JPS5936638A - (12z)-labda-12,-14-dien-17alpha-oic acid and agent for improving taste and flavor of tobacco composed thereof - Google Patents

Abstract

NEW MATERIAL:(12Z)-Labda-12,14-dien-17alpha-oic acid of formula I . USE:An agent for improving the taste and flavor of tobacco. It is extremely effective to improve the taste and flavor of tobacco and suppress its irritation. The improving effect can be achieved by diluting the compound with a solvent such as ethanol, and adding to the raw material of tobacco in an amount of 0.01- 30ppm (W/W), preferably 0.1-1ppm. CONSTITUTION:A microbial strain JTS-162 (FERM-P No.5702) is inoculated in a medium containing abienol, and cultured aerobically at 28 deg.C. After about 12hr, a chelate inhibitor such as alpha,alpha'-dipyridyl, etc. is added to the medium, and the cultivation is continued. The preferable conversion effect can be attained by the cultivation for about 45-50hr. After the conversion, the culture liquid is extracted with an organic solvent, and the solvent is distilled off to obtain the conversion product, which is eluted and fractionated with a solvent using a silica gel column to obtain the compound of formula.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention provides a microbial transformation of avienol (1
The present invention relates to 2Z)-labda-12,14-diene-17α-oic aracid and a tobacco flavor improver comprising the compound.

In recent years, the preference for cigarettes has been shifting towards products with a lighter flavor and richer aroma, and with this trend, the leaf tobacco used in product cigarettes is often lighter in flavor and contains less nicotine. It's starting to happen. 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 flavorings suitable for such purposes by microbial conversion of certain compounds. 53-107
The description can be found in No. 482, No. 53-107498, etc.

From this point of view, the present inventor conducted research with the aim of obtaining useful tobacco barbs by converting avienol with microorganisms, and found that by applying a type of microorganism to avienol under certain conditions, As a converted product, we have discovered a novel compound that is extremely unique in improving tobacco flavor and taste and suppressing irritation compared to avienol, and have accomplished the present invention.

That is, the present invention is an odor improver for tobacco, which comprises a compound represented by the following formula (1) and a compound represented by the above formula (1).

The compound represented by the formula (1) is (12Z)-labda-1
2-diene-17-α-oic acid ((12Z
)-1abda-12,14-dien-1? -a
-oic acid) (hereinafter referred to as compound (1)).

Avienol used as a substrate for microbial conversion in the present invention is a known compound represented by the formula (It), and is known as an essential oil component of Turkish leaf tobacco, etc. However, it is preferable to use abienol alone, and the compound of the present invention can be obtained by microbial conversion of niahienol. An example of the manufacturing method will be explained. First, the microorganism JTS-162 strain (Feikoken Bibori No. 5702) was inoculated into a medium containing avianol, cultured aerobically at 28°C, and after about 12 hours, chelate inhibition of α, αI-dipyridyl, etc. After adding the agent, the mixture is further cultured aerobically at 28°C for about 36 to 60 hours. The most favorable conversion effect is obtained by culturing at a temperature of about 45 to 5 degrees. He extracted the culture solution after conversion using organic solvents such as ethyl vinegar and ethyl ether.
The solvent is distilled off under reduced rE to obtain the converted product. This conversion product was eluted using a silica gel column with a solvent such as a hexane-ethyl acetate mixture, and
;-J: Make 7 Wi and add 7 compound (1).

1. Morphological properties (1) It is a bacillus, and the cell morphology changes as the culture progresses. At the early stage of feeding, cells elongate and form branch schools. After 12 to 14 hours of culture, the cells undergo regular 4) breakage, and then become short rod-shaped. Size 4: J #f (0,6~0.8) x (5~15) # at the beginning of feeding, after division (
0,6~o,8)x(o,B~1.8)μ.

(2) Gram staining S positive (3) Acid-fastness: Positive 5- (4) Spore formation ability: None 5) Motility: None 2, Chemical composition analysis (11!ff Spore wall 0 constituents main Amino acid 4: t meso
-diaminopimelic acid.

