JPS60214894A - Method of epoxide production from cycloolefin utilizing microorganism - Google Patents

Abstract

PURPOSE:A microorganism in Nocardia, capable of producing epoxide is allowed to act on cycloolefin in the presence of a water-insoluble organic solvent under aerobic conditions to enable high-yield production of the corresponding expoxide. CONSTITUTION:A microorganism in Nocardia, capable of producing epoxide, is allowed to a cycloolefin in the prescence of a water-insoluble organic solvent selected from paraffins of 5-17C, olefins of 10-18C, halogenated paraffins of 9- 16C and alkylbenzenes bearing side chains of 6-15C under aerobic conditions to produce the corresponding epoxide. The resultant epoxide is isolated and collected.

Description

DETAILED DESCRIPTION OF THE INVENTION Field of Industrial Application The present invention relates to a method for producing a corresponding epoxide from a cycloolefin using microorganisms.

Epoxide compounds are used for synthetic resins, surfactants, pharmaceuticals, yk
It is widely used as a raw material or intermediate for the production of millet and other organic chemical products.

The method of producing epoxide using microorganisms is as follows:
Genus Nocardia + Myobacterium
Genus + Methylococcus Genus - Methyl
osinulI@, Pseudomonag genus.

Corynebaaterium genus @Msthylob
aot@r1um IA or Candlda genus g'3
It is known that epoxides from linear olefins or aromatic olefins are produced by microorganisms such as
The reproduction of corresponding cyclic epoxides from cycloolefins has not yet been reported.

OBJECTS OF THE INVENTION The present invention was made based on the knowledge that epoxide-producing bacteria belonging to the genus Pycnocardia, which are endemic microorganisms, can also produce d-type epoxides from cycloolefins. The present invention aims to provide a method for preparing epoxides corresponding to cyclorefins for FIJ using microorganisms belonging to the genus Nocardia and having epoxide-producing ability.

I will take a closer look at this investigation.

Structure of the Invention The exclusive enemy of the structure of the present invention is the epoxide-producing group belonging to the genus Nocardia.
w Prompt l'a punishment 4-r: In F, react with cycloolefin under aerobic conditions to produce the corresponding epoxide,
The purpose is to separate and collect the obtained epoxide.

The microorganism used in the present invention belongs to the genus Tucartia, and may be exemplified by J. corallina.

This bacterium is FE in the Agency of Industrial Science and Technology's Microbial Technology 1νI兇Rr.
With the acceptance of RM-P-4094, Showa 1” 1952 6
the law of nature! The mycological properties of the ingredients are described in detail in Japanese Patent Publication No. 1983-40.

In the present invention, the cycloolefin (hereinafter referred to as F raw material olefin) used as a reaction substrate for producing an epoxide tank using the above-mentioned microorganisms has a carbon number of 6 to 8.
cycloolefins, namely cyclohexene, cycloheptene, and cyclooctene.

and alkylated versions thereof and cyclopentenes, such as l-methylcyclopentene.

Examples include 1-methylcyclohexene, 3-methylcyclohexene, 4-methylcyclohexene, etc.

In the present invention, these raw material olefins may be used alone or as a compound as a reaction substrate.

In the present invention, the waterwheel-based organic fill (hereinafter simply referred to as organic fill) to be present when carrying out epoxidation using microorganisms listed in the above-mentioned family IIX family, oleophylphyte (hereinafter simply referred to as "organic fill"), has a charcoal number of 9 to 9. Paraffin having 17 carbon atoms, carbon number 10 to 18, IT T olefin, carbon number 9
A carefully selected organic solvent consisting of a halogenated paraffin having a chain length of 6 to 16, and an alkyl benzene having a side chain of 6 to 15 in chain length; Alternatively, it can also be used as a mixture of 24i1 or more.

To congratulate these organic lubricants, the number of carbon atoms is 9.
~17 (l-9 of paraffin with 2)
rn is about 20-25% of the kerosene fraction of petroleum.
1 (i.e., the fraction with a temperature of about 160°C to 350°C is hydrodesulfurized, separated and recovered using zeolite (molecular sheep), etc.) 6 is commonly used as a raw material for soft detergents.

Among the upper tr2ha5 fins, those with a large number of carbon atoms have a higher effect of promoting epoxidation, and those with 12 to 16 carbon atoms are particularly preferred. Incidentally, if the number of carbon atoms is less than 9, the promoting effect of epoxidation cannot be observed, and even if the force is greater than 17, the promoting effect decreases, and in addition, it will solidify at room temperature, which is not practical. . In addition, the isoparaffins of the upper self-paraffins coexist with the normal paraffins in the above-mentioned fraction6, and can be separated from the normal back-in by IE distillation, but they cannot be actually removed. It is convenient to use it as a mixture with normal paraffin.In addition, those with short chain lengths such as methyl and ethyl side chains are common, but isome2fins having 12 to 16 carbon atoms are epoxidized. Favorable for promotion.

