JPH03145429A - Bacteriostatic agent - Google Patents

Abstract

PURPOSE: To obtain a bacteriostatic agent suitable for suppressing poisonous bacteria in foods, cosmetics, etc., by compounding a specific glucopyrid as active component. CONSTITUTION: This bacteriostatic agent contains glucopyrid consisting of one kind or more of 2,3,4-tri-o-acylglucose of the formula as active component. A compound of the formula is a glucopyrid derived from upper cuticule of Lycopersicon pennellii Corr. a wild species of tomato which mainly distributes on extremely dried west slopes at low altitude in the Andes in Peru and secretes viscous secretion on the surface of its leaves, stokes, etc. The component is obtained by bringing parts of leaves, stokes, etc., to contact with an organic solvent such as chloroform or hexane and extracting them. Losing no activity by the influences of glucoses such as starch, the agent bacteriostatic is suitable for suppressing flat sour germs in hermetic container-packed foods or drinks which is kept at a warm temperature.

Description

[Detailed Description of the Invention] [Industrial Application Field] The present invention relates to the wild tomato Lycoversicon pennelly.
The present invention relates to a bacteriostatic agent containing gel colipid derived from the upper cuticle of Corycus as an active ingredient and suitable for suppressing harmful bacteria in foods, cosmetics, etc.

[Conventional technology] Ensuring the long-term shelf life of various products such as foods and cosmetics makes it possible to extend product storage and distribution periods, expand distribution areas and sales channels, and expand the range of choices in consumption timing. shall be.

Therefore, various techniques have been studied and implemented to obtain good storage stability of various products.

For example, in order to reduce the quality of products due to the growth and activity of various microorganisms, various sterilization methods can be applied to products, and bacteriostatic agents can be used.

These technologies are appropriately selected and used in consideration of their impact on product prices and the like.

[Three Problems to be Solved by the Invention] Many compounds have been developed and used as bacteriostatic agents to suppress the growth and activity of various microorganisms and prevent the quality of products derived from these microorganisms from deteriorating. Many of these can be obtained through chemical synthesis.

However, in products that come into direct contact with animals and humans, as well as various foods and beverages, there is an increasing demand for the use of bacteriostatic agents made of naturally derived substances.

On the other hand, various foods and beverages packaged in sealed containers such as cans, bottles, and retort pouches are being actively commercialized, and there is a desire to develop more suitable bacteriostatic agents for these products.

For example, in foods and drinks such as coffee drinks, soups, and shiruko that are sold heated to around 55°C,
Various bacteriostatic agents are used to prevent flat sour spoilage caused by heat-resistant spore-forming bacteria, so-called flat sour bacteria. However, with conventional bacteriostatic agents,
In some cases, the effects of these foods may be reduced due to the effects of sugars such as starch that are normally included in these foods.

The present invention was created in view of the problems with conventional bacteriostatic agents as described above, and can be used in a wide range of applications.
Among these, the object is to provide a bacteriostatic agent that can provide practical effects even in the presence of sugars.

Another object of the present invention is to provide a bacteriostatic agent that can respond to the recent growing trend towards naturally derived substances.

[Means for Solving the Problems] The bacteriostatic agent of the present invention contains glucolipid extracted from Lycopersicon benelli Corr, a type of wild tomato, as its active ingredient. .

The glucolipid is an L-cuticular lipid component of the above-mentioned wild tomato, that is, a sticky lipid component (secreted to the surface of leaves, stems, etc.) contained in the upper cuticle or secreted from the upper cuticle (secreted to the surface of leaves, stems, etc.). It can be obtained from the plant by extraction using an organic solvent. The glucolipid contains one or more types of 2,3.4-)so-0-acylglucose represented by the following formula (I).

(However, R1, R2, R3 are each independently 3 to 3
Represents a fatty acid selected from the group consisting of alkanoic acids and alkenoic acids containing 0 carbon atoms and optionally having side chains. ) Lycoversicon benelli containing the above glucolipids
Koru is Solanum Benery Koru (Saranu Koru).
It is also known as "M and Renga Yuzuki" (Nami 2), and is mainly distributed in the low-altitude, extremely dry western slopes of the Andes Mountains in Peru. Secretes a sticky exudate on the surface.

For details, see Japanese Patent Application Laid-Open No. 61-263994, for example.
As disclosed in the Publication No. 893, and as described in line 6 from the bottom of the upper left column on page 893 to line 3 of the lower left column on the same page, by receiving such kind of distribution from various organizations, the human resources can be improved. You can also directly handle things that grow wild in Peru.

To obtain a component containing glucolipid derived from Lycoversicon benelli col, which is the active ingredient of the bacteriostatic agent of the present invention,
Lipid extraction procedures with suitable organic solvents can be used.

