JPS5929177B2 - Manul derivatives and tobacco flavor improvers comprising the derivatives - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a novel compound manur derivative and a tobacco flavor improver comprising the derivative.

In recent years, the preference for cigarettes has been shifting towards lighter-tasting, richer-flavored tobacco products.As a result, the leaf tobacco used in product cigarettes is often lighter in flavor and contains less nicotine. I'm getting used to it.

However, such raw leaf tobacco generally lacks aroma and flavor, so it is necessary to add various flavoring agents to improve the aroma and taste of the product. On the other hand, research is being conducted to produce flavorants suitable for such purposes through microbial transformation of certain compounds, such as:
Cigarette flavors produced by microbial conversion of ionone compounds include JP-A-53-12498 and JP-A-53-107482.
No. 53-107498, etc.

From this point of view, the present inventors conducted research with the aim of obtaining a useful tobacco flavoring agent by converting manur with microorganisms, and found that it was possible to apply a certain type of microorganism to manur under certain conditions. As a result, we have discovered a new compound that is extremely effective as a converting substance in improving tobacco odor and suppressing irritation, leading to the present invention. That is,
The present invention relates to a compound represented by the following formula (I) (wherein R is -CH
Represents O or -COOH.

) and a tobacco flavor improver comprising a manur derivative represented by the above formula (1). The compound represented by formula (1) includes the following two types of compounds depending on the difference in R.

R-1 CHO: 13-beta-hydroxylafda-8
(17)・14-diene-18-r. (13-β-
Hydroxy-1abda-8(17)・14-Di
ene-18-al) (hereinafter referred to as compound (1-1)).

) R=-COOH: 13-betahydroxy-rhabda-8(17). (13-β-HydrOxy-1abda-8(17
)・14-Diene-18-01cacid) (hereinafter referred to as compound (1-2)).

) The manur used as a substrate for microbial conversion in the present invention is a known compound represented by the formula (I
ne (Daclydium bifOrme)], and is known as a synthetic raw material for fragrances.

Next, the method for producing the compound of the present invention, ie, the method by microbial conversion of manur, will be explained step by step.

First, the JTS-162 strain (Feikoken Bacterial Serial No. 5702) was inoculated into a medium containing manur and cultured aerobically at 28°C. After one day, a chelate inhibitor such as αα7-dipyridyl was added and further grown at 28°C. Perform aerobic culture. After the converted culture solution is extracted with an organic solvent such as ethyl acetate or ethyl ether, the solvent is distilled off under reduced pressure to obtain a converted product. Compound (1-1) and compound (1
-2) respectively. The microorganism JTS-162 used in the present invention is a microorganism isolated from the soil in Yokohama city, and its mycological properties are as follows.

1. Morphological properties (1) It is a bacillus, and the cell morphology changes as the culture progresses.

At the beginning of culture, cells elongate and branch. Culture 12
In ~14 hours, the cells undergo irregular division and then become rod-shaped. The size is (0.6-0.8) × at the early stage of culture.
After (5-15) μm division, (0.6-0.8)×(0.
8 to 1.8) μm. (2) Gram staining: Positive (3) Spore acidity: Positive (4) Spore forming ability: None (5) Motility: None 2. Chemical composition analysis (1) The main amino acid constituting the cell wall is MesO-diaminopimelic acid.

(2) The content of guanine + cytosine in DNA is 62,
It is 5 mol%.

3. Culture findings (1) Broth agar plate culture (28°C, 6 days incubation): growth is rather slow, colony shape is circular, diameter is 1.5-2 mm,
The periphery is wavy, the surface is smooth, and the color is white, changing color as the culture progresses.

The color of the medium does not change. (2) Meat juice agar slant culture (28
℃, 4-day culture): Growth is rather slow.

The shape and color tone are the same as those for broth agar plate culture. (3) Meat juice liquid medium (cultured at 28°C for 6 days). The medium is not very cloudy.

A fungal film slowly forms on the surface and then settles to become sediment. When cultured with shaking, it shows uniform growth. (4
) Meat juice gelatin puncture culture (28°C, 6 weeks culture): No liquefaction.

Bacterial cells grow on the surface. (5) Litmus milk (28℃,
Cultivation for 6 weeks): Produces acid slowly.

4 Physiological properties (1) Growth conditions: 20-30℃ is the optimum temperature for growth, pH is 6
．． The optimum value is 5 to 8.5; it cannot grow under anaerobic conditions.

