JPS5810078B2 - Novel 2,6,10-trimethyl-1-pentadecanol produced by microorganisms and its production method - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to norpristane represented by the structural formula
:2,6,10-triphthyl-1-pentadecane 2,6
・10-trimethylpentadecane)
Alcohol derivative 2,6,10-trimethyl-1-pentadecanol (2,6,10-trimethyl-
1-pentadecanol) and its production method.

2,6,10-trimethyl-1-pentadecanol is
Vitamin E by leading to ketones by chemical methods
and can be used as a raw material for the synthesis of homologs to vitamins.

Furthermore, since this compound itself is a higher alcohol, it is a useful substance that can also be used as a surfactant.

Norpristan is an isoprenoid hydrocarbon with many methyl side chains and is less easily decomposed by microorganisms than linear hydrocarbons, so there are few studies on it.

To date, the only example in which the production of oxidized derivatives of norpristan was observed by culturing microorganisms using turpristan as the main carbon source is the terminal oxidation of carboxylic acids, i.e., 2, 6, 10, by a strain of Mycobacterium. ~Production of trimethylpentadecanoic acid and 6,10,14-trimethylpentadecanoic acid (Lipids, vol, 11,
p72 (1976)) is known.

However, there is no known example of producing 2,6,10-)riftyl-1-pentadecanol having an alcohol group generated by oxidation of 2,6,10-trimethylpentadecane.

Therefore, the present inventors conducted research on a method of oxidizing norpristan using microorganisms, conducted a wide range of microorganism searches, and discovered a strain with such properties.
As a result of intensive research into a method for effectively producing 2,6,10-trimethyl-1-pentadecanol, the present invention has been completed.

That is, the present invention involves culturing microorganisms newly isolated by the present inventors from soil in a nutrient medium containing norpristan as the main carbon source, and producing novel 2,6,10-trimethyl-1 from the culture.
- A method for producing the derivative is provided, which comprises separating and recovering pentadecanol.

The strain BPM1613 (Feikoken Bacterial Serial No. 1609) belonging to the genus Nocardia, which the present inventors newly isolated from soil, is one example that can be most advantageously used in the method of the present invention. Its mycological properties are as follows.

The color display is based on the "Color Standard J" published by the Japan Color Research Institute.

A. Cell morphology As shown in the culture characteristics below, this strain exhibits a characteristic orange to pink color in most media.

Young vegetative cells grow like hyphae, and branching is rarely observed.

In old cultures, hyphal rupture occurs and bacilli (0.4 to 0.6
X1.8 to 2.4μ).

Durham staining positive.

No flagella. It is not stained by acid-fast bacterial staining using the Twill and Nielsen methods.

No epiphyte of aerial mycelia was observed. B. Properties on various media (1) Sucrose-nitrate agar medium (30° C.) Slight growth, colony color pink, no diffusible pigment.

(2) Glucose-asparagine agar medium (30°C):
No growth.

(3) Glycerin-asparagine agar medium (30°C):
Growth is weak, colony color pink, no diffusible pigment.

(4) Starch agar medium (30°C): No growth.

(5) Tyrosine agar medium (30°C): weak growth, colony color grayish white, no diffusible pigment.

(6) Nutrient agar medium (30°C): medium growth, colony color orange, no diffusible pigment.

(7) Yeast/malt agar medium (30°C): Good growth,
Settlement color orange, no diffusible pigments.

(8) Oatmil agar medium (30°C): Medium growth, colony color orange, no diffusible pigment.

(9) Calcium-maleide agar medium (27°C): Medium growth, pink colony color.

(10) Juicy agar slope (27℃): Medium growth, pale pink color of the village.

(11) Egg-albumin slope (27°C): Growth is weak, colony color white.

(12) Potato slice medium (27°C): medium growth,
The color of the village is pale orange.

(13) Carrot section medium (27°C): medium growth, pale pink colony color.

C0 Physiological properties (1) Growth temperature range (nutrient agar slope): 20-42℃ (2) Liquefaction of gelatin: Negative (3) Hydrolysis of starch: Negative (4) Coagulation/peptonization of skim milk: Negative ( 5) Lidomus milk: No change 6) Melanin-like pigment production: Negative (7) Nitrate ring group: Positive (8) L-arabinose, D-xylose, D-glucose, D-7 lactose, sucrose, D- Mannitol, glycerin, lactose, D-galactose, D
- No acid or gas formation from mannose, maltose, trehalose or starch.

(9) Catalase test: Negative (10) Production of indole: Negative (11) Production of hydrogen sulfide: Negative, assimilation of each carbon source (on Purido Mu Gotlieb agar medium, 30°C, 7 days) L -arabinose (+), D-xylose (10), D-
Glucose (++), D-Fructose (++), Sucrose (++), Inositol (+), L-Rhamnose (-), Raffinose (+), D-Mannitol (+
), (++): Moderate growth, (+): Slight growth, (-): No growth. This strain was also tested using the method of Arai et al. (Journal of
General Applied Microbiol
ogy, 9°119 (1963): The Actin
omycetales.

The Jena International Sy
mposium on Taxonomy1273(1
When we measured the infrared absorption spectrum of the bacterial cells cultured in Glycerol Kelner Morton medium according to 968), we found absorption bands characteristic of the genus Nocardia (■: C,
E type, ■: C type, ■: C type, ■: D type) were given.

