JP2003300900A - Calcium absorption promoter, promoter of crystal growth of calcium phosphate, and food and drink and fodder containing them - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a calcium absorption promoter promoting the absorption by inhibiting the crystallization of calcium in the intestinal tract, and a promoter of crystal growth of calcium phosphate effective for promoting recalcification of a tooth by promoting the crystal growth of the calcium phosphate in the oral cavity, and to provide a food and drink, and a fodder containing the promoter. <P>SOLUTION: The calcium-absorption promoter and/or the agent for promoting crystal growth of the calcium phosphate contains a cultured product or an extracellular cultured product of one or more kinds of lactic acid bacteria belonging to the genera selected from Lactococcus, Lactobacillus, Enterococcus and Streptococcus as an active ingredient. <P>COPYRIGHT: (C)2004,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a novel use of a lactic acid bacterium culture, and more specifically, a calcium absorption promoter and calcium phosphate containing, as an active ingredient, a lactic acid bacterium culture having a calcium crystallization inhibitory action. The present invention relates to foods and drinks containing a crystal growth promoter and an intracellular culture.

[0002]

2. Description of the Related Art It is said that calcium is the most abundant mineral in the human body, 99% of which is used for bone and tooth formation, and the remaining 1% is the activity of various enzymes. It plays an important role in life activities such as expression, muscle contraction, cell excitement sedation or blood coagulation.

Although it is such an important calcium,
Looking at the intake, it is said that the required amount for the Japanese is 600 mg per adult per day, but according to the 1998 National Nutrition Survey Results Report by the Ministry of Health and Health, the actual intake The actual amount is less than the required amount of 568 mg. Insufficient intake of calcium causes osteoporosis,
It is known to cause serious illness such as high blood pressure, which has become a social problem. Furthermore, calcium ingested in the gastrointestinal tract as food is absorbed into the blood from the intestine by a complicated mechanism, but the absorption rate of calcium salts and calcium agents in the intestine is 50% or less, and more than half is absorbed. There is also a report that it is excreted outside the body. Therefore, a substance that enhances the absorption of calcium in the intestinal tract is being developed.

As one of them, casein phosphopeptide (CPP) has been developed. CPP is a phosphopeptide obtained in a hydrolyzed product by allowing trypsin to act on casein, and binds to calcium to form a soluble complex. Therefore, it is considered that calcium is solubilized by suppressing the precipitation of calcium in an aqueous solution and the absorption rate of calcium is increased (Japan Food Science, Vol. 1, pp. 21-32 [1990]). JP-A-58-170440; JP-A-7-241172). However, since CPP is an enzymatic decomposition product of casein, it is necessary to enzymatically react the raw material, casein, and the peptide, which is a by-product of enzymatic decomposition, has a bitter taste. It has some problems such as the necessity of separating the peptides sufficiently, and the price is very expensive.

[0005] Poly-L-glutamic acid also acts to increase the absorption rate of calcium in the intestinal tract (Biosci. Biotech. Bioche
m., 58, 1662-1665 [1994]).
However, it is not allowed as a food additive because it is a synthetic product and is not used for safety reasons. In addition, poly produced by microorganisms
-γ-glutamic acid (JP-A-3-30648)
The calcium crystallization inhibitory activity is low and the viscosity of the solution is extremely high, which makes it inconvenient to handle.

As a substance that promotes absorption of calcium, a peptide derived from bone (JP-A-4-16165) and butyric acid as a basic component (JP-A-4-1083).
No. 60), but they have not been put to practical use because of problems in production and use.

Furthermore, the present inventors have found that a protein obtained from bacterial cells of a bacterium inhabiting Lake Vanda in Antarctica has an effect of suppressing calcium crystallization and thereby promoting absorption of calcium. However, it has not been put to practical use (Japanese Patent Laid-Open No. 2000-239180).

