionine, etc.), nitrates (e.g. sodium nitrate, ammonium nitrate, etc.), ammonium salts (e.g. ammonium chloride,
ammonium phosphate, etc.), and inorganic salts such as sulfates, carbonates, etc. of magnesium, cobalt, potassium, sodium, calcium [-], etc.
Phosphates, chlorides, etc. are used as appropriate. The medium is usually preferably a liquid medium, and the culture is preferably carried out under aerobic conditions with shaking or aeration. The pH of the medium is PH6-8, and the culture temperature is 20-40℃.
More preferably 24°C to 32°C, culture time 2 to 8
It is appropriate to do this for several days. When foaming occurs in liquid culture, antifoaming agents such as silicone oil, vegetable oil, and surfactants (eg, polypropylene glycol P-2000, Adekanol LG-126, etc.) are used as appropriate. For quantitative determination of γ-fluoro-L-threonine, a microbial assay method commonly used for measuring antibacterial active substances is applied. That is, as a test bacterium, for example, Pseudomonas aeruginosa MB2835 is used, and as a test medium, a synthetic medium that does not contain L-threonine and L-serine, such as Davis minimal agar medium [Davis et al., Journal of Bacteriology (J. Bioassay methods such as the cup method and paper disc method using γ-fluoro-L-threonine as a standard substance can be used. Streptomyces Cattleya, which is given as a specific example of the γ-fluoro-L-threonine-producing bacterium in the present invention, simultaneously accumulates the antibiotics thienamycin and N-acetylenamycin in the medium through fermentation. These antibiotics also have a growth inhibiting effect on the test bacteria in the above quantitative method. However, by adjusting the pH of the γ-fluoro-L-threonine fermentation broth containing thienamycin and N-acetyl thienamycin to 50°C for 30 minutes, γ-fluoro-L-threonine itself was removed. It was found that thienamycin and N-acetylthienamycin were not affected, but were substantially degraded to the extent that they did not affect the above quantitative method. This fact shows that when performing the assay by adding 0.1% (W/V) L-threonine to the assay medium,
This is also evidenced by the fact that no inhibition circles are observed. If the above pretreatment is performed, thienamycin and N
-Even if the sample also contains acetylthienamycin, it is possible to detect γ-fluoro-
Only L-threonine can be measured. γ-Fluoro-L-threonine can be collected from the culture solution after completion of the culture by utilizing its physical and chemical properties in accordance with the method normally used for collecting water-soluble amphoteric fermentation products. . γ-Fluoro-L-threonine is mainly present in the liquid part of the culture medium. After the cultivation is completed, the bacterial cells and other solid parts are removed by over-operation using diatomaceous earth as an aid or by centrifugation, and the γ-fluoro-L-threonine present in the liquid or supernatant is extracted and purified. . γ-Fluoro-L-threonine takes advantage of its amphoteric nature and adsorbs the culture solution or supernatant onto a cation exchange resin or anion exchange resin, and then absorbs it with an appropriate acid, base, salt [-] or any of them. It is eluted with a buffer consisting of a combination of Further, impurities can also be removed by treating a solution of γ-fluoro-L-threonine with an ion exchange resin that does not adsorb γ-fluoro-L-threonine. Instead of ion exchange resin, use ion exchange cellulose, ion exchange Cephadex, ion exchange membrane,
Ion exchange liquids and other ion exchangers can of course also be used. Furthermore, before or after the adsorption/desorption treatment using these ion exchangers, it is possible to remove the precipitate of impurities produced by adding lower alcohols such as methanol or acetone to the γ-fluoro-L-threonine-containing material, and Purification purity can be increased by precipitating and collecting fluoro-L-threonine. Furthermore, the culture solution, ion exchanger treatment solution or γ-fluoro-L-threonine solution is mixed with activated carbon or porous adsorption resin such as Amberlite XAD.
-2, XAD-4, XAD-7 (manufactured by Rohm and Haas) or Diaion HP-10, HP-20,
By treating with HP-50 (manufactured by Mitsubishi Kasei Corporation) or the like, adsorbable impurities can be removed and purification purity can be increased. Furthermore, since γ-fluoro-L-threonine is highly soluble in water, it is not extracted by organic solvents that are substantially immiscible with water, such as ether, chloroform, benzene, ethyl acetate, butyl acetate, butanol, methyl ethyl ketone, and methyl isobutyl ketone. , treatment with these solvents can be used if necessary to remove contaminants. In addition, γ-fluoro-L-threonine is used as a salt that is sparingly soluble in water due to its basic and acidic properties.
It can also be precipitated and collected using conventional methods. Examples of such salts include common salts of amino acids, such as metal salts such as sodium, potassium, calcium, magnesium, and aluminum, organic amine salts such as N,N'-dibenzylethylenediamine, and procaine, hydrochloric acid, and nitric acid. , inorganic acid salts such as sulfuric acid, organic acid salts such as formic acid, acetic acid, and lactic acid, sulfonic acid salts such as methanesulfonic acid, isethionic acid, and p-toluenesulfonic acid, and amine acid salts such as glutamic acid, aspartic acid, lysine, and arginine. Examples include. Furthermore, in order to obtain highly pure γ-fluoro-L-threonine, adsorption column chromatography using silica gel, alumina, activated carbon, etc. as a carrier, or gel using Sephadex (manufactured by Pharmacia) or Biogel (manufactured by Biorad), etc. The past is done. γ-Fluoro-L-threonine can be crystallized from water, but it is very soluble in water and almost insoluble in alcohol, so by dissolving it in a small amount of water and adding alcohol, γ- Crystals of fluoro-L-threonine can be obtained. The physicochemical properties of γ-fluoro-L-threonine thus obtained are as described above. Next, the biological properties of the new substance γ-fluoro-L-threonine will be described. Table 2 shows the in vitro MIC of γ-fluoro-L-threonine against various test bacteria.