The above properties are described in Bergey's Manual of Deter.
(minative Bacteriology) 7th edition or The Actinomycete by Waxman
When examined using Volume 2 of S., it was found that it belongs to the genus Nocardia.

Next, to describe the present invention in more detail, any carbon source can be used as the culture source as long as the above-mentioned norpristan is used as the main carbon source.

In addition, as nitrogen sources, potassium nitrate, sodium nitrate,
In addition to nitrates such as ammonium nitrate, ammonium salts such as ammonium chloride, ammonium sulfate, and ammonium phosphate, ammonia, or organic nitrogen compounds such as urea, peptone, and corn sweep liquor can be used.

In addition, as an inorganic salt, potassium phosphate, sodium phosphate, magnesium sulfate, iron, manganese, etc. are used as necessary. 3 Also, as a growth promoting factor, vitamins, amino acids, yeast extract containing them, corn steep liquor, etc. , wort, etc. can be used, but nothing is acceptable.

The pH of the medium is usually good on the alkaline side of 7 to 10.

A suitable culture temperature is 20 to 40°C, particularly preferably around 30°C.

The culture is carried out under aerobic conditions such as aerated agitation culture for 3 to 5 days.

In addition, the culture concentration of norpristan, which is the main raw material (main carbon source), may be at any concentration as long as it can be used by this strain, but it is generally used in the range of 1 to 10% of the culture solution. Ru.

After the cultivation is completed, the oxidized product can be extracted and collected depending on the substance's solubility in various solvents.

All known methods such as adsorption, elution, dissolution, and distillation based on this property can be used in the present invention.

One example is as follows: After extracting the culture with a solvent such as ether, benzene, or chloroform under acidic conditions,
The oxidation products can be fractionated and collected by applying the extract to a silica gel chromatograph and developing it with a solvent such as hexane, petroleum ether, benzene, acetone, methanol, ether, etc. alone or in combination.

If necessary, the same chromatography may be repeated for further purification.

According to the above method, a strain belonging to the genus Nocardia can be cultured in a medium containing norpristan as the main carbon source, and novel 2,6,10-trimethyl-1-pentadecanol can be isolated and collected from the medium. However, the main raw material (main carbon source), norpristan, is known as a component of petroleum and shale oil, and shale oil, which is attracting attention as an alternative energy source to petroleum in the future, has been developed and industrialized, and thiourea can be extracted from shale oil etc. If it can be extracted in large quantities by the adduct method, it will become possible to supply it at low cost, and it may become possible to economically produce the derivative by the method of the present invention.

The physical and chemical properties of 2,6,10-trimethyl-1-pentadecanol are as follows.

It is a colorless, neutral oily substance with a molecular weight of 270.50 (CI8H380) and soluble in ethyl ether, acetone, ethyl acetate, benzene, hexane, etc.

Infrared absorption spectrum of this compound (absorption characteristic of aliphatic alcohols having a side chain at the α position is 3330 cm-1
and 1031035C) are shown in Figure 1, 'H
-Nuclear magnetic resonance absorption spectrum (a δ3.42 doublet signal is observed) is shown in FIG.

In addition, the mass spectrum (Fig. 3) of the compound was measured, and M
10-H20252 was obtained.

From these results, this compound is a novel 2, 6, 10-
It was confirmed that it was trimethyl-1-pentadecanol.

The embodiments of the present invention will be described below with reference to Examples, but the present invention is not limited to the Examples.

Example Norpristan 3.0%, NaNO30.50%, KH
2PO40.15%, Na2HP040.15%, Mg
SO4・7H200,05%, FeSO47H200,
001%, CaCl22H200,001%, yeast extract 0.2%, pH 7.2, and shaken in advance at 30°C for 2 days with the same medium (but with 0.5% normal paraffin instead of norpristan). A seed culture solution of a strain belonging to the genus Nocaldea (BPM1613, Kaikoken Bacteria No. 1609) obtained by culturing was inoculated at a ratio of 8% (volume), and this was spread over 500 m
Shaking culture was performed at 30° C. for 5 days in a 1-volume shoulder flask.

After completion of the culture, the culture was extracted with ether under acidic sulfuric acid (PH2), and after distilling off the solvent, it was applied to a silica gel column, and hexane and benzene were eluted in this order.

2,6,10-trimethyl-1-pentadecanol is
Norpristan was eluted followed by benzene, yielding 320 m9 from 11 cultures.

[Brief explanation of drawings]

Figure 1 shows the infrared absorption spectrum of 2,6,10-trimethyl-1-pentadecanol, and Figure 2 shows the infrared absorption spectrum of 2,6,10-trimethyl-1-pentadecanol.
Figure 3 shows the mass hector of 2,6,10-trimethyl-1-pentadecanol.

Claims (1)

[Scope of Claims] 1 2,6,10-)rifthyl-1-pentadecanol represented by the structural formula. 2. 2.6. Using a microorganism characterized by culturing a strain belonging to the genus Nocardia in a medium containing norprimemethane as the main carbon source, and separating and collecting 26,10-trimethyl-1-pentadecanol from the medium. 10-trimethyl-
Method for producing 1-pentadecanol.