WO2013085009A1 - Food and drink with high lactobacillus gasseri survivability, and method for producing same - Google Patents
Food and drink with high lactobacillus gasseri survivability, and method for producing same Download PDFInfo
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- WO2013085009A1 WO2013085009A1 PCT/JP2012/081702 JP2012081702W WO2013085009A1 WO 2013085009 A1 WO2013085009 A1 WO 2013085009A1 JP 2012081702 W JP2012081702 W JP 2012081702W WO 2013085009 A1 WO2013085009 A1 WO 2013085009A1
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- lactic acid
- lactobacillus
- gasseri
- food
- acid bacteria
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- 235000013305 food Nutrition 0.000 title claims abstract description 57
- 241000186606 Lactobacillus gasseri Species 0.000 title claims abstract description 34
- 238000004519 manufacturing process Methods 0.000 title claims description 20
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims abstract description 188
- 241000894006 Bacteria Species 0.000 claims abstract description 148
- 239000004310 lactic acid Substances 0.000 claims abstract description 94
- 235000014655 lactic acid Nutrition 0.000 claims abstract description 94
- 235000013361 beverage Nutrition 0.000 claims abstract description 46
- 238000003860 storage Methods 0.000 claims abstract description 19
- 241000186605 Lactobacillus paracasei Species 0.000 claims description 41
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 28
- 239000008103 glucose Substances 0.000 claims description 28
- 235000020183 skimmed milk Nutrition 0.000 claims description 22
- 238000002156 mixing Methods 0.000 claims description 18
- 240000006024 Lactobacillus plantarum Species 0.000 claims description 17
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims description 17
- 229940072205 lactobacillus plantarum Drugs 0.000 claims description 17
- 241000186679 Lactobacillus buchneri Species 0.000 claims description 13
- 238000000034 method Methods 0.000 claims description 8
- 235000015140 cultured milk Nutrition 0.000 claims description 5
- 230000008569 process Effects 0.000 claims description 3
- 230000002035 prolonged effect Effects 0.000 abstract 1
- 239000002609 medium Substances 0.000 description 38
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- 239000008267 milk Substances 0.000 description 10
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- 238000012216 screening Methods 0.000 description 10
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- 244000199885 Lactobacillus bulgaricus Species 0.000 description 4
- 235000013960 Lactobacillus bulgaricus Nutrition 0.000 description 4
- 229930006000 Sucrose Natural products 0.000 description 4
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 4
- 235000013861 fat-free Nutrition 0.000 description 4
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- 230000004151 fermentation Effects 0.000 description 4
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- 230000036541 health Effects 0.000 description 4
- 230000036542 oxidative stress Effects 0.000 description 4
- 238000011218 seed culture Methods 0.000 description 4
- 241000193830 Bacillus <bacterium> Species 0.000 description 3
- 238000012790 confirmation Methods 0.000 description 3
- 238000000151 deposition Methods 0.000 description 3
- 229940004208 lactobacillus bulgaricus Drugs 0.000 description 3
- 230000007774 longterm Effects 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 230000020477 pH reduction Effects 0.000 description 3
- 101001076687 Lactobacillus gasseri Inulosucrase Proteins 0.000 description 2
- VLSOAXRVHARBEQ-UHFFFAOYSA-N [4-fluoro-2-(hydroxymethyl)phenyl]methanol Chemical compound OCC1=CC=C(F)C=C1CO VLSOAXRVHARBEQ-UHFFFAOYSA-N 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
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- 102000016938 Catalase Human genes 0.000 description 1
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- 235000013958 Lactobacillus casei Nutrition 0.000 description 1
- 241000186673 Lactobacillus delbrueckii Species 0.000 description 1
- 241001662087 Lactobacillus gasseri ATCC 33323 = JCM 1131 Species 0.000 description 1
- 240000002605 Lactobacillus helveticus Species 0.000 description 1
- 235000013967 Lactobacillus helveticus Nutrition 0.000 description 1
- 241001468157 Lactobacillus johnsonii Species 0.000 description 1
- 241000218588 Lactobacillus rhamnosus Species 0.000 description 1
- 244000057717 Streptococcus lactis Species 0.000 description 1
- 235000014897 Streptococcus lactis Nutrition 0.000 description 1
- 241000194020 Streptococcus thermophilus Species 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- LFYJSSARVMHQJB-QIXNEVBVSA-N bakuchiol Chemical compound CC(C)=CCC[C@@](C)(C=C)\C=C\C1=CC=C(O)C=C1 LFYJSSARVMHQJB-QIXNEVBVSA-N 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 210000005056 cell body Anatomy 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
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- 235000021001 fermented dairy product Nutrition 0.000 description 1
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- 239000000796 flavoring agent Substances 0.000 description 1
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- 239000001963 growth medium Substances 0.000 description 1
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
- A23C9/1234—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt characterised by using a Lactobacillus sp. other than Lactobacillus Bulgaricus, including Bificlobacterium sp.
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/145—Gasseri
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
Definitions
- the present invention relates to a method for producing a food or drink product having a high survival rate of Lactobacillus gasseri (hereinafter sometimes simply referred to as Lactobacillus gasseri or gasseri). More specifically, the present invention relates to a method for producing a food or drink in which the viability of gasseri is increased by mixing gasseri and other specific lactic acid bacteria. Moreover, it is related with the food / beverage products containing the gasseri microbe manufactured by the said manufacturing method.
- lactic acid bacteria and bifidobacteria contribute to pH reduction and flavor improvement due to lactic acid fermentation. It has become clear to show activity.
- Foods that claim to be functional by useful microorganisms such as lactic acid bacteria and bifidobacteria, mainly yogurt and lactic acid bacteria beverages, are increasing, and it is expected that demand will continue to increase with increasing interest in health.
- the number of viable bacteria that can assure the functionality of the food is specified. That is, it is essential that a certain number of bacteria survive within the shelf life of the food. Moreover, even if it is food other than food for specified health use, it is very important for quality control to maintain the number of viable bacteria in food. However, depending on the bacterial species, it may be easily killed during storage under the influence of the environment in food, such as pH and osmotic pressure, and the maintenance of the number of viable bacteria is often difficult.
- bifidobacteria are known to be easily affected by the oxygen content and pH in foods, and the following methods are used to improve the survival of the bacteria.
- a method of adding a cell-free extract of catalase-positive microorganisms that are harmless to hygiene to foods in order to prevent a decrease in the number of bifidobacteria during storage of liquid or paste-like foods is known (see Patent Document 1).
- a method for producing fermented milk in which Bifidobacteria and Lactococcus lactis subsp. Lactis are mixed and cultured is known.
- gasseri bacteria as lactic acid bacteria used in foods as useful microorganisms in the same manner as bifidobacteria. Since gasseri bacteria can be easily killed depending on the environmental conditions in the food, improvement of survival is required in the same way as bifidobacteria.
- As a method for improving the viability of gasseri bacteria it is known that the viability when stored at a low temperature is increased by mixed culture with a lactic acid strain having high antioxidant ability (see Non-Patent Document 1). Yes.
- Table 1 shows the results of examining the effect of elimination of oxidative stress on the storage viability of gasseri bacteria.
- Table 1 shows that L-cysteine hydrochloride, which is a reducing agent, is used to suspend rinsing cells of gasseri in 1% glucose and 0.3 M lactic acid buffer solution at pH 3.7 equivalent to that of lactic acid bacteria beverages. It is the result of having stored at 10 degreeC and measuring the number of gasseri bacteria (CFU / g) about the case where it does not add when it adds.
- This invention makes it a subject to provide the manufacturing method of the low pH and high sugar concentration food / beverage products with which the survival property of gasseri bacteria is favorable after a long-term storage.
- a method for producing a food or drink containing Lactobacillus gasseri The pH of the food or drink is 3.5 to 4.5, and the sugar concentration is 5% to 15%.
- the said manufacturing method including the process of mixing Lactobacillus gasseri (Lactobacillus gasseri) and specific lactic acid bacteria.
- Specific lactic acid bacteria when added to a pH 4.0 medium containing 3% (w / w) reduced skim milk and 10% (w / w) glucose and left to stand at 10 ° C. for 21 days, the pH of the medium is adjusted. Lactic acid bacteria that can be reduced by 0.1 or more.
- the Lactobacillus paracasei is SBT0327 (NITE ABP-1129), SBT2105 (NITE P-1130), SBT2203 (NITE ABP-1131), SBT2215 (NITE ABP-1132), SBT11408 (NITE ABP-11325)
- the Lactobacillus plantarum is any one of SBT1534 (FERM ABP-11518), SBT0624 (FERM P-11920), ATCC 43199, ATCC8014, and the Lactobacillus buchnerii is SBT2028 (FERM P-11921).
- the low pH and high sugar concentration food / beverage products in which the survival property of gasseri bacteria is very high even after long-term storage can be provided. Therefore, the foodstuff which can fully maintain and exhibit the function of gasseri bacteria which are useful microorganisms can be provided.
- the present invention adds a low pH, high sugar-concentrated food containing gasseri bacteria to 3% (w / w) reduced skim milk, 10% (w / w) glucose at pH 4.0 at 10 ° C. It is related with the manufacturing method of food-drinks including the process of adding and mixing the specific lactic acid bacteria which can reduce pH 0.1 or more when left still for days.
- the present invention is a method for producing a food or drink containing Lactobacillus gasseri, characterized in that a specific lactic acid bacterium is mixed with a gasseri bacterium in a food having a low pH and a high sugar concentration.
- the low pH of the food / beverage product of the present invention is 3.5 to 4.5
- the high sugar concentration is 5% to 15%, preferably pH 3.6 to 4.2
- the sugar concentration is 10%.
- -15% more preferably pH 3.7-4.0, sugar concentration 12% -14%, most preferably pH 3.8, sugar concentration 12.7%.
- the food / beverage products of this invention should just be the food / beverage products of the said low pH and high sugar concentration, are not limited to milk, dairy products, and fermented milk products, A vegetable drink, a fruit drink, etc. are included. Among them, fermented milk and lactic acid bacteria beverages defined by a ministerial ordinance concerning the component specifications of milk and dairy products are preferable.
- the gasseri bacterium in the present invention may be any one belonging to Lactobacillus gasseri, and SBT2055 (FERM BP-10953), SBT2056 (FERM BP-11038), SBT0274 (FERM BP-11039), SBT1703. (FERM P-17785), SBT10239 (FERM P-16639), SBT10241 (FERM P-17786) and the like.
- the specific lactic acid bacterium to be mixed with the gasseri bacterium may be a lactic acid bacterium that improves the viability of the gasseri bacterium by mixing with the gasseri bacterium, and as such a lactic acid bacterium, 3% (w / w)
- Lactic acid bacteria having such properties are added to a pH 4.0 medium containing 3% (w / w) reduced skim milk and 10% (w / w) glucose, and left to stand at 10 ° C. for 21 days. It is obtained by screening lactic acid bacteria capable of lowering the pH of the medium by 0.1 or more, specifically, Lactobacillus paracasei (hereinafter sometimes referred to as Lactobacillus paracasei), Lactobacillus paracasei. Plantarum (Lactobacillus plantarum, hereinafter referred to as Lactobacillus plantarum) and Lactobacillus buchneri (hereinafter, also referred to as Lactobacillus buchneri).
- Lactobacillus paracasei Lactobacillus paracasei
- Plantarum Lactobacillus plantarum
- Lactobacillus buchneri hereinafter, also referred to as Lactobacillus buchneri
- the number of specific lactic acid bacteria to be mixed with Lactobacillus gasseri is larger.
- the number of specific lactic acid bacteria to be mixed with Lactobacillus gasseri is larger.
- the survival improvement effect of gasseri is considered to correlate with metabolic activity under low temperature, pH, and high sugar concentration stress conditions.
- a 3% (w / w) reduced skim milk having a pH of 4.0, and lowering the pH by a certain level under 10% (w / w) glucose conditions was used as an index for screening.
- Specific lactic acid bacteria to be mixed with Lactobacillus gasseri include Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus buchneri, etc., but 3% (w / w) reduced skim milk and 10% ( w / w) Particularly limited as long as it is a lactic acid bacterium capable of lowering the pH of the medium by 0.1 or more when it is added to a pH 4.0 medium containing glucose and statically cultured at 10 ° C. for 21 days. is not.
- Lactobacillus paracasei examples include SBT0327 (NITE ABP-1129), SBT2105 (NITE P-1130), SBT2203 (NITE ABP-1131), SBT2215 (NITE ABP-1132), SBT11408 (NITE ABP-113)
- Lactobacillus plantarum examples include SBT1534 (FERM ABP-11518), SBT0624 (FERM P-11920), ATCC43199, ATCC8014, Lactobacillus buchneri, SBT2028 (FERM P-11192), JCM1115, and the like.
- examples of the step of mixing gasseri and specific lactic acid bacteria include the following forms. (1) After adding and fermenting gasseri bacteria to an unfermented milk medium, a specific lactic acid bacterium is added to the fermented product. (2) A specific lactic acid bacterium is added to an unfermented milk medium and fermented, and then a gasseri bacterium is added to the fermented product. (3) Gasseri bacteria and specific lactic acid bacteria are fermented simultaneously or sequentially with unfermented milk medium. (4) Gasseri bacteria and specific lactic acid bacteria are added simultaneously or sequentially to foods and drinks other than fermented lactic acid bacteria.
- Another embodiment of the present invention is a food or drink product having a pH of 3.5 to 4.5 and a sugar concentration of 5 to 15%, which includes gasseri bacteria and specific lactic acid bacteria obtained by the above production method.
- the survival number of gasseri bacteria in the food and drink of the present invention is much higher than when not mixed with specific lactic acid bacteria, for example, 10 times in 1 week, 1,000 to 10,000 times in 2 weeks, 1 in 3 weeks. , 10,000 to 10,000 times different.
- the number of surviving bacteria in food and drink is 1 ⁇ 10 6 CFU / g or more after storage for 21 days after addition of gasseri bacteria.
- Test Example 1 Screening of lactic acid bacteria mixed with gasseri bacteria (1) A screening test for lactic acid bacteria mixed with gasseri bacteria in the food and drink of the present invention was conducted.
- Test method Nonfat dry milk and glucose are dissolved in water to prepare a medium containing 3% (w / w) reduced skim milk and 10% (w / w) glucose, and the medium is sterilized at 115 ° C. for 20 minutes. did. Lactic acid sterilized by filtration was added to the sterilized medium to adjust the pH to about 4.0 to prepare a screening medium.
- the screening medium was inoculated with 3% each of seed cultures cultured in milk medium (described in Table 2), and the pH was measured when left at 10 ° C. for 21 days.
- the strain to be selected was selected.
- Test results Table 2 shows the results of the initial bacterial count of each lactic acid bacterium and the pH after storage. According to this, the pH of the medium to which Lactobacillus paracasei was added decreased by 0.1 or more.
- Lactobacillus paracasei was selected as a bacterium to be mixed with gasseri.
- Test Example 2 Screening of lactic acid bacteria mixed with gasseri bacteria (1)
- the medium when added to a pH 4.0 medium containing 3% (w / w) reduced skim milk and 10% (w / w) glucose, the medium was allowed to stand for 21 days at 10 ° C.
- Lactobacillus paracasei ATCC 25598, Lactobacillus plantarum ATCC 43199, Lactobacillus plantarum ATCC 8014, and Lactobacillus buchneri JCM1115 were selected.
- Table 3 shows the initial bacterial count of each lactic acid bacterium and the pH after storage. The screening was performed by the same method as in Test Example 1.
- Example 1 Test for confirming improvement in survival of gasseri by mixing with Lactobacillus paracasei
- the bacteria selected in the screening test of test example 1 are mixed with gasseri, and the survival of gasseri Confirmed to improve.
- (1) Preparation of lactic acid bacteria beverage of the present invention Nonfat dry milk and glucose are dissolved in water to prepare a medium containing 16% (w / w) reduced skim milk and 3% (w / w) glucose. Sterilized at 120 ° C. for 120 minutes.
- Lactic acid bacteria beverages of the present invention 1 to 5 having a pH of 3.8 and a sugar concentration of 12.7%, obtained by mixing 36 g of the sugar solution obtained above, 9 g of a fermented Lactobacillus paracasei product, and 0.5 g of S. cerevisiae SBT2055 Manufactured.
- (2) Preparation of control lactic acid bacteria beverage A control lactic acid bacteria beverage containing only gasseri bacteria and not mixed with Lactobacillus paracasei was produced. A quasi-fermented product was prepared by adding lactic acid sterilized by filtration to the sterilizing medium (1) and adjusting the pH to about 3.7.
- a control lactic acid bacteria beverage was prepared by mixing 9 g of the pseudo-fermented product, 36 g of the sugar solution of the above (1), and 0.5 g of concentrated bacteria of gasseri bacteria SBT2055. (3) Survival rate measurement test and results The lactic acid bacteria beverages and control lactic acid bacteria beverages of the products 1 to 5 of the present invention were each stored in a plastic container at 10 ° C. for 21 days, and live bacteria of gasseri and Lactobacillus paracasei Numbers were measured over time. The survival rate was obtained by calculating the number of viable bacteria on the 21st day by the viable cell count on the 0th day (the same applies in the following tests). The results are shown in Table 4.
- Example 2 Confirmation test for improvement of survival of gasseri by mixing with Lactobacillus paracasei When the number of Lactobacillus paracasei was changed and mixed with gasseri, and dead cells of Lactobacillus paracasei The effect on the viability of gasseri was investigated for the case of mixing with gasseri.
- (1) Preparation of lactic acid bacteria beverage of the present invention Nonfat dry milk and glucose are dissolved in water to prepare a medium containing 16% (w / w) reduced skim milk and 3% (w / w) glucose. Sterilized at 120 ° C. for 120 minutes.
- Lactic acid sterilized by filtration was added to the sterilized medium to adjust the pH to about 4.0 to prepare a pseudo-fermented product.
- Lactobacillus paracasei SBT11408 washed cells are suspended in a pseudo-fermented product, and three types of Lactobacillus paracasei-added pseudo-fermented products with different numbers of paracasei by diluting 10 times, 100 times, and 1,000 times 3 was prepared. Further, the washed cells of gasseri bacterium SBT2055 were suspended in 12.6% reduced skim milk and used. A 3% (w / w) isomerized liquid sugar / 14% (w / w) white sugar solution was prepared in water and sterilized at 121 ° C.
- the lactic acid bacteria drink (control 2) was manufactured using what sterilized the lactobacillus paracasei addition pseudo fermented product which contains about 1 * 10 ⁇ 7 > CFU / g of Lactobacillus paracasei for 30 minutes at 80 degreeC or more.
- (3) Survival rate measurement test and results The lactic acid bacteria beverages and control lactic acid bacteria beverages (Controls 1 and 2) of the products 5 to 7 of the present invention were respectively stored in plastic containers for 21 days at 10 ° C. The viable count of Bacillus paracasei was measured over time. The results are shown in Table 5.
- Example 3 Confirmation test for improvement of survival of gasseri by mixing with Lactobacillus paracasei at a sugar concentration of 5-15% in food and drink (1)
- Preparation of lactic acid bacteria beverage of the present invention Nonfat dry milk and glucose A 16% (w / w) reduced skim milk medium was prepared by dissolving in water, and the medium was sterilized at 115 ° C. for 20 minutes. Lactic acid sterilized by filtration was added to the sterilized medium to adjust the pH to about 3.5 to prepare a pseudo-fermented product. Lactobacillus paracasei SBT11408 washed cells were suspended in the pseudofermented product to prepare a lactobacillus paracasei-added pseudofermented product.