【table】

[Table] The MIC of γ-fluoro-L-threonine is 50 μg/ml or more in nutrient medium, and its antibacterial effect is extremely weak, but in Davis minimal medium it is 0.2 μg/ml against many test bacteria. ml or less. By adding various amino acids individually to a synthetic medium, γ-fluoro-
Table 3 shows the results of examining the antibacterial activity of L-threonine.

【table】

[Table] When L-serine and L-threonine are added, γ
- Since the antibacterial activity of fluoro-L-threonine disappears, γ-fluoro-L-threonine is
It is an antimetabolite of serine and L-threonine. In mouse infection treatment experiments, E.
The PD ₅₀ was 77 mg/Kg against Coli, but it was ineffective against Streptococcus aureus. LD ₅₀ for DDY mice (♀) is 320mg/
Kg(iv). Subsequently, according to Examples, the target compound γ-
An example of the production of fluoro-L-threonine is shown below.
Since the properties of -fluoro-L-threonine have been clarified by the present invention, various methods for producing γ-fluoro-L-threonine can be considered based on these properties. Therefore, the present invention is not limited to the examples, and of course, modification means of the examples may be used, as well as known means based on the properties of γ-fluoro-L-threonine revealed by the present invention. production·
It covers all extraction, concentration, and purification methods. Example 1 A tube containing a freeze-dried culture of Streptomyces Cattleya NRRL8057 was opened aseptically, and the contents were
ml of sterile physiological saline, and then the whole amount was mixed with A medium (30 g/ml of Syucross, 15 g/ml of Daistainer's Soluble, 5 g/yeast extract).
g/, corn gluten meal 5g/, PH7.0)
Inoculate a 300 ml Erlenmeyer flask containing 60 ml. This type of flask is incubated at 28° C. for 72 hours in a rotary shaker at 220 rpm. This culture solution is mixed with a 10% glycerol solution at a ratio of 1:1, divided into tubes, and stored as frozen vegetative bacteria (FVM) in a -85°C freezer. Melt this FVM at 30℃ and add 1.2ml to 60ml of A medium.
Inoculate a 300 ml Erlenmeyer flask containing This flask was cultured in a rotary shaker at 220 rpm for 48 hours, and 2 ml of this seed culture was poured into B medium (30 g of glycerin, 10 g of Detailer's Solve, 23 g of corn steep liquor,
Cottonseed meal 7.5g/, Glycine 1.5g/,
CaHPO ₄・2H ₂ O0.5g/, CoCl・6H ₂ O0.002
The cells were transplanted into each flask containing 25 ml of a medium containing 25 ml of a medium containing 0.01 g/g/, MgSO ₄ .7H ₂ O 0.01 g/, PH 7.5) as a basal medium, and each component shown in Table 4. This flask was placed in a rotary shaker at 220 rpm.
After culturing at 28° C. for 6 days, the culture solution was centrifuged at 3000 rpm for 10 minutes, and the γ-fluoro-L-threonine content of the broth supernatant was measured. The relationship between the added components and the production of γ-fluoro-L-threonine is shown in Table 4. None of the added ingredients showed antibacterial activity in the above-mentioned measurement method at the amounts tested.