Incidentally, the olefin having 1 t3 to 18 carbon atoms may be a low polymer or oligomer C of propylene or butylene, and those commercially available as sample millet may also be used. Generally, it is a linear or less branched monoolefin.

However, olefins with fewer than 8 carbon atoms do not have the effect of accelerating epoxidation, and those with more carbon atoms have a lower effect, and in addition, the viscosity decreases, making them impractical.

The 3-halogenated paraffins having the -C-containing system Nos. 9 to 16 as Itokiichi agents include chlorinated LIQ U VC, brominated paraffin, denyl chloride, JA towndecyl, dodecyl chloride, and tottesyl chloride. Mushibi Tetradecyl.

Decyl bromide. Embodies unnon, dodecyl bromide, tetradecyl bromide. Includes decyl bromide.

In addition, the number of carbon atoms is 9. Even if c9 is less or more than 16, the effect of promoting epoxidation cannot be seen.

For a long time, argylbenzene having a chain length of 6 to 15 has been used as an intermediate for ymN hard or soft detergents. It has it in the side chain.

If the chain length is outside the above range of 6 to 15, the effect of promoting epoxidation is either nil or so low that it is not practical.

In the present invention, by allowing the above-mentioned Taniari+S solvent to exist in the system, it becomes possible to collect the epoxide as shown in the examples below.

In the present invention, in order to cause the epoxide-producing bacteria belonging to Nocardia camphorina to act on the raw material olefin in the presence F of the organic m agent, for example, (a) a Streptococcus spp. A method of reacting by aerobically contacting with the presence of an organic m-agent, (b) combining the above reaction mixture with the raw material olefin and the 4R machine bath agent, A. The system has been repeated for a long time, and the history includes a method of adding growth-promoting substances and cultivating it under aerobic conditions in nutrient-rich soil. , 3- for ia
It is good.

In the method of (a) above, in which the 4a bacterial cells are brought into contact with the raw material olefin in an aerobic manner in the presence of an organic solvent,
First, as a carbon source 4) A'JIj, for example, glucose.

Sucrose, ammonium chloride, starch hydrolyzate, carbohydrates such as propane, butane, dodecane, tetradecane, and other substances with high bacterial growth effects such as acetic acid are used, and ammonium tetraride, ammonium sulfate, and , Phosphorus+! j
! Ammonium, ammonium nitrate.

Nitrogen compounds such as urea, aqueous ammonia, amino acids and other assimilated organic compounds, potassium phosphate.

Ren 1 - skin sodium, 1 accent magnesium, manganese sulfate + 1Ilfc g ferrous, ferric chloride, calcium chloride.

Epoxide-producing bacteria of the genus Nocardia are added to a medium containing focal salts such as manganese chloride and, if necessary, growth-promoting substances such as vitamin M, mother extract, and corn staple liquor. The microorganisms are inoculated and cultured under aerobic conditions F to proliferate the microorganisms.

To the bacterial cell culture thus obtained, a suspension of bacterial cells isolated from the culture, or an immobilized bacterial cell,
The raw material olefin and the organic solvent are all added together, and air, oxygen, and an oxygen-containing gas such as an oxygen-enriched gas are supplied for reaction.

The ratio of the raw material olefin and organic solvent to the bacterial cell-containing aqueous acid such as the bacterial cell culture solution or bacterial cell suspension may vary depending on the type of organic solvent, but the raw material olefin C is usually 0. 1-25 vol/voL%, preferably 0.5-5 vot/vot%, and 1-200 vat/voL% for organic solvents.

Preferably it is 10-100 vot/maoL%.

The reaction... has a pI of 5-9, preferably 6-8 (in the region 20
The treatment is carried out at a temperature of ~50°C, preferably 25-45°C, for 1-6 days. The reaction can be carried out at a trace concentration pressure or by carrying out an increased pressure F to improve the productivity of epoxide. In addition, during the reaction, the bacterial cells were present in I14, q.
The epoxide production 72M activity of the bacterial cells can be maintained or increased by appropriately adding a water source and other components. The reaction can be carried out either in 11 J component force mode or in continuous mode. In the case of a batch reaction method, the raw material olefin can be fed in its entirety at the start of the reaction, or it can be fed continuously or intermittently during the reaction.

In the above reaction, the epoxide C4) produced is mainly present in the aiso bath agent phase, so it can be separated and collected by applying the phase fraction nW and extraction method for distillation.

Next, the method according to J-on 4 of 14iJfj Yamaguchi) is a method in which all the raw material olefin and organic solvent are added during the bacterial growth in method 2 above, and epoxide is produced in one step. Bottle conditions (pH, temperature, pressure, etc.) The reaction conditions, reaction force formula, and separation and collection method described in (a) above can be similarly used for the culture force formula and the separation and collection of the produced epoxide.

The epoxide obtained by the present invention can be used in a wide variety of conventionally known applications as mentioned above.

Hereinafter, the present invention will be explained in more detail with reference to Examples.