- washed cells of gasseri bacterium SBT2055 were suspended in 12.6% reduced skim milk and used.
- sugar solutions aqueous glucose solutions having final concentrations of 5%, 10%, and 15% were prepared and sterilized at 121 ° C. for 15 minutes.
- a control lactic acid bacteria beverage was prepared by mixing at 15%.
- Survival rate measurement test and results The lactic acid bacteria beverages and the control lactic acid bacteria beverages of the products 8 to 10 of the present invention were each stored in a plastic container at 10 ° C. for 21 days, and live bacteria of gasseri and Lactobacillus paracasei Numbers were measured over time. The results are shown in Table 6.
- Lactobacillus paracasei to which 1 ⁇ 10 7 CFU / g or more is added and the lactic acid bacteria beverage of the present invention having a glucose concentration of 5% to 15% improves the survival of Lactobacillus gasseri, and the lactobacilli after storage for 21 days
- the viable count of gasseri was 1 ⁇ 10 6 CFU / g or more. The effect was particularly remarkable when the glucose concentration was 10% or more.
- Example 4 Skim milk powder and glucose were dissolved in water to prepare a medium containing 16% (w / w) reduced skim milk and 3% (w / w) glucose, and the medium was sterilized at 95 ° C. for 120 minutes. 3% (v / v) of Lactobacillus buchneri SBT2028 seed culture was added to the sterilized medium cooled to 37 ° C. and cultured at 37 ° C. until the pH reached about 4.0 to obtain a fermented Lactobacillus buchneri. Further, a 3% (w / w) isomerized liquid sugar / 14% (w / w) white sugar solution was prepared in water and sterilized at 121 ° C.
- Lactobacillus gasseri 1.9 ⁇ 10 9 CFU / g
- Lactobacillus bucheneri 4.8 ⁇ 10 8 CFU / g added Lactobacillus gasseri viable count after storage for 21 days is 5. It was 0 ⁇ 10 6 CFU / g.
- Example 5 Skim milk powder and glucose were dissolved in water to prepare a medium containing 16% (w / w) reduced skim milk and 3% (w / w) glucose, and the medium was sterilized at 95 ° C. for 120 minutes.
- 3% (v / v) each of seed cultures of Lactobacillus plantarum SBT1534 and Lactobacillus gasseri SBT10241 were added to a sterilized medium cooled to 37 ° C., and fermented at 37 ° C. until the pH reached about 4.0. .
- a 3% (w / w) isomerized liquid sugar / 14% (w / w) white sugar solution was prepared in water and sterilized at 121 ° C.
- Lactobacillus gasseri is 2.0 ⁇ 10 9 CFU / g and Lactobacillus plantarum is 5.4 ⁇ 10 8 CFU / g.
- the number of viable Lactobacillus gasseri after storage for 21 days is 12 ⁇ 10 6 CFU / g.
- Example 6 Confirmation test for improvement of survival of gasseri by mixing with Lactobacillus paracasei, Lactobacillus plantarum or Lactobacillus buchneri The bacteria selected in the screening test of Test Example 2 above It mixed and it confirmed that the survival property of gasseri bacteria improved.
- a lactic acid bacteria beverage of the present invention and a control lactic acid bacteria beverage were prepared in the same manner as in Example 1.
- Lactobacillus gasseri SBT2055 and the standard strain Lactobacillus gasseri JCM1131 are used as gasseri bacteria, and Lactobacillus paracasei ATCC 25598, Lactobacillus plantarum ATCC 43199, Bacillus plantarum ATCC 8014 and Lactobacillus buchneri JCM1115 were used.
- a control lactic acid bacteria beverage a lactic acid bacteria beverage using Lactobacillus bulgaricus (NBRC P-13953) as a bacterium mixed with gasseri was also prepared. About the prepared lactic acid bacteria drink, the survival rate was measured about the gasseri bacteria and the mixed bacteria by the same method as Example 1. The results are shown in Table 7.
- the lactic acid bacteria beverages of the present invention in which gasseri and Lactobacillus paracasei, Lactobacillus plantarum, and Lactobacillus buchnerii were mixed were markedly different from the control lactic acid bacteria beverages.
- the number of surviving bacteria was high, and the number of viable bacteria of Lactobacillus gasseri after storage for 21 days was 1 ⁇ 10 6 CFU / g or more in all of the products 11 to 18 of the present invention.
- Lactobacillus bulgaricus was mixed, the viability of gasseri was not improved, and the survival rate of Lactobacillus gasseri after storage for 21 days was 0.00006% or less.
- the low pH and high sugar concentration food / beverage products in which the survival property of gasseri bacteria is very high even after long-term storage can be provided. Therefore, the foodstuff which can fully maintain and exhibit the function of gasseri bacteria which are useful microorganisms can be provided.
- Lactobacillus gasseri SBT2055 Name and address of the depository institution that deposited the biological material National Institute of Advanced Industrial Science and Technology Patent Biological Deposit Center 1-Chuo 1-chome, Tsukuba City, Ibaraki Prefecture, Japan 6 (zip code 305-8566) Date of deposit of biological materials at Loi depository organization March 27, 1996 (original deposit date) February 26, 2008 (Date of transfer to deposit under the Budapest Treaty by original deposit) Deposit number FERM BP-10953 assigned by the depository in Thailand for deposit (2) Lactobacillus gasseri SBT2056 (A) Name and address of the depositary that deposited the biological material Date of deposit of the biological material at the same depository in Loi above (1) April 22, 1986 (original deposit date) October 9, 2008 (Date of transfer to deposit under the Budapest Treaty by original deposit) Deposit number FERM BP-11038 assigned by the depository in Hai for the deposit (3) Lactobacillus gasseri SBT0274 The name and address of the depository that
- Pseudoplantarum SBT0624 (A) Name and address of the depositary that deposited the biological material Date of deposit of the biological material at the same depository in (4) above The deposit number assigned by the depositary institution on December 20, 1990 FERM P-11920 (14) Lactobacillus buchneri SBT2028 (A) Name and address of the depositary that deposited the biological material Date of deposit of the biological material at the same depository in (4) above The deposit number assigned by the depositary institution on December 20, 1990 FERM P-11921
Abstract
This invention addresses the problem of providing a low pH, high sugar concentration food and drink with good Lactobacillus gasseri survivability even after prolonged storage, which can be obtained by adding a specific lactic acid bacteria to a low pH, high sugar concentration food and beverage containing Lactobacillus gasseri. The specific lactic acid bacteria refers to a lactic acid bacteria capable of reducing pH to a constant value in a low pH, high sugar concentration condition.
Description
本発明は、ラクトバチルス・ガセリ（Lactobacillus gasseri、以下単にラクトバチルス・ガセリ、ガセリ菌ということがある。）の生残性の高い飲食品の製造方法に関する。さらに詳しくは、ガセリ菌と他の特定の乳酸菌を混合することでガセリ菌の生残性を高めた飲食品の製造方法に関する。また、当該製造方法によって製造されたガセリ菌を含む飲食品に関する。
The present invention relates to a method for producing a food or drink product having a high survival rate of Lactobacillus gasseri (hereinafter sometimes simply referred to as Lactobacillus gasseri or gasseri). More specifically, the present invention relates to a method for producing a food or drink in which the viability of gasseri is increased by mixing gasseri and other specific lactic acid bacteria. Moreover, it is related with the food / beverage products containing the gasseri microbe manufactured by the said manufacturing method.
乳酸菌およびビフィズス菌は、多くの発酵食品において、乳酸発酵によるｐH低下、風味改善に寄与している一方、生きたままの状態で腸まで到達させることで、整腸効果、免疫賦活など様々な生理活性を示すことが明らかになってきている。ヨーグルトや乳酸菌飲料を中心に乳酸菌・ビフィズス菌等の有用微生物による機能性をうたった食品は増えており、健康への関心の高まりとともに今後も需要は増大していくと考えられる。
In many fermented foods, lactic acid bacteria and bifidobacteria contribute to pH reduction and flavor improvement due to lactic acid fermentation. It has become clear to show activity. Foods that claim to be functional by useful microorganisms such as lactic acid bacteria and bifidobacteria, mainly yogurt and lactic acid bacteria beverages, are increasing, and it is expected that demand will continue to increase with increasing interest in health.
特定保健用食品として承認されるためには、試験に基づいた科学的根拠を提示することが求められる。したがって、有用微生物を用いる特定保健用食品に関しては、その食品が掲げる機能性が保証されるだけの生菌数を規定することになる。すなわち、当該食品の賞味期限内に一定以上の菌数が生き残ることが必須である。また、特定保健用食品以外の食品であっても、食品中の生菌数を維持することは品質管理上、非常に重要である。
しかしながら、菌種によっては食品中の環境、例えばｐHや浸透圧等の影響を受けて保存中に容易に死滅していく場合があり、生菌数の維持管理は困難な場合が多い。 To be approved as a food for specified health use, it is required to provide scientific evidence based on the test. Therefore, for the food for specified health use that uses useful microorganisms, the number of viable bacteria that can assure the functionality of the food is specified. That is, it is essential that a certain number of bacteria survive within the shelf life of the food. Moreover, even if it is food other than food for specified health use, it is very important for quality control to maintain the number of viable bacteria in food.
However, depending on the bacterial species, it may be easily killed during storage under the influence of the environment in food, such as pH and osmotic pressure, and the maintenance of the number of viable bacteria is often difficult.
しかしながら、菌種によっては食品中の環境、例えばｐHや浸透圧等の影響を受けて保存中に容易に死滅していく場合があり、生菌数の維持管理は困難な場合が多い。 To be approved as a food for specified health use, it is required to provide scientific evidence based on the test. Therefore, for the food for specified health use that uses useful microorganisms, the number of viable bacteria that can assure the functionality of the food is specified. That is, it is essential that a certain number of bacteria survive within the shelf life of the food. Moreover, even if it is food other than food for specified health use, it is very important for quality control to maintain the number of viable bacteria in food.
However, depending on the bacterial species, it may be easily killed during storage under the influence of the environment in food, such as pH and osmotic pressure, and the maintenance of the number of viable bacteria is often difficult.
例えば、ビフィズス菌は食品中の酸素含量やｐHの影響を受けやすいことが知られており、その生残性を改善するために以下に挙げる方法がとられている。液状またはのり状食品におけるビフィズス菌の保存中の菌数低下を防止するために、衛生上無害なカタラーゼ陽性微生物の無細胞抽出液を食品に添加する方法（特許文献１参照）が知られている。また、ビフィズス菌とラクトコッカス・ラクティス・サブスピーシーズ・ラクティス（Lactococcus lactis subsp. lactis）を混合培養する発酵乳の製造方法（特許文献２参照）が知られている。
For example, bifidobacteria are known to be easily affected by the oxygen content and pH in foods, and the following methods are used to improve the survival of the bacteria. A method of adding a cell-free extract of catalase-positive microorganisms that are harmless to hygiene to foods in order to prevent a decrease in the number of bifidobacteria during storage of liquid or paste-like foods is known (see Patent Document 1). . In addition, a method for producing fermented milk in which Bifidobacteria and Lactococcus lactis subsp. Lactis are mixed and cultured (see Patent Document 2) is known.
また、ビフィズス菌と同様に有用微生物として食品に用いられる乳酸菌として、ガセリ菌がある。ガセリ菌についても、食品中の環境条件によっては容易に死滅していくため、ビフィズス菌と同様に生残性の改善が求められている。ガセリ菌の生残性を改善する方法としては、抗酸化能が高い乳酸菌株との混合培養によって、低温で保存した場合の生残性が上昇すること（非特許文献１参照）が知られている。
In addition, there are gasseri bacteria as lactic acid bacteria used in foods as useful microorganisms in the same manner as bifidobacteria. Since gasseri bacteria can be easily killed depending on the environmental conditions in the food, improvement of survival is required in the same way as bifidobacteria. As a method for improving the viability of gasseri bacteria, it is known that the viability when stored at a low temperature is increased by mixed culture with a lactic acid strain having high antioxidant ability (see Non-Patent Document 1). Yes.
しかしながら、発酵乳を含む食品中ではｐＨ低下、浸透圧、酸化ストレス等が複合的に作用するため、単に抗酸化能が高い乳酸菌株と混合培養するという酸化ストレスの解消だけではガセリ菌の生残性を十分に改善することはできなかった。
ここで、ガセリ菌の保存生残性に対する酸化ストレスの解消の影響を調べた結果を表１に示す。表１は、乳酸菌飲料と同等のｐＨ３．７において、１％グルコース、０．３Ｍ乳酸緩衝液にガセリ菌の洗浄菌体を懸濁し、酸化ストレス解消のため還元剤であるＬ－システイン塩酸塩を添加した場合と添加しなかった場合について、１０℃で保存してガセリ菌数（ＣＦＵ／ｇ）を測定した結果である。
本結果より、ガセリ菌の生残性は、Ｌ－システイン塩酸塩の添加によって酸化ストレスが解消された場合であっても、酸化ストレスを改善していない場合に比べれば若干改善したものの、２１日間保存後の生残数は２．６×１０３ ＣＦＵ／ｇにすぎず、ガセリ菌の有用微生物としての効果を発揮するためには満足できるものではなかった。 However, in foods containing fermented milk, pH reduction, osmotic pressure, oxidative stress, etc. act in a complex manner. The sex could not be improved sufficiently.
Here, Table 1 shows the results of examining the effect of elimination of oxidative stress on the storage viability of gasseri bacteria. Table 1 shows that L-cysteine hydrochloride, which is a reducing agent, is used to suspend rinsing cells of gasseri in 1% glucose and 0.3 M lactic acid buffer solution at pH 3.7 equivalent to that of lactic acid bacteria beverages. It is the result of having stored at 10 degreeC and measuring the number of gasseri bacteria (CFU / g) about the case where it does not add when it adds.
From this result, even when the oxidative stress was eliminated by the addition of L-cysteine hydrochloride, the viability of Gasseri was improved slightly compared to the case where the oxidative stress was not improved, but for 21 days. The number of survivors after storage was only 2.6 × 10 3 CFU / g, which was not satisfactory for exhibiting the effect of gasseri as a useful microorganism.
ここで、ガセリ菌の保存生残性に対する酸化ストレスの解消の影響を調べた結果を表１に示す。表１は、乳酸菌飲料と同等のｐＨ３．７において、１％グルコース、０．３Ｍ乳酸緩衝液にガセリ菌の洗浄菌体を懸濁し、酸化ストレス解消のため還元剤であるＬ－システイン塩酸塩を添加した場合と添加しなかった場合について、１０℃で保存してガセリ菌数（ＣＦＵ／ｇ）を測定した結果である。
本結果より、ガセリ菌の生残性は、Ｌ－システイン塩酸塩の添加によって酸化ストレスが解消された場合であっても、酸化ストレスを改善していない場合に比べれば若干改善したものの、２１日間保存後の生残数は２．６×１０３ ＣＦＵ／ｇにすぎず、ガセリ菌の有用微生物としての効果を発揮するためには満足できるものではなかった。 However, in foods containing fermented milk, pH reduction, osmotic pressure, oxidative stress, etc. act in a complex manner. The sex could not be improved sufficiently.
Here, Table 1 shows the results of examining the effect of elimination of oxidative stress on the storage viability of gasseri bacteria. Table 1 shows that L-cysteine hydrochloride, which is a reducing agent, is used to suspend rinsing cells of gasseri in 1% glucose and 0.3 M lactic acid buffer solution at pH 3.7 equivalent to that of lactic acid bacteria beverages. It is the result of having stored at 10 degreeC and measuring the number of gasseri bacteria (CFU / g) about the case where it does not add when it adds.
From this result, even when the oxidative stress was eliminated by the addition of L-cysteine hydrochloride, the viability of Gasseri was improved slightly compared to the case where the oxidative stress was not improved, but for 21 days. The number of survivors after storage was only 2.6 × 10 3 CFU / g, which was not satisfactory for exhibiting the effect of gasseri as a useful microorganism.
本発明は、長期保存後もガセリ菌の生残性が良好な低ｐH、高糖濃度飲食品の製造方法を提供することを課題とする。
This invention makes it a subject to provide the manufacturing method of the low pH and high sugar concentration food / beverage products with which the survival property of gasseri bacteria is favorable after a long-term storage.
以下、本発明の構成を示す。
（１）ラクトバチルス・ガセリ（Lactobacillus gasseri）を含む飲食品の製造方法であって、
飲食品のｐＨが３．５～４．５、糖濃度が５％～１５％であり、
ラクトバチルス・ガセリ（Lactobacillus gasseri）と特定の乳酸菌を混合する工程を含む前記製造方法。
特定の乳酸菌；３％（ｗ／ｗ）還元脱脂乳および１０％（ｗ／ｗ）グルコース を含むｐＨ４．０の培地に添加して１０℃において２１日間静置培養したときに該培地のｐＨを０．１以上低下させることができる乳酸菌。
（２）前記特定の乳酸菌が、ラクトバチルス・パラカゼイ(Lactobacillus paracasei)、ラクトバチルス・プランタラム（Lactobacillus plantarum）またはラクトバチルス・ブフネリ（Lactobacillus buchneri）である上記（１）に記載の飲食品の製造方法。
（３）前記ラクトバチルス・パラカゼイがＳＢＴ０３２７（ＮＩＴＥ ＡＢＰ－１１２９）、ＳＢＴ２１０５（ＮＩＴＥ Ｐ－１１３０）、ＳＢＴ２２０３（ＮＩＴＥ ＡＢＰ－１１３１）、ＳＢＴ２２１５（ＮＩＴＥ ＡＢＰ－１１３２）、ＳＢＴ１１４０８（ＮＩＴＥ ＡＢＰ－１１３３）、ＡＴＣＣ２５５９８のいずれかであり、前記ラクトバチルス・プランタラムがＳＢＴ１５３４（ＦＥＲＭ ＡＢＰ－１１５１８）、ＳＢＴ０６２４（ＦＥＲＭ Ｐ－１１９２０）、ＡＴＣＣ４３１９９、ＡＴＣＣ８０１４のいずれかであり、前記ラクトバチルス・ブフネリがＳＢＴ２０２８（ＦＥＲＭ Ｐ‐１１９２１）またはＪＣＭ１１１５である請求項１または２に記載の飲食品の製造方法。
（４）前記飲食品が、発酵乳および乳酸菌飲料である上記（１）～（３）のいずれかに記載の飲食品の製造方法。
（５）ラクトバチルス・ガセリ（Lactobacillus gasseri）および特定の乳酸菌を含む、ｐＨが３．５～４．５、糖濃度が５％～１５％の飲食品；
特定の乳酸菌；１０℃において３％（ｗ／ｗ）還元脱脂乳および１０％（ｗ／ｗ）グルコース を含むｐＨ４．０の培地に添加して２１日間静置培養したときに当該培地のｐＨを０．１以上低下させることができる乳酸菌。
（６）２１日間保存後のラクトバチルス・ガセリの生菌数が１×１０６ＣＦＵ／ｇ以上である上記（５）に記載の飲食品。 The configuration of the present invention will be described below.
(1) A method for producing a food or drink containing Lactobacillus gasseri,
The pH of the food or drink is 3.5 to 4.5, and the sugar concentration is 5% to 15%.