Parts of the plant used for extracting the components include, for example, the epidermal parts of leaves, stems, pedicels, buds, and fruits, and among these, leaves and stems are easier to use.

For the extraction operation using an organic solvent, a known operation for lipid extraction can be appropriately selected and used.

For example, by contacting parts such as leaves and stems of the above plants with an organic solvent such as chloroform, hexane, or methylene chloride,
The glucolipid-containing component can be obtained by recovering the component extracted into the organic solvent through an operation such as evaporation of the organic solvent.

Note that in this extraction operation, the conditions may be set in consideration of the volume, the quality of the extracted components obtained, and the like.

For example, it is preferable to set conditions in which contamination with lipids such as chlorophyll found in the deep layers of plants is unlikely to occur.

The extracted components obtained through the above extraction operation are usually 89-
It contains 95% by weight of the above glucolipids, 5-10% by weight of alkanes, and up to about 1% of terpenoids.

The extracted component thus obtained can be used as it is as an active ingredient of the static agent of the present invention.

Furthermore, various fractions and purified products obtained by fractionating and purifying the extracted components as necessary may be used as the active ingredients of the bacteriostatic agent of the present invention.

These extracted components, various fractions, and purified products can be obtained in various forms such as liquid, paste, and semi-solid by fractionation or special methods, but any of them can be used as the bacteriostatic material of the present invention. It can be used as an active ingredient in drugs.

Regarding these extraction, fractionation, and purification operations, please refer to JP-A-61-2
6:1994.

Whether or not the glucolipid is present in these extracted components, various fractions, and purified products is disclosed in, for example, JP-A No. 61-2.
Publication No. 63994, page 898, upper left column, line 3 - same r! This can be confirmed by analysis using the thin layer chromatography method described in the 6th line of the lower left column.

In addition, glucolipid consisting of one or more of the compounds represented by the general formula (I) shown above can be obtained, for example, by fractionating polar components and non-polar components from the above-mentioned organic solvent extracted components by an appropriate method. It can be obtained as a polar component of

Further, the composition of the obtained glucolipid can be determined by analysis by various methods such as high performance liquid chromatography.

The antistatic agent of the present invention may be used by being mixed with the product at an appropriate stage in the preparation of the product to which it is applied.

For example, for pastry products such as sausages, kamaboko, and yokan, and cream-like products, the bacteriostatic agent of the present invention is added to the raw material mixture and then the product is manufactured by operations such as molding. Bacteriostatic agents are available.

In addition, for solid materials such as those contained in side dishes, the bacteriostatic agent of the present invention in liquid or paste form may be used as is, or the bacteriostatic agent of the present invention may be dissolved or mixed in an appropriate solvent. It can be used by mixing it with a solid material and applying it to its surface.

The blending ratio of the bacteriostatic agent of the present invention is 0.0005% by weight or more, preferably o
, oot weight% or more.

The upper limit of its blending amount takes into consideration the miscibility with various products, the solubility of the product in liquid form, or the effect on the taste and aroma of the product if it is a food or drink. All you have to do is decide.

For example, when used for liquid or semi-solid products, or paste products, about 3% by weight, preferably 2% by weight.
It can be added in an amount of up to about 6%, and in products using an emulsifier, it can be added in an amount of up to about 6%.

In addition, when applying to a solid material as described above, an amount that can be applied effectively to the surface of the solid material may be used.

The bacteriostatic agent of the present invention can be used in products in various fields such as foods, beverages, cosmetics, paints, and wall materials. In particular, since its effect is not reduced by the influence of sugars such as starch, it is suitable, for example, as a bacteriostatic agent for flat sours in foods and beverages in sealed containers that are maintained in a heated state.

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

Reference Example A glucolipid containing a plurality of 2,3,4-tri-o-acylglucoses represented by the above formula (I) was prepared from the leaves and stems of Lycoversicon benelli col according to the method described in JP-A-61-263994. (Composition) was obtained as follows.

First, the leaves and stems of Lycoversicon benelli col were immersed in chloroform for about 1 minute, and the components extracted into the chloroform were recovered by evaporating the chloroform.

Next, the obtained extract was dissolved in methanol, and the obtained solution was cooled. At that time, the crystalline material containing alkanes generated in the methanol solution was filtered off from the methanol solution, and then the methanol solution was refluxed with activated carbon to clarify it.

Water was further added to the decolorized methanol solution to obtain an aqueous methanol solution. The amount of water added at this time was such that the volume ratio of methanol to water in the aqueous methanol solution was about 4/1 (methanol/water).

The resulting aqueous methanol solution was extracted once with hexane.

After the hexane extraction process, the remaining aqueous methanol solution was diluted with water. The amount of water added at that time was such that the volume ratio of methanol to water in the diluted aqueous methanol solution obtained was methanol/water = approximately 271.