By the way, human tooth enamel is a crystal of hydroxyapatite composed of calcium phosphate. In the human oral cavity, decalcification (early caries) in which the enamel calcium phosphate dissolves and remineralization that crystallizes as before occurs, maintaining a stable state. However, when tooth decay bacteria adhere to the surface of teeth and produce plaques and acids, decalcification proceeds further, resulting in tooth decay. With the aim of preventing dental caries, the development of new food materials that promote remineralization in order to repair early dental caries is awaited.

JP-A-10-158178 discloses that a fermented product using a lactic acid bacterium belonging to Lactobacillus and / or Streptococcus has an action as a mineral absorption promoter. However, this fermented product only has the effect of promoting the absorption of calcium, and it is not mentioned that it also has the effect of promoting the growth of calcium phosphate crystals.

[0010]

DISCLOSURE OF THE INVENTION The present invention solves the above-mentioned problems of the prior art, suppresses the crystallization of calcium in the intestinal tract, promotes its absorption, and grows calcium phosphate crystal growth of bones and teeth. The present invention also aims to provide a calcium absorption promoting / calcium phosphate crystal growth promoting agent for preventing early caries and osteoporosis by promoting the above, and foods and drinks and feed containing the same.

[0011]

[Means for Solving the Problems] As a result of intensive studies to solve the above problems, the present inventors have found that Lactococcus spp.
Proteins extracted from a culture and / or extracellular culture of one or more lactic acid bacteria selected from the group consisting of actobacillus, Enterococcus and Streptococcus, and calcium crystallization inhibitory action and calcium phosphate crystal growth The present invention was completed based on the findings that they have both a promoting action. Therefore, it has become possible to provide a calcium absorption promoting / calcium phosphate crystal growth promoting agent that promotes calcium absorption in the intestinal tract and promotes calcium phosphate crystal growth in bones and teeth.

Further, the present invention provides foods and drinks having the above-mentioned two different effects by incorporating the above calcium absorption promoter / calcium phosphate crystal growth promoter into the foods and drinks. Moreover, it is a food or drink and a feed in which calcium is further added to them.

The present invention also relates to the genus Lactococcus, Lactoba
Disclosed is a calcium phosphate crystal growth promoter containing, as an active ingredient, a culture and / or extracellular culture of one or more lactic acid bacteria selected from the group consisting of the genus cillus, the genus Enterococcus, and the genus Streptococcus.

Further, the present invention provides foods and drinks having the above calcium phosphate crystal growth promoting effect and feed. Moreover, it is a food or drink and a feed in which calcium is further added to them.

[0015]

BEST MODE FOR CARRYING OUT THE INVENTION To explain the present invention in detail, the present inventors have found that Lactococcus spp., Lactobacillus isolated from fermented butter and cheese containing a large amount of calcium.
It was confirmed that lactic acid bacteria belonging to the genera Genus Enterococcus and Streptococcus have a calcium crystallization inhibitory effect. Among these lactic acid bacteria, especially Lactococcus lactis and Lacto
bacillus acidophilus, Lactobacillus brevis, Lacto
bacillus delbrueckii, Lactobacillus helveticus,
Lactobacillus phamnosus, Enterococcus durans, St
reptococcus salivarius, Lactobacillus casei
It was a lactic acid bacterium with a strong calcium crystallization inhibitory effect and was a preferable bacterium.

In the present invention, as a method for preparing a culture obtained from lactic acid bacteria, the above lactic acid bacteria are pre-cultured in 803 medium or 804 medium designated by IFO (Fermentation Research Institute), and then 0.1% ( W / V) Yeast Pepton containing CaCl ₂
e (YP) medium, Acetate (AM) medium, or Lactobacillu
The culture was obtained by culturing in s (NEW) medium at 30 ° C. or 37 ° C. until it reached a stationary phase, and the culture was further centrifuged to separate the cells and the culture supernatant.

As a method for obtaining an extracellular culture, a generally used method can be appropriately used. In the present invention, the culture supernatant is suction-filtered with a 0.45 μm filter to limit the filtrate. External filtration or polyethylene glycol (PE
G 20,000), followed by treatment with distilled water against dialysis three times.