The said manufacturing method including the process of mixing Lactobacillus gasseri (Lactobacillus gasseri) and specific lactic acid bacteria.
Specific lactic acid bacteria; when added to a pH 4.0 medium containing 3% (w / w) reduced skim milk and 10% (w / w) glucose and left to stand at 10 ° C. for 21 days, the pH of the medium is adjusted. Lactic acid bacteria that can be reduced by 0.1 or more.
(2) The method for producing a food or drink according to (1) above, wherein the specific lactic acid bacterium is Lactobacillus paracasei, Lactobacillus plantarum, or Lactobacillus buchneri. .
(3) The Lactobacillus paracasei is SBT0327 (NITE ABP-1129), SBT2105 (NITE P-1130), SBT2203 (NITE ABP-1131), SBT2215 (NITE ABP-1132), SBT11408 (NITE ABP-11325) The Lactobacillus plantarum is any one of SBT1534 (FERM ABP-11518), SBT0624 (FERM P-11920), ATCC 43199, ATCC8014, and the Lactobacillus buchnerii is SBT2028 (FERM P-11921). ) Or JCM1115. The method for producing a food or drink according to claim 1 or 2.
(4) The method for producing a food or drink according to any one of (1) to (3), wherein the food or drink is fermented milk and a lactic acid bacteria beverage.
(5) A food or drink containing Lactobacillus gasseri and a specific lactic acid bacterium having a pH of 3.5 to 4.5 and a sugar concentration of 5 to 15%;
Specific lactic acid bacteria: when added to a pH 4.0 medium containing 3% (w / w) reduced skim milk and 10% (w / w) glucose at 10 ° C. Lactic acid bacteria that can be reduced by 0.1 or more.
(6) The food or drink according to (5) above, wherein the viable cell count of Lactobacillus gasseri after storage for 21 days is 1 × 10 6 CFU / g or more.
（１）ラクトバチルス・ガセリ（Lactobacillus gasseri）を含む飲食品の製造方法であって、
飲食品のｐＨが３．５～４．５、糖濃度が５％～１５％であり、
ラクトバチルス・ガセリ（Lactobacillus gasseri）と特定の乳酸菌を混合する工程を含む前記製造方法。
特定の乳酸菌；３％（ｗ／ｗ）還元脱脂乳および１０％（ｗ／ｗ）グルコース を含むｐＨ４．０の培地に添加して１０℃において２１日間静置培養したときに該培地のｐＨを０．１以上低下させることができる乳酸菌。
（２）前記特定の乳酸菌が、ラクトバチルス・パラカゼイ(Lactobacillus paracasei)、ラクトバチルス・プランタラム（Lactobacillus plantarum）またはラクトバチルス・ブフネリ（Lactobacillus buchneri）である上記（１）に記載の飲食品の製造方法。
（３）前記ラクトバチルス・パラカゼイがＳＢＴ０３２７（ＮＩＴＥ ＡＢＰ－１１２９）、ＳＢＴ２１０５（ＮＩＴＥ Ｐ－１１３０）、ＳＢＴ２２０３（ＮＩＴＥ ＡＢＰ－１１３１）、ＳＢＴ２２１５（ＮＩＴＥ ＡＢＰ－１１３２）、ＳＢＴ１１４０８（ＮＩＴＥ ＡＢＰ－１１３３）、ＡＴＣＣ２５５９８のいずれかであり、前記ラクトバチルス・プランタラムがＳＢＴ１５３４（ＦＥＲＭ ＡＢＰ－１１５１８）、ＳＢＴ０６２４（ＦＥＲＭ Ｐ－１１９２０）、ＡＴＣＣ４３１９９、ＡＴＣＣ８０１４のいずれかであり、前記ラクトバチルス・ブフネリがＳＢＴ２０２８（ＦＥＲＭ Ｐ‐１１９２１）またはＪＣＭ１１１５である請求項１または２に記載の飲食品の製造方法。
（４）前記飲食品が、発酵乳および乳酸菌飲料である上記（１）～（３）のいずれかに記載の飲食品の製造方法。
（５）ラクトバチルス・ガセリ（Lactobacillus gasseri）および特定の乳酸菌を含む、ｐＨが３．５～４．５、糖濃度が５％～１５％の飲食品；
特定の乳酸菌；１０℃において３％（ｗ／ｗ）還元脱脂乳および１０％（ｗ／ｗ）グルコース を含むｐＨ４．０の培地に添加して２１日間静置培養したときに当該培地のｐＨを０．１以上低下させることができる乳酸菌。
（６）２１日間保存後のラクトバチルス・ガセリの生菌数が１×１０６ＣＦＵ／ｇ以上である上記（５）に記載の飲食品。 The configuration of the present invention will be described below.
(1) A method for producing a food or drink containing Lactobacillus gasseri,
The pH of the food or drink is 3.5 to 4.5, and the sugar concentration is 5% to 15%.
The said manufacturing method including the process of mixing Lactobacillus gasseri (Lactobacillus gasseri) and specific lactic acid bacteria.
Specific lactic acid bacteria; when added to a pH 4.0 medium containing 3% (w / w) reduced skim milk and 10% (w / w) glucose and left to stand at 10 ° C. for 21 days, the pH of the medium is adjusted. Lactic acid bacteria that can be reduced by 0.1 or more.
(2) The method for producing a food or drink according to (1) above, wherein the specific lactic acid bacterium is Lactobacillus paracasei, Lactobacillus plantarum, or Lactobacillus buchneri. .
(3) The Lactobacillus paracasei is SBT0327 (NITE ABP-1129), SBT2105 (NITE P-1130), SBT2203 (NITE ABP-1131), SBT2215 (NITE ABP-1132), SBT11408 (NITE ABP-11325) The Lactobacillus plantarum is any one of SBT1534 (FERM ABP-11518), SBT0624 (FERM P-11920), ATCC 43199, ATCC8014, and the Lactobacillus buchnerii is SBT2028 (FERM P-11921). ) Or JCM1115. The method for producing a food or drink according to claim 1 or 2.
(4) The method for producing a food or drink according to any one of (1) to (3), wherein the food or drink is fermented milk and a lactic acid bacteria beverage.
(5) A food or drink containing Lactobacillus gasseri and a specific lactic acid bacterium having a pH of 3.5 to 4.5 and a sugar concentration of 5 to 15%;
Specific lactic acid bacteria: when added to a pH 4.0 medium containing 3% (w / w) reduced skim milk and 10% (w / w) glucose at 10 ° C. Lactic acid bacteria that can be reduced by 0.1 or more.
(6) The food or drink according to (5) above, wherein the viable cell count of Lactobacillus gasseri after storage for 21 days is 1 × 10 6 CFU / g or more.
本発明によれば、長期間保存後もガセリ菌の生残性が極めて高い、低ｐＨ、高糖濃度飲食品を提供することができる。したがって、有用微生物であるガセリ菌の機能を十分に維持し発揮できる食品を提供することができる。
ADVANTAGE OF THE INVENTION According to this invention, the low pH and high sugar concentration food / beverage products in which the survival property of gasseri bacteria is very high even after long-term storage can be provided. Therefore, the foodstuff which can fully maintain and exhibit the function of gasseri bacteria which are useful microorganisms can be provided.
上記課題に鑑み検討を重ねた結果、低ｐＨ、高糖濃度飲食品において特定の乳酸菌をガセリ菌と混合することで、飲食品保存中の生残性を高めることができることを見出し、本発明を完成させるに至った。
すなわち、本発明は、ガセリ菌を含む低ｐＨ、高糖濃度食品に、１０℃においてｐＨ４．０の３％（ｗ／ｗ）還元脱脂乳、１０％（ｗ／ｗ）グルコースに添加して２１日間静置したときにｐＨを０．１以上低下させることができる特定乳酸菌を添加して混合する工程を含む、飲食品の製造方法に関する。
本発明はラクトバチルス・ガセリ（Lactobacillus gasseri）を含む飲食品の製造方法であって、低ｐH、高糖濃度の食品中で特定の乳酸菌をガセリ菌と混合することを特徴とする。
本発明の飲食品の低ｐＨとは３．５～４．５であり、高糖濃度とは５％～１５％であり、好ましくはｐＨ３．６～４．２であり、糖濃度は１０％～１５％であり、さらに好ましくはｐＨ３．７～４．０であり、糖濃度は１２％～１４％であり、もっとも好ましくはｐH３．８、糖濃度１２．７％である。
本発明の飲食品は、上記低ｐH、高糖濃度の飲食品であればよく、乳、乳製品、発酵乳製品に限定されず、野菜飲料、果実飲料等をも含む。そのうちでも好ましくは、乳および乳製品の成分規格等に関する省令で定められた発酵乳および乳酸菌飲料である。 As a result of repeated investigations in view of the above problems, it has been found that by mixing a specific lactic acid bacterium with gasseri bacteria in a low pH, high sugar concentration food and drink, the survival of the food and drink can be increased, and the present invention is It came to complete.
That is, the present invention adds a low pH, high sugar-concentrated food containing gasseri bacteria to 3% (w / w) reduced skim milk, 10% (w / w) glucose at pH 4.0 at 10 ° C. It is related with the manufacturing method of food-drinks including the process of adding and mixing the specific lactic acid bacteria which can reduce pH 0.1 or more when left still for days.
The present invention is a method for producing a food or drink containing Lactobacillus gasseri, characterized in that a specific lactic acid bacterium is mixed with a gasseri bacterium in a food having a low pH and a high sugar concentration.
The low pH of the food / beverage product of the present invention is 3.5 to 4.5, the high sugar concentration is 5% to 15%, preferably pH 3.6 to 4.2, and the sugar concentration is 10%. -15%, more preferably pH 3.7-4.0, sugar concentration 12% -14%, most preferably pH 3.8, sugar concentration 12.7%.
The food / beverage products of this invention should just be the food / beverage products of the said low pH and high sugar concentration, are not limited to milk, dairy products, and fermented milk products, A vegetable drink, a fruit drink, etc. are included. Among them, fermented milk and lactic acid bacteria beverages defined by a ministerial ordinance concerning the component specifications of milk and dairy products are preferable.
すなわち、本発明は、ガセリ菌を含む低ｐＨ、高糖濃度食品に、１０℃においてｐＨ４．０の３％（ｗ／ｗ）還元脱脂乳、１０％（ｗ／ｗ）グルコースに添加して２１日間静置したときにｐＨを０．１以上低下させることができる特定乳酸菌を添加して混合する工程を含む、飲食品の製造方法に関する。
本発明はラクトバチルス・ガセリ（Lactobacillus gasseri）を含む飲食品の製造方法であって、低ｐH、高糖濃度の食品中で特定の乳酸菌をガセリ菌と混合することを特徴とする。
本発明の飲食品の低ｐＨとは３．５～４．５であり、高糖濃度とは５％～１５％であり、好ましくはｐＨ３．６～４．２であり、糖濃度は１０％～１５％であり、さらに好ましくはｐＨ３．７～４．０であり、糖濃度は１２％～１４％であり、もっとも好ましくはｐH３．８、糖濃度１２．７％である。
本発明の飲食品は、上記低ｐH、高糖濃度の飲食品であればよく、乳、乳製品、発酵乳製品に限定されず、野菜飲料、果実飲料等をも含む。そのうちでも好ましくは、乳および乳製品の成分規格等に関する省令で定められた発酵乳および乳酸菌飲料である。 As a result of repeated investigations in view of the above problems, it has been found that by mixing a specific lactic acid bacterium with gasseri bacteria in a low pH, high sugar concentration food and drink, the survival of the food and drink can be increased, and the present invention is It came to complete.
That is, the present invention adds a low pH, high sugar-concentrated food containing gasseri bacteria to 3% (w / w) reduced skim milk, 10% (w / w) glucose at pH 4.0 at 10 ° C. It is related with the manufacturing method of food-drinks including the process of adding and mixing the specific lactic acid bacteria which can reduce pH 0.1 or more when left still for days.
The present invention is a method for producing a food or drink containing Lactobacillus gasseri, characterized in that a specific lactic acid bacterium is mixed with a gasseri bacterium in a food having a low pH and a high sugar concentration.
The low pH of the food / beverage product of the present invention is 3.5 to 4.5, the high sugar concentration is 5% to 15%, preferably pH 3.6 to 4.2, and the sugar concentration is 10%. -15%, more preferably pH 3.7-4.0, sugar concentration 12% -14%, most preferably pH 3.8, sugar concentration 12.7%.
The food / beverage products of this invention should just be the food / beverage products of the said low pH and high sugar concentration, are not limited to milk, dairy products, and fermented milk products, A vegetable drink, a fruit drink, etc. are included. Among them, fermented milk and lactic acid bacteria beverages defined by a ministerial ordinance concerning the component specifications of milk and dairy products are preferable.
本発明におけるガセリ菌は、ラクトバチルス・ガセリ（Lactobacillus gasseri）に属するものであればいずれでもよく、ＳＢＴ２０５５（ＦＥＲＭ ＢＰ－１０９５３）、ＳＢＴ２０５６（ＦＥＲＭ ＢＰ－１１０３８）、ＳＢＴ０２７４（ＦＥＲＭ ＢＰ－１１０３９）、ＳＢＴ１７０３（ＦＥＲＭ Ｐ－１７７８５）、ＳＢＴ１０２３９（ＦＥＲＭ Ｐ－１６６３９）、ＳＢＴ１０２４１（ＦＥＲＭ Ｐ－１７７８６）等が挙げられる。
The gasseri bacterium in the present invention may be any one belonging to Lactobacillus gasseri, and SBT2055 (FERM BP-10953), SBT2056 (FERM BP-11038), SBT0274 (FERM BP-11039), SBT1703. (FERM P-17785), SBT10239 (FERM P-16639), SBT10241 (FERM P-17786) and the like.
本発明において、ガセリ菌と混合する特定の乳酸菌は、ガセリ菌と混合することでガセリ菌の生残性を向上させる乳酸菌であればよく、そのような乳酸菌としては、３％（ｗ／ｗ）還元脱脂乳および１０％（ｗ／ｗ）グルコースを含むｐＨ４．０の培地に添加して１０℃で２１日間静置培養したときに当該培地のｐＨを０．１以上低下させることができる乳酸菌が挙げられる。そのような性質を有する乳酸菌は、３％（ｗ／ｗ）還元脱脂乳および１０％（ｗ／ｗ）グルコースを含むｐＨ４．０の培地に添加して１０℃で２１日間静置培養したときに当該培地のｐＨを０．１以上低下させることができる乳酸菌をスクリーニングすることにより得られ、具体的にはラクトバチルス・パラカゼイ(Lactobacillus paracasei、以下ラクトバチルス・パラカゼイということがある。）、ラクトバチルス・プランタラム（Lactobacillus plantarum、以下ラクトバチルス・プランタラムということがある。）、ラクトバチルス・ブフネリ（Lactobacillus buchneri、以下ラクトバチルス・ブフネリということがある。）等が挙げられる。後述する実施例２で示したように、添加するパラカゼイ菌の菌数を変えた場合に、代謝に伴うｐＨ低下率とガセリ菌の生残性が反比例した。よって、ラクトバチルス・ガセリと混合する特定の乳酸菌数は多い方が好ましいが、例えば、ガセリ菌に対して０．００１％以上、好ましくは０．０１％以上混合することにより、混合したガセリ菌の生残性を向上させることができる。また死菌体では効果がみられなかったことから、ガセリ菌の生残性向上効果は低温、ｐＨ、高糖濃度ストレス条件下における代謝活性と相関すると考えられる。そこでｐＨ４．０の３％（ｗ／ｗ）還元脱脂乳、１０％（ｗ／ｗ）グルコース条件で一定以上ｐＨを低下させることをスクリーニングの指標とした。
なお、ラクトバチルス・ガセリと混合する特定の乳酸菌としては、ラクトバチルス・パラカゼイ、ラクトバチルス・プランタラム、ラクトバチルス・ブフネリ等が挙げられるが、３％（ｗ／ｗ）還元脱脂乳および１０％（ｗ／ｗ）グルコースを含むｐＨ４．０の培地に添加して１０℃において２１日間静置培養したときに該培地のｐＨを０．１以上低下させることができる乳酸菌であれば特に限定されるものではない。
ラクトバチルス・パラカゼイとしては、例えば、ＳＢＴ０３２７（ＮＩＴＥ ＡＢＰ－１１２９）、ＳＢＴ２１０５（ＮＩＴＥ Ｐ－１１３０）、ＳＢＴ２２０３（ＮＩＴＥ ＡＢＰ－１１３１）、ＳＢＴ２２１５（ＮＩＴＥ ＡＢＰ－１１３２）、ＳＢＴ１１４０８（ＮＩＴＥ ＡＢＰ－１１３３）、ＡＴＣＣ２５５９８、前記ラクトバチルス・プランタラムとしては、ＳＢＴ１５３４（ＦＥＲＭ ＡＢＰ－１１５１８）、ＳＢＴ０６２４（ＦＥＲＭ Ｐ－１１９２０）、ＡＴＣＣ４３１９９、ＡＴＣＣ８０１４、ラクトバチルス・ブフネリとしては、ＳＢＴ２０２８（ＦＥＲＭ Ｐ－１１９２１）、ＪＣＭ１１１５等が挙げられる。 In the present invention, the specific lactic acid bacterium to be mixed with the gasseri bacterium may be a lactic acid bacterium that improves the viability of the gasseri bacterium by mixing with the gasseri bacterium, and as such a lactic acid bacterium, 3% (w / w) A lactic acid bacterium capable of lowering the pH of the medium by 0.1 or more when added to a pH 4.0 medium containing reduced skim milk and 10% (w / w) glucose and left to stand at 10 ° C. for 21 days. Can be mentioned. Lactic acid bacteria having such properties are added to a pH 4.0 medium containing 3% (w / w) reduced skim milk and 10% (w / w) glucose, and left to stand at 10 ° C. for 21 days. It is obtained by screening lactic acid bacteria capable of lowering the pH of the medium by 0.1 or more, specifically, Lactobacillus paracasei (hereinafter sometimes referred to as Lactobacillus paracasei), Lactobacillus paracasei. Plantarum (Lactobacillus plantarum, hereinafter referred to as Lactobacillus plantarum) and Lactobacillus buchneri (hereinafter, also referred to as Lactobacillus buchneri). As shown in Example 2 to be described later, when the number of paracasei bacteria to be added was changed, the pH reduction rate accompanying metabolism and the viability of gasseri bacteria were inversely proportional. Therefore, it is preferable that the number of specific lactic acid bacteria to be mixed with Lactobacillus gasseri is larger. For example, by mixing 0.001% or more, preferably 0.01% or more of gasseri bacteria, Survivability can be improved. In addition, since no effect was observed in dead cells, the survival improvement effect of gasseri is considered to correlate with metabolic activity under low temperature, pH, and high sugar concentration stress conditions. Therefore, a 3% (w / w) reduced skim milk having a pH of 4.0, and lowering the pH by a certain level under 10% (w / w) glucose conditions was used as an index for screening.
Specific lactic acid bacteria to be mixed with Lactobacillus gasseri include Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus buchneri, etc., but 3% (w / w) reduced skim milk and 10% ( w / w) Particularly limited as long as it is a lactic acid bacterium capable of lowering the pH of the medium by 0.1 or more when it is added to a pH 4.0 medium containing glucose and statically cultured at 10 ° C. for 21 days. is not.
