JPS6038116B2 - Method for producing phenylphenols - Google Patents

Description

[Detailed description of the invention] The present invention relates to a method for producing phenylphenols.

O-phenylphenol is useful as an antifungal agent, and m-phenylphenol is a substance useful as an intermediate raw material for agricultural chemicals.

Chemical synthesis methods and fermentation methods are known as methods for producing these phenylphenols.

For example, Tokusekki No. 49-42652 and No. 49-35365 are known as methods for chemically synthesizing o-phenylphenol, and U.S. Patent No. 3,363,008 is known as a method for chemically synthesizing m-phenylphenol. However, both have the disadvantage that the process is complicated. On the other hand, as a fermentation method, Zuglea ramigera (Znug1 female arem
Pseudomonas butida (Pseudomonas butida)
How to use Expe
nmentia, 27, 1173 (1971)), but none of them have a sufficient yield.

The present inventors have developed a method for producing phenylphenols that eliminates the above-mentioned drawbacks.
We investigated the use of biphenyl, which is produced in large quantities during oil refining.

First, after searching for biphenyl-assimilating bacteria in the soil of various places, biphenyl conversion products were isolated from the cultures of these assimilating bacteria, and their structures were determined. As a result, Pseudomonas (PseMom) was isolated from soil in southern Chiba Prefecture.
Onas) It was discovered that a new strain of counterfeit bacteria produced a significant amount of phenylphenol, and the present invention was completed. The present invention provides a method for producing phenylphenols, which comprises culturing Pseudomonas SOLO4 needles in a medium containing biphenyl, and collecting phenylphenols from the culture. In the present invention, o-phenylphenol and m-phenylphenol are produced by fermentation, and it is possible to collect and purify them. The phenylphenols-producing bacteria used in the method of the present invention may be any strain as long as it has the ability to assimilate biphenyl and produce a sufficient amount of phenylphenols to be collected into the culture. Pseudomonas SOIO43 Vermilion, which the present inventors isolated from soil in southern Chiba Prefecture, is suitable.

The mycological properties of Pseudomonas SOIO43 beads are as shown below. a Morphological properties {1} shape Acampus size 0.5 to 0.6 '5-Gram staining negative'6} Acid-fastness None b Cultural properties [1} Broth agar plate: Poor growth, circular, surface smooth, with ridges, surrounding all green, content homogeneous, yellow, moist light (21) Broth agar plate: Poor growth , Thread-like paste gravy liquid Poor growth, slightly cloudy, no film, some sediment [4) Meat juice gelatin puncture Poor growth, no liquid '5} Litmus milk Acidic, litmus reduction, unchanged c
Physiological properties '1} Reduction of nitrate None '2} Denitrification reaction Negative {3'M old test negative ■ VP test Negative '5} Indole formation None Tsukuda Hydrogen sulfide generation weak '7} Starch hydrolyzate None' 8) Citric acid availability None' 9) Ammonium salt, yes Nitrate availability 'IQ pigment formation None (11) Urease negative (12) Oxidase positive (13 Catalase negative (1 heart growth temperature (°C)) 15-41 ( Optimal 30-37)
PH 5.1-8.3 (optimal 6-7.5) (15 Attitude towards oxygen Aerobic (16) ○-F test Oxidative (17 Generation of acid from boiling substances See table below and generation of gas No formation Table-1: pH 6.2-6.8 (18) Utilization of carbon compounds Refer to the table below-2 Based on the above mycological properties, the Manual of Determination of Purge Itaminative bacteriology −
(Bargey's Manual of De
te plate inative Bacteriology) No. 7
As a result of examining the differences with known strains according to the standards of the 8th edition and the 8th edition, it was recognized that it belongs to the genus Pseudomonas, but no similar strains were found within the same genus.

Based on the above results, the present inventors recognized this strain as a new strain and named it Pseudomonas SOIO43. This strain is kept as Pseudomonas SGI043 beads (Choken Bacteria Collection No. 4471) at the Institute of Microbial Technology, Agency of Industrial Science and Technology.
For culturing the microorganisms used in the present invention, carbon sources commonly used as culture medium components can be used, but it is optimal to use piphenyl as the sole or main carbon source.

