JPS58150527A - Abienol derivative and flavor and taste improver for tobacco consisting of said derivative - Google Patents

Abstract

NEW MATERIAL:An abienol derivative expressed by formulaI(R is -CH2OH-, -COOH or -COOCH3). USE:A flavor and teste improver and irritation suppressant for tobacco capable of harmonizing well with the original flavor, suppressing the irritation, mildening the flavor and making the effect last. PROCESS:A microorganism JTS 131 strain (FERM No. 6303) is inoculated into and cultivated in a culture medium containing an abienol expressed by formula II (a material oil component of Abies balsamea, etc.) aerobically at 28 deg.C for 12hr, and a chelating inhibitor, e.g. alpha,alpha'-dipyridyl-ethylenediaminetetraacetic acid, etc. is added thereto. The strain is further cultivated aerobically to carry out the converting reaction. After cultivating for about 45-50hr, the culture fluid is extracted with ethyl acetate, etc. and the solvent is then distilled off under reduced pressure to give the aimed compound expressed by formulaI.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an aroma improver for tobacco ζ, which is comprised of novel compounds, an abienol derivative and a scissors derivative.

In recent years, the preference for cigarettes has been shifting towards products with a lighter flavor and richer aroma, but as a result, the leaf tobacco used in product cigarettes is often lighter in flavor and has a lower nicotine content. It's starting to look like this. however,
Since such JI S9 and ζ generally lack aroma and taste, it is necessary to add various flavoring agents to improve the aroma and taste of the product.

On the other hand, research has been conducted to produce fragrances suitable for such purposes by so-called microbial conversion of certain camphor compounds. No. 53-124911, Special Publication No. 66-421109, No. 56-42898, 1i
The description is given in ils6-5'4299 and the like.

From this perspective, the present inventors conducted research with the aim of obtaining a useful tobacco flavoring agent by microbial conversion of avienol. The inventors have discovered a new compound that is extremely effective in improving the aroma and taste of tobacco and suppressing irritation, and have accomplished the present invention. That is, an object of the present invention is to provide a novel compound effective in improving the flavor and aroma of tobacco, and is a tobacco flavor improver comprising a compound represented by the following formula (1).

Due to the general difference, the compound represented by the formula (1) is
It includes the following three new compounds.

R=CHjOH: (127,) Humiliation Loveda-12, 1
4-shear38-t-ru, ((12Z)-1abda
-12,14-dian-18-ol), (0)R=-COOH: (12Z)-rub p
--12,14-Jin-18-Oic Arashido,
((12ZL)-1abda-12,14-dien
-18-oicacid), hereinafter referred to as compound 2).

((12Z)-1abda-12,14-dien-
18-oie acid methyliter)
, (hereinafter referred to as compound 3).

Avienol used as a substrate for microbial conversion in the present invention is a known compound represented by the formula (1).
Pulsamea (Canada-Fir Balsam) (Abie)
seblsamea (Canada*fir*bal
It is known as a lumber component such as sam)).

Next, an example of the production of avienol, a compound of the present invention, by microbial conversion will be explained step by step.

First, microorganism JT8-131 was added to a medium containing Avienol 1.
strain (Choken Soyori No. 6303) was inoculated and cultured aerobically at 28°C, and after 12 hours, αl-dipyridyl, ethyle/
The most favorable conversion effect can be obtained by adding a chelate inhibitor such as diaminetetraacetic acid (hereinafter referred to as gDTA) and culturing aerobically for about 36 to 60 hours at 28 DEG C. for about 45 to 50 hours. After conversion, use the Tomo culture solution.
After extraction with an organic solvent such as ethyl acetate or ethyl ether, the solvent is distilled off under reduced pressure to obtain a converted product. Compounds 1.2 and 3 are purified, respectively, by bathing out the conversion product with a solvent such as a hexa/-ethyl acetate mixture using a phosphoric gel column and evaporating.

