WO2014163123A1 - 豆乳発酵物 - Google Patents
豆乳発酵物 Download PDFInfo
- Publication number
- WO2014163123A1 WO2014163123A1 PCT/JP2014/059786 JP2014059786W WO2014163123A1 WO 2014163123 A1 WO2014163123 A1 WO 2014163123A1 JP 2014059786 W JP2014059786 W JP 2014059786W WO 2014163123 A1 WO2014163123 A1 WO 2014163123A1
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- WIPO (PCT)
- Prior art keywords
- lactic acid
- strain
- soymilk
- fermented
- bulgaricus
- Prior art date
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- 235000013322 soy milk Nutrition 0.000 title claims abstract description 194
- 239000000126 substance Substances 0.000 title abstract 6
- 241000194020 Streptococcus thermophilus Species 0.000 claims abstract description 103
- 229930182843 D-Lactic acid Natural products 0.000 claims abstract description 65
- JVTAAEKCZFNVCJ-UWTATZPHSA-N D-lactic acid Chemical compound C[C@@H](O)C(O)=O JVTAAEKCZFNVCJ-UWTATZPHSA-N 0.000 claims abstract description 65
- 229940022769 d- lactic acid Drugs 0.000 claims abstract description 64
- 239000007858 starting material Substances 0.000 claims abstract description 63
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 154
- 241000186660 Lactobacillus Species 0.000 claims description 104
- 229940039696 lactobacillus Drugs 0.000 claims description 104
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Images
Classifications
-
- 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
- A23C9/127—Fermented milk preparations; Treatment using microorganisms or enzymes using microorganisms of the genus lactobacteriaceae and other microorganisms or enzymes, e.g. kefir, koumiss
-
- 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
- A23C11/00—Milk substitutes, e.g. coffee whitener compositions
- A23C11/02—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins
- A23C11/10—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins
- A23C11/103—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins containing only proteins from pulses, oilseeds or nuts, e.g. nut milk
- A23C11/106—Addition of, or treatment with, microorganisms
-
- 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
- A23C9/13—Fermented milk preparations; Treatment using microorganisms or enzymes using additives
-
- 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
- A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
-
- 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
- A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
- A23L11/60—Drinks from legumes, e.g. lupine drinks
- A23L11/65—Soy drinks
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
- C12P7/56—Lactic acid
-
- 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
- A23V2400/137—Delbrueckii
-
- 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
- A23V2400/21—Streptococcus, lactococcus
- A23V2400/249—Thermophilus
Definitions
- the present invention relates to a soymilk fermented product obtained by fermenting soymilk with lactic acid bacteria.
- Soybean also called field meat
- soymilk is a milky beverage obtained by elution of protein and other ingredients from the soybean with hot water, etc., and removing fiber. It has been attracting attention as a low cholesterol, high protein health material.
- fermenting soy milk with lactic acid bacteria which are also attracting attention as health materials, is also expected to have a healthy function due to the synergistic effect of soybeans and lactic acid bacteria.
- the unpleasant odor peculiar to soybeans, the soy odor, the fermentation odor and pickle odor due to lactic acid bacteria are problems, and it is difficult to penetrate the market.
- a method for improving flavor by fermentation with two types of lactic acid bacteria, Lactobacillus plantarum and Lactococcus lactis see Patent Document 2
- Lactobacillus acidophilus or Lactobacillus casei
- Bulgaricus see Patent Document 3
- Lactobacillus delbruecki subsp see Patent Document 3
- Streptococcus thermophilus and lac including a method of fermenting soy milk with one strain selected from Bulgaricus and Lactobacillus acidophilus and three lactic acid bacteria of Lactobacillus casei and Streptococcus thermophilus (see Patent Document 4)
- Various varieties that promote lactic acid fermentation and improve flavor in soy milk by adding different bacterial species to Bacillus delbruecki, subspice sheath bulgaricus, or replacing these bacterial species with other bacterial species There is a report.
- a method of fermenting soymilk by adding sugar, lactic acid bacteria and protease simultaneously to soy milk has also been reported (see Patent Document 5).
- JP-A-63-7743 JP-A-5-184320 Japanese Patent Laid-Open No. 10-201415 JP 2011-167190 A JP-A-7-147898
- lactic acid bacteria makes the production and management of each strain extremely complicated, making it difficult to maintain the quality of production lots, preparing food ingredients other than soybean-derived foods, and inoculating lactic acid bacteria.
- processing complexity is added, such as adding an enzyme.