As for the method for obtaining the intracellular culture, it can be prepared by using a generally used cell disruption method. For example, a method using a pressure-type cell disruption device such as a French press or an X-press, a mechanical grinding method using a ball mill or a dyno mill, an ultrasonic treatment method, a method using a homogenizer, a freeze-thaw method, an osmotic treatment method, rhizotium. An enzyme treatment method using, etc. can be mentioned, but they can also be combined. In the present invention, since the active ingredient of the culture of lactic acid bacteria having a calcium crystallization inhibitory effect is considered to be a protein, considering stability of the protein and extraction efficiency, pressure-type cell disruption, mechanical grinding, It is preferable to use a physical disruption method such as sonication or a homogenizer. Nucleic acid may be removed from the above-mentioned intracellular culture by using protamine sulfate, streptomycin sulfate or the like, if necessary. Further, the culture is optionally subjected to salting out with ammonium sulfate or the like, precipitation with an organic solvent such as ethanol, fractionation by the isoelectric focusing method, ion exchange, adsorption,
It may be purified by gel filtration, hydrophobicity, or chromatography such as affinity chromatography, or subjected to a dialysis or concentration process.

[Table 1] to [Table 5] show 803 medium, 804 medium, YP (yeast peptone: Y) used in the present invention.
The composition of east Peptone) medium, AM (acetate: Acetate) medium and NEW (Lactobacillus) medium is shown.

[0020]

[Table 1]

[0021]

[Table 2]

[0022]

[Table 3]

[0023]

[Table 4]

[0024]

[Table 5]

The culture of the lactic acid bacterium of the present invention obtained by the above method is capable of suppressing the crystallization of calcium at a protein concentration of 0.5 to 10.0 μg / ml and further promoting the crystal growth of calcium phosphate. Was recognized,
It was found to be useful not only as a calcium absorption promoter but also as a crystal growth promoter for calcium phosphate.

As a result, the culture of the lactic acid bacterium of the present invention is used for the prevention and treatment of osteoporosis for the purpose of promoting calcium absorption, and for promoting the remineralization of teeth for the purpose of crystal growth of calcium phosphate to promote the initial treatment. It can be used for the prevention and treatment of tooth decay.

As the calcium absorption enhancer and the calcium phosphate crystal growth agent of the present invention, the culture prepared by the above method may be used as it is, but generally, it is dissolved or dispersed in a suitable liquid carrier, or a suitable liquid carrier is used. Mix with powder carrier or adsorb to powder carrier, and if desired, further add emulsifier, dispersant, suspending agent, spreading agent, permeating agent, wetting agent, stabilizer, etc. It can be used as a calcium absorption enhancer or a calcium phosphate crystal growth agent in the form of a preparation such as an agent, a capsule, a tablet, and a powder.

As the food and drink containing the calcium absorption promoter and the calcium phosphate crystal growth agent of the present invention, various foods and drinks, for example, soft drinks, fruit juice drinks, fermented drinks and drinks such as milk, chewing gum, Confectionery such as candy, tablets, gummy jelly, biscuits and chocolate, frozen desserts such as ice cream and frozen desserts, dairy products such as yogurt and cheese, meat products such as ham and sausage, fish paste products such as kamaboko and chikuwa, bread and hot cakes. , Various side dishes, pudding, soup, etc.

Further, pet foods such as dog foods and cat foods of the present invention, and feeds such as livestock feeds are feeds containing the above calcium absorption promoter and calcium phosphate crystal growth agent.

When the calcium absorption promoter and the calcium phosphate crystal growth agent of the present invention are used especially for the purpose of promoting absorption of calcium, they are blended with a raw material containing calcium, for example, dairy products such as milk, yogurt and cheese. However, when calcium is not contained, the effect of promoting absorption of calcium can be further promoted by blending it with calcium. In any case, it is desirable that the ratio be such that the weight of the calcium crystallization inhibitor (protein) is 0.4% or more of the weight of calcium.