Examples of Lactobacillus paracasei include SBT0327 (NITE ABP-1129), SBT2105 (NITE P-1130), SBT2203 (NITE ABP-1131), SBT2215 (NITE ABP-1132), SBT11408 (NITE ABP-113) Examples of the Lactobacillus plantarum include SBT1534 (FERM ABP-11518), SBT0624 (FERM P-11920), ATCC43199, ATCC8014, Lactobacillus buchneri, SBT2028 (FERM P-11192), JCM1115, and the like. .
なお、ラクトバチルス・ガセリと混合する特定の乳酸菌としては、ラクトバチルス・パラカゼイ、ラクトバチルス・プランタラム、ラクトバチルス・ブフネリ等が挙げられるが、３％（ｗ／ｗ）還元脱脂乳および１０％（ｗ／ｗ）グルコースを含むｐＨ４．０の培地に添加して１０℃において２１日間静置培養したときに該培地のｐＨを０．１以上低下させることができる乳酸菌であれば特に限定されるものではない。
ラクトバチルス・パラカゼイとしては、例えば、ＳＢＴ０３２７（ＮＩＴＥ ＡＢＰ－１１２９）、ＳＢＴ２１０５（ＮＩＴＥ Ｐ－１１３０）、ＳＢＴ２２０３（ＮＩＴＥ ＡＢＰ－１１３１）、ＳＢＴ２２１５（ＮＩＴＥ ＡＢＰ－１１３２）、ＳＢＴ１１４０８（ＮＩＴＥ ＡＢＰ－１１３３）、ＡＴＣＣ２５５９８、前記ラクトバチルス・プランタラムとしては、ＳＢＴ１５３４（ＦＥＲＭ ＡＢＰ－１１５１８）、ＳＢＴ０６２４（ＦＥＲＭ Ｐ－１１９２０）、ＡＴＣＣ４３１９９、ＡＴＣＣ８０１４、ラクトバチルス・ブフネリとしては、ＳＢＴ２０２８（ＦＥＲＭ Ｐ－１１９２１）、ＪＣＭ１１１５等が挙げられる。 In the present invention, the specific lactic acid bacterium to be mixed with the gasseri bacterium may be a lactic acid bacterium that improves the viability of the gasseri bacterium by mixing with the gasseri bacterium, and as such a lactic acid bacterium, 3% (w / w) A lactic acid bacterium capable of lowering the pH of the medium by 0.1 or more when added to a pH 4.0 medium containing reduced skim milk and 10% (w / w) glucose and left to stand at 10 ° C. for 21 days. Can be mentioned. Lactic acid bacteria having such properties are added to a pH 4.0 medium containing 3% (w / w) reduced skim milk and 10% (w / w) glucose, and left to stand at 10 ° C. for 21 days. It is obtained by screening lactic acid bacteria capable of lowering the pH of the medium by 0.1 or more, specifically, Lactobacillus paracasei (hereinafter sometimes referred to as Lactobacillus paracasei), Lactobacillus paracasei. Plantarum (Lactobacillus plantarum, hereinafter referred to as Lactobacillus plantarum) and Lactobacillus buchneri (hereinafter, also referred to as Lactobacillus buchneri). As shown in Example 2 to be described later, when the number of paracasei bacteria to be added was changed, the pH reduction rate accompanying metabolism and the viability of gasseri bacteria were inversely proportional. Therefore, it is preferable that the number of specific lactic acid bacteria to be mixed with Lactobacillus gasseri is larger. For example, by mixing 0.001% or more, preferably 0.01% or more of gasseri bacteria, Survivability can be improved. In addition, since no effect was observed in dead cells, the survival improvement effect of gasseri is considered to correlate with metabolic activity under low temperature, pH, and high sugar concentration stress conditions. Therefore, a 3% (w / w) reduced skim milk having a pH of 4.0, and lowering the pH by a certain level under 10% (w / w) glucose conditions was used as an index for screening.
Specific lactic acid bacteria to be mixed with Lactobacillus gasseri include Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus buchneri, etc., but 3% (w / w) reduced skim milk and 10% ( w / w) Particularly limited as long as it is a lactic acid bacterium capable of lowering the pH of the medium by 0.1 or more when it is added to a pH 4.0 medium containing glucose and statically cultured at 10 ° C. for 21 days. is not.
Examples of Lactobacillus paracasei include SBT0327 (NITE ABP-1129), SBT2105 (NITE P-1130), SBT2203 (NITE ABP-1131), SBT2215 (NITE ABP-1132), SBT11408 (NITE ABP-113) Examples of the Lactobacillus plantarum include SBT1534 (FERM ABP-11518), SBT0624 (FERM P-11920), ATCC43199, ATCC8014, Lactobacillus buchneri, SBT2028 (FERM P-11192), JCM1115, and the like. .
本発明の飲食品の製造方法において、ガセリ菌および特定の乳酸菌を混合する工程としては、以下の形態等が挙げられる。
（１）ガセリ菌を未発酵の乳培地に添加して発酵させた後に当該発酵物に特定の乳酸菌を添加する。
（２）特定の乳酸菌を未発酵の乳培地に添加して発酵させた後に当該発酵物にガセリ菌を添加する。
（３）ガセリ菌および特定の乳酸菌を未発酵の乳培地に同時あるいは順次添加して発酵させる。
（４）乳酸菌発酵物以外の飲食品にガセリ菌および特定乳酸菌を同時あるいは順次添加する。
上記いずれの混合形態で得られた飲食品であっても本発明のガセリ菌生残性向上の効果は得られる。上記形態のうち、好ましくは（２）および（４）である。また、発酵後の発酵乳製品に添加する場合、発酵に使用する乳酸菌として、通常よく使用されるラクトバチルス・デルブリュッキ・サブスピーシーズ・ブルガリクス（Lactobacillus delbrueckii subsp. bulgaricus）やストレプトコッカス・サーモフィルス(Streptococcus thermophilus)等が例示できる。 In the method for producing a food or drink of the present invention, examples of the step of mixing gasseri and specific lactic acid bacteria include the following forms.
(1) After adding and fermenting gasseri bacteria to an unfermented milk medium, a specific lactic acid bacterium is added to the fermented product.
(2) A specific lactic acid bacterium is added to an unfermented milk medium and fermented, and then a gasseri bacterium is added to the fermented product.
(3) Gasseri bacteria and specific lactic acid bacteria are fermented simultaneously or sequentially with unfermented milk medium.
(4) Gasseri bacteria and specific lactic acid bacteria are added simultaneously or sequentially to foods and drinks other than fermented lactic acid bacteria.
Even if it is the food / beverage products obtained by any said mixed form, the effect of the gasseri microbe survival improvement of this invention is acquired. Of the above forms, (2) and (4) are preferred. When added to fermented dairy products after fermentation, Lactobacillus delbrueckii subsp. Bulgaricus or Streptococcus thermophilus (Lactobacillus delbrueckii subsp. Bulgaricus) commonly used as the lactic acid bacteria used for fermentation ) Etc. can be illustrated.
（１）ガセリ菌を未発酵の乳培地に添加して発酵させた後に当該発酵物に特定の乳酸菌を添加する。
（２）特定の乳酸菌を未発酵の乳培地に添加して発酵させた後に当該発酵物にガセリ菌を添加する。
（３）ガセリ菌および特定の乳酸菌を未発酵の乳培地に同時あるいは順次添加して発酵させる。
（４）乳酸菌発酵物以外の飲食品にガセリ菌および特定乳酸菌を同時あるいは順次添加する。
上記いずれの混合形態で得られた飲食品であっても本発明のガセリ菌生残性向上の効果は得られる。上記形態のうち、好ましくは（２）および（４）である。また、発酵後の発酵乳製品に添加する場合、発酵に使用する乳酸菌として、通常よく使用されるラクトバチルス・デルブリュッキ・サブスピーシーズ・ブルガリクス（Lactobacillus delbrueckii subsp. bulgaricus）やストレプトコッカス・サーモフィルス(Streptococcus thermophilus)等が例示できる。 In the method for producing a food or drink of the present invention, examples of the step of mixing gasseri and specific lactic acid bacteria include the following forms.
(1) After adding and fermenting gasseri bacteria to an unfermented milk medium, a specific lactic acid bacterium is added to the fermented product.
(2) A specific lactic acid bacterium is added to an unfermented milk medium and fermented, and then a gasseri bacterium is added to the fermented product.
(3) Gasseri bacteria and specific lactic acid bacteria are fermented simultaneously or sequentially with unfermented milk medium.
(4) Gasseri bacteria and specific lactic acid bacteria are added simultaneously or sequentially to foods and drinks other than fermented lactic acid bacteria.
Even if it is the food / beverage products obtained by any said mixed form, the effect of the gasseri microbe survival improvement of this invention is acquired. Of the above forms, (2) and (4) are preferred. When added to fermented dairy products after fermentation, Lactobacillus delbrueckii subsp. Bulgaricus or Streptococcus thermophilus (Lactobacillus delbrueckii subsp. Bulgaricus) commonly used as the lactic acid bacteria used for fermentation ) Etc. can be illustrated.
本発明の他の形態は、上記の製造方法により得られたガセリ菌および特定の乳酸菌を含む、ｐＨが３．５～４．５、糖濃度が５％～１５％の飲食品である。本発明の飲食品におけるガセリ菌の生残数は、特定乳酸菌と混合しない場合にくらべて格段に高く、例えば１週間で１０倍、２週間で１，０００～１０，０００倍、３週間で１，０００～１０，０００倍も異なる。飲食品における生残菌数が、ガセリ菌添加後２１日間保存後に１×１０６ＣＦＵ／ｇ以上である。
Another embodiment of the present invention is a food or drink product having a pH of 3.5 to 4.5 and a sugar concentration of 5 to 15%, which includes gasseri bacteria and specific lactic acid bacteria obtained by the above production method. The survival number of gasseri bacteria in the food and drink of the present invention is much higher than when not mixed with specific lactic acid bacteria, for example, 10 times in 1 week, 1,000 to 10,000 times in 2 weeks, 1 in 3 weeks. , 10,000 to 10,000 times different. The number of surviving bacteria in food and drink is 1 × 10 6 CFU / g or more after storage for 21 days after addition of gasseri bacteria.
以下、飲食品として発酵乳製品を例に本発明をさらに詳細に説明するが、本発明はこれらに限定されるものではない。
Hereinafter, the present invention will be described in more detail using fermented milk products as examples of foods and drinks, but the present invention is not limited to these.
〔試験例１〕ガセリ菌と混合する乳酸菌のスクリーニング（１）
本発明の飲食品においてガセリ菌と混合する乳酸菌のスクリーニング試験を行った。
（１）試験方法
脱脂粉乳とグルコースを水に溶解し、３％（ｗ／ｗ）還元脱脂乳、１０％（ｗ／ｗ）グルコースを含む培地を調製し、当該培地を１１５℃で２０分間殺菌した。殺菌した培地にろ過滅菌した乳酸を添加してｐＨを約４．０に調整してスクリーニング用培地を調製した。前記スクリーニング用培地に、乳培地で培養したシードカルチャー（表２に記載）を各３％接種して、１０℃にて２１日間静置したときのｐＨを測定し、ｐＨを０．１以上低下させる菌株を選択した。
（２）試験結果
各乳酸菌の初発菌数および保存後のｐＨの結果を表２に示す。これによればラクトバチルス・パラカゼイを添加した培地のｐＨは０．１以上低下した。これに対してラクトバチルス・ブルガリクス、ラクトバチルス・ヘルベティカス、ラクトバチルス・ジョンソニー、ラクトバチルス・ラムノーサスの各基準株では、０．１以上のｐＨ低下は認められなかった。したがって、この中からは、ガセリ菌と混合する菌としてラクトバチルス・パラカゼイを選択した。 [Test Example 1] Screening of lactic acid bacteria mixed with gasseri bacteria (1)
A screening test for lactic acid bacteria mixed with gasseri bacteria in the food and drink of the present invention was conducted.
(1) Test method Nonfat dry milk and glucose are dissolved in water to prepare a medium containing 3% (w / w) reduced skim milk and 10% (w / w) glucose, and the medium is sterilized at 115 ° C. for 20 minutes. did. Lactic acid sterilized by filtration was added to the sterilized medium to adjust the pH to about 4.0 to prepare a screening medium. The screening medium was inoculated with 3% each of seed cultures cultured in milk medium (described in Table 2), and the pH was measured when left at 10 ° C. for 21 days. The strain to be selected was selected.
(2) Test results Table 2 shows the results of the initial bacterial count of each lactic acid bacterium and the pH after storage. According to this, the pH of the medium to which Lactobacillus paracasei was added decreased by 0.1 or more. On the other hand, in each of the reference strains of Lactobacillus bulgaricus, Lactobacillus helveticus, Lactobacillus johnsonii, and Lactobacillus rhamnosus, no pH decrease of 0.1 or more was observed. Therefore, Lactobacillus paracasei was selected as a bacterium to be mixed with gasseri.
本発明の飲食品においてガセリ菌と混合する乳酸菌のスクリーニング試験を行った。
（１）試験方法
脱脂粉乳とグルコースを水に溶解し、３％（ｗ／ｗ）還元脱脂乳、１０％（ｗ／ｗ）グルコースを含む培地を調製し、当該培地を１１５℃で２０分間殺菌した。殺菌した培地にろ過滅菌した乳酸を添加してｐＨを約４．０に調整してスクリーニング用培地を調製した。前記スクリーニング用培地に、乳培地で培養したシードカルチャー（表２に記載）を各３％接種して、１０℃にて２１日間静置したときのｐＨを測定し、ｐＨを０．１以上低下させる菌株を選択した。
（２）試験結果
各乳酸菌の初発菌数および保存後のｐＨの結果を表２に示す。これによればラクトバチルス・パラカゼイを添加した培地のｐＨは０．１以上低下した。これに対してラクトバチルス・ブルガリクス、ラクトバチルス・ヘルベティカス、ラクトバチルス・ジョンソニー、ラクトバチルス・ラムノーサスの各基準株では、０．１以上のｐＨ低下は認められなかった。したがって、この中からは、ガセリ菌と混合する菌としてラクトバチルス・パラカゼイを選択した。 [Test Example 1] Screening of lactic acid bacteria mixed with gasseri bacteria (1)
A screening test for lactic acid bacteria mixed with gasseri bacteria in the food and drink of the present invention was conducted.
(1) Test method Nonfat dry milk and glucose are dissolved in water to prepare a medium containing 3% (w / w) reduced skim milk and 10% (w / w) glucose, and the medium is sterilized at 115 ° C. for 20 minutes. did. Lactic acid sterilized by filtration was added to the sterilized medium to adjust the pH to about 4.0 to prepare a screening medium. The screening medium was inoculated with 3% each of seed cultures cultured in milk medium (described in Table 2), and the pH was measured when left at 10 ° C. for 21 days. The strain to be selected was selected.
(2) Test results Table 2 shows the results of the initial bacterial count of each lactic acid bacterium and the pH after storage. According to this, the pH of the medium to which Lactobacillus paracasei was added decreased by 0.1 or more. On the other hand, in each of the reference strains of Lactobacillus bulgaricus, Lactobacillus helveticus, Lactobacillus johnsonii, and Lactobacillus rhamnosus, no pH decrease of 0.1 or more was observed. Therefore, Lactobacillus paracasei was selected as a bacterium to be mixed with gasseri.
〔試験例２〕ガセリ菌と混合する乳酸菌のスクリーニング（１）
試験例１の他に、３％（ｗ／ｗ）還元脱脂乳および１０％（ｗ／ｗ）グルコース を含むｐＨ４．０の培地に添加して１０℃において２１日間静置培養したときに該培地のｐＨを０．１以上低下させることができる乳酸菌のスクリーニングを行ったところ、ラクトバチルス・パラカゼイＡＴＣＣ２５５９８、ラクトバチルス・プランタラムＡＴＣＣ４３１９９、ラクトバチルス・プランタラムＡＴＣＣ８０１４、ラクトバチルス・ブフネリＪＣＭ１１１５が選定された。各乳酸菌の初発菌数および保存後のｐＨは表３に示すとおりである。なお、スクリーニングは試験例１と同様の方法によって実施した。 [Test Example 2] Screening of lactic acid bacteria mixed with gasseri bacteria (1)
In addition to Test Example 1, when added to a pH 4.0 medium containing 3% (w / w) reduced skim milk and 10% (w / w) glucose, the medium was allowed to stand for 21 days at 10 ° C. As a result, Lactobacillus paracasei ATCC 25598, Lactobacillus plantarum ATCC 43199, Lactobacillus plantarum ATCC 8014, and Lactobacillus buchneri JCM1115 were selected. Table 3 shows the initial bacterial count of each lactic acid bacterium and the pH after storage. The screening was performed by the same method as in Test Example 1.
試験例１の他に、３％（ｗ／ｗ）還元脱脂乳および１０％（ｗ／ｗ）グルコース を含むｐＨ４．０の培地に添加して１０℃において２１日間静置培養したときに該培地のｐＨを０．１以上低下させることができる乳酸菌のスクリーニングを行ったところ、ラクトバチルス・パラカゼイＡＴＣＣ２５５９８、ラクトバチルス・プランタラムＡＴＣＣ４３１９９、ラクトバチルス・プランタラムＡＴＣＣ８０１４、ラクトバチルス・ブフネリＪＣＭ１１１５が選定された。各乳酸菌の初発菌数および保存後のｐＨは表３に示すとおりである。なお、スクリーニングは試験例１と同様の方法によって実施した。 [Test Example 2] Screening of lactic acid bacteria mixed with gasseri bacteria (1)
In addition to Test Example 1, when added to a pH 4.0 medium containing 3% (w / w) reduced skim milk and 10% (w / w) glucose, the medium was allowed to stand for 21 days at 10 ° C. As a result, Lactobacillus paracasei ATCC 25598, Lactobacillus plantarum ATCC 43199, Lactobacillus plantarum ATCC 8014, and Lactobacillus buchneri JCM1115 were selected. Table 3 shows the initial bacterial count of each lactic acid bacterium and the pH after storage. The screening was performed by the same method as in Test Example 1.