Motozono strain JTS-131 is an avianol-converted strain isolated from soil, 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 early stage of culture, the cells elongate and form branch cells. After 12 to 14 hours of culture, the cells develop irregular divisions, and then the cells become short culm-shaped. At the beginning of culture, the size is (0
,6~0.8)X(5~15)/J, after division is (0,6
~0.8)X(1,2~18)μ.

(2) Durham staining: Positive (3) Acid-fastness: Positive (4) Spore-forming ability: None 5) Motility: None 2. Chemical composition analysis (1) The main amino acids constituting the cell wall are meso-diaminobimeli/acid It is.

(2) The content of kuanin tentonno in DNA is 63.0
It's morchi.

3. Culture findings (1) Broth agar plate culture (28°C, 4 days culture): Growth is rather slow. Colonies are circular in shape, 1 to 2 cm in diameter, mucoid, and yellowish in color. The color of the medium does not change.

(2) Juicy agar slant culture (28°C, 4 days culture): Growth is rather slow and shows mucoid shape. The color tone is the same as the broth agar plate culture.

(3) Broth liquid culture (28°C, 6 days culture): The medium is not very cloudy. A bacterial film slowly forms on the surface and then settles to become sediment.

When cultured with shaking, it shows uniform growth.

(4) Broth gelatin perforated sea bream culture (28°C, 6 weeks culture):
Does not liquefy. Bacterial cells grow in a membranous and mucoid shape on the surface.

(5) Ridmus Fist Milk (28°C, 6 weeks culture):
alkali.

4 Physiological properties (1) Growth conditions: 25-35℃ is the optimum temperature for growth, DH is 6
The optimum value is 5-8.0, and it cannot grow under anaerobic conditions.

(2) Auxotrophy: None 3) Nitrate reduction: None 4) Dep/P/ hydrolysis: None 5) Citric acid utilization: Positive (6) Urease: Positive (7) Oxidase: Negative ( 8) Catalase: Positive (9) Pigment production: None Qo) O-F test: Fermentative Qt) Methyl red test: Negative (12) V/P test: Negative (13) Inodole production, none 14) Below Production of acid and gas from sugars Acid Gas I L-arabinose + -2D-xylose - 3D-glucose 10 - 41J-mannose 10 -5D-fructose + 6 D-galactose + -7 maltose
187, sugar 10 - acid gas 9 lactose - 10) levalose + o D-fulvit + 12 D-mannit + 13 wild boar y) 114 glycerin / 115 starch - +・・・・・・・・・・・・・Generation −・・・・・・・・・
... Not produced (15) Grows using the following compounds as vertical sources.

Baralai/1 Pyruvate, phenol.

(16) Decomposition of Aesculin: Positive (17) Decomposition of Zwie/60, two positive (18) Decomposition of Tyroshi/Positive (19) Assimilation of Avienol and Sclareol: Positive or higher results indicate Inoternashi, Naru Journal Op. Bacteriology (Interna)
tional Journal of Systema
ticBacteriology) pages 356, 198
Approved 1iita of Bac
terial Namea) and a report by M. Gutsdofello et al.
Interoational Pacteriology
Jourgal of &ystamaticBact
99, 1977] and other literature, the present strain JTS-131 was isolated from Rhodocoeeua erythropolis (Rhodocoeeua erythropolis).
It was identified as "thro-polis".

Next, a more specific explanation will be given using manufacturing examples.

[Production example] L-slant medium (pH 7) consisting of 1% Bactotrypto (manufactured by Difco, USA), yeast extract O5, 0.5% sodium chloride, 0.1% glucose, and 1.5% agar. , 2) was prepared in a test tube, inoculated with JT8-131 strain, and statically cultured at 28° C. for 3 days. This was used as a seed cell.

Next, (NH,) Go So, 2tXK, HPO, 2t
, Me2O.