The effective dose of the calcium absorption enhancer of the present invention is difficult to define in a general manner, but it is not particularly limited when taken orally and exerts a calcium absorption enhancer effect in the intestinal tract. 0.08 mg / k
g / day or more is appropriate, preferably 0.1-100 mg
/ Kg / day.

Similarly, in the case of promoting remineralization of teeth for the purpose of promoting calcium phosphate crystal growth, 0.0
It is preferable to take 1 to 100 mg / kg / day.

[0033]

EXAMPLES The present invention will be described in more detail with reference to Examples and Test Examples. [Example 1] Preparation of culture of Lactococcus lactis IFO 3427 strain Lactococcus lactis IFO 3427 strain was pre-cultured in IFO-designated 804 medium at 30 ° C for 48 hours to prepare YP medium (5 l).
Main culture was carried out at 30 ° C. until the stationary phase was reached. The bacterial cells were removed, the culture supernatant was suction filtered with a 0.45 μm filter, then concentrated by ultrafiltration to 200 ml, dialyzed against distilled water 3 times at 4 ° C., and Lactococcus lactis IF.
24.3 mg of O 3427 strain culture was obtained.

[Example 2] Lactococcus lactis IFO 342
Preparation of 7-strain culture Lactococcus lactis obtained by the method described in Example 1
A solution of IFO 3427 strain culture was added to Super Q Toyopearl (Su
Fractionation was performed by anion chromatography (2.6 × 20 cm) of perQ-TOYOPEAL 650M (Tosoh Corp.), and the obtained active fraction was further superose 12 (Superose 12).
(Manufactured by Amersham)) Fractionated by gel filtration chromatography (1.6 × 60 cm), and the obtained active fraction is L
Actococcus lactis IFO 3427 strain culture was used. SDS-polyacrylamide gel electrophoresis shows a molecular weight of 34.0
It turned out to be 00.

[Example 3] Lactobacillus rhamnosus IF
Preparation of O 12521 Strain Culture By the method described in Example 1, Lactobacillus rhamnosus IFO
The 12521 strain was precultured in an IFO-designated 804 medium at 30 ° C. for 48 hours, and then precultured in an AM medium (5 l) at 30 ° C. until a stationary phase was reached. The bacterial cells were collected, washed twice with 10 mM Tris-hydrochloric acid buffer solution (pH 8.7), and washed with about 2 times 0.5 volume.
It was suspended in an aqueous solution of M in EDTA (pH 8.0) and sonicated for 10 minutes in a cycle of 30 seconds. After that, the supernatant was dialyzed against distilled water at 4 ° C to obtain Lactobacillus rhamnosus I
The FO 12521 strain culture was used.

Test Example 1 Measurement of Calcium Crystallization Inhibitory Effect First, a calcium crystallization inhibitor was examined in order to increase the calcium concentration in the intestinal tract and enhance the absorbability of calcium.

A reaction in which CaCO ₃ is precipitated from the reaction of an aqueous sodium hydrogen carbonate solution (NaHCO ₃ aqueous solution) and an aqueous calcium chloride solution (CaCl ₂ aqueous solution) (NaHCO ₃ + CaCl ₂ → CaCO ₃ +
HCl + NaCl) was used to determine the effect of suppressing calcium crystallization (Biochem. Biophys. Res. Comm., 110).
(1), p69-74, 1983).

That is, Lactococcu was added to an aqueous sodium hydrogencarbonate solution (1.5 ml) adjusted to 20 mM and pH 8.7.
30 μl of the s lactis IFO 3427 strain culture was added, and well stirred with a stirrer. After that, an aqueous calcium chloride solution (1.5 ml) adjusted to 20 mM and pH 8.7 was added,
The reaction was carried out at 25 ° C. During the reaction process, wavelength 570nm
The absorbance at was measured over time. The result is shown in FIG.