- the problem to be solved by the present invention is that only when Streptococcus thermophilus and Lactobacillus delbrukis subspice bulgaricus (Lactobacillus delbrueckis subbulis. Even if it is used as a lactic acid bacteria mixed starter, soy milk, which is a raw material, is not subjected to processing such as enzyme treatment or extraction treatment, and without adding raw materials such as sugar sources, it is equivalent to yogurt fermented with milk.
- the object is to provide a fermented soymilk product having a flavor and smooth physical properties.
- Streptococcus thermophilus which is a lactic acid bacterium for yogurt defined by CODEX
- Lactobacillus delbruecki subspice bulgaricus In a lactic acid bacteria mixed starter, when combined with Streptococcus thermophilus capable of accumulating 0.4 g / L or more of fructose in soy milk, D-lactic acid of 0.4 g / L or more can be produced when combined with the Streptococcus thermophilus.
- the present invention when the present invention is (1) inoculated and cultured in soy milk, inoculated into Streptococcus thermophilus that can accumulate 0.4 g / L or more fructose in the fermented product and soy milk,
- a fermented soymilk product obtained by using a mixed starter of lactic acid bacteria containing Lactobacillus delbruecki subspice bulgaricus capable of accumulating 0.4 g / L or more of D-lactic acid in the fermented product. It is.
- the present invention also includes (2) Streptococcus thermophilus capable of accumulating 0.4 g / L or more of fructose in the fermented product when inoculated and cultured in soy milk, and inoculated and cultured in soy milk.
- the present invention provides a mixed starter for lactic acid bacteria comprising Lactobacillus delbrucky subspice bulgaricus capable of accumulating 0.4 g / L or more of D-lactic acid in a fermented product.
- soymilk is used as a raw material, and even if only Streptococcus thermophilus and Lactobacillus delbruecki subspice bulgaricus are used as a lactic acid bacteria mixed starter, the soymilk as a raw material is subjected to enzyme treatment, extraction treatment, etc. It is possible to provide a fermented soymilk product that is as excellent in flavor and physical properties as yoghurt that is obtained by subjecting milk to lactic acid fermentation without processing and without adding a raw material such as a sugar source. Since the fermented yogurt-like soymilk of the present invention does not give an unpleasant odor peculiar to fermented soybeans, it can be expected to spread as a health material having an excellent function due to the synergistic effect of soybeans and lactic acid bacteria.
- FIG. 3 is a view showing the amount of D-lactic acid in the obtained fermented soymilk.
- FIG. 3 is a graph showing the amount of D-lactic acid in fermented soymilk.
- a soymilk membrane permeate obtained by adding 1.0% (w / v) fructose to a solution obtained by permeating unadjusted soymilk through an MF ultrafiltration membrane was obtained at 42 ° C.
- FIG. 3 is a diagram showing the amount of D-lactic acid produced in a fermentation broth obtained by fermentation with S. bulgaricus alone at 42 ° C. for 24 hours.
- FIG. 3 is a graph showing the amount of D-lactic acid produced in a fermented soymilk obtained by fermentation at 42 ° C. for 24 hours.
- the fermented soymilk of the present invention can be obtained by fermenting soymilk with only two types of lactic acid bacteria, Streptococcus thermophilus and Lactobacillus delbrucky subspice bulgaricus, without adding a special processing such as enzyme treatment.
- soy milk used in the present invention any soy milk obtained by a conventional method may be used, and commercially available soy milk may be used.
- a liquid obtained by steaming and grinding beans that have been swollen by immersing exfoliated soybeans or defatted soybeans in water and using a liquid obtained by removing okara from the liquids And more preferable from the viewpoint of texture.
- the JAS standard defines soybean solid content of 8.0% or more as soy milk, prepared soybean milk is defined as 6.0% or more, and soy milk beverage is defined as 4.0% or more. It is not particularly limited.
- the Streptococcus thermophilus used in the present invention can be grown in soy milk, and when grown alone in soy milk, it decomposes sucrose in soy milk to produce glucose and fructose, of which fructose is 0.4 g / L. It is required to have the property of accumulating at the above concentration.
- Lactobacillus delbruecki subspice bulgaricus cannot assimilate sucrose in soy milk as a sugar source, but Streptococcus thermophilus assimilate them by breaking down sucrose into glucose and fructose Lactobacillus delbruecki subsp. Bulgaricus can also grow.
- Streptococcus thermophilus takes in its own degradation product as a sugar source while decomposing sucrose, but if a strain with a particularly high degradability is selected, the decomposed sugar accumulates in the fermented product. And in the fermented soymilk in which the amount of accumulated fructose exceeds 0.4 g / L, Lactobacillus delbruecki subsp. Bulgaricus grows well and produces various metabolites during the growth process. Can play a big role in shaping the flavor of yogurt.