-7H, o 0.2f, CaC1m +12H! O
0,2f, Fe3O4*7H100,01f, Hm
A liquid medium (pH 7,0) consisting of 1 L of 300 ml was placed in an Erlenmeyer flask and sterilized at 121°C for 15 minutes. Avienol IF in powdered form was added to this sterilized wave body culture medium K powder, and a 10-centiaxial 60 aqueous solution (surfactant, manufactured by Kanto Kagaku Co., Ltd.) was added. Said OL-slant medium 1
The duty of JT8-131 stock III! The I body was suspended in 5-ml of OJ*Cw/Rim* salt water and inoculated into the above-mentioned liquid medium. Then, using a rotary shaker, 210
After 12 hours of culturing at 28° C., 10 mM of αl-dipyridyl was added as a chelate inhibitor, and the culture was continued for an additional 48 hours. A culture containing a conversion product of a and enol was obtained by this culture, and Compound 2 and Compound 3 were isolated from this culture by the following operation.

Separately, lomM/Ic of CKEDTA, a substitute for at'-lysyl, was added as a chelate inhibitor, and the mixture was uniformly cultured to fractionate Compound-1.

That is, the culture (txd'-dipyridyl and EDT
11 g/pxt of ethyl acetate was added as a solvent to an equal volume mixture of the culture containing A), and extraction with stirring was performed twice. The extracts were combined and the solvent was removed under reduced pressure to give 0.95%
A conversion product of F was obtained. Next, a column was prepared using 70R 7Lica gel (Wako Gel C-200, manufactured by Wako Pure Chemical Industries, Ltd.), and the conversion product was mixed with hexane:ethyl acetate (
It was eluted at a ratio of 7:3, v/v) and fractionated into 5-fold fractions using a fraction collector. By subjecting the frucne containing each compound to the above-mentioned column treatment again and purifying it by distilling off the solvent, Compound 1, which matches the spectral data shown below, was obtained. Compound 2 was mixed with O,xOr (yield 5t) and compound 3 was mixed with 0.10f (
A yield of 5% was obtained in each case. Next, spectral data of Compound 1, Compound 2, and Compound 3 of the present invention are shown.

Compound 1 Molecular formula: CoHhOt”C-NMR [CDC4, TMS), δ (ppm):
1 s, 5(yl, 17.6(P), 18.0(f)
, 19.9 (p), 20.0 (f), 23.2 (U, 2
43(pka5.2(勺, 37.6(8), 38.
9(8), aj7(i), 43.6 (4BB with i
(4).

62.1(j), 'yi, 5(z), 74.2(a),
113.6 (i), 130.5 (j included 133.7 (d)
, 134.1(a), lRem-”:3320.294
0.2860.1460°1385.1045.895
゜MSmma: 306 (M”), 291,288.
258.237.211.181, 135.95.81
．． 55.43° Compound 2 Molecular formula: 2Cゎ'a Os .

C-NM'R (CDCz, TMs]acppm): 1
L9 (p l 6.3 (Jl 17.6 (J), 1
9.9(J), 19. (A), 23.18 23
．． 9(P), 36.8(4,3&3(gl 39.3
(J), 43.4(double 47.1(sχ50.2(4,
62,0 (J included 718 (4 included 113.4 (χ included 1
jO, 5 (dχ133.8 (j entry 134.0 (pass 183
．． 18>.

IRcm: 3400.2930.2860,]7
00.1690.1455.1385.1260.90
0. MSmma: 305 (MV) (s), 302
．． 287.273.2Q.

241, 221.175.134.119.81.55
．． 40゜Compound 3 Molecular formula: C,,)1uOa"C-NMkLCCDCl,,TMS) a (pp
m) :1! 5.9 (1 person 16.4ω ball 17.
6 (i included 19.9 f# included 23.0 (χ included 23.1
(S24,1(y entered 3s,7(- entered 38.3(sχ
39-3('') x 43.6(i), 47.5(s)
,.

