JP3789632B2 - Liquid preparation containing sporic lactic acid bacteria - Google Patents

Description

【００１６】
【表２】

[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a liquid preparation containing sporic lactic acid bacteria.
[0002]
[Prior art and problems to be solved by the invention]
Recently, there has been an increasing demand for “health-oriented foods aimed at improving the constitution”, in particular, products aimed at improving the intestinal environment. For example, certain types of lactic acid bacteria or bifidobacteria improve gut microbiota when continuously ingested, thereby accelerating research to promote health. However, lactic acid bacteria generally have a short life, and their survival rate is extremely low, especially under dry conditions. In addition, in order to obtain a health promoting effect, it is necessary to reach the intestinal tract by removing the influence of gastric juice and bile acids. Therefore, many manufacturers are beginning to adopt acid-resistant lactic acid bacteria and introduction methods using enteric capsules.
On the other hand, sporic lactic acid bacteria (Bacillus coagulans), unlike general lactic acid bacteria, are extremely resistant to drying, heat and acid under certain conditions, and germinate and proliferate actively in the intestine. It is applied to various foods.
However, the spore lactic acid bacteria are excellent in heat resistance and the like if the spores remain as mature spores without germination, and once germinate and germinate spores (the germinated spores are further transformed into vegetative cells), Heat resistance, chemical resistance, etc. are lost.
Therefore, as a result of various investigations on the stabilization of spore lactic acid bacteria (maintaining mature spores without germinating spores), they are extremely stable in a powder state and should maintain a certain number of bacteria for more than one year. It has been known. However, no method has been proposed for stably maintaining in an aqueous system.
That is, even if the sporic lactic acid bacteria are stored in physiological saline or an aqueous sugar solution, the number of death sterilization increases with time, and it cannot be applied to various beverages.
Furthermore, when the spores of spore lactic acid bacteria are added to the solution, the low stability of the spores has been a major problem in the product design of foods and pharmaceuticals. For example, under low pH conditions, there has been a problem that after spore germination, vegetative cells grow and then die.
[0003]
[Means for Solving the Problems]
As a result of intensive studies to solve the above problems, the present inventors have controlled the osmotic pressure of the liquid under low pH conditions, thereby suppressing spore germination in the liquid and maintaining the spore morphology stably. The present invention has been completed.
That is, the present invention
A liquid preparation containing spore lactic acid bacteria characterized by satisfying the following 1) to 4) and storing at 50 ° C. for 2 weeks and having a remaining spore ratio of 12.2 % or more.
1) Contain at least 1.0 × 10 ³ spores / ml of Bacillus coagulans spores
2) Osmotic pressure should be in the range of 0.699 to 1.5 Osmol / kg.
3) The pH is in the range of 2.7 to 4.5.
4) Containing reduced maltose starch syrup and / or sucrose
DETAILED DESCRIPTION OF THE INVENTION
Hereinafter, the present invention will be described in detail.
Spore germination is induced by divalent metal ions such as dissolved calcium ions and magnesium ions when water enters from the spore surface (bacterial anatomy: written by Shoji Mizushima and Shinichiro Miura, Kodansha Scientific) Publishing, 1979). The present inventors have found a method of suppressing germination by increasing the osmotic pressure around the spores, that is, by increasing the pressure applied to the surface of the spores, thereby reducing the entry of water from the outside. Strictly speaking, osmotic pressure is a definition that can be handled with dilute solutions. In the case of a liquid preparation with a high solute concentration, it seems that it is difficult to apply osmotic pressure at first glance, but the present inventors have a positive correlation between the osmotic pressure and the spore residual rate of spore lactic acid bacteria. I found out. Since the osmotic pressure is proportional to the number of molecules (solutes) dissolved in water, there is a high relationship between the osmotic pressure and the amount of solute.
In order to increase the osmotic pressure of the solution, water-soluble components such as salts and saccharides may be added. Examples of the salts include salts containing metals such as sodium, potassium, magnesium, and calcium, milk minerals, and organic acid salts. Examples of the saccharide include monosaccharides such as glucose, disaccharides such as sucrose, oligosaccharides, polysaccharides, and sugar alcohols. Proteins, amino acids, peptides, and the like can also be used.
As the oligosaccharide, fructooligosaccharide, isomaltooligosaccharide, xylo-oligosaccharide, dairy oligosaccharide and the like can be used.
Examples of the sugar alcohol include erythritol, xylitol, maltitol, sorbitol and the like. Sugar alcohol is not assimilated by Bacillus coagulans and has a germination inhibitory effect during storage, which is preferable.
[0005]
In the present invention, the osmotic pressure was measured with an osmometer (model name OM802) using the freezing point depression method of Vogel (Germany).
In the present invention, the osmotic pressure of the solution needs to be in the range of 0.3 to 1.5 Osmol / kg, and preferably in the range of 0.4 to 1.0 Osmol / kg. If it is 0.3 Osmol / kg or more, the stability of the spore is improved. However, if it exceeds 1.5 Osmol / kg, the balance of the taste as a liquid is lost and the value of the product is lost.
[0006]