2 Auxotrophy: None 3 Nitrate reduction: Positive 4 Starch hydrolysis: None 5 Citric acid utilization: Positive 6 Urease: Positive 7 Oxidase: Positive 8 Catalase: Positive 9 Pigment production: None C[0) O- F test: Fermentative (auto) methyl red test: Negative (12) V-P test: Negative (auto) Production of indole: None A4) Production of acid and gas from the following sugars (auto) The following compounds are converted to carbon L-alanine, paraffin, pyruvate grown as a source of 10 pionic acid.

(auto) Degradation of esculin: Positive (auto) Degradation of Tween 60: Positive (auto) Decomposition of tyrosine: Negative (auto) Assimilation of avienol, sclareol, and manul: From the results above, Virgies Manual.
Of Determinative Bacteriology (Ber)
geyεManualOfDetermina. tiv
eBacterlOlOgy), 8th edition (1974)
Based on this, the present strain JTS-162 was identified as Nocardia restricta.

Next, a manufacturing example will be described in more detail. Production example Glucose 1.0%, Monosodium glutamate 0.5%,
K2HPO4O. l%, MgSO4・7H200.02
%, KClO. A MSG slant medium (PH7.2) consisting of Ol%, yeast extract 0.5%, and agar 1.5% was prepared in a test tube, and the JTS-162 strain was inoculated into it.
The cells were cultured at 28°C for 3 days and used as a seed culture.

Then, (NH4)2S042y. K2HP0427,
MgSO4・7H200.27, CaCl2・2H20
0.0017, FeSO4・7H200.017, H2
A liquid medium (PH 7.0) consisting of Oll was placed in a 31-volume Erlenmeyer flask and sterilized at 121°C for 15 minutes.

Add Manul (commercial product) 1y and 1% Tw to the sterilized medium.
10 mj of an aqueous solution of een6O (surfactant, manufactured by Kanto Kagaku Co., Ltd.) was added. JTS-1 for 1 bottle of MSG slant medium
The inoculum of 62 strains was added to 5 mj of sterilized physiological saline (0.8
%, W/V) and the aforementioned liquid medium 11? ．． was inoculated. 210 rpm 128℃ using a rotary shaker
Cultured for 24 hours with αα5 as a chelate inhibitor.
-Dipyridyl was added at a concentration of 10mM/ml and cultured for an additional 48 hours to obtain a conversion product. Compound (1-1) and compound (1-2) were obtained from this culture. That is, 500 m of ethyl acetate was added to the culture at a time.
The mixture was added one by one and extracted with stirring twice.

The extracts were combined and the solvent was distilled off under reduced pressure to obtain a conversion of 0.857. Next, a column was prepared using 507 silica gel (manufactured by Mallinckrodt, 100 mesh), and the conversion product was eluted with hexane:ethyl acetate (6:4, V/V), and collected in 8 ml portions using a fraction collector. . The fractions containing each compound were purified again by the silica gel column treatment described above. The solvent was distilled off to give compound (11) 0.117 (yield 11%) and compound (1
-2) was obtained in 0.10y (yield 10%). The results of spectrometry of each compound were as follows. Compound (1-1): Compound (1-2): Molecular formula: C2OH32O3 To use the compound of the present invention as a tobacco flavor improver, dilute it to an appropriate concentration with a solvent such as ethanol or ethylene glycol, and prepare the product. 0.01-30 for tobacco raw material
The effect is exhibited by adding ppm (w/w).

The types of tobacco to which the compounds of the present invention can be effectively applied are not particularly limited, and are effective not only for tobacco obtained through cultivation, but also for recycled tobacco, pipe tobacco, etc. manufactured from tobacco scraps. . EXAMPLES Hereinafter, the effects of the present invention will be specifically explained with reference to Examples.

Example 1 Compound (1-1) or Compound (1
-2) After dissolving 0.5 watts of each in 3 ml of ethanol and spraying and adding them, roll them into paper, and compare the rolls of the above-mentioned shredded tobacco without the addition of this compound. The results were compared using the two-point discrimination method.

Claims (1)

[Claims] 1. A manul derivative represented by the general formula (I). ▲There are mathematical formulas, chemical formulas, tables, etc.▼ (R in the formula is -CHO
, or -COOH. ). 2. A tobacco flavor improver comprising a manur derivative represented by the general formula (I). ▲There are mathematical formulas, chemical formulas, tables, etc.▼(I) (R in the formula represents -CHO or -COOH).