Example I Nocardia aorallna B-276 (
Institute of Microbial Technology, Agency of Industrial Science and Technology Deposit number FIRM-P
-4094), 3 platinum loops of 1NBG medium (Oxiid Co., Ltd. 6 Lapurenko "Pauta"-' 110g, Bacteriological Begton e lo L glucose 10), 5g of sodium chloride, tap water added, bound at 1000, and IN caustic soda aqueous solution. After setting the temperature to 7.5 degrees, the liquid medium J100-, which had been heat-sterilized in an autoclave at 120°C for 15 minutes, was inoculated into a Sakalo flask with a volume of fc500-, and then heated at 30°C.
It was incubated for 48 hours (150 times/min). The bacterial cells produced by this cultivation were added to 0.01 mol of enemy buffer (
After washing once with a reaction medium (pH 7.5) and then once with a reaction medium shown in F, resuspend in the reaction medium shown in F so that the dry cell mass is 3.3 m/-. 1 suspension was prepared.

To the reaction medium *HPO+ 1.74 g 1 SO, 7 to 20 L, 50 (I FeSO4@7&0 50 m9 deionized water 1 t pH 8,0 (PI (adjusted with 2N aqueous solution) as above) 20ml of the obtained It bacterial suspension, 0.1ml of the various raw material olefins shown in Table 1, and 2x of each organic solvent shown in Table 1 were placed in a 500ml flask and the flask was stoppered. 9 Next, culture was carried out at 30°C with reciprocating shaking at 150 times/min, and 501 nt of ether was added to the flask during the 24-hour period, and the ether extract was analyzed. ~100 mesh filled inner diameter 3 pieces, length 2
A Hitachi model 163 ionizing flame gas chromatograph equipped with a glass column of 200 m was used. The products formed during the reaction were measured using a gas chromatograph, and the retention time and 'J! The mass spectrum measured by the amount of L analysis was compared with the retention time and mass spectrum of a standard sample, and furthermore, it was confirmed that the product was hydrolyzed under hydrochloric acid acidity, and it was confirmed that it was the corresponding epoxide.

Table 1 shows the amount of epoxide produced corresponding to the type of W, raw olefin and organic solvent. The amount of epoxide produced was determined by gas chromatography.

In addition, as a comparative example, the results were similarly analyzed for cases in which the above organic solvent was not added and cases in which other organic solvents were used. At the same time, the first letter was added.

As seen in Table 1, it is possible to significantly improve epoxide aggregation by including a specific chemical agent in the system according to the present invention.゛Example 2 In Example 1, except that 2 to 20 ml of the normal paraffin mixture separated from the gas oil fraction was used as the raw material A reflex/chlorohexene total 0.57 L and 41 U g agent. The reaction was carried out with one ridge of the same hand as described in Example 1, and the result was also! The resulting product was analyzed in the same manner as in Example 1. The results are shown in Table 2.

As an example of the rx#-1 ratio, Table 2 also shows the results of a similar analysis in the case where the above-mentioned normal paraffin mixture was not added.

g2 Table 11, l:) n-CH~20 wt%+n Cu~
28wt%.

n-C1B~25w%+nC14~l8wt
9f; *Rho CI 3-5 wt%, J)-yohi n C
16-4wt% IMF≧I'm happy to see that it's the second garment. [In addition, it is necessary to have the Inno system in the system, so that the generation of Liebokizod will be directed towards the person.

1・1・'X1i)\ Japan 1 Koji 51 Song Dynasty Ceremony
, Embedded 'January Hatake, Hiro Toyote, Nikoichi Shochu 2, 1982 [January 411, Mr. Kazuo Wakasugi, Commissioner of the Patent Office ■, Indication of the case, 1982 Patent Application No. 7195 t1 No. 2, Title of the invention: Microorganisms Method for producing epoxide from Tsuku I-1 oresoin using 3, relationship with the case of the person making the amendment Patent applicant name: Japan Mining Co., Ltd. 4, agent address: 2-7-7 Higashi-Shinbashi, Minato-ku, Tokyo Shinbashi Kokusai Building 5, the day of the amendment order.'' - 6, Supplement 7, Supplement) 3. Supplement I [Details of Contents P: shall be amended as shown in F.

(1) In Table 150, 14-methylhexene-1 is corrected to 14-methylhexene-1 for the highest Fti in the ``Raw material olefin'' column to 1.4-methyl-11-hexene.

Claims (3)

[Claims] (1) Microorganisms belonging to the genus Tucartia that have an epoxide-producing ability were allowed to act on cycloolefins under aerobic conditions F in the presence of a water mill bath agent to produce the corresponding epoxide. , the obtained epoxide was divided into 1 t.
, Take II! v1 Blue method of making epoxide with all the characteristics. (2) Water-insoluble Mla bath additives include paraffins having 9 to 17 carbon atoms, olefins having 10 to 18 carbon atoms, halogenated paraffins having 9 to 16 carbon atoms, and chain lengths of 6 to 15 carbon atoms. 15 selected from the group consisting of argylpenocene having -+- side chains. (3) Epoxide raw Mr+B belonging to the genus Tucartia
The microorganism with
dla coralllna).