〔実施例１〕ラクトバチルス・パラカゼイとの混合によるガセリ菌の生残性の向上確認試験
上記試験例１のスクリーニング試験で選択された菌をガセリ菌と混合して、ガセリ菌の生残性が向上することを確認した。
（１）本発明の乳酸菌飲料の調製
脱脂粉乳とグルコースを水に溶解し、１６％（ｗ／ｗ）還元脱脂乳、３％（ｗ／ｗ）グルコースを含む培地を調製し、当該培地を９５℃で１２０分間殺菌した。３７℃まで冷却した殺菌培地にラクトバチルス・パラカゼイＳＢＴ０３２７、ＳＢＴ２１０５、ＳＢＴ２２０３、ＳＢＴ２２１５、ＳＢＴ１１４０８のシードカルチャーをそれぞれ３％（ｖ／ｖ）添加して３７℃でｐＨ３．７程度になるまで培養し、ラクトバチルス・パラカゼイ発酵物を得た。
また、３％（ｗ／ｗ）異性化液糖・１４％（ｗ／ｗ）上白糖溶液を水で調製し、１２１℃１５分間滅菌して糖液（糖濃度１６％）とした。
上記で得られた糖液３６ｇ、ラクトバチルス・パラカゼイ発酵物９ｇおよびガセリ菌ＳＢＴ２０５５濃縮菌体０．５ｇを混合し、ｐＨ３．８、糖濃度１２．７％である本発明１～５の乳酸菌飲料を製造した。
（２）対照乳酸菌飲料の調製
ガセリ菌のみが含まれ、ラクトバチルス・パラカゼイと混合していない対照乳酸菌飲料を製造した。前記（１）の殺菌培地にろ過滅菌した乳酸を添加してｐＨを約３．７に調整したものを擬似発酵物とした。擬似発酵物９ｇ、前記（１）の糖液３６ｇ、およびガセリ菌ＳＢＴ２０５５の濃縮菌体０．５ｇを混合し対照乳酸菌飲料を製造した。
（３）生残率の測定試験および結果
本発明品１～５の乳酸菌飲料および対照乳酸菌飲料について、それぞれプラスチック容器に入れて１０℃で２１日間保存し、ガセリ菌およびラクトバチルス・パラカゼイの生菌数を経時的に測定した。生残率は、２１日目の生菌数を０日目の生菌数で序して求めた（以下の試験において同じ）。結果を表４に示す。
本結果から明らかなように、ガセリ菌とラクトバチルス・パラカゼイを混合させた本発明品１～５の乳酸菌飲料では、いずれもラクトバチルス・パラカゼイと混合させずにガセリ菌のみが含まれる対照乳酸菌飲料に比べて著しくガセリ菌の生残菌数が高く、２１日間保存後のラクトバチルス・ガセリの生菌数は本発明品１～５の全てにおいて１×１０６ＣＦＵ／ｇ以上であった。
以上より、３％（ｗ／ｗ）還元脱脂乳および１０％（ｗ／ｗ）グルコース を含むｐＨ４．０の培地に添加して１０℃において２１日間静置培養したときに該培地のｐＨを０．１以上低下させることができる乳酸菌、と混合することによりガセリ菌の生残性が飛躍的に向上することが明らかとなった。 [Example 1] Test for confirming improvement in survival of gasseri by mixing with Lactobacillus paracasei The bacteria selected in the screening test of test example 1 are mixed with gasseri, and the survival of gasseri Confirmed to improve.
(1) Preparation of lactic acid bacteria beverage of the present invention Nonfat dry milk and glucose are dissolved in water to prepare a medium containing 16% (w / w) reduced skim milk and 3% (w / w) glucose. Sterilized at 120 ° C. for 120 minutes. 3% (v / v) seed cultures of Lactobacillus paracasei SBT0327, SBT2105, SBT2203, SBT2215, and SBT11408 were added to a sterilized medium cooled to 37 ° C, and cultured at 37 ° C until the pH reached about 3.7. A Bacillus paracasei fermentation product was obtained.
Further, a 3% (w / w) isomerized liquid sugar / 14% (w / w) white sugar solution was prepared in water and sterilized at 121 ° C. for 15 minutes to obtain a sugar liquid (sugar concentration: 16%).
Lactic acid bacteria beverages of the present invention 1 to 5 having a pH of 3.8 and a sugar concentration of 12.7%, obtained by mixing 36 g of the sugar solution obtained above, 9 g of a fermented Lactobacillus paracasei product, and 0.5 g of S. cerevisiae SBT2055 Manufactured.
(2) Preparation of control lactic acid bacteria beverage A control lactic acid bacteria beverage containing only gasseri bacteria and not mixed with Lactobacillus paracasei was produced. A quasi-fermented product was prepared by adding lactic acid sterilized by filtration to the sterilizing medium (1) and adjusting the pH to about 3.7. A control lactic acid bacteria beverage was prepared by mixing 9 g of the pseudo-fermented product, 36 g of the sugar solution of the above (1), and 0.5 g of concentrated bacteria of gasseri bacteria SBT2055.
(3) Survival rate measurement test and results The lactic acid bacteria beverages and control lactic acid bacteria beverages of the products 1 to 5 of the present invention were each stored in a plastic container at 10 ° C. for 21 days, and live bacteria of gasseri and Lactobacillus paracasei Numbers were measured over time. The survival rate was obtained by calculating the number of viable bacteria on the 21st day by the viable cell count on the 0th day (the same applies in the following tests). The results are shown in Table 4.
As is apparent from these results, the lactic acid bacteria beverages of the present invention products 1 to 5 mixed with gasseri bacteria and Lactobacillus paracasei all contain only gasseri bacteria without being mixed with lactobacilli paracasei. remarkably high survival bacterial count of gasseri bacteria, the viable cell count of Lactobacillus gasseri after storage for 21 days were 1 × 10 6 CFU / g or more in all of the inventive products 1-5 compared to.
From the above, when the culture medium was added to a pH 4.0 medium containing 3% (w / w) reduced skim milk and 10% (w / w) glucose and left to stand at 10 ° C. for 21 days, the pH of the medium was reduced to 0. It was clarified that the viability of gasseri bacteria was dramatically improved by mixing with lactic acid bacteria that can be reduced by 1 or more.
上記試験例１のスクリーニング試験で選択された菌をガセリ菌と混合して、ガセリ菌の生残性が向上することを確認した。
（１）本発明の乳酸菌飲料の調製
脱脂粉乳とグルコースを水に溶解し、１６％（ｗ／ｗ）還元脱脂乳、３％（ｗ／ｗ）グルコースを含む培地を調製し、当該培地を９５℃で１２０分間殺菌した。３７℃まで冷却した殺菌培地にラクトバチルス・パラカゼイＳＢＴ０３２７、ＳＢＴ２１０５、ＳＢＴ２２０３、ＳＢＴ２２１５、ＳＢＴ１１４０８のシードカルチャーをそれぞれ３％（ｖ／ｖ）添加して３７℃でｐＨ３．７程度になるまで培養し、ラクトバチルス・パラカゼイ発酵物を得た。
また、３％（ｗ／ｗ）異性化液糖・１４％（ｗ／ｗ）上白糖溶液を水で調製し、１２１℃１５分間滅菌して糖液（糖濃度１６％）とした。
上記で得られた糖液３６ｇ、ラクトバチルス・パラカゼイ発酵物９ｇおよびガセリ菌ＳＢＴ２０５５濃縮菌体０．５ｇを混合し、ｐＨ３．８、糖濃度１２．７％である本発明１～５の乳酸菌飲料を製造した。
（２）対照乳酸菌飲料の調製
ガセリ菌のみが含まれ、ラクトバチルス・パラカゼイと混合していない対照乳酸菌飲料を製造した。前記（１）の殺菌培地にろ過滅菌した乳酸を添加してｐＨを約３．７に調整したものを擬似発酵物とした。擬似発酵物９ｇ、前記（１）の糖液３６ｇ、およびガセリ菌ＳＢＴ２０５５の濃縮菌体０．５ｇを混合し対照乳酸菌飲料を製造した。
（３）生残率の測定試験および結果
本発明品１～５の乳酸菌飲料および対照乳酸菌飲料について、それぞれプラスチック容器に入れて１０℃で２１日間保存し、ガセリ菌およびラクトバチルス・パラカゼイの生菌数を経時的に測定した。生残率は、２１日目の生菌数を０日目の生菌数で序して求めた（以下の試験において同じ）。結果を表４に示す。
本結果から明らかなように、ガセリ菌とラクトバチルス・パラカゼイを混合させた本発明品１～５の乳酸菌飲料では、いずれもラクトバチルス・パラカゼイと混合させずにガセリ菌のみが含まれる対照乳酸菌飲料に比べて著しくガセリ菌の生残菌数が高く、２１日間保存後のラクトバチルス・ガセリの生菌数は本発明品１～５の全てにおいて１×１０６ＣＦＵ／ｇ以上であった。
以上より、３％（ｗ／ｗ）還元脱脂乳および１０％（ｗ／ｗ）グルコース を含むｐＨ４．０の培地に添加して１０℃において２１日間静置培養したときに該培地のｐＨを０．１以上低下させることができる乳酸菌、と混合することによりガセリ菌の生残性が飛躍的に向上することが明らかとなった。 [Example 1] Test for confirming improvement in survival of gasseri by mixing with Lactobacillus paracasei The bacteria selected in the screening test of test example 1 are mixed with gasseri, and the survival of gasseri Confirmed to improve.
(1) Preparation of lactic acid bacteria beverage of the present invention Nonfat dry milk and glucose are dissolved in water to prepare a medium containing 16% (w / w) reduced skim milk and 3% (w / w) glucose. Sterilized at 120 ° C. for 120 minutes. 3% (v / v) seed cultures of Lactobacillus paracasei SBT0327, SBT2105, SBT2203, SBT2215, and SBT11408 were added to a sterilized medium cooled to 37 ° C, and cultured at 37 ° C until the pH reached about 3.7. A Bacillus paracasei fermentation product was obtained.
Further, a 3% (w / w) isomerized liquid sugar / 14% (w / w) white sugar solution was prepared in water and sterilized at 121 ° C. for 15 minutes to obtain a sugar liquid (sugar concentration: 16%).
Lactic acid bacteria beverages of the present invention 1 to 5 having a pH of 3.8 and a sugar concentration of 12.7%, obtained by mixing 36 g of the sugar solution obtained above, 9 g of a fermented Lactobacillus paracasei product, and 0.5 g of S. cerevisiae SBT2055 Manufactured.
(2) Preparation of control lactic acid bacteria beverage A control lactic acid bacteria beverage containing only gasseri bacteria and not mixed with Lactobacillus paracasei was produced. A quasi-fermented product was prepared by adding lactic acid sterilized by filtration to the sterilizing medium (1) and adjusting the pH to about 3.7. A control lactic acid bacteria beverage was prepared by mixing 9 g of the pseudo-fermented product, 36 g of the sugar solution of the above (1), and 0.5 g of concentrated bacteria of gasseri bacteria SBT2055.
(3) Survival rate measurement test and results The lactic acid bacteria beverages and control lactic acid bacteria beverages of the products 1 to 5 of the present invention were each stored in a plastic container at 10 ° C. for 21 days, and live bacteria of gasseri and Lactobacillus paracasei Numbers were measured over time. The survival rate was obtained by calculating the number of viable bacteria on the 21st day by the viable cell count on the 0th day (the same applies in the following tests). The results are shown in Table 4.
As is apparent from these results, the lactic acid bacteria beverages of the present invention products 1 to 5 mixed with gasseri bacteria and Lactobacillus paracasei all contain only gasseri bacteria without being mixed with lactobacilli paracasei. remarkably high survival bacterial count of gasseri bacteria, the viable cell count of Lactobacillus gasseri after storage for 21 days were 1 × 10 6 CFU / g or more in all of the inventive products 1-5 compared to.
From the above, when the culture medium was added to a pH 4.0 medium containing 3% (w / w) reduced skim milk and 10% (w / w) glucose and left to stand at 10 ° C. for 21 days, the pH of the medium was reduced to 0. It was clarified that the viability of gasseri bacteria was dramatically improved by mixing with lactic acid bacteria that can be reduced by 1 or more.
〔実施例２〕ラクトバチルス・パラカゼイとの混合によるガセリ菌の生残性の向上確認試験
ラクトバチルス・パラカゼイの菌数を変更してガセリ菌と混合した場合、およびラクトバチルス・パラカゼイの死菌体をガセリ菌と混合した場合について、ガセリ菌の生残性に対する影響を調べた。
（１）本発明の乳酸菌飲料の調製
脱脂粉乳とグルコースを水に溶解し、１６％（ｗ／ｗ）還元脱脂乳、３％（ｗ／ｗ）グルコースを含む培地を調製し、当該培地を９５℃で１２０分間殺菌した。前記殺菌培地にろ過滅菌した乳酸を添加してｐＨを約４．０に調整して、擬似発酵物を調製した。擬似発酵物にラクトバチルス・パラカゼイＳＢＴ１１４０８洗浄菌体を懸濁するとともに、１０倍、１００倍、１，０００倍希釈することでパラカゼイ菌数の異なる３種類のラクトバチルス・パラカゼイ添加擬似発酵物１～３を調製した。またガセリ菌ＳＢＴ２０５５の洗浄菌体を１２．６％還元脱脂乳で懸濁して使用した。
３％（ｗ／ｗ）異性化液糖・１４％（ｗ／ｗ）上白糖溶液を水で調製し、１２１℃１５分間滅菌して糖液（糖濃度１６％）とした。
前記擬似発酵物１～３を９ｇ、糖液３６ｇ、およびガセリ菌ＳＢＴ２０５５洗浄菌体０．５ｇを混合し、ｐＨ３．８、糖濃度１２．７％のラクトバチルス・パラカゼイ菌数の異なる本発明５～７の乳酸菌飲料を製造した。
（２）対照乳酸菌飲料の調製
前記（１）の擬似発酵物９ｇ、糖液３６ｇ、およびガセリ菌ＳＢＴ２０５５洗浄菌体０．５ｇを混合し、ラクトバチルス・パラカゼイを含まない乳酸菌飲料（対照１）を製造した。
また、ラクトバチルス・パラカゼイを１×１０７ＣＦＵ／ｇ程度含むラクトバチルス・パラカゼイ添加擬似発酵物を８０℃以上で３０分間殺菌したものを用いて乳酸菌飲料（対照２）を製造した。
（３）生残率の測定試験および結果
本発明品５～７の乳酸菌飲料および対照乳酸菌飲料（対照１、２）について、それぞれプラスチック容器に入れて１０℃で２１日間保存し、ガセリ菌およびラクトバチルス・パラカゼイの生菌数を経時的に測定した。結果を表５に示す。
本結果からラクトバチルス・パラカゼイを１×１０７ＣＦＵ／ｇ以上を添加した本発明の乳酸菌飲料においてガセリ菌の生残性が顕著に向上することがわかった。また、ｐＨ４．０においても本発明は効果を発揮するが、ラクトバチルス・パラカゼイ死菌体では効果がみられないことが明らかである。 [Example 2] Confirmation test for improvement of survival of gasseri by mixing with Lactobacillus paracasei When the number of Lactobacillus paracasei was changed and mixed with gasseri, and dead cells of Lactobacillus paracasei The effect on the viability of gasseri was investigated for the case of mixing with gasseri.
(1) Preparation of lactic acid bacteria beverage of the present invention Nonfat dry milk and glucose are dissolved in water to prepare a medium containing 16% (w / w) reduced skim milk and 3% (w / w) glucose. Sterilized at 120 ° C. for 120 minutes. Lactic acid sterilized by filtration was added to the sterilized medium to adjust the pH to about 4.0 to prepare a pseudo-fermented product. Lactobacillus paracasei SBT11408 washed cells are suspended in a pseudo-fermented product, and three types of Lactobacillus paracasei-added pseudo-fermented products with different numbers of paracasei by diluting 10 times, 100 times, and 1,000 times 3 was prepared. Further, the washed cells of gasseri bacterium SBT2055 were suspended in 12.6% reduced skim milk and used.
A 3% (w / w) isomerized liquid sugar / 14% (w / w) white sugar solution was prepared in water and sterilized at 121 ° C. for 15 minutes to give a sugar liquid (sugar concentration 16%).
9 g of the simulated fermented products 1 to 3, 36 g of sugar solution, and 0.5 g of washed cell of S. gasseri SBT2055 were mixed, and the present invention 5 having a different number of Lactobacillus paracasei bacteria having a pH of 3.8 and a sugar concentration of 12.7%. -7 lactic acid bacteria beverages were produced.
(2) Preparation of Control Lactic Acid Beverage Beverage 9 g of the pseudo-fermented product of (1), sugar solution 36 g, and gasseri SBT2055 washed cell body 0.5 g were mixed, and a lactic acid bacteria beverage containing no Lactobacillus paracasei (Control 1) Manufactured.
Moreover, the lactic acid bacteria drink (control 2) was manufactured using what sterilized the lactobacillus paracasei addition pseudo fermented product which contains about 1 * 10 < 7 > CFU / g of Lactobacillus paracasei for 30 minutes at 80 degreeC or more.
(3) Survival rate measurement test and results The lactic acid bacteria beverages and control lactic acid bacteria beverages (Controls 1 and 2) of the products 5 to 7 of the present invention were respectively stored in plastic containers for 21 days at 10 ° C. The viable count of Bacillus paracasei was measured over time. The results are shown in Table 5.
From these results, it was found that the viability of gasseri bacteria was significantly improved in the lactic acid bacteria beverage of the present invention to which 1 × 10 7 CFU / g or more of Lactobacillus paracasei was added. Moreover, although this invention exhibits an effect also in pH 4.0, it is clear that an effect is not seen with a Lactobacillus paracasei dead cell.
ラクトバチルス・パラカゼイの菌数を変更してガセリ菌と混合した場合、およびラクトバチルス・パラカゼイの死菌体をガセリ菌と混合した場合について、ガセリ菌の生残性に対する影響を調べた。
（１）本発明の乳酸菌飲料の調製
脱脂粉乳とグルコースを水に溶解し、１６％（ｗ／ｗ）還元脱脂乳、３％（ｗ／ｗ）グルコースを含む培地を調製し、当該培地を９５℃で１２０分間殺菌した。前記殺菌培地にろ過滅菌した乳酸を添加してｐＨを約４．０に調整して、擬似発酵物を調製した。擬似発酵物にラクトバチルス・パラカゼイＳＢＴ１１４０８洗浄菌体を懸濁するとともに、１０倍、１００倍、１，０００倍希釈することでパラカゼイ菌数の異なる３種類のラクトバチルス・パラカゼイ添加擬似発酵物１～３を調製した。またガセリ菌ＳＢＴ２０５５の洗浄菌体を１２．６％還元脱脂乳で懸濁して使用した。
３％（ｗ／ｗ）異性化液糖・１４％（ｗ／ｗ）上白糖溶液を水で調製し、１２１℃１５分間滅菌して糖液（糖濃度１６％）とした。
前記擬似発酵物１～３を９ｇ、糖液３６ｇ、およびガセリ菌ＳＢＴ２０５５洗浄菌体０．５ｇを混合し、ｐＨ３．８、糖濃度１２．７％のラクトバチルス・パラカゼイ菌数の異なる本発明５～７の乳酸菌飲料を製造した。
（２）対照乳酸菌飲料の調製
前記（１）の擬似発酵物９ｇ、糖液３６ｇ、およびガセリ菌ＳＢＴ２０５５洗浄菌体０．５ｇを混合し、ラクトバチルス・パラカゼイを含まない乳酸菌飲料（対照１）を製造した。
また、ラクトバチルス・パラカゼイを１×１０７ＣＦＵ／ｇ程度含むラクトバチルス・パラカゼイ添加擬似発酵物を８０℃以上で３０分間殺菌したものを用いて乳酸菌飲料（対照２）を製造した。
（３）生残率の測定試験および結果
本発明品５～７の乳酸菌飲料および対照乳酸菌飲料（対照１、２）について、それぞれプラスチック容器に入れて１０℃で２１日間保存し、ガセリ菌およびラクトバチルス・パラカゼイの生菌数を経時的に測定した。結果を表５に示す。
本結果からラクトバチルス・パラカゼイを１×１０７ＣＦＵ／ｇ以上を添加した本発明の乳酸菌飲料においてガセリ菌の生残性が顕著に向上することがわかった。また、ｐＨ４．０においても本発明は効果を発揮するが、ラクトバチルス・パラカゼイ死菌体では効果がみられないことが明らかである。 [Example 2] Confirmation test for improvement of survival of gasseri by mixing with Lactobacillus paracasei When the number of Lactobacillus paracasei was changed and mixed with gasseri, and dead cells of Lactobacillus paracasei The effect on the viability of gasseri was investigated for the case of mixing with gasseri.