The IJ diluted aqueous methanol solution was further extracted with hexane at least twice.

The hexane-extracted fractions obtained by the above operations were combined into one, which was further washed with water.

Finally, hexane was evaporated from the washed hexane fraction, and the resulting product was further dried to obtain a colorless pasty to solid non-bitter glucolipid (composition).

R1 of the formula (I) is added to the obtained glucolipid (composition).
, R2, and R3 were analyzed according to a conventional method, and the molar ratio of each fatty acid group to the total fatty acid groups contained in the glucolipid was determined.

The results are shown below.

Content of each fatty acid group relative to all R1, R2, R3; mol% 1-C4 (2-methylpropanoic acid) group 30.1a-C
, (3-methylbutanoic acid) group 4.1i-C5(2-
methylbutanoic acid) group 2.4i-C,. (8-methylnonanoic acid) group 23.6n-C,. (decanoic acid) group
9.4n-C+2 (dodecanoic acid) group
1.9 Example 1 Clostridium, a representative strain of flat sour bacteria
Thermoaceticum ((:IosLridiumth
The bacteriostatic effect of the glucolipid (composition) obtained in Reference Example against C. ermoaceticum was investigated by the following procedure.

First, use a modified TGC medium with the composition shown below (manufactured by Rasui).
Iron (III) ammonium citrate was added to the mixture to give a concentration of 0.05% by weight to prepare a modified TGC medium containing iron (III) ammonium citrate (cm-TGC medium).

Modified TGC medium composition: Peptone 17.0g Soy bebutone 3.0g Glucose
6.0g sodium chloride
2.5g agar
0.7g Sodium thioglycolate 0.5
gL-lecithin 0.25g sodium sulfite O, 1g water
! 1 (pH 7,
σ±0.1) Next, a medium was prepared by adding various concentrations of glucolipid (composition) obtained in the reference example to this cm-T GC medium, and 17ff11 of each medium was added to one type of medium. Each sample was dispensed into two test tubes (16 x 150 mm, with screw caps).

The medium in each test tube was sterilized by autoclaving at +21'C for 40 minutes, and while each medium was still liquid, 1.0+5ffi of Clostridium thermoaceticum spore solution was added to each medium until the final spore count was 9. 3×10′ pieces were added. Next, each test tube was tightly sealed with a cap and then left to stand at a temperature of 65°C.

In addition, the spores contained in the spore solution were removed using a hot water bath.
It was activated in advance by treatment at 0°C for 15 minutes.

After 2 weeks, the colored state of each test tube was visually observed, and the growth of the bacteria was determined by the blackening of the medium due to the reaction between H2S produced as the bacteria grew and Fe added to the medium. It was determined whether As a result, the minimum growth inhibiting concentration of glucolipid (composition) was found to be 50 ppm.

Example 2 The bacteriostatic effect of glucolipid (composition) obtained in Reference Example against the following eight types of harmful bacteria typically found in various side dishes was investigated.

■Escherichia coli ■Pseudomonas fluorescens
■Bacillus cereus
s) ■Lactobacillus plantarum (Lactobacillus LL!!!)■
Leuconostoc mesenteroides
stoc mesenteroides)■Micrococcus luteus■Staphylococcus aureus (SLah Iococcus aureus)
) ■ Streptococcus faecalis (SLre Lococcus faecalis
) First, a cell culture solution in the range from the logarithmic growth phase to the stationary phase was prepared from each cryopreserved strain of each of the above bacteria.

In addition, the blank cultures of ■, ■, ■, ■, and ■ are carried out on SCD medium (
(manufactured by Daigo), and the uncultivated cultures of ■, ■, and ■ were carried out in B) II medium (manufactured by DI FCO), and the cultivation time and temperature ranged from 30°C for 8 to 24 hours. The conditions suitable for this were selected.

Next, solutions were prepared by adding various concentrations of the glucolipid (composition) obtained in the reference example to the SCD medium, and agar was added to each solution! , 5% by weight/volume, and poured into a 90 mm diameter Petri dish to obtain plates containing glucolipid (composition) at various concentrations.

Furthermore, on each of these agar media, one platinum loop of the blank bacterial cell culture solution obtained previously was streaked, and the growth of the bacteria (at 30°C) was observed over time, and 2. 3. After 7 days, the minimum growth-inhibiting concentration of glucolipid (composition) for the sample (■) was determined.

Furthermore, in place of ``Kan'' in ``■'', each of the bacteria in ``■'' to ``■'' was used individually, and in ``Kan'' in ``■'' to ``■'', BHI medium was used.
The same operations as above were repeated, except that the culture conditions were set to the optimal growth conditions for each bacteria, to determine the minimum growth-inhibiting concentration for each bacteria.