When a control (distilled water) is added,
A rapid increase in absorbance was observed by about 200 seconds,
The maximum value was shown after 0 seconds, and the reaction was completed. On the other hand, Lactoc
When the occus lactis IFO 3427 strain culture (final protein concentration 0.8 μg / ml) was added, no increase in absorbance was observed and calcium crystallization was completely suppressed.

Further, this extract was treated with proteinase K (Pro
teinase K), a decrease in calcium crystallization inhibitory activity was observed, suggesting that the center of activity was a protein.

[Test Example 2] Comparison of calcium crystallization inhibitory effect of various lactic acid bacterium cultures Based on the results of Test Example 1, the inhibition rate was calculated from the absorbance after 200 seconds of reaction according to the following [Formula 1]. , YP medium, AM
According to the extraction method of Example 1 or Example 3, various lactic acid bacteria cultured in the medium and NEW medium were subjected to a comparative test of the calcium crystallization inhibitory effect of the prepared culture (final protein concentration was 0.8 μg / ml). . The results are shown in [Table 6].

[0042]

[Equation 1]

[0043]

[Table 6]

From these results, L cultured in YP medium was
A particularly strong effect was observed on the extracellular culture of actococcus lactis IFO 3427.

[Test Example 3] Comparative Calcium Crystallization Inhibitory Effect of Various Microorganism Cultures According to the method of Test Example 2, the lactic acid bacterium of the present invention, Lactococcus la.
ctis IFO 3427 (YP medium, extracellular), Lactobacillus he
lveticus IFO 15019 (AM medium, extracellular), Streptococcu
s salivarius IFO 13957 (YP medium, extracellular), the bacterium and yeast derived from Antarctica described in Japanese Patent Laid-Open No. 2000-239181 by the present inventors already known to have a calcium crystallization inhibitory action. The cultures of No. 1 were comparatively examined for the effect of suppressing calcium crystallization. The results are shown in [Table 7].
Shown in.

[0046]

[Table 7]

As can be seen from these results, the culture of lactic acid bacteria of the present invention has a significantly higher effect of suppressing calcium crystallization than those known in the related art. Therefore, proteins obtained from these lactic acid bacteria also have an effect of suppressing calcium crystallization. Is expected to be a novel protein having

[Test Example 4] Lactococcus lactis IFO 342
7. Properties of extracellular culture of 7 Therefore, the properties of the extracellular culture of Lactococcus lactis IFO 3427 of the present invention were examined. The results are shown in [Table 8].
Shown in.

[0049]

[Table 8]

When the test lactic acid bacterium culture was treated with Proteinase K, the calcium crystallization-inhibiting effect was found to be low, so it is presumed that the calcium crystallization-inhibiting substance is a protein.

[Test Example 5] Calcium crystals with other substances
Comparison of anti-aging effect Then, a Lactococcus lactis IFO 3427 strain culture (0.
8 μg / ml) and other substances, namely casein phosphopep
Cide (CPP III) (2.0 μg / ml), Poly-L-glutami
Acid (2.0 μg / ml), poly-γ-glutamic acid (2.
0 μg / ml), EDTA: ethylenediaminetetraacetic acid (1.5
× 10⁻⁴M), citric acid (0.4 × 10⁻⁴ M), Hoss
Vitin (15 μg / ml), Heparin (15 μg / ml), co
Androitin sulfate C (15 μg / ml), albumin (10
μg / ml), lactoferrin (10 μg / ml), lipase
(10 μg / ml), trypsinogen (10 μg / ml),
α-chymotrypsinogen A (10 μg / ml), α-amyla
For enzyme (10 μg / ml) and esterase (10 μg / ml)
Then, the effect of suppressing calcium crystallization was comparatively examined. That
The results are shown in [Table 9].