50.4(jχ 51.8(P included 62.2(-(χ
7 4.2(s), 113.4←su, 130.5(
j), iaa, s (stock 133.9 (4,1789 (+
s'.

IRcm 1:3300,2940.1725.171
0.1460.1440.1245.900°MSmma: 334 (M”), 3]9.316.
303.275.239.205.179.161.1
From the results of spectrum data of 21.55.43° or more, it was confirmed that Compound 1, Compound 2, and Compound 3 each had a chemical structure represented by the above formula (1).

When compound 1, compound 2, and compound 3 of the present invention are added to cigarettes, they harmonize well with the original tobacco aroma, suppress stimulation, mellow the aroma, and have a long-lasting effect.
It has been found that it has many excellent effects, including less dissipation during the cigarette manufacturing process.

In order to use the compound of the present invention as an fc hato flavor improver, it is diluted to an appropriate concentration with a solvent such as ethanol or ethylene glycol, and added to a concentration of 0.01 to
30 ppm (W/W) preferably 0.1 to 1.0 p
The 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 Vibe cigarettes manufactured using tobacco tobacco as a raw material. is also valid.

Hereinafter, the effects of the present invention will be specifically explained using examples.

Example 1: Compound 1, Compound 2, or Compound 3 produced by the running method as shown in the above production example was added to 100 tons of shredded tobacco for "Cherry", a product of the Japan Monopoly Corporation, just before rolling, in 3-ethanol. Spray so that each dissolves to 0.5 ppm.
After the addition, the tobacco was rolled into paper rolls, and the odor and taste when smoked were compared using a two-point discrimination method using a roll of the above-mentioned shredded tobacco without the addition of the present compound as a control.

The evaluation by 20 specialized sensory test panels was poor, as shown in Tables 1, 2, and 3.

Table 1 Note) Flavored product: Compound 1 added. The number riJilL- indicates the number of friends, and the mark * indicates a significant difference between samples with a risk rate of 1%. Table 2 Note) Flavored product: Compound 2 added. The numbers are good, nine people.
Headquarters indicates that there is a significant difference between the samples with a risk of 541.

#I3 table Note) Flavored product: Compound 3 added. The number is the number of people who said it was good.

* indicates that there is a significant difference between the samples with a risk factor of 1-.

Extract tobacco scrap tobacco with hot water at 100°C, separate the water-soluble part and the insoluble part after death, beat the water-insoluble part, and add 1 part of the dry weight to this.
The kraft pulp of No. 5 was added and the mixed product was formed into a thin paper, and the above water-soluble portion was added back to the thin paper. 1 compound @l, compound 2 or compound 3 as 3-
After dissolving it in ethanol and spraying it to a concentration of 1.0 ppm K, it was punched and rolled into paper, and the punched and rolled pieces without the addition of the present compound were used as a control to detect odor, oxidation, and irritation. About the two-point identification method! We compared the taste. The evaluations by 20 specialized sensory test panels were as shown in Tables 4, S, and 6.

Table 4 Note) Flavored product: Compound 1 added. The number is the number of people who said it was good.

* indicates that there is a significant difference between samples with a criticality factor of 1φ.

Table 5 Note) Flavored product: Compound 2 added. Number of people who thought the numbers were good: 0
* indicates a significant difference between samples with a risk rate of 1◆0 Table 6 Note) Flavored product: Compound 3 added. The numbers are the number of Arashii people.

*mark O indicates that there is a significant difference between samples with a risk rate of 1% Patent applicant: Japan Monopoly Corporation

Claims (1)

[Claims] 1. 84 avianol compounds represented by the general formula (1). During wiping 8- is -CH*0) L-COOH or If- 0
A good tobacco product consisting of an avienol derivative represented by 0)2-beni formula (1) representing -CHs! I! Ambiguity improver. (The ship in the formula is -CCO2L-COOH or - to -0-CH
(representation of beauty)