Table 1 shows the results obtained by the above operations.

Table 1 As is clear from the results in Table 1, the glucolipid (composition) obtained in the reference example has a particularly excellent bacteriostatic effect against bacteria of the genus Bacillus, and also has a particularly excellent bacteriostatic effect on bacteria of the genus Bacillus. It was also found to have the effect of inhibiting growth and delaying the start of growth of bacteria.

Furthermore, since the amount of bacteria used in the above test was much higher than the amount of bacteria that would be mixed into the prepared food during normal handling, it is expected that the effect would be even higher than the results shown in Table 1 in normal handling of prepared dishes. can.

Example 3 To a mixture obtained by mixing 50 kg of milk, 15 kg of skim milk, 910 kg of milk, 1 kg of coffee extract, and 60 kg of water, 0.1 kg of glucolipid (composition) obtained in Reference Example was added.
g and mixed well.

Next, this mixture was homogenized according to a conventional method, preheated at 80° C. for 10 minutes, and then heated at 200 sI1. Place it in a large capacity can and seal it.

Furthermore, the can containing the mixture was sterilized at 120° C. for 30 minutes.

The thus obtained canned coffee milk beverage was heated at 45-70°C.
It was left standing at a high temperature.

After 15 days had passed, each can was opened and the presence or absence of growth of heat-resistant bacteria in the coffee milk beverage was examined by the following method.

Test method; cm-T G described in Example 1 dispensed into test tubes (+6 x 150 aun, with screw cap)
Add 1 ml of coffee milk drink taken from the can to C medium 5IIlj2, and add 2 ml of liquid paraffin to the medium.
After layering 12, seal the test tube with a cap,
This was left standing under temperature condition F of 65°C. After 2 weeks, the culture medium was observed for black discoloration, and if black discoloration occurred, it was marked as "
It was determined that there was growth of heat-resistant bacteria.

As a result, none of the 500 cans extracted as samples was determined to have "growth of heat-resistant bacteria."

In addition, the target preparation 5 that does not contain tomato libid
Seven of the 00 cans were determined to have "growth of heat-resistant bacteria."

Example 4 Powdered Zarashi-an 50kg, sugar 80kg, salt 1.5k
g, 400 kl of water (and then further added 0.5 kg of glucolipid (composition) obtained in the reference example,
The mixture was mixed under heating at 80°C.

The resulting mixture was sealed in a can with a capacity of 200 mIt, and sterilized at 120° C. for 20 minutes.

45 to 70 sterilized canned shiruko obtained by the above procedure
It was left standing at a high temperature of ℃.

After 15 days had passed, each can was opened and the presence or absence of growth of heat-resistant bacteria in the contents was examined in the same manner as in Example 3.

In addition, preparation 1 not containing tomato lipid was used as a control.
,000 cans, 22 cans were determined to have "growth of heat-resistant bacteria."

Example 5 700 g of matshu potatoes, 50 g of sliced cucumber, 50 g of carrots boiled after slicing, 50 g of chopped onion
g and mayonnaise 150g (14g vinegar, 1 salad oil
Potato salad was obtained by mixing Glucolipid (composition) Ig obtained in Reference Example with 20g of glucolipid (composition) obtained in Reference Example.

The obtained potato salad was stored at a temperature of 10°C,
Ten panelists monitored the change in taste over time.

Taste evaluation was performed according to the following criteria, and the average of the scores (flavor scores) given by 10 people was determined.

The results are shown in Table 2.

Note that a potato salad obtained in the same manner as above without adding glucolipid was stored under the same conditions and was used as a control.

Flavor score criteria; 5... Fresh and good flavor 4... Slightly low flavor) 3... Slightly degraded odor (rancid odor) 2... Significantly degraded odor (rancid odor) [Effects of the invention] The present invention provides a bacteriostatic agent that can be used in a wide range of applications.

The sedative agent of the present invention is useful for foods, cosmetics, etc., and in particular, since the saccharide effect is not affected by the presence of sugars such as starch, it can be suitably used for products containing saccharides.

Moreover, the sedative agent of the present invention contains as an active ingredient an ingredient extracted and separated from plants, fractionated and purified as necessary, and can meet the demand for a sedative agent derived from natural products.

The Blunt Cell Research

Claims (1)

[Claims] 1) 2,3,4-tri- represented by the following general formula (I)
A bacteriostatic agent characterized by containing glucolipid consisting of one or more o-acylglucoses as an active ingredient. ▲There are mathematical formulas, chemical formulas, tables, etc.▼ (I) [However, R^1, R^2, and R^3 each independently contain 3 to 30 carbon atoms and have a side chain. represents a fatty acid selected from the group consisting of alkanoic acids and alkenoic acids. ]