[0052]

[Table 9]

As shown in [Table 9], when the Lactococcus lactis IFO 3427 strain culture (final protein concentration 0.8 μg / ml) prepared according to the method of Example 1 was added, the crystallization inhibition rate was 100%. When compared with other substances, casein phosphopeptide (CPP III) known as a calcium solubilizer is 62.8%, poly-L-glutamic acid (manufactured by Sigma; No. P-4886, sodium). (Salt) was 59.4%, and poly-γ-glutamic acid extracted from natto and precipitated with ethanol was purified to give an inhibition rate of 49.0%. 1 for each chelating agent such as EDTA or citric acid
The inhibition rates were 4.4% and 24.5%, and phosvitin, which was reported to inhibit crystallization of calcium carbonate, had an inhibition rate of 30.1%. In addition, no significant crystallization inhibitory effect was observed even when phosphoproteins and enzymes were added. The protein extracted from the Lactococcus lactis IFO 3427 strain of the present invention markedly inhibited calcium crystallization at a low concentration.

[Test Example 6] Comparison of Calcium Phosphate Crystal Growth Promoting Effect Next, according to the method of Example 2, various lactic acid bacteria cultivated in various media were examined for their effects on the calcium phosphate crystal growth. The reaction test was carried out by adding 1.5 ml of the culture prepared according to the method of Example 2 diluted with 0.05 M imidazole buffer (pH 7.0) to 6 mg of dicalcium phosphate (DCPD), 37 24 at ℃
It was carried out by stirring for an hour. The imidazole buffer solution was used as a blank. The reaction solution after 0 hours and 24 hours was filtered with a 0.45 μm filter, and the phosphoric acid content of the filtrate was measured with Phospha C-Test Wako to determine the calcium phosphate crystal growth promoting action rate. The result is
It shows in [Table 10]. The result was determined by the relative activity with the blank.

[0055]

[Table 10]

All the cultures of lactic acid bacteria of the present invention showed a strong calcium phosphate crystal growth promoting action.

Even when compared with phosvitin, albumin, lactoferrin, lipase and trypsinogen, which have a relatively calcium phosphate crystal growth promoting action, Lactococcus la
The ctisIFO 3427 strain culture was approximately twice as effective.

Hereinafter, examples of foods and drinks containing the calcium absorption promoter and the calcium phosphate crystal growth promoter of the present invention will be described. In addition, the compounding amount was described by weight%.

Example 4 Chewing gum was prepared according to the following formulation.

[0060]

[Table 11]

Example 5 Chewing gum was prepared according to the following formulation.

[0062]

[Table 12]

Example 6 Tablet confectionery was prepared according to the following formulation.

[0064]

[Table 13]

Example 7 Chocolate was prepared according to the following formulation.

[0066]

[Table 14]

Example 8 A beverage was prepared according to the following formulation.

[0068]

[Table 15]

Example 9 A beverage was prepared according to the following formulation.

[0070]

[Table 16]

[Example 10] An ice cream was prepared according to the following formulation.

[0072]

[Table 17]

[Example 11] A dog food was prepared according to the following formulation.

[0074]

[Table 18]

Example 12 A capsule was prepared according to the following formulation.

[0076]

[Table 19]

The above components were uniformly mixed, and the mixed powder was filled in a hard capsule.

Example 13 An injection was prepared according to the following formulation.

[0079]

[Table 20]

The above mixed solution was filtered through a membrane filter and then sterilized and filtered again. The filtrate was aseptically dispensed into a vial, filled with nitrogen gas, and then sealed to obtain an injection.

Example 14 Tablets were prepared according to the following formulation.

[0082]

[Table 21]

The above ingredients were uniformly mixed, and the mixed powder was compressed into tablets to give tablets of 200 mg each.

Fine particles for direct hitting No. 209 (20% magnesium aluminometasilicate, 30% corn starch, 50% lactose)
%)

[Example 15] A syrup was prepared according to the following formulation.

[0086]

[Table 22]

The extracted protein was completely dissolved in purified water, and a single syrup was added and mixed to obtain a syrup agent.

Example 16 A candy was prepared according to the following formulation.