(2) Kumquat of guanine + cytosine in DNA is 62.5
It is mole%.

3.Culture findings (1) Broth agar plate culture (28℃, 6 days culture): Growth is slightly slow.Connie shape is circular, diameter is 1.5-2 cm.
The riverside is wavy 9 The surface is smooth.

The color is white and turns brown as the culture progresses. The color of the medium does not change.

(2) Meat juice agar slant culture (cultured at 28°C for 4 days): Growth is rather slow. The shape and 9 colors are the same as those for meat juice agar plate culture.

(3) Meat juice liquid culture (28°C, 6 days culture) The coffin does not come out very well. Bacteria M%' are slowly formed on the surface and then settle to become sediment. When cultured successfully, it shows uniform growth.

6- (4) Meat juice gelatin puncture culture (28°C, 6 weeks culture) Bacterial cells grew on the surface of the liquefied gelatin.

(5) Lidomus milk (cultured at 28°C for 6 weeks):
Slowly produces acid.

4 Physiological characteristics (1) Growth row A11: The optimum temperature for growth is 20-30°C, the optimum pH is 66-85, and it cannot grow under amorous conditions.

(2) Auxotrophy: None 3) Reduction of 61'I ugly salt: Positive (4) Hydrolysis of starch: None 5) Utilization of citric liquor: Positive (6) Urease: Positive (Kaoxidase: 1) Damage (8) Catalase: Positive (9) Pigment production; None Ifυ O-)゛Test: Fermentative 01) Methyl leatothist: Negative (121V-P test: Negative 03) Indole production; None 04) From less sugars! tJ Hygas production acid gas 1, L-arabinose 10-2, D-xylose + 3, D-glucose + -4, D-
Mannose 10-5, D-fructose
+ 6 D-galactose 10 -7, malt
Sugar 18 Sucrose
+ - 9, Lactose -10, Trehalose + 11 D-Sorvit + 12 D-Mannit + 13 Inosit + 14 Glycerin + 15 Starch - + ...Produced - - Not produced a9 Use the following compounds as carbon sources Grow.

D, L-alanine, paraffin, pyruvic acid, propionic acid.

(g) Aesculin minute wr: Positive α Tween 60 decomposition: Positive (kn) Chiron decomposition: Negative (b) Avienol, sclareol, and manul assimilation: From the results above positive, Purge Aids Manual of Q de Terminal Bacteriology (B@rg@ys
'Manual of Determinatlv*
Bactsriology), 8th edition (1974), this strain JTS-162 was used as a strain of Nocardia φ restricta.
was identified.

Next, a more specific explanation will be given with reference to manufacturing examples.

(Manufacturing example) Bacto Trypton (manufactured by Difco, USA)
A well-known L-slant medium (PH 7,2) consisting of 1% yeast extract, 05% yeast extract, 0.5% sodium chloride, 0 glucose, and 1.5% IS 1 agar was prepared in a test tube, and JT8- Approximately 10 gold loops of 162 stocks were inoculated,
It was statically cultured at 28°C for 3 days and used as a cocoon.

9- 1 then (NH,), 8042 t, K,) IP
042t.

Mfso4-7H,00,2f, CaCl2-2H
,00,2f , Pe5o4*7H,OO,01f
, H,01t+kfg ground 1 (mer7.o
) into a rectangular flask and sterilized at 121°C for 15 minutes. Powdered Avienol 1f and 1% Tween 60 aqueous solution (surfactant,
(manufactured by Kanto Kagaku Co., Ltd.) 1 (added jmt. The above L-
Add 5 ml of the inoculum of JT8-162 strain for one slant medium.
The cells were sterilized in 0.8% physiological saline and inoculated into the liquid medium described above. Then, using a rotary shaker, 210
After culturing at 28°C for 12 hours, α, α′-dipyridyl was added as a chelate inhibitor to l Q g (m
ole/ml was added, and the culture was continued for an additional 48 hours. A culture containing the conversion product of avianol was obtained by this culture, and the following operation was performed from this culture to obtain the compound (
1) was fractionated.