- the metabolism of Lactobacillus delbrucky subspice bulgaricus is important for fermenting soymilk like yogurt, but D-lactic acid produced by the metabolism is produced in Streptococcus thermophilus that produces only L-lactic acid. Therefore, it can be used as an easily measurable index indicating the metabolic activity of Lactobacillus delbruecki, subspice bulgaricus.
- the fermented product in which the amount of D-lactic acid accumulates 0.4 g / L or more in the fermented product has a particularly good flavor as a result of active production of metabolites by Lactobacillus delbruecki, subspice bulgaricus. I found out.
- Lactic acid bacteria having the above properties can be selected from lactic acid bacteria used in the production of milk yogurt and similar lactic acid bacteria in the environment.
- all generally widely used means can be used.
- 0.4 g / L or more of fructose is accumulated in soy milk by inducing mutation using ultraviolet light, inducing mutation using a drug, or subculture in a medium containing soy milk as a main component. It is possible to obtain Streptococcus thermophilus and Lactobacillus delbruecki subspice bulgaricus which show a high production amount of D-lactic acid of 0.4 g / L or more in soymilk when cultured at the same time.
- Lactic acid bacteria that produce D-lactic acid are Lactobacillus delbruecki subspice bulgaricus, but cannot assimilate sucrose, which is the main component of sugar in soy milk, and cannot grow alone, It can be killed.
- many Streptococcus thermophilas can assimilate sucrose and grow to some extent in soy milk.
- the lactic acid bacteria decompose sucrose to produce glucose and fructose, which results in Lactobacillus delbrucky subspice bulgaricus. The inventors have found that, when they coexist, they can be utilized and gradually grown.
- Lactobacillus delbruecki, subspice bulgaricus In order to grow Lactobacillus delbruecki, subspice bulgaricus, it may be possible to supplement soymilk with sugar, but supplementation with sugar alone may not produce a sufficient amount of D-lactic acid. Because of the compatibility of the two strains other than the sugar complementation relationship, Lactobacillus is an active metabolite that can produce 0.4 g / L or more of D-lactic acid when cultured with Streptococcus thermophilus that accumulates fructose. It is important to select Derbrucky Subspices bulgaricus.
- the lactic acid bacteria used in the present invention can be obtained by selection or breeding as described above.
- a combination of Streptococcus thermophilus K1580 strain and Lactobacillus delbrukee subspice bulgaricus K1581 strain A combination of Streptococcus thermophilus K1584 and Lactobacillus delbruecki subsp.
- Bulgaricus K1585, Streptococcus thermophilus Danisco strain isolated from mixed starter YO-MIX499) and Lactobacillus delbrukee subspice bulgaricus A combination of Danisco Co.
- strains strains isolated from mixed starter YO-MIX505, 511, 863, and the like are applicable. Since the fermented soymilk of the present invention can be obtained by using a lactic acid bacterium having the above properties, it is not limited to the strains described in the Examples.
- soymilk with lactic acid bacteria it can be carried out based on a generally known method for producing yogurt or lactic acid bacteria beverages, such as Streptococcus thermophilus 1 ⁇ 10 4 to 1 ⁇ 10 8 cfu / mL, More preferably, 1 ⁇ 10 5 to 1 ⁇ 10 7 cfu / mL, and Lactobacillus delbruecki subspice bulgaricus 1 ⁇ 10 2 to 1 ⁇ 10 7 cfu / mL, more preferably 1 ⁇ 10 3 to
- the soy milk is inoculated to 1 ⁇ 10 5 cfu / mL and fermented at a fermentation temperature of 30 to 45 ° C., more preferably 37 to 42 ° C. within a range of 4 to 24 hours. Is not to be done.
- the fermented yoghurt-like soymilk of the present invention thus obtained is characterized by containing 0.4 g / L or more of D-lactic acid, and metabolism of Streptococcus thermophilus and Lactobacillus delbruecki subspice bulgaricus Both of them are flourishing, showing the amount of lactic acid that could not be obtained by using commercially available lactic acid bacteria mixed starters as they are, and having yogurt-like flavor and physical properties having aromatic components derived from each bacterial species It becomes fermented soymilk. Moreover, since it is possible to ferment without adding raw materials other than soybean origin, it is not influenced by the taste derived from an auxiliary raw material. Furthermore, the fermented soymilk of the present invention has high versatility, and can be applied not only to yogurt-like foods but also to any products related to soybean-related foods and drinks such as seasonings, cheese, and beverage forms.