[0089]

[Table 23]

Example 17 Biscuits were prepared according to the following formulation.

[0091]

[Table 24]

The above materials were mixed to form a dough, which was spread, molded, and then baked in an oven to produce a biscuit.

Example 18 Chewing gum was prepared according to the following formulation.

[0094]

[Table 25]

Example 19 Chewing gum was prepared according to the following formulation.

[0096]

[Table 26]

[Example 20] Tablet confectionery was prepared according to the following formulation.

[0098]

[Table 27]

Example 21 Chocolate was prepared according to the following formulation.

[0100]

[Table 28]

Example 22 A beverage was prepared according to the following formulation.

[0102]

[Table 29]

Example 23 A beverage was prepared according to the following formulation.

[0104]

[Table 30]

[Example 24] An ice cream was prepared according to the following formulation.

[0106]

[Table 31]

Example 25 A feed for laying chickens was prepared according to the following formulation.

[0108]

[Table 32]

Example 26 A capsule was prepared according to the following formulation.

[0110]

[Table 33]

The above components were uniformly mixed, and the mixed powder was filled into hard capsules.

[Example 27] An injection was prepared according to the following formulation.

[0113]

[Table 34]

The above mixed solution was filtered through a membrane filter and then subjected to sterilization filtration again, and the filtered solution was aseptically dispensed into a vial, filled with nitrogen gas, and then sealed to obtain an injection.

Example 28 A tablet was prepared according to the following formulation.

[0116]

[Table 35]

The above components were uniformly mixed, and the mixed powder was compressed into tablets to give tablets of 200 mg each.

Fine granules for direct hits No.209 (20% magnesium aluminometasilicate, 30% corn starch, 50 lactose)
%)

[Example 29] A syrup was prepared according to the following formulation.

[0120]

[Table 36]

The extracted protein was completely dissolved in purified water, and a single syrup was added and mixed to obtain a syrup agent.

[0122]

INDUSTRIAL APPLICABILITY The culture of the lactic acid bacterium of the present invention is remarkably excellent in calcium absorption promoting action and calcium phosphate crystal growth promoting action as compared with the substances known in the prior art, and further widely used in dairy products. Since it is a substance derived from lactic acid bacteria, it is extremely safe. In addition, foods and drinks containing the same exhibit excellent calcium absorption promoting effect and calcium phosphate crystal growth promoting effect.

[Brief description of drawings]

Figure 1: Lactococcus lactis on calcium crystallization
IFO 3427 strain culture (final protein concentration 0.8 μg /
(ml) is the graph which examined the influence.

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) A61K 35/74 A61P 3/14 A61P 1/02 19/10 3/14 43/00 105 19/10 A61K 37 / 02 43/00 105 A23L 2/00 F (72) Inventor Junichi Mori 1-33, Mikiteicho, Higashi-Osaka City, Osaka Prefecture 702 (72) Inventor Yoji Saeki 1-4-4 Haruno, Saitama City Urban Mirai Higashi-Omiya 3 No. 806 F term (reference) 2B150 AA06 AB20 AC05 AC06 AC07 DH04 DH14 4B017 LC03 LK01 LK15 LK21 LP01 4B018 LB08 MD20 MD86 ME05 MF01 4C084 AA02 AA03 BA05 CA04 MA16 MA23 MA34 MA35 MA37B A17 ZA21 MA47 MA21 MA47 MA67 MA14 MA14 AA01 AA02 BC55 BC56 BC57 BC61 CA32 MA16 MA23 MA34 MA35 MA37 MA47 MA66 NA14 ZA67 ZA97 ZB21 ZC21

Claims (24)