In the present invention, the pH of the liquid preparation needs to be in the range of 2.7 to 4.5, preferably 3.2 to 4.0, particularly preferably 3.3 to 3.8. If the pH is within the above range, spore germination is relatively unlikely and high-temperature sterilization is not required, so that a savory liquid agent can be obtained.
[0007]
The liquid preparation of the present invention can be applied to liquid foods and pharmaceuticals. Examples of foods include lactic acid bacteria beverages, soft drinks, jelly-like beverages, soups, desserts such as pudding and jelly, potions, fruit juices, vegetable juices, ice creams, and emulsified foods. In pharmaceuticals, it is used for liquids, syrups, dry syrups, emulsions, extracts, decoctions and the like that are generally expressed as drinks. However, the liquid form is not limited to this.
[0008]
Distribution of the foods and pharmaceuticals is limited to three types: frozen, chilled, and room temperature, and it is desired that beverages be stored at these temperatures. This product contains sporic lactic acid bacteria and may die if frozen. For this reason, storage is preferably chilled and normal temperature. However, in the case of distribution and storage in a chilled state, running costs related to transportation and storage become large, so that storage at room temperature is preferable in consideration of industrial usefulness.
[0009]
In the present invention, for example, assuming a solution of 1 liter per bottle, when Bacillus Coagulanse spores are mixed in the solution to a concentration of 1.0 × 10 ³ cells / ml, the net spore count is 1.0 × 10 ⁶ cells / ml. Become a book. By including this amount, the effect of improving the intestinal environment can be obtained. Preferably, it is blended at a concentration of 1.0 × 10 ⁴ to 1.0 × 10 ⁸ cells / ml, particularly preferably 1.0 × 10 ⁵ to 1.0 × 10 ⁷ cells / ml.
The liquid preparation of the present invention has a spore residual rate of 1% or more after storage at 50 ° C. for 2 weeks.
In the present invention, the spore survival rate was measured as follows.
The beverage base formulation was exposed to 85 ° C for 30 minutes and sterilized. To this, a spore suspension of spore lactic acid bacteria previously heated at 85 ° C. for 10 minutes was added at 1.0 × 10 ³ cells / ml or more under aseptic conditions. This was stored in a thermostatic chamber at 50 ° C., and the residual rate after 2 weeks was measured. In other words, 0.5 ml each was sampled after 2 weeks, diluted with 4.5 ml of sterilized 50 mM phosphate buffer (pH 7), heated at 80 ° C. for 30 minutes, cooled, and applied to BCP agar medium (Nissui Pharmaceutical Co., Ltd.) After spraying and culturing at 50 ° C. for 2 days, the spore survival rate was determined from the number of colonies formed.
Storage at 50 ° C for 2 weeks is an accelerated test and corresponds to about 1 year and 4 months when stored at room temperature. The spore residual rate after storage at room temperature for 6 months is preferably 1% or more, particularly 10% or more, more preferably 50% or more.
[0010]
In the present invention, the sporic lactic acid bacteria are not limited to commercially available products, but those newly isolated from nature, or those that have been selected or mutated from commercially available products can be used.
[0011]
An antioxidant can be added to the liquid preparation of the present invention for the purpose of suppressing oxidative degradation of components that occur during storage. Among these, fat-soluble antioxidants such as vitamin E and β-carotene are preferable because they have a spore stabilization effect.
[0012]
Moreover, addition of fermented milk is desirable for the liquid agent of this invention from flavor improvement. Fermented milk is prepared by fermenting a raw material containing one or more of skim milk powder, whole milk powder, raw milk and milk protein using Lactobacillus and / or Bifidobacterium. Fermented milk includes types such as sterilized fermented milk, fermented milk containing live bacteria, and fermented milk (brown fermented milk) containing Maillard reaction products, among which the use of sterilized fermented milk is preferable. Moreover, the fermentation liquid and fermentation extract which used the whey component as a raw material can also be used.
[0013]
In addition, flavoring agents, yogurt flavors, fruit flavors, stabilizers, polysaccharides such as pectin and the like can be added to the liquid preparation of the present invention.
[0014]
【Example】
Hereinafter, the present invention will be described more specifically with reference to examples.
Examples 1-7, Comparative Examples 1-2
To the solution having the formulation shown in Table 1, sporic lactic acid bacteria isolated from Lacris (registered trademark, manufactured by Sankyo Co., Ltd.) powder were added in the amounts shown in Table 1. The pH (adjusted with lactic acid and citric acid) and osmotic pressure are also shown in Table 1.
Table 1 also shows the results of measuring the residual ratio of spores after storage at 50 ° C. for 2 weeks for this solution.
Example 8, Comparative Examples 3-4
The test was conducted in the same manner as in the above example except that the liquid formulation having the formulation shown in Table 2 was used. The results are shown in Table 2.
[0015]
[Table 1]

[0016]
[Table 2]

Claims (4)

A liquid preparation containing a spore lactic acid bacterium characterized by satisfying the following 1) to 4) and storing for 2 weeks at 50 ° C. and having a spore residual rate of 12.2 % or more.
1) Contain at least 1.0 × 10 ³ spores / ml of Bacillus coagulans spores
2) Osmotic pressure should be in the range of 0.699 to 1.5 Osmol / kg.
3) The pH is in the range of 2.7 to 4.5.
4) Containing reduced maltose starch syrup and / or sucrose   The liquid preparation according to claim 1, comprising a sugar alcohol.   The liquid preparation according to claim 1 or 2, comprising an antioxidant. Fermented milk is contained, The liquid agent in any one of Claims 1-3 characterized by the above-mentioned.