- Lactic acid bacteria mixed starter As a lactic acid bacteria mixed starter, commercially available lactic acid bacteria mixed starter (manufactured by Danisco, Streptococcus thermophilus strain and Lactobacillus strain) A freeze-dried product containing both the Delbrucky, Subspice sheath, and Bulgaricus strains (hereinafter sometimes referred to as “commercially mixed starter”) was used. Various commercially available mixed starters exist depending on the properties of the Streptococcus thermophilus strain and Lactobacillus delbruecki subspice sheath bulgaricus strain.
- YO-MIX300, YO-MIX305, YO-MIX496, YO-MIX499, YO-MIX505, YO-MIX511, YO-MIX863, YO-MIX883, YO-MIX885, YO-MIX401, YO-MIX421, YO -MIX495 and YO-MIX601 both are product numbers.
- Streptococcus thermophilus Only commercially available Streptococcus thermophilus strains from commercially available starters (both including two strains of Streptococcus thermophilus and Lactobacillus delbruxii subspice bulgaricus) Isolated.
- the isolated strains are Streptococcus thermophilus (St.) 45 strain, St. 40 strains, St. 505, St. 511, St. 863, St. 883 strain, St. 885, St. 300 shares, St. 305 strain, St. 495, St. 496 strain, St. 499 strain, St. 187, St. 205, St. 207 strain, St. 208, St. 211, St. 401, St. 421, St. 492 strain, St. 601 shares.
- inoculum of 1% of the inoculum of the isolate is inoculated into soymilk and cultured at 37 to 45 ° C. for 12 to 24 hours.
- 0.4 g / L or more of fructose is added.
- Streptococcus thermophilus strains that can be accumulated were selected and designated as selected strains. Specifically, Streptococcus thermophilus (St.) 40 strain, St. 511 strain, St863 strain, St. 885, St. 496 strain, St. 499 strain, St. 187, St. 205, St. 401, St. 421, St. 492 strain, St. 601 strains were selected.
- Streptococcus thermophilus St. Co., Ltd. owned by the NIZO food research company based in the Netherlands. 2333 strain and Lactobacillus delbruecki subspice bulgaricus (Lactobacillus delbrueckii subsp. Bulgaricus) Lb. 185 strains were inoculated into unadjusted soymilk (manufactured by Kikkoman Soy Foods) and subcultured. The soymilk fermented product was repeatedly inoculated into soymilk at a ratio of 1% (v / v) and cultured at 37-45 ° C. for 12-24 hours. Subculturing was carried out for about 700 generations, and the Streptococcus thermophilus strain isolated from the obtained soymilk fermented product was designated as K1580 strain.
- Streptococcus thermophilus St. owned by NIZO food research, Inc. 131 strain and Lactobacillus delbruecki subspice bulgaricus (bacteria Lactobacillus delbrueckii subsp. Bulgaricus) Lb. 194 strains were inoculated into unadjusted soymilk (manufactured by Kikkoman Soy Foods) and subcultured. The soymilk fermented product was repeatedly inoculated into soymilk at a ratio of 1% (v / v) and cultured at 37-45 ° C. for 12-24 hours. The Streptococcus thermophilus strain isolated from the obtained fermented soymilk was subcultured for about 700 generations, and was designated as K1584 strain.
- Lactobacillus delbruecki subspicis bulgaricus Only the Lactobacillus delbrucky subspices bulgaricus strain was isolated from the commercial starter. The isolated strain is based on Lactobacillus delbruecki subsp. Bulgaricus (Lb.) 300 strain, Lb. 305 strain, Lb. 401 strain, Lb. Lb. 421, Lb. 492 strain, Lb. 495 strain, Lb. 496 strain, Lb. 499 strain, Lb. 505 strain, Lb. 511, Lb. 601 strain, Lb. 863, Lb. 883 strain, Lb. There were 885 strains.
- Streptococcus thermophilus St. owned by NIZO food research. 2333 strain and Lactobacillus bulgaricus Lb. 185 was inoculated into unadjusted soy milk (manufactured by Kikkoman Soy Foods) and subcultured. The soymilk fermented product was repeatedly inoculated into soymilk at a ratio of 1% (v / v) and cultured at 37-45 ° C. for 12-24 hours. Subculturing was carried out for about 700 generations, and the Lactobacillus delbruecki subspice sheath bulgaricus strain isolated from the fermented soymilk was designated as K1581 strain.
- Streptococcus thermophilus St. which is also owned by NIZO food research.