[Claims] 1. A genus Lactococcus, Lactobacillus, Ente
extracted from a culture of one or more lactic acid bacteria selected from the group consisting of genus rococcus and genus Streptococcus,
A calcium absorption promoting / calcium phosphate crystal growth promoting agent comprising a protein having both a calcium crystallization suppressing effect and a calcium phosphate crystal growth promoting effect as an active ingredient. 2. Lactococcus, Lactobacillus, Ente
An active ingredient of a protein extracted from an extracellular culture of one or more lactic acid bacteria selected from the group consisting of genus rococcus and genus Streptococcus and having both calcium crystallization inhibitory action and calcium phosphate crystal growth promoting action A calcium absorption-promoting / calcium phosphate crystal growth-promoting agent. 3. The culture is Lactococcus lactis IFO 3
The calcium absorption promoting / calcium phosphate crystal growth promoting agent according to claim 1 or 2, which is a 427 strain culture. 4. The calcium absorption promoting / calcium phosphate crystal growth promoting agent according to claim 3, wherein the Lactococcus lactis IFO 3427 strain culture is cultured in yeast peptone medium. 5. The culture is Lactobacillus brevis IFO
The culture according to claim 1 or 2, which is a 13110 strain culture.
The calcium absorption promoter / calcium phosphate crystal growth promoter described. 6. The calcium absorption promoting / calcium phosphate crystal growth promoting agent according to claim 5, wherein the Lactobacillus brevis IFO 13110 strain culture is cultured in a Lactobacillus medium. 7. The culture is Lactobacillus rhamnosus
The calcium absorption promoting / calcium phosphate crystal growth promoting agent according to claim 1 or 2, which is a culture of IFO 12521 strain. 8. The Lactobacillus rhamnosus IFO 1252
The calcium absorption promoting / calcium phosphate crystal growth promoting agent according to claim 7, wherein one strain culture is cultured in an acetate medium. 9. The culture is an Enterococcus durans IFO
It is a 13131 strain culture, The claim 1 or 2 characterized by the above-mentioned.
The calcium absorption promoter / calcium phosphate crystal growth promoter described. 10. The Enterococcus durans IFO 13131
The calcium absorption promoting / calcium phosphate crystal growth promoting agent according to claim 9, wherein the strain culture is cultured in yeast peptone medium. 11. The culture is Streptococcus salivari.
3. A culture of us IFO 13957 strain.
Alternatively, the calcium absorption promoting / calcium phosphate crystal growth promoting agent according to item 2. 12. The Streptococcus salivarius IFO 1
The calcium absorption promoting / calcium phosphate crystal growth promoting agent according to claim 11, wherein the 3957 strain culture is cultured in yeast peptone medium. 13. The culture is Lactobacillus casei Su
The calcium absorption promoter / calcium phosphate crystal growth promoter according to claim 1 or 2, which is a culture of bsp. casei JCM 1134 strain. 14. The Lactobacillus casei Subsp. Cas
The calcium absorption promoting / calcium phosphate crystal growth promoting agent according to claim 13, wherein the culture of the ei JCM 1134 strain is cultured in yeast peptone medium. 15. A food or drink comprising the calcium absorption promoting / calcium phosphate crystal growth promoting agent according to claim 1. 16. A feed comprising the calcium absorption promoting / calcium phosphate crystal growth promoting agent according to any one of claims 1 to 14. 17. The food or drink according to claim 15, further comprising calcium. 18. The feed according to claim 16, which further contains calcium. 19. Lactococcus, Lactobacillus, En
A calcium phosphate crystal growth promoter, which comprises a culture of one or more lactic acid bacteria selected from the group consisting of the genus terococcus and the genus Streptococcus as an active ingredient. 20. Lactococcus, Lactobacillus, En
A calcium phosphate crystal growth promoter, which comprises an extracellular culture of one or more lactic acid bacteria selected from the group consisting of the genus terococcus and the genus Streptococcus as an active ingredient. 21. A food or drink containing the calcium phosphate crystal growth promoter according to claim 19 or 20. 22. A feed comprising the calcium phosphate crystal growth promoting agent according to claim 19 or 20. 23. The food or drink according to claim 21, further comprising calcium. 24. The feed according to claim 22, further comprising calcium.