(1) Preparation of lactic acid bacteria beverage of the present invention Nonfat dry milk and glucose are dissolved in water to prepare a medium containing 16% (w / w) reduced skim milk and 3% (w / w) glucose. Sterilized at 120 ° C. for 120 minutes. Lactic acid sterilized by filtration was added to the sterilized medium to adjust the pH to about 4.0 to prepare a pseudo-fermented product. Lactobacillus paracasei SBT11408 washed cells are suspended in a pseudo-fermented product, and three types of Lactobacillus paracasei-added pseudo-fermented products with different numbers of paracasei by diluting 10 times, 100 times, and 1,000 times 3 was prepared. Further, the washed cells of gasseri bacterium SBT2055 were suspended in 12.6% reduced skim milk and used.
A 3% (w / w) isomerized liquid sugar / 14% (w / w) white sugar solution was prepared in water and sterilized at 121 ° C. for 15 minutes to give a sugar liquid (sugar concentration 16%).
9 g of the simulated fermented products 1 to 3, 36 g of sugar solution, and 0.5 g of washed cell of S. gasseri SBT2055 were mixed, and the present invention 5 having a different number of Lactobacillus paracasei bacteria having a pH of 3.8 and a sugar concentration of 12.7%. -7 lactic acid bacteria beverages were produced.
(2) Preparation of Control Lactic Acid Beverage Beverage 9 g of the pseudo-fermented product of (1), sugar solution 36 g, and gasseri SBT2055 washed cell body 0.5 g were mixed, and a lactic acid bacteria beverage containing no Lactobacillus paracasei (Control 1) Manufactured.
Moreover, the lactic acid bacteria drink (control 2) was manufactured using what sterilized the lactobacillus paracasei addition pseudo fermented product which contains about 1 * 10 < 7 > CFU / g of Lactobacillus paracasei for 30 minutes at 80 degreeC or more.
(3) Survival rate measurement test and results The lactic acid bacteria beverages and control lactic acid bacteria beverages (Controls 1 and 2) of the products 5 to 7 of the present invention were respectively stored in plastic containers for 21 days at 10 ° C. The viable count of Bacillus paracasei was measured over time. The results are shown in Table 5.
From these results, it was found that the viability of gasseri bacteria was significantly improved in the lactic acid bacteria beverage of the present invention to which 1 × 10 7 CFU / g or more of Lactobacillus paracasei was added. Moreover, although this invention exhibits an effect also in pH 4.0, it is clear that an effect is not seen with a Lactobacillus paracasei dead cell.
〔実施例３〕飲食品中の糖濃度が５～１５％におけるラクトバチルス・パラカゼイとの混合によるガセリ菌の生残性の向上確認試験
（１）本発明の乳酸菌飲料の調製
脱脂粉乳とグルコースを水に溶解し、１６％（ｗ／ｗ）還元脱脂乳培地を調製し、当該培地を１１５℃で２０分間殺菌した。前記殺菌培地にろ過滅菌した乳酸を添加してｐＨを約３．５に調整して、擬似発酵物を調製した。擬似発酵物にラクトバチルス・パラカゼイＳＢＴ１１４０８洗浄菌体を懸濁し、ラクトバチルス・パラカゼイ添加擬似発酵物を調製した。また、ガセリ菌ＳＢＴ２０５５の洗浄菌体を１２．６％還元脱脂乳で懸濁して使用した。糖液は終濃度５％、１０％、１５％となるグルコース水溶液をそれぞれ調製し、１２１℃で１５分間殺菌した。ラクトバチルス・パラカゼイ添加擬似発酵物９ｇ、糖液３６ｇ、およびガセリ菌ＳＢＴ２０５５洗浄菌体０．５ｇを混合し、グルコース濃度の異なる本発明品８～１０の乳酸菌飲料を製造した。
（２）対照乳酸菌飲料の調製
前記（１）の擬似発酵物９ｇ、ガセリ菌ＳＢＴ２０５５洗浄菌体０．５ｇ、および糖液は終濃度５％、１０％。１５％となるように混合して対照乳酸菌飲料を製造した。
（３）生残率の測定試験および結果
本発明品８～１０の乳酸菌飲料および対照乳酸菌飲料について、それぞれプラスチック容器に入れて１０℃で２１日間保存し、ガセリ菌およびラクトバチルス・パラカゼイの生菌数を経時的に測定した。結果を表６に示す。
ラクトバチルス・パラカゼイを１×１０７ＣＦＵ／ｇ以上添加した、グルコース濃度５％～１５％の本発明の乳酸菌飲料は、ラクトバチルス・ガセリの生残性が向上し、２１日間保存後のラクトバチルス・ガセリの生菌数は１×１０６ＣＦＵ／ｇ以上であった。特にグルコース濃度１０％以上の場合に効果が顕著であった。
[Example 3] Confirmation test for improvement of survival of gasseri by mixing with Lactobacillus paracasei at a sugar concentration of 5-15% in food and drink (1) Preparation of lactic acid bacteria beverage of the present invention Nonfat dry milk and glucose A 16% (w / w) reduced skim milk medium was prepared by dissolving in water, and the medium was sterilized at 115 ° C. for 20 minutes. Lactic acid sterilized by filtration was added to the sterilized medium to adjust the pH to about 3.5 to prepare a pseudo-fermented product. Lactobacillus paracasei SBT11408 washed cells were suspended in the pseudofermented product to prepare a lactobacillus paracasei-added pseudofermented product. Further, the washed cells of gasseri bacterium SBT2055 were suspended in 12.6% reduced skim milk and used. As the sugar solutions, aqueous glucose solutions having final concentrations of 5%, 10%, and 15% were prepared and sterilized at 121 ° C. for 15 minutes. Lactobacillus paracasei added pseudo-fermented product 9 g, sugar solution 36 g, and gasseri bacteria SBT2055 washed cells 0.5 g were mixed to produce lactic acid bacteria beverages of the present invention products 8 to 10 having different glucose concentrations.
(2) Preparation of Control Lactic Acid Beverage 9 g of the simulated fermented product of (1) above, 0.5 g of the washed Bacterium SBT2055 cell, and sugar solution have final concentrations of 5% and 10%. A control lactic acid bacteria beverage was prepared by mixing at 15%.
(3) Survival rate measurement test and results The lactic acid bacteria beverages and the control lactic acid bacteria beverages of the products 8 to 10 of the present invention were each stored in a plastic container at 10 ° C. for 21 days, and live bacteria of gasseri and Lactobacillus paracasei Numbers were measured over time. The results are shown in Table 6.
Lactobacillus paracasei to which 1 × 10 7 CFU / g or more is added and the lactic acid bacteria beverage of the present invention having a glucose concentration of 5% to 15% improves the survival of Lactobacillus gasseri, and the lactobacilli after storage for 21 days The viable count of gasseri was 1 × 10 6 CFU / g or more. The effect was particularly remarkable when the glucose concentration was 10% or more.
（１）本発明の乳酸菌飲料の調製
脱脂粉乳とグルコースを水に溶解し、１６％（ｗ／ｗ）還元脱脂乳培地を調製し、当該培地を１１５℃で２０分間殺菌した。前記殺菌培地にろ過滅菌した乳酸を添加してｐＨを約３．５に調整して、擬似発酵物を調製した。擬似発酵物にラクトバチルス・パラカゼイＳＢＴ１１４０８洗浄菌体を懸濁し、ラクトバチルス・パラカゼイ添加擬似発酵物を調製した。また、ガセリ菌ＳＢＴ２０５５の洗浄菌体を１２．６％還元脱脂乳で懸濁して使用した。糖液は終濃度５％、１０％、１５％となるグルコース水溶液をそれぞれ調製し、１２１℃で１５分間殺菌した。ラクトバチルス・パラカゼイ添加擬似発酵物９ｇ、糖液３６ｇ、およびガセリ菌ＳＢＴ２０５５洗浄菌体０．５ｇを混合し、グルコース濃度の異なる本発明品８～１０の乳酸菌飲料を製造した。
（２）対照乳酸菌飲料の調製
前記（１）の擬似発酵物９ｇ、ガセリ菌ＳＢＴ２０５５洗浄菌体０．５ｇ、および糖液は終濃度５％、１０％。１５％となるように混合して対照乳酸菌飲料を製造した。
（３）生残率の測定試験および結果
本発明品８～１０の乳酸菌飲料および対照乳酸菌飲料について、それぞれプラスチック容器に入れて１０℃で２１日間保存し、ガセリ菌およびラクトバチルス・パラカゼイの生菌数を経時的に測定した。結果を表６に示す。
ラクトバチルス・パラカゼイを１×１０７ＣＦＵ／ｇ以上添加した、グルコース濃度５％～１５％の本発明の乳酸菌飲料は、ラクトバチルス・ガセリの生残性が向上し、２１日間保存後のラクトバチルス・ガセリの生菌数は１×１０６ＣＦＵ／ｇ以上であった。特にグルコース濃度１０％以上の場合に効果が顕著であった。
[Example 3] Confirmation test for improvement of survival of gasseri by mixing with Lactobacillus paracasei at a sugar concentration of 5-15% in food and drink (1) Preparation of lactic acid bacteria beverage of the present invention Nonfat dry milk and glucose A 16% (w / w) reduced skim milk medium was prepared by dissolving in water, and the medium was sterilized at 115 ° C. for 20 minutes. Lactic acid sterilized by filtration was added to the sterilized medium to adjust the pH to about 3.5 to prepare a pseudo-fermented product. Lactobacillus paracasei SBT11408 washed cells were suspended in the pseudofermented product to prepare a lactobacillus paracasei-added pseudofermented product. Further, the washed cells of gasseri bacterium SBT2055 were suspended in 12.6% reduced skim milk and used. As the sugar solutions, aqueous glucose solutions having final concentrations of 5%, 10%, and 15% were prepared and sterilized at 121 ° C. for 15 minutes. Lactobacillus paracasei added pseudo-fermented product 9 g, sugar solution 36 g, and gasseri bacteria SBT2055 washed cells 0.5 g were mixed to produce lactic acid bacteria beverages of the present invention products 8 to 10 having different glucose concentrations.
(2) Preparation of Control Lactic Acid Beverage 9 g of the simulated fermented product of (1) above, 0.5 g of the washed Bacterium SBT2055 cell, and sugar solution have final concentrations of 5% and 10%. A control lactic acid bacteria beverage was prepared by mixing at 15%.
(3) Survival rate measurement test and results The lactic acid bacteria beverages and the control lactic acid bacteria beverages of the products 8 to 10 of the present invention were each stored in a plastic container at 10 ° C. for 21 days, and live bacteria of gasseri and Lactobacillus paracasei Numbers were measured over time. The results are shown in Table 6.
Lactobacillus paracasei to which 1 × 10 7 CFU / g or more is added and the lactic acid bacteria beverage of the present invention having a glucose concentration of 5% to 15% improves the survival of Lactobacillus gasseri, and the lactobacilli after storage for 21 days The viable count of gasseri was 1 × 10 6 CFU / g or more. The effect was particularly remarkable when the glucose concentration was 10% or more.
〔実施例４〕
脱脂粉乳とグルコースを水に溶解し、１６％（ｗ／ｗ）還元脱脂乳、３％（ｗ／ｗ）グルコースを含む培地を調製し、当該培地を９５℃で１２０分間殺菌した。３７℃まで冷却した殺菌培地にラクトバチルス・ブフネリＳＢＴ２０２８のシードカルチャーを３％（ｖ／ｖ）添加して３７℃でｐＨ４．０程度になるまで培養し、ラクトバチルス・ブフネリ発酵物を得た。
また、３％（ｗ／ｗ）異性化液糖・１４％（ｗ／ｗ）上白糖溶液を水で調製し、１２１℃１５分間滅菌して糖液（糖濃度１６％）とした。
上記で得られた糖液３６ｇ、ラクトバチルス・ブフネリ発酵物９ｇおよびガセリ菌ＳＢＴ１７０３濃縮菌体０．５ｇを混合し、ｐＨ３．８、糖濃度１２．７％である本発明の乳酸菌飲料を製造した。
ラクトバチルス・ガセリが１．９×１０９ＣＦＵ／ｇ、ラクトバチルス・ブフネリが４．８×１０８ＣＦＵ／ｇ 添加された条件で２１日間保存後のラクトバチルス・ガセリの生菌数は５．０×１０６ＣＦＵ／ｇであった。 Example 4
Skim milk powder and glucose were dissolved in water to prepare a medium containing 16% (w / w) reduced skim milk and 3% (w / w) glucose, and the medium was sterilized at 95 ° C. for 120 minutes. 3% (v / v) of Lactobacillus buchneri SBT2028 seed culture was added to the sterilized medium cooled to 37 ° C. and cultured at 37 ° C. until the pH reached about 4.0 to obtain a fermented Lactobacillus buchneri.
Further, a 3% (w / w) isomerized liquid sugar / 14% (w / w) white sugar solution was prepared in water and sterilized at 121 ° C. for 15 minutes to obtain a sugar liquid (sugar concentration: 16%).
36 g of the sugar solution obtained above, 9 g of the fermented Lactobacillus buchnerii and 0.5 g of concentrated Bacteria SBT1703 were mixed to produce a lactic acid bacteria beverage of the present invention having a pH of 3.8 and a sugar concentration of 12.7%. .
Lactobacillus gasseri 1.9 × 10 9 CFU / g, Lactobacillus bucheneri 4.8 × 10 8 CFU / g added Lactobacillus gasseri viable count after storage for 21 days is 5. It was 0 × 10 6 CFU / g.
脱脂粉乳とグルコースを水に溶解し、１６％（ｗ／ｗ）還元脱脂乳、３％（ｗ／ｗ）グルコースを含む培地を調製し、当該培地を９５℃で１２０分間殺菌した。３７℃まで冷却した殺菌培地にラクトバチルス・ブフネリＳＢＴ２０２８のシードカルチャーを３％（ｖ／ｖ）添加して３７℃でｐＨ４．０程度になるまで培養し、ラクトバチルス・ブフネリ発酵物を得た。
また、３％（ｗ／ｗ）異性化液糖・１４％（ｗ／ｗ）上白糖溶液を水で調製し、１２１℃１５分間滅菌して糖液（糖濃度１６％）とした。
上記で得られた糖液３６ｇ、ラクトバチルス・ブフネリ発酵物９ｇおよびガセリ菌ＳＢＴ１７０３濃縮菌体０．５ｇを混合し、ｐＨ３．８、糖濃度１２．７％である本発明の乳酸菌飲料を製造した。
ラクトバチルス・ガセリが１．９×１０９ＣＦＵ／ｇ、ラクトバチルス・ブフネリが４．８×１０８ＣＦＵ／ｇ 添加された条件で２１日間保存後のラクトバチルス・ガセリの生菌数は５．０×１０６ＣＦＵ／ｇであった。 Example 4
Skim milk powder and glucose were dissolved in water to prepare a medium containing 16% (w / w) reduced skim milk and 3% (w / w) glucose, and the medium was sterilized at 95 ° C. for 120 minutes. 3% (v / v) of Lactobacillus buchneri SBT2028 seed culture was added to the sterilized medium cooled to 37 ° C. and cultured at 37 ° C. until the pH reached about 4.0 to obtain a fermented Lactobacillus buchneri.
Further, a 3% (w / w) isomerized liquid sugar / 14% (w / w) white sugar solution was prepared in water and sterilized at 121 ° C. for 15 minutes to obtain a sugar liquid (sugar concentration: 16%).
36 g of the sugar solution obtained above, 9 g of the fermented Lactobacillus buchnerii and 0.5 g of concentrated Bacteria SBT1703 were mixed to produce a lactic acid bacteria beverage of the present invention having a pH of 3.8 and a sugar concentration of 12.7%. .
Lactobacillus gasseri 1.9 × 10 9 CFU / g, Lactobacillus bucheneri 4.8 × 10 8 CFU / g added Lactobacillus gasseri viable count after storage for 21 days is 5. It was 0 × 10 6 CFU / g.
〔実施例５〕
脱脂粉乳とグルコースを水に溶解し、１６％（ｗ／ｗ）還元脱脂乳、３％（ｗ／ｗ）グルコースを含む培地を調製し、当該培地を９５℃で１２０分間殺菌した。３７℃まで冷却した殺菌培地にラクトバチルス・プランタラムＳＢＴ１５３４およびラクトバチルス・ガセリＳＢＴ１０２４１のシードカルチャーをそれぞれ３％（ｖ／ｖ）ずつ添加して、３７℃でｐＨ４．０程度になるまで発酵させた。
また、３％（ｗ／ｗ）異性化液糖・１４％（ｗ／ｗ）上白糖溶液を水で調製し、１２１℃１５分間滅菌して糖液（糖濃度１６％）とした。
上記で得られた糖液３６ｇ、ラクトバチルス・プランタラムおよびラクトバチルス・ガセリの発酵物９．５ｇを混合し、ｐＨ３．８、糖濃度１２．７％である本発明の乳酸菌飲料を製造した。
ラクトバチルス・ガセリが２．０×１０９ＣＦＵ／ｇ、ラクトバチルス・プランタラムが５．４×１０８ＣＦＵ／ｇ 添加された条件で２１日間保存後のラクトバチルス・ガセリの生菌数は１２×１０６ＣＦＵ／ｇであった。 Example 5
Skim milk powder and glucose were dissolved in water to prepare a medium containing 16% (w / w) reduced skim milk and 3% (w / w) glucose, and the medium was sterilized at 95 ° C. for 120 minutes. 3% (v / v) each of seed cultures of Lactobacillus plantarum SBT1534 and Lactobacillus gasseri SBT10241 were added to a sterilized medium cooled to 37 ° C., and fermented at 37 ° C. until the pH reached about 4.0. .
Further, a 3% (w / w) isomerized liquid sugar / 14% (w / w) white sugar solution was prepared in water and sterilized at 121 ° C. for 15 minutes to obtain a sugar liquid (sugar concentration: 16%).
36 g of the sugar solution obtained above and 9.5 g of a fermented product of Lactobacillus plantarum and Lactobacillus gasseri were mixed to produce a lactic acid bacteria beverage of the present invention having a pH of 3.8 and a sugar concentration of 12.7%.
Lactobacillus gasseri is 2.0 × 10 9 CFU / g and Lactobacillus plantarum is 5.4 × 10 8 CFU / g. The number of viable Lactobacillus gasseri after storage for 21 days is 12 × 10 6 CFU / g.