That is, 50 Omt of ethyl acetate was added as a solvent to the culture at a time, and extraction with stirring was performed twice. The extracts were combined and the solvent was distilled off under reduced pressure to give 0.8 Of of the converted product. Next, 50V silica gel (Wako Gel C- manufactured by Wako Pure Chemical Industries, Ltd.)
200) to make a column and convert/f: the product -
・Imasan: Elute with ethyl acetate (525, m/v),
3. 5mt each with fraction collector. ) flu
It's L. The fraction containing the compound (summer) was subjected to column treatment 3311 as described above, the solvent was distilled off, and fI!
Compound (1) is 0.301f by J FJ.

Next, spectral data of the compound qIA(1) of the present invention is shown.

Go to the molecular formula “Cto”at.

1”c-NMR (CDC1, TM8) δ (ppm
): is, 9(q).

16.3(Q), t7. a(t), 19.9(Q), 2
31(t).

24, o(t), a6. o(a), 47.2(d), s
o, 2(t).

62.2(d), 74.7(!l), 113.5(t)
, 1B0.7(s) * 13:'l-9(')
s 13319 (s) t 183-7 (s
).

IR□I-': 3400 t 2930 s 16
85, 138551245.900°λis m/z: 320 (M'-), a 0
2, 2 g 7, 274.

236.221.179, 161.148, 134, 1
From the spectral data of 19.84,43° or more, compound (1) was found to be the above-mentioned (1
) It was confirmed that it has the chemical structure represented by the formula.

When the compound (1) of the present invention is added to cigarettes, it harmonizes well with the original tobacco aroma, suppresses irritation, and mellows the aroma, and has a long-lasting effect and has a low number of digits during the cigarette manufacturing process. It was found to have many excellent effects.

In order to use the compound of the present invention as a tobacco flavor improver, it is diluted with a solvent such as ethanol or ethylene glycol to an appropriate concentration, and 0.01 to 3
0 ppm (w/w), preferably 0.1-lp
This effect is exhibited by adding pm. The types of tobacco to which the compounds of the present invention can be effectively applied are not particularly limited, and include not only tobacco obtained through cultivation, but also recycled tobacco and pipe tobacco manufactured from tobacco scraps. .

EXAMPLES Hereinafter, the effects of the present invention will be specifically explained with reference to Examples.

Example 1゜11 before rolling NfJ Monopoly Public Corporation trade name "Cherry" 100 tons of chopped river tobacco were dissolved in 3 mt of ethanol with the compound (1) prepared by the method shown in the above production example. The odor and taste when smoked were compared using a two-point discrimination method using paper rolls containing 0.5 ppm of fuguri and using the above-mentioned shredded tobacco rolls without compound (1) as a control. . The evaluations by the 20 expert panelists for sensory testing were as shown in Table 1.

Table 1? , E) Numbers indicate the number of people. Chubu has a risk rate of 1
Significant difference σ1 between samples in %. Show that something is true.

13- Example 2 Waste tobacco was extracted with hot water at 100°C and separated into a water-soluble part and an insoluble part, and then the water-insoluble part was beaten and 1 part of the dry weight was
Example 1 was applied to the regenerated sheet tobacco 100f made by molding a mixture to which 5% of kraft pulp was added into thin paper and returning the above-mentioned water-soluble portion to the thin paper. The same compound (1) was dissolved in 3 mt of ethanol to give a concentration of 1.0 ppm.
After spraying and adding the compound (1), the sheet was cut into 91 pieces and rolled into paper, and the aroma, taste, and irritation were evaluated using the two-point discrimination method using the above-mentioned sheet without the addition of compound (1), which was cut into 9 rolls, as a control. compared. The evaluations by the 20 expert panelists for sensory testing were as shown in Table 2.

2nd barley 14- Note) The number of people who assumed that the letter P was white. The Chubu area has a risk rate of 1%, indicating that there is a significant difference from the trial number.

Qt! “m Applicant Japan Monopoly Corporation 15- 223-

Claims (1)

[Claims] 1. A compound represented by formula (1). 2. A tobacco flavor improver comprising a compound represented by formula (1).