The nitrogen source is not particularly limited, but potassium nitrate, ammonium nitrate, ammonium sulfate, ammonium chloride, ammonium phosphorus, polybeptone, etc. can be used. Also, as inorganic salts, disodium phosphate,
Potassium phosphate, etc. can be used, and magnesium sulfate, calcium chloride, ferrous sulfate, etc. can be used as the collecting metal. Furthermore, a surfactant, an antifoaming agent, etc. may be added as necessary. Appropriate culture methods include methods using a liquid column, such as the shake-face culture method and the deep aeration strain culture method. It is appropriate that the culture temperature is 25 to 35 oo, the culture pH is around neutral, and the number of days of culture is 2 to 5 days. After completion of the culture, the accumulated phenylphenols can be extracted and purified from the solution obtained by operations such as centrifugation.

For purification, solvent extraction, column chromatography,
Means such as neutralization, concentration, and crystallization can be carried out alone or in appropriate combinations. The purification method and physicochemical properties of phenylphenols are described below.

B. Purification method After the culture is completed, the culture solution is centrifuged to obtain a supernatant fraction, and after adjusting the pH to 7, extraction is performed with a solvent such as ethyl acetate.

After drying the solvent layer with Na2S04, it is concentrated to dryness. The residue was subjected to silica gel column chromatography and eluted with benzene. As a result, it is divided into o-phenylphenol elution fraction and m-phenylphenol elution fraction,
After each fraction is concentrated to dryness, it is recrystallized in n-hexane to obtain phenylphenols having the physicochemical properties shown below. (b) Physical properties {a).

-Phenylphenol '1' Elemental analysis value Actual value C: 84.7%, H: 5.9%, N: 0% Theoretical value C: 84.7%, H: 5.9%, 0:9. 4%■
Molecular weight mass vector m/e maximum value 170 Theoretical value 170 '3} Infrared absorption spectrum (KB Yodase method) absorption peak (arc-1) 3520 (1OH), 15851195 750 {b) m-phenylphenol' 1) Actual elemental analysis values C: 85.8%, H: 5.8%, N: 0% Theoretical values C: 84.7%, H: 5.9%, 0: 9.4%'2
} Molecular weight mass vector m/e maximum value 17o Theoretical value 170 '3} Absorption peak (inhibition 1) of infrared absorption spectrum (KBr tablet method) 3320 (1OH), 1430, 1100, 755 Based on the above analytical data, the following It was confirmed that they were 0-phenylphenol and m-phenylphenol shown by the structural formulas.

The present invention aims to make effective use of biphenyl, which is an unused fraction of petroleum, and moreover allows the target phenylphenols to be easily produced. It can be produced with high yield through a process that is easy to use.

The phenylphenols obtained by the present invention are useful as antifungal agents, concentrated drugs, and intermediate raw materials for pharmaceuticals. Next, the present invention will be explained in detail with reference to examples. Example 1 Pseudomonas bacteria SGI043 beads (Choken Bacteria No. 4471) were mixed with 2% piphenyl and 0 ammonium nitrate.
．． 2%, disodium phosphate (IZk salt) 1%, potassium phosphate 0.55%, magnesium sulfate (heptahydrate) 0
．． 02%, calcium chloride (dihydrate) 0.001%, ferrous sulfate (heptahydrate) 0.0001%, yeast extract 0.0
1%, corn staple liquor - pH containing 0.01%
7.0 medium 50 secretion (500' modified flask with protrusion)
The cells were plated and rotary cultured for 3 days at 30 qo.

After the culture was completed, the culture solution 420' was centrifuged, and the obtained sterilized culture solution was used as a vessel 7, and then extracted with ethyl acetate. The ethyl acetate layer was dehydrated with sodium sulfate and then concentrated to dryness.
The resulting residue was subjected to silica gel column chromatography and eluted with benzene. Through this elution operation, o-phenylphenol and m-phenylphenol are eluted in this order. After each fraction was concentrated, it was recrystallized from n-hexane. In this way, a mixture of o-phenylphenol 410 and P'm-phenylphenol 130 was obtained in crystalline form.

Example 2 The same procedure as in Example 1 was repeated except that potassium nitrate was used instead of ammonium nitrate.

When the production amounts of 0-phenylphenol and m-phenylphenol were measured by gas chromatography, the former was 1300 females per evening and the latter was 410 females per evening.

Claims (1)

[Claims] 1 Pseudomonas strain SG1043 (Feikoken Bacterial Collection No. 44
71) in a medium containing biphenyl, and collecting phenylphenols from the culture.