脱脂粉乳とグルコースを水に溶解し、１６％（ｗ／ｗ）還元脱脂乳、３％（ｗ／ｗ）グルコースを含む培地を調製し、当該培地を９５℃で１２０分間殺菌した。３７℃まで冷却した殺菌培地にラクトバチルス・プランタラムＳＢＴ１５３４およびラクトバチルス・ガセリＳＢＴ１０２４１のシードカルチャーをそれぞれ３％（ｖ／ｖ）ずつ添加して、３７℃でｐＨ４．０程度になるまで発酵させた。
また、３％（ｗ／ｗ）異性化液糖・１４％（ｗ／ｗ）上白糖溶液を水で調製し、１２１℃１５分間滅菌して糖液（糖濃度１６％）とした。
上記で得られた糖液３６ｇ、ラクトバチルス・プランタラムおよびラクトバチルス・ガセリの発酵物９．５ｇを混合し、ｐＨ３．８、糖濃度１２．７％である本発明の乳酸菌飲料を製造した。
ラクトバチルス・ガセリが２．０×１０９ＣＦＵ／ｇ、ラクトバチルス・プランタラムが５．４×１０８ＣＦＵ／ｇ 添加された条件で２１日間保存後のラクトバチルス・ガセリの生菌数は１２×１０６ＣＦＵ／ｇであった。 Example 5
Skim milk powder and glucose were dissolved in water to prepare a medium containing 16% (w / w) reduced skim milk and 3% (w / w) glucose, and the medium was sterilized at 95 ° C. for 120 minutes. 3% (v / v) each of seed cultures of Lactobacillus plantarum SBT1534 and Lactobacillus gasseri SBT10241 were added to a sterilized medium cooled to 37 ° C., and fermented at 37 ° C. until the pH reached about 4.0. .
Further, a 3% (w / w) isomerized liquid sugar / 14% (w / w) white sugar solution was prepared in water and sterilized at 121 ° C. for 15 minutes to obtain a sugar liquid (sugar concentration: 16%).
36 g of the sugar solution obtained above and 9.5 g of a fermented product of Lactobacillus plantarum and Lactobacillus gasseri were mixed to produce a lactic acid bacteria beverage of the present invention having a pH of 3.8 and a sugar concentration of 12.7%.
Lactobacillus gasseri is 2.0 × 10 9 CFU / g and Lactobacillus plantarum is 5.4 × 10 8 CFU / g. The number of viable Lactobacillus gasseri after storage for 21 days is 12 × 10 6 CFU / g.
〔実施例６〕ラクトバチルス・パラカゼイ、ラクトバチルス・プランタラムまたはラクトバチルス・ブフネリとの混合によるガセリ菌の生残性の向上確認試験
上記試験例２のスクリーニング試験で選択された菌をガセリ菌と混合して、ガセリ菌の生残性が向上することを確認した。
実施例１と同様の方法により、本発明の乳酸菌飲料及び対照乳酸菌飲料を調製した。なお、ガセリ菌としては、ラクトバチルス・ガセリＳＢＴ２０５５と基準株であるラクトバチルス・ガセリＪＣＭ１１３１を使用し、また、ガセリ菌に混合する菌として、ラクトバチルス・パラカゼイＡＴＣＣ２５５９８、ラクトバチルス・プランタラムＡＴＣＣ４３１９９、ラクトバチルス・プランタラムＡＴＣＣ８０１４、ラクトバチルス・ブフネリＪＣＭ１１１５を使用した。また、対照乳酸菌飲料として、ガセリ菌に混合する菌として、ラクトバチルス・ブルガリクス （ＮＢＲＣ Ｐ‐１３９５３）を使用した乳酸菌飲料も調製した。
調製した乳酸菌飲料について、実施例１と同様の方法により、ガセリ菌及び混合した菌について生残率を測定した。結果を表７に示す。 [Example 6] Confirmation test for improvement of survival of gasseri by mixing with Lactobacillus paracasei, Lactobacillus plantarum or Lactobacillus buchneri The bacteria selected in the screening test of Test Example 2 above It mixed and it confirmed that the survival property of gasseri bacteria improved.
A lactic acid bacteria beverage of the present invention and a control lactic acid bacteria beverage were prepared in the same manner as in Example 1. Note that Lactobacillus gasseri SBT2055 and the standard strain Lactobacillus gasseri JCM1131 are used as gasseri bacteria, and Lactobacillus paracasei ATCC 25598, Lactobacillus plantarum ATCC 43199, Bacillus plantarum ATCC 8014 and Lactobacillus buchneri JCM1115 were used. In addition, as a control lactic acid bacteria beverage, a lactic acid bacteria beverage using Lactobacillus bulgaricus (NBRC P-13953) as a bacterium mixed with gasseri was also prepared.
About the prepared lactic acid bacteria drink, the survival rate was measured about the gasseri bacteria and the mixed bacteria by the same method as Example 1. The results are shown in Table 7.
上記試験例２のスクリーニング試験で選択された菌をガセリ菌と混合して、ガセリ菌の生残性が向上することを確認した。
実施例１と同様の方法により、本発明の乳酸菌飲料及び対照乳酸菌飲料を調製した。なお、ガセリ菌としては、ラクトバチルス・ガセリＳＢＴ２０５５と基準株であるラクトバチルス・ガセリＪＣＭ１１３１を使用し、また、ガセリ菌に混合する菌として、ラクトバチルス・パラカゼイＡＴＣＣ２５５９８、ラクトバチルス・プランタラムＡＴＣＣ４３１９９、ラクトバチルス・プランタラムＡＴＣＣ８０１４、ラクトバチルス・ブフネリＪＣＭ１１１５を使用した。また、対照乳酸菌飲料として、ガセリ菌に混合する菌として、ラクトバチルス・ブルガリクス （ＮＢＲＣ Ｐ‐１３９５３）を使用した乳酸菌飲料も調製した。
調製した乳酸菌飲料について、実施例１と同様の方法により、ガセリ菌及び混合した菌について生残率を測定した。結果を表７に示す。 [Example 6] Confirmation test for improvement of survival of gasseri by mixing with Lactobacillus paracasei, Lactobacillus plantarum or Lactobacillus buchneri The bacteria selected in the screening test of Test Example 2 above It mixed and it confirmed that the survival property of gasseri bacteria improved.
A lactic acid bacteria beverage of the present invention and a control lactic acid bacteria beverage were prepared in the same manner as in Example 1. Note that Lactobacillus gasseri SBT2055 and the standard strain Lactobacillus gasseri JCM1131 are used as gasseri bacteria, and Lactobacillus paracasei ATCC 25598, Lactobacillus plantarum ATCC 43199, Bacillus plantarum ATCC 8014 and Lactobacillus buchneri JCM1115 were used. In addition, as a control lactic acid bacteria beverage, a lactic acid bacteria beverage using Lactobacillus bulgaricus (NBRC P-13953) as a bacterium mixed with gasseri was also prepared.
About the prepared lactic acid bacteria drink, the survival rate was measured about the gasseri bacteria and the mixed bacteria by the same method as Example 1. The results are shown in Table 7.
本結果から明らかなように、ガセリ菌とラクトバチルス・パラカゼイ、ラクトバチルス・プランタラム、ラクトバチルス・ブフネリを混合させた本発明品の乳酸菌飲料では、いずれも対照乳酸菌飲料に比べて著しくガセリ菌の生残菌数が高く、２１日間保存後のラクトバチルス・ガセリの生菌数は本発明品１１～１８の全てにおいて１×１０6ＣＦＵ／ｇ以上であった。
一方、ラクトバチルス・ブルガリクスを混合した場合は、ガセリ菌の生残性の向上は見られず、21日間保存後のラクトバチルス・ガセリの生存率は0.00006％以下となった。
以上より、３％（ｗ／ｗ）還元脱脂乳および１０％（ｗ／ｗ）グルコースを含むｐＨ４．０の培地に添加して１０℃において２１日間静置培養したときに該培地のｐＨを０．１以上低下させることができる乳酸菌、と混合することによりガセリ菌の生残性が飛躍的に向上することが明らかとなった。 As is apparent from these results, the lactic acid bacteria beverages of the present invention in which gasseri and Lactobacillus paracasei, Lactobacillus plantarum, and Lactobacillus buchnerii were mixed were markedly different from the control lactic acid bacteria beverages. The number of surviving bacteria was high, and the number of viable bacteria of Lactobacillus gasseri after storage for 21 days was 1 × 10 6 CFU / g or more in all of the products 11 to 18 of the present invention.
On the other hand, when Lactobacillus bulgaricus was mixed, the viability of gasseri was not improved, and the survival rate of Lactobacillus gasseri after storage for 21 days was 0.00006% or less.
From the above, when added to a pH 4.0 medium containing 3% (w / w) reduced skim milk and 10% (w / w) glucose and left to stand at 10 ° C. for 21 days, the pH of the medium was reduced to 0. It was clarified that the viability of gasseri bacteria was dramatically improved by mixing with lactic acid bacteria that can be reduced by 1 or more.
一方、ラクトバチルス・ブルガリクスを混合した場合は、ガセリ菌の生残性の向上は見られず、21日間保存後のラクトバチルス・ガセリの生存率は0.00006％以下となった。
以上より、３％（ｗ／ｗ）還元脱脂乳および１０％（ｗ／ｗ）グルコースを含むｐＨ４．０の培地に添加して１０℃において２１日間静置培養したときに該培地のｐＨを０．１以上低下させることができる乳酸菌、と混合することによりガセリ菌の生残性が飛躍的に向上することが明らかとなった。 As is apparent from these results, the lactic acid bacteria beverages of the present invention in which gasseri and Lactobacillus paracasei, Lactobacillus plantarum, and Lactobacillus buchnerii were mixed were markedly different from the control lactic acid bacteria beverages. The number of surviving bacteria was high, and the number of viable bacteria of Lactobacillus gasseri after storage for 21 days was 1 × 10 6 CFU / g or more in all of the products 11 to 18 of the present invention.
On the other hand, when Lactobacillus bulgaricus was mixed, the viability of gasseri was not improved, and the survival rate of Lactobacillus gasseri after storage for 21 days was 0.00006% or less.
From the above, when added to a pH 4.0 medium containing 3% (w / w) reduced skim milk and 10% (w / w) glucose and left to stand at 10 ° C. for 21 days, the pH of the medium was reduced to 0. It was clarified that the viability of gasseri bacteria was dramatically improved by mixing with lactic acid bacteria that can be reduced by 1 or more.
本発明によれば、長期間保存後もガセリ菌の生残性が極めて高い、低ｐH、高糖濃度飲食品を提供することができる。したがって、有用微生物であるガセリ菌の機能を十分に維持し発揮できる食品を提供することができる。
ADVANTAGE OF THE INVENTION According to this invention, the low pH and high sugar concentration food / beverage products in which the survival property of gasseri bacteria is very high even after long-term storage can be provided. Therefore, the foodstuff which can fully maintain and exhibit the function of gasseri bacteria which are useful microorganisms can be provided.
[寄託生物材料への言及]
（１）Lactobacillus gasseri ＳＢＴ２０５５
イ 当該生物材料を寄託した寄託機関の名称及び住所
独立行政法人 産業技術総合研究所 特許生物寄託センター
日本国茨城県つくば市東１丁目１番地１中央第６（郵便番号305-8566）
ロ イの寄託機関に生物材料を寄託した日付
平成８年３月２７日（原寄託日）
平成２０年２月２６日（原寄託によりブタペスト条約に基づく寄託への移管日）
ハ イの寄託機関が寄託について付した受託番号
ＦＥＲＭ ＢＰ－１０９５３
（２）Lactobacillus gasseri ＳＢＴ２０５６
イ 当該生物材料を寄託した寄託機関の名称及び住所
上記（１）に同じ
ロ イの寄託機関に生物材料を寄託した日付
昭和６１年４月２２日（原寄託日）
平成２０年１０月９日（原寄託によりブタペスト条約に基づく寄託への移管日）
ハ イの寄託機関が寄託について付した受託番号
ＦＥＲＭ ＢＰ－１１０３８
（３）Lactobacillus gasseri ＳＢＴ０２７４
イ 当該生物材料を寄託した寄託機関の名称及び住所
上記（１）に同じ
ロ イの寄託機関に生物材料を寄託した日付
平成１２年３月１５日（原寄託日）
平成２０年１０月９日（原寄託によりブタペスト条約に基づく寄託への移管日）
ハ イの寄託機関が寄託について付した受託番号
ＦＥＲＭ ＢＰ－１１０３９
（４）Lactobacillus gasseri ＳＢＴ１７０３
イ 当該生物材料を寄託した寄託機関の名称及び住所
工業技術院生命工学技術研究所
日本国茨城県つくば市東１丁目１番3号（郵便番号305-8566）
ロ イの寄託機関に生物材料を寄託した日付
平成１２年３月１５日
ハ イの寄託機関が寄託について付した受託番号
ＦＥＲＭ Ｐ－１７７８５
（５）Lactobacillus gasseri ＳＢＴ１０２３９
イ 当該生物材料を寄託した寄託機関の名称及び住所
上記（４）に同じ
ロ イの寄託機関に生物材料を寄託した日付
平成１０年２月１７日
ハ イの寄託機関が寄託について付した受託番号
ＦＥＲＭ Ｐ－１６６３９
（６）Lactobacillus gasseri ＳＢＴ１０２４１
イ 当該生物材料を寄託した寄託機関の名称及び住所
上記（４）に同じ
ロ イの寄託機関に生物材料を寄託した日付
平成１２年３月１５日
ハ イの寄託機関が寄託について付した受託番号
ＦＥＲＭ Ｐ－１７７８６
（７）Lactobacillus paracasei ＳＢＴ０３２７
イ 当該生物材料を寄託した寄託機関の名称及び住所
独立行政法人 製品評価技術基盤機構 特許微生物寄託センター
日本国千葉県木更津市かずさ鎌足2-5-8（郵便番号292-0818）
ロ イの寄託機関に生物材料を寄託した日付
２０１１年８月１８日
２０１２年１２月６日（原寄託によりブタペスト条約に基づく寄託への移管日）
ハ イの寄託機関が寄託について付した受託番号（受領番号）
ＮＩＴＥ ＡＢＰ－１１２９
（８）Lactobacillus paracasei ＳＢＴ２１０５
イ 当該生物材料を寄託した寄託機関の名称及び住所
上記（７）に同じ
ロ イの寄託機関に生物材料を寄託した日付
２０１１年８月１８日
ハ イの寄託機関が寄託について付した受託番号
ＮＩＴＥ Ｐ－１１３０
（９）Lactobacillus paracasei ＳＢＴ２２０３
イ 当該生物材料を寄託した寄託機関の名称及び住所
上記（７）に同じ
ロ イの寄託機関に生物材料を寄託した日付
２０１１年８月１８日
２０１２年１２月６日（原寄託によりブタペスト条約に基づく寄託への移管日）
ハ イの寄託機関が寄託について付した受託番号（受領番号）
ＮＩＴＥ ＡＢＰ－１１３１
（１０）Lactobacillus paracasei ＳＢＴ２２１５
イ 当該生物材料を寄託した寄託機関の名称及び住所
上記（７）に同じ
ロ イの寄託機関に生物材料を寄託した日付
２０１１年８月１８日
２０１２年１２月６日（原寄託によりブタペスト条約に基づく寄託への移管日）
ハ イの寄託機関が寄託について付した受託番号(受領番号)
ＮＩＴＥ ＡＢＰ－１１３２
（１１）Lactobacillus paracasei ＳＢＴ１１４０８
イ 当該生物材料を寄託した寄託機関の名称及び住所
上記（７）に同じ
ロ イの寄託機関に生物材料を寄託した日付
２０１１年８月１８日
２０１２年１２月６日（原寄託によりブタペスト条約に基づく寄託への移管日）
ハ イの寄託機関が寄託について付した受託番号(受領番号)
ＮＩＴＥ ＡＢＰ－１１３３
（１２）Lactobacillus plantarum ＳＢＴ１５３４
イ 当該生物材料を寄託した寄託機関の名称及び住所
上記（４）に同じ
ロ イの寄託機関に生物材料を寄託した日付
平成３年７月１０日
２０１２年１２月６日（原寄託によりブタペスト条約に基づく寄託への移管日。国内寄託番号ＦＥＲＭ Ｐ－１２３５１より移管）
ハ イの寄託機関が寄託について付した受託番号(受領番号)
ＦＥＲＭ ＡＢＰ－１１５１８
（１３）Lactobacillus casei subsp. pseudoplantarum ＳＢＴ０６２４
イ 当該生物材料を寄託した寄託機関の名称及び住所
上記（４）に同じ
ロ イの寄託機関に生物材料を寄託した日付
平成２年１２月２０日
ハ イの寄託機関が寄託について付した受託番号
ＦＥＲＭ Ｐ－１１９２０
（１４）Lactobacillus buchneri ＳＢＴ２０２８
イ 当該生物材料を寄託した寄託機関の名称及び住所
上記（４）に同じ
ロ イの寄託機関に生物材料を寄託した日付
平成２年１２月２０日
ハ イの寄託機関が寄託について付した受託番号
ＦＥＲＭ Ｐ－１１９２１ [Reference to deposited biological materials]
(1) Lactobacillus gasseri SBT2055
Name and address of the depository institution that deposited the biological material National Institute of Advanced Industrial Science and Technology Patent Biological Deposit Center 1-Chuo 1-chome, Tsukuba City, Ibaraki Prefecture, Japan 6 (zip code 305-8566)
Date of deposit of biological materials at Loi depository organization March 27, 1996 (original deposit date)
February 26, 2008 (Date of transfer to deposit under the Budapest Treaty by original deposit)
Deposit number FERM BP-10953 assigned by the depository in Thailand for deposit
(2) Lactobacillus gasseri SBT2056
(A) Name and address of the depositary that deposited the biological material Date of deposit of the biological material at the same depository in Loi above (1) April 22, 1986 (original deposit date)
October 9, 2008 (Date of transfer to deposit under the Budapest Treaty by original deposit)
Deposit number FERM BP-11038 assigned by the depository in Hai for the deposit
(3) Lactobacillus gasseri SBT0274
The name and address of the depository that deposited the biological material The date on which the biological material was deposited at the same depository in (1) above March 15, 2000 (original deposit date)
October 9, 2008 (Date of transfer to deposit under the Budapest Treaty by original deposit)
Deposit number FERM BP-11039 assigned by the depository in Thailand for deposit
(4) Lactobacillus gasseri SBT1703
The name and address of the depository institution where the biological material was deposited 1-3, Higashi 1-chome, Tsukuba City, Ibaraki, Japan, Institute of Biotechnology, National Institute of Advanced Industrial Science and Technology (zip code 305-8566)
Date of deposit of biological material at the depository in Loi March 15, 2000 Deposit number FERM P-17785 attached to the depository by the depository in Hai
(5) Lactobacillus gasseri SBT10239
(A) Name and address of the depositary that deposited the biological material Date of deposit of the biological material at the same depository in the same (4) above February 17, 1998 The deposit number assigned by the depositary of hi to the deposit FERM P-16639
(6) Lactobacillus gasseri SBT10241
(A) Name and address of the depositary that deposited the biological material Date of deposit of the biological material at the same depository in (4) above March 15, 2000 The deposit number assigned by the depositary of hi FERM P-17786
(7) Lactobacillus paracasei SBT0327
(I) Name and address of the depository that deposited the biomaterial The National Institute of Technology and Evaluation, Patent Microorganism Deposit Center 2-5-8 Kazusa Kamashiri, Kisarazu City, Chiba Prefecture, Japan (zip code 292-0818)
Date of deposit of biological materials at Loi depository August 18, 2011 December 6, 2012 (Date of transfer to original deposit under the Budapest Treaty)
The deposit number (receipt number) assigned by the depositary in Japan for the deposit
NITE ABP-1129
(8) Lactobacillus paracasei SBT2105
(A) Name and address of the depository that deposited the biological material Date of deposit of the biological material at the same depository in the above (7) August 18, 2011 Deposit number assigned by the depositary institution for high NITE P-1130
(9) Lactobacillus paracasei SBT2203
(A) Name and address of the depositary that deposited the biological material Date of depositing the biological material at the same depository in (7) above August 18, 2011 December 6, 2012 (according to the Budapest Treaty by the original deposit) Transfer date to deposit based on)
The deposit number (receipt number) assigned by the depositary in Japan for the deposit
NITE ABP-1131
(10) Lactobacillus paracasei SBT2215
(A) Name and address of the depositary that deposited the biological material Date of depositing the biological material at the same depository in (7) above August 18, 2011 December 6, 2012 (according to the Budapest Treaty by the original deposit) Transfer date to deposit based on)