[Table] Example 2 Streptomyces cattleya NRRL8057
Inoculate 1.2 ml of FVM into a 300 ml Erlenmeyer flask containing 60 ml of A medium. this flask
Incubate at 28°C for 48 hours in a rotary shaker at 220 rpm. Add 10ml each of this first stage seed culture to A medium.
Transplant into 5 flasks containing 200 ml,
Culture at 28°C for 24 hours in a reciprocating pump at 170 strokes/min to obtain a second-stage seed culture. Collect this second stage seed culture and use A medium 130
Inoculate a 200 stainless steel tank containing
This tank was cultured at 160 rpm, 100/min, 28°C for 30 hours. Third stage seed culture solution obtained in this way 13
C medium (glycerin 20g/, Daystaylor's Soluble 20g/, corn staple liquor)
15g/, cottonseed meal 5g/, soybean casein 3g/
, Sodium succinate 1g/, CaHPO ₄・
2H ₂ O2.5g/, CoCl ₂・6H ₂ O0.01g/, KM
-750.5g/PH7.5) Transfer to a 200 stainless steel tank containing 130. This tank at 160rpm,
100/min, 30℃ (0→100h) then 28℃
(100→162 hours) for 162 hours to obtain a culture solution 120 containing 142 μg/ml of γ-fluoro-L-threonine. Add 4.8 kg of supernatant (Hyflo Super Cell) to 120 of the culture solution, and add 20 of the filtered solution to Dowex 1 x 2 (HCO ₃ ) 4
The solution was passed through the column and washed with water from Step 6. Amberlite IR-120 for active flow liquid and washing water meter 10.35L
Add (H)1.35, stir for 20 minutes, pack into a column, and add 2.6
After washing with water, elute with 2N ammonia water,
470 ml of active fraction was obtained. After concentration and acetone treatment,
11.1 g of brown-gray precipitate was obtained. 10.0 g of this was dissolved in 40 ml of water, passed through a 500 ml activated carbon column (activated carbon for Wako chromatography), and then developed with 2000 ml of water. The active fractions were combined and concentrated to dryness to obtain 2.45 g of solid. Amberlite after dissolving in 10ml of water
The solution was passed through a 225 ml CG-50 (NH ₄ ⁺ ) column and developed with water. 88 ml of the active fraction was concentrated to dryness, dissolved in 4 ml of water, added with 6 ml of ethanol, and allowed to stand at 5°C overnight to obtain 740 mg of γ-fluoro-L-threonine in the form of colorless needle-like crystals.
I got it. Example 3 The third stage seed culture 13 obtained in the same manner as in Example 2 was mixed with D medium (20 g of glycerin/, 20 g of daytailer's soluble/, 15 g of cornstarch liquor, 5 g of cottonseed meal/, Sodium succinate 1g/, KF0.12g/, CaHPO ₄ .
2H ₂ O2.5g/, CoCl ₂・6H ₂ O0.01g/, KM
Transfer to a 200 stainless steel tank containing -750.5g/, PH7.5) 130. this tank
Cultured for 114 hours at 160 rpm, 100/min, and 30°C, and 114 μg/g of γ-fluoro-L-threonine/
A culture medium containing 120 ml was obtained. Add 3 kg of supernatant (Hyflo Super Cell) to 120 L of culture solution,
Amberlite IRA-401S
(OH) The solution was passed through a 15 L column. After washing the column with 24 L of water, the active fraction obtained by elution with N-hydrochloric acid water was passed through a 16 L column of Amberlite IR-120 (H ⁺ ). After washing the column with 24 L of water, it was eluted with 2N aqueous ammonia, and the active fraction was concentrated under reduced pressure to about 1 L. Activated carbon concentrate 50%
After decolorizing with g, the solution was passed through a 3.7 L activated carbon column (purified Shirasagi for chromatography), and then developed with water. 1.17 L of active fractions were collected and concentrated under reduced pressure to obtain 54 g of oily syrup. This syrup was dissolved in 200 ml of water and passed through a 4.8 L column of Amberlite CG-50 (NH ₄ ⁺ ), and the column was developed with water. 720 ml of the active fraction was concentrated under reduced pressure to obtain 20 g of syrupy residue. Next, acetonitrile as a developing solvent:
Separation and purification was performed by silica gel column chromatography (Wacoal Gel W-200, 2L) using water (9:1:1). The γ-fluoro-L-threonine elution fractions were collected and concentrated under reduced pressure to obtain 8.02 g of pale yellow powder. This powder was repeatedly crystallized twice from an ethanol-water system to obtain 3.0 g of γ-fluoro-L-threonine as colorless needle-like crystals.

[Brief explanation of drawings]

FIG. 1 shows the infrared absorption curve of the novel γ-fluoro-L-threonine measured in potassium bromide tablets. Figure 2 shows the absorption curve of the ¹ H-NMR spectrum measured using the internal standard DSS in heavy water (90MHz).

Claims (1)

[Claims] 1 Formula: L-threo-2-amino-4-fluoro-3-hydroxybutyric acid or its salts. 2 L-threo-2 belonging to the genus Streptomyces
- Amino-4-fluoro-3-hydroxybutyric acid-producing bacteria were cultured in a fluorine compound-containing medium, and L-threo-2-amino-4-fluoro-
L-threo-2-amino-4-fluoro-3 characterized by separating and collecting 3-hydroxybutyric acid
- A method for producing hydroxybutyric acid or its salts. 3 L-threo-2 belonging to the genus Streptomyces
3. The production method according to claim 2, wherein the -amino-4-fluoro-3-hydroxybutyric acid producing bacterium is Streptomyces cattleya. 4 The fluorine compound used in the production medium is inorganic fluorine compounds such as ammonium fluoride, alkali metal salts of fluorine, alkaline earth metal salts of fluorine, or organic fluorine compounds such as monofluoroacetic acid, m-fluoro- DL-phenylalanine,
p-fluoro-DL-phenylalanine, 4-
Fluoroglutamic acid is used alone or in combination, and the amount used is relative to the culture medium.
The manufacturing method according to claim 2, wherein the content is 0.001 to 0.5% (W/V).