The deposit number (receipt number) assigned by the depositary in Japan for the deposit
NITE ABP-1132
(11) Lactobacillus paracasei SBT11408
(A) Name and address of the depositary that deposited the biological material Date of depositing the biological material at the same depository in (7) above August 18, 2011 December 6, 2012 (according to the Budapest Treaty by the original deposit) Transfer date to deposit based on)
The deposit number (receipt number) assigned by the depositary in Japan for the deposit
NITE ABP-1133
(12) Lactobacillus plantarum SBT1534
(A) Name and address of the depositary that deposited the biological material Date of deposit of the biological material at the same depository in (4) above July 10, 1991 December 6, 2012 (the Budapest Treaty by the original deposit) Date of transfer to deposit based on US (transferred from domestic deposit number FERM P-12351)
The deposit number (receipt number) assigned by the depositary in Japan for the deposit
FERM ABP-11518
(13) Lactobacillus casei subsp. Pseudoplantarum SBT0624
(A) Name and address of the depositary that deposited the biological material Date of deposit of the biological material at the same depository in (4) above The deposit number assigned by the depositary institution on December 20, 1990 FERM P-11920
(14) Lactobacillus buchneri SBT2028
(A) Name and address of the depositary that deposited the biological material Date of deposit of the biological material at the same depository in (4) above The deposit number assigned by the depositary institution on December 20, 1990 FERM P-11921
（１）Lactobacillus gasseri ＳＢＴ２０５５
イ 当該生物材料を寄託した寄託機関の名称及び住所
独立行政法人 産業技術総合研究所 特許生物寄託センター
日本国茨城県つくば市東１丁目１番地１中央第６（郵便番号305-8566）
ロ イの寄託機関に生物材料を寄託した日付
平成８年３月２７日（原寄託日）
平成２０年２月２６日（原寄託によりブタペスト条約に基づく寄託への移管日）
ハ イの寄託機関が寄託について付した受託番号
ＦＥＲＭ ＢＰ－１０９５３
（２）Lactobacillus gasseri ＳＢＴ２０５６
イ 当該生物材料を寄託した寄託機関の名称及び住所
上記（１）に同じ
ロ イの寄託機関に生物材料を寄託した日付
昭和６１年４月２２日（原寄託日）
平成２０年１０月９日（原寄託によりブタペスト条約に基づく寄託への移管日）
ハ イの寄託機関が寄託について付した受託番号
ＦＥＲＭ ＢＰ－１１０３８
（３）Lactobacillus gasseri ＳＢＴ０２７４
イ 当該生物材料を寄託した寄託機関の名称及び住所
上記（１）に同じ
ロ イの寄託機関に生物材料を寄託した日付
平成１２年３月１５日（原寄託日）
平成２０年１０月９日（原寄託によりブタペスト条約に基づく寄託への移管日）
ハ イの寄託機関が寄託について付した受託番号
ＦＥＲＭ ＢＰ－１１０３９
（４）Lactobacillus gasseri ＳＢＴ１７０３
イ 当該生物材料を寄託した寄託機関の名称及び住所
工業技術院生命工学技術研究所
日本国茨城県つくば市東１丁目１番3号（郵便番号305-8566）
ロ イの寄託機関に生物材料を寄託した日付
平成１２年３月１５日
ハ イの寄託機関が寄託について付した受託番号
ＦＥＲＭ Ｐ－１７７８５
（５）Lactobacillus gasseri ＳＢＴ１０２３９
イ 当該生物材料を寄託した寄託機関の名称及び住所
上記（４）に同じ
ロ イの寄託機関に生物材料を寄託した日付
平成１０年２月１７日
ハ イの寄託機関が寄託について付した受託番号
ＦＥＲＭ Ｐ－１６６３９
（６）Lactobacillus gasseri ＳＢＴ１０２４１
イ 当該生物材料を寄託した寄託機関の名称及び住所
上記（４）に同じ
ロ イの寄託機関に生物材料を寄託した日付
平成１２年３月１５日
ハ イの寄託機関が寄託について付した受託番号
ＦＥＲＭ Ｐ－１７７８６
（７）Lactobacillus paracasei ＳＢＴ０３２７
イ 当該生物材料を寄託した寄託機関の名称及び住所
独立行政法人 製品評価技術基盤機構 特許微生物寄託センター
日本国千葉県木更津市かずさ鎌足2-5-8（郵便番号292-0818）
ロ イの寄託機関に生物材料を寄託した日付
２０１１年８月１８日
２０１２年１２月６日（原寄託によりブタペスト条約に基づく寄託への移管日）
ハ イの寄託機関が寄託について付した受託番号（受領番号）
ＮＩＴＥ ＡＢＰ－１１２９
（８）Lactobacillus paracasei ＳＢＴ２１０５
イ 当該生物材料を寄託した寄託機関の名称及び住所
上記（７）に同じ
ロ イの寄託機関に生物材料を寄託した日付
２０１１年８月１８日
ハ イの寄託機関が寄託について付した受託番号
ＮＩＴＥ Ｐ－１１３０
（９）Lactobacillus paracasei ＳＢＴ２２０３
イ 当該生物材料を寄託した寄託機関の名称及び住所
上記（７）に同じ
ロ イの寄託機関に生物材料を寄託した日付
２０１１年８月１８日
２０１２年１２月６日（原寄託によりブタペスト条約に基づく寄託への移管日）
ハ イの寄託機関が寄託について付した受託番号（受領番号）
ＮＩＴＥ ＡＢＰ－１１３１
（１０）Lactobacillus paracasei ＳＢＴ２２１５
イ 当該生物材料を寄託した寄託機関の名称及び住所
上記（７）に同じ
ロ イの寄託機関に生物材料を寄託した日付
２０１１年８月１８日
２０１２年１２月６日（原寄託によりブタペスト条約に基づく寄託への移管日）
ハ イの寄託機関が寄託について付した受託番号(受領番号)
ＮＩＴＥ ＡＢＰ－１１３２
（１１）Lactobacillus paracasei ＳＢＴ１１４０８
イ 当該生物材料を寄託した寄託機関の名称及び住所
上記（７）に同じ
ロ イの寄託機関に生物材料を寄託した日付
２０１１年８月１８日
２０１２年１２月６日（原寄託によりブタペスト条約に基づく寄託への移管日）
ハ イの寄託機関が寄託について付した受託番号(受領番号)
ＮＩＴＥ ＡＢＰ－１１３３
（１２）Lactobacillus plantarum ＳＢＴ１５３４
イ 当該生物材料を寄託した寄託機関の名称及び住所
上記（４）に同じ
ロ イの寄託機関に生物材料を寄託した日付
平成３年７月１０日
２０１２年１２月６日（原寄託によりブタペスト条約に基づく寄託への移管日。国内寄託番号ＦＥＲＭ Ｐ－１２３５１より移管）
ハ イの寄託機関が寄託について付した受託番号(受領番号)
ＦＥＲＭ ＡＢＰ－１１５１８
（１３）Lactobacillus casei subsp. pseudoplantarum ＳＢＴ０６２４
イ 当該生物材料を寄託した寄託機関の名称及び住所
上記（４）に同じ
ロ イの寄託機関に生物材料を寄託した日付
平成２年１２月２０日
ハ イの寄託機関が寄託について付した受託番号
ＦＥＲＭ Ｐ－１１９２０
（１４）Lactobacillus buchneri ＳＢＴ２０２８
イ 当該生物材料を寄託した寄託機関の名称及び住所
上記（４）に同じ
ロ イの寄託機関に生物材料を寄託した日付
平成２年１２月２０日
ハ イの寄託機関が寄託について付した受託番号
ＦＥＲＭ Ｐ－１１９２１ [Reference to deposited biological materials]
(1) Lactobacillus gasseri SBT2055
Name and address of the depository institution that deposited the biological material National Institute of Advanced Industrial Science and Technology Patent Biological Deposit Center 1-Chuo 1-chome, Tsukuba City, Ibaraki Prefecture, Japan 6 (zip code 305-8566)
Date of deposit of biological materials at Loi depository organization March 27, 1996 (original deposit date)
February 26, 2008 (Date of transfer to deposit under the Budapest Treaty by original deposit)
Deposit number FERM BP-10953 assigned by the depository in Thailand for deposit
(2) Lactobacillus gasseri SBT2056
(A) Name and address of the depositary that deposited the biological material Date of deposit of the biological material at the same depository in Loi above (1) April 22, 1986 (original deposit date)
October 9, 2008 (Date of transfer to deposit under the Budapest Treaty by original deposit)
Deposit number FERM BP-11038 assigned by the depository in Hai for the deposit
(3) Lactobacillus gasseri SBT0274
The name and address of the depository that deposited the biological material The date on which the biological material was deposited at the same depository in (1) above March 15, 2000 (original deposit date)
October 9, 2008 (Date of transfer to deposit under the Budapest Treaty by original deposit)
Deposit number FERM BP-11039 assigned by the depository in Thailand for deposit
(4) Lactobacillus gasseri SBT1703
The name and address of the depository institution where the biological material was deposited 1-3, Higashi 1-chome, Tsukuba City, Ibaraki, Japan, Institute of Biotechnology, National Institute of Advanced Industrial Science and Technology (zip code 305-8566)
Date of deposit of biological material at the depository in Loi March 15, 2000 Deposit number FERM P-17785 attached to the depository by the depository in Hai
(5) Lactobacillus gasseri SBT10239
(A) Name and address of the depositary that deposited the biological material Date of deposit of the biological material at the same depository in the same (4) above February 17, 1998 The deposit number assigned by the depositary of hi to the deposit FERM P-16639
(6) Lactobacillus gasseri SBT10241
(A) Name and address of the depositary that deposited the biological material Date of deposit of the biological material at the same depository in (4) above March 15, 2000 The deposit number assigned by the depositary of hi FERM P-17786
(7) Lactobacillus paracasei SBT0327
(I) Name and address of the depository that deposited the biomaterial The National Institute of Technology and Evaluation, Patent Microorganism Deposit Center 2-5-8 Kazusa Kamashiri, Kisarazu City, Chiba Prefecture, Japan (zip code 292-0818)
Date of deposit of biological materials at Loi depository August 18, 2011 December 6, 2012 (Date of transfer to original deposit under the Budapest Treaty)
The deposit number (receipt number) assigned by the depositary in Japan for the deposit
NITE ABP-1129
(8) Lactobacillus paracasei SBT2105
(A) Name and address of the depository that deposited the biological material Date of deposit of the biological material at the same depository in the above (7) August 18, 2011 Deposit number assigned by the depositary institution for high NITE P-1130
(9) Lactobacillus paracasei SBT2203
(A) Name and address of the depositary that deposited the biological material Date of depositing the biological material at the same depository in (7) above August 18, 2011 December 6, 2012 (according to the Budapest Treaty by the original deposit) Transfer date to deposit based on)
The deposit number (receipt number) assigned by the depositary in Japan for the deposit
NITE ABP-1131
(10) Lactobacillus paracasei SBT2215
(A) Name and address of the depositary that deposited the biological material Date of depositing the biological material at the same depository in (7) above August 18, 2011 December 6, 2012 (according to the Budapest Treaty by the original deposit) Transfer date to deposit based on)
The deposit number (receipt number) assigned by the depositary in Japan for the deposit
NITE ABP-1132
(11) Lactobacillus paracasei SBT11408
(A) Name and address of the depositary that deposited the biological material Date of depositing the biological material at the same depository in (7) above August 18, 2011 December 6, 2012 (according to the Budapest Treaty by the original deposit) Transfer date to deposit based on)
The deposit number (receipt number) assigned by the depositary in Japan for the deposit
NITE ABP-1133
(12) Lactobacillus plantarum SBT1534
(A) Name and address of the depositary that deposited the biological material Date of deposit of the biological material at the same depository in (4) above July 10, 1991 December 6, 2012 (the Budapest Treaty by the original deposit) Date of transfer to deposit based on US (transferred from domestic deposit number FERM P-12351)
The deposit number (receipt number) assigned by the depositary in Japan for the deposit
FERM ABP-11518
(13) Lactobacillus casei subsp. Pseudoplantarum SBT0624
(A) Name and address of the depositary that deposited the biological material Date of deposit of the biological material at the same depository in (4) above The deposit number assigned by the depositary institution on December 20, 1990 FERM P-11920
(14) Lactobacillus buchneri SBT2028
(A) Name and address of the depositary that deposited the biological material Date of deposit of the biological material at the same depository in (4) above The deposit number assigned by the depositary institution on December 20, 1990 FERM P-11921
Claims (6)
- ラクトバチルス・ガセリ（Lactobacillus gasseri）を含む飲食品の製造方法であって、
飲食品のｐＨが３．５～４．５、糖濃度が５％～１５％であり、
ラクトバチルス・ガセリ（Lactobacillus gasseri）と特定の乳酸菌を混合する工程を含む前記製造方法。
特定の乳酸菌；３％（ｗ／ｗ）還元脱脂乳および１０％（ｗ／ｗ）グルコースを含むｐＨ４．０の培地に添加して１０℃において２１日間静置培養したときに該培地のｐＨを０．１以上低下させることができる乳酸菌。 A method for producing a food or drink containing Lactobacillus gasseri,
The pH of the food or drink is 3.5 to 4.5, and the sugar concentration is 5% to 15%.
The said manufacturing method including the process of mixing Lactobacillus gasseri (Lactobacillus gasseri) and specific lactic acid bacteria.
Specific lactic acid bacteria: when added to a pH 4.0 medium containing 3% (w / w) reduced skim milk and 10% (w / w) glucose and left to stand at 10 ° C. for 21 days, the pH of the medium was adjusted. Lactic acid bacteria that can be reduced by 0.1 or more. - 前記特定の乳酸菌が、ラクトバチルス・パラカゼイ(Lactobacillus paracasei)、ラクトバチルス・プランタラム（Lactobacillus plantarum）またはラクトバチルス・ブフネリ（Lactobacillus buchneri）である請求項１に記載の飲食品の製造方法。 The method for producing a food or drink according to claim 1, wherein the specific lactic acid bacterium is Lactobacillus paracasei, Lactobacillus plantarum, or Lactobacillus buchneri.
- 前記ラクトバチルス・パラカゼイがＳＢＴ０３２７（ＮＩＴＥ ＡＢＰ－１１２９）、ＳＢＴ２１０５（ＮＩＴＥ Ｐ－１１３０）、ＳＢＴ２２０３（ＮＩＴＥ ＡＢＰ－１１３１）、ＳＢＴ２２１５（ＮＩＴＥ ＡＢＰ－１１３２）、ＳＢＴ１１４０８（ＮＩＴＥ ＡＢＰ－１１３３）、ＡＴＣＣ２５５９８のいずれかであり、前記ラクトバチルス・プランタラムがＳＢＴ１５３４（ＦＥＲＭ ＡＢＰ－１１５１８）、ＳＢＴ０６２４（ＦＥＲＭ Ｐ－１１９２０）、ＡＴＣＣ４３１９９、ＡＴＣＣ８０１４のいずれかであり、前記ラクトバチルス・ブフネリがＳＢＴ２０２８（ＦＥＲＭ Ｐ－１１９２１）またはＪＣＭ１１１５である請求項１または２に記載の飲食品の製造方法。 The Lactobacillus paracasei is SBT0327 (NITE ABP-1129), SBT2105 (NITE P-1130), SBT2203 (NITE ABP-1131), SBT2215 (NITE ABP-1132), SBT11408 (NITE ABP-1253) The Lactobacillus plantarum is SBT1534 (FERM ABP-11518), SBT0624 (FERM P-11920), ATCC 43199, or ATCC8014, and the Lactobacillus buchneri is SBT2028 (FERM P-111921) or JCM1115 The method for producing a food or drink according to claim 1 or 2.
- 前記飲食品が、発酵乳および乳酸菌飲料である請求項１～３いずれかに記載の飲食品の製造方法。 The method for producing a food or drink according to any one of claims 1 to 3, wherein the food or drink is fermented milk and a lactic acid bacteria beverage.
- ラクトバチルス・ガセリ（Lactobacillus gasseri）および特定の乳酸菌を含む、ｐＨが３．５～４．５、糖濃度が５％～１５％の飲食品；
特定の乳酸菌；１０℃において３％（ｗ／ｗ）還元脱脂乳および１０％（ｗ／ｗ）グルコース を含むｐＨ４．０の培地に添加して２１日間静置培養したときに当該培地のｐＨを０．１以上低下させることができる乳酸菌。 A food or drink containing Lactobacillus gasseri and a specific lactic acid bacterium having a pH of 3.5 to 4.5 and a sugar concentration of 5 to 15%;
Specific lactic acid bacteria: when added to a pH 4.0 medium containing 3% (w / w) reduced skim milk and 10% (w / w) glucose at 10 ° C. Lactic acid bacteria that can be reduced by 0.1 or more. - ２１日間保存後のラクトバチルス・ガセリの生菌数が１×１０６ＣＦＵ／ｇ以上である請求項５に記載の飲食品。
The food / beverage product according to claim 5, wherein the viable count of Lactobacillus gasseri after storage for 21 days is 1 × 10 6 CFU / g or more.
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| JP2013548301A JP6084576B6 (en) | 2011-12-06 | 2012-12-06 | Food / beverage products with high survival of gasseri bacteria and method for producing the same |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH10229819A (en) * | 1997-02-17 | 1998-09-02 | Yakult Honsha Co Ltd | Bifidobacteria-fermented milk and its producing method |
| JP2008005811A (en) * | 2006-06-30 | 2008-01-17 | Snow Brand Milk Prod Co Ltd | Lactobacillus growth promoter and survivability improver |
| JP2008199905A (en) * | 2007-02-16 | 2008-09-04 | Snow Brand Milk Prod Co Ltd | Improving agent for survivability of lactic acid bacterium |
| JP5076029B2 (en) * | 2010-07-30 | 2012-11-21 | 株式会社明治 | Lactic acid bacteria with metabolic syndrome improvement effect |
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2012
- 2012-12-06 WO PCT/JP2012/081702 patent/WO2013085009A1/en active Application Filing
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH10229819A (en) * | 1997-02-17 | 1998-09-02 | Yakult Honsha Co Ltd | Bifidobacteria-fermented milk and its producing method |
| JP2008005811A (en) * | 2006-06-30 | 2008-01-17 | Snow Brand Milk Prod Co Ltd | Lactobacillus growth promoter and survivability improver |
| JP2008199905A (en) * | 2007-02-16 | 2008-09-04 | Snow Brand Milk Prod Co Ltd | Improving agent for survivability of lactic acid bacterium |
| JP5076029B2 (en) * | 2010-07-30 | 2012-11-21 | 株式会社明治 | Lactic acid bacteria with metabolic syndrome improvement effect |
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| JP6084576B2 (en) | 2017-02-22 |
| JPWO2013085009A1 (en) | 2015-04-27 |
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