- 131 and Lactobacillus delbruecki subspice bulgaricus (Lactobacillus delbrueckii subsp. Bulgaricus) Lb. 194 was inoculated into unadjusted soymilk (Kikkoman Soy Foods) and subcultured.
- the soymilk fermented product was repeatedly inoculated into soymilk at a ratio of 1% (v / v) and cultured at 37-45 ° C. for 12-24 hours.
- Lactobacillus delbruecki subspice sheath bulgaricus strain isolated from the obtained soymilk fermented product was designated as K1585 strain.
- St. 131 and Lb The Lactobacillus delbruecki subspices bulgaricus strain obtained from 185 subcultures was designated as K1583 strain.
- component-unadjusted soy milk manufactured by Kikkoman Soy Foods
- a composition comprising a lactic acid bacteria mixed starter composed of Streptococcus thermophilus strain and Lactobacillus delbrucky subspice sheath bulgaricus strain was added and fermented at 42 ° C. for 24 hours. After the fermentation was completed, the amount of D-lactic acid in the obtained fermented soymilk was measured by the enzyme electrode method (Oji Scientific Co., Ltd., Biosensor BF-5).
- FIG. 1 shows fermentation of soy milk obtained by fermentation of various Streptococcus thermophilus strains and a breeding strain, Lactobacillus delbruecki subspice sheath bulgaricus K1581 strain, and a lactic acid bacteria mixed starter. It is a figure which shows the result of having measured the amount of D-lactic acid in a thing.
- FIG. 2 shows that, instead of Lactobacillus delbruecki subspice sheath bulgaricus K1581 strain, Lactobacillus delbruecki subspice sheath bulgaricus K1585 strain, which is also a breeding strain, is used as a lactic acid bacteria mixed starter.
- FIG. 2 is a diagram showing the results of measuring the amount of D-lactic acid in the soymilk fermented product obtained by fermenting soymilk.
- FIG. 3 shows a Streptococcus thermophilus K1580 strain, which is a breeding strain, and each Lactobacillus delbruecki subspice sheath bulgaricus strain isolated from a commercially available starter.
- FIG. 3 is a graph showing the results of measuring the amount of D-lactic acid in a fermented soymilk obtained by fermentation under the same conditions.
- both the Streptococcus thermophilus strain and the Lactobacillus delbruecki subspice sheath bulgaricus strain were obtained by fermentation using a commercially available lactic acid bacteria mixed starter as it is.
- the fermented product not much D-lactic acid is produced, whereas by substituting both the Streptococcus thermophilus strain and the Lactobacillus delbrucky subspice sheath bulgaricus strain with selected or bred strains, 0.
- a soymilk fermented product containing 4 g / L or more of D-lactic acid can be obtained.
- lactic acid bacteria mixed starter composed of lactic acid bacteria bred in the present invention (Streptococcus thermophilus K1580 strain and Lactobacillus delbruecki subspice sheath bulgaricus strain K1581 strain)
- further D-lactic acid is used.
- a large amount of fermented soymilk can be obtained (see FIG. 1).
- Lactobacillus delbruecki subspice sheath bulgaricus strain selected and bred and difference in metabolic ability in soy milk membrane permeation of Lactobacillus delbruecki subspice sheath bulgaricus strain isolated from commercial starter
- soy milk membrane permeate is obtained by removing insoluble proteins and lipids from macromolecules from soy milk, and it is considered that there are no significant differences in free amino acids, soluble peptides, sugars, minerals, etc. in soy milk.
- the amount of D-lactic acid produced by the Lactobacillus delbruecki subspice sheath bulgaricus strain bred in the present invention was significantly higher than that of the commercially available starter and ATCC 11842 strain.
- the acidic soymilk membrane permeate a marked difference was observed in the amount of D-lactic acid produced between the commercial starter and the breeding strain compared to the unadjusted soymilk membrane permeate.
- the Lactobacillus delbruecki subspice sheath bulgaricus strain isolated from a commercially available starter produces a low amount of D-lactic acid, especially under low pH conditions (pH 5.0), whereas Thus, it was confirmed that about 3 times or more of D-lactic acid was produced.
- pH is lowered by the growth of the thermophilus strain first, and then the Lactobacillus delbrukee subspice sheath bulgaricus strain grows.
- Some of the isolates derived from commercial starters are Lb. 505 and Lb. 3 and the like, strains that produced 0.4 g / L or more of D-lactic acid as a selected strain in FIG. 3 are also included, and in FIGS. 5 and 6, D-lactic acid is higher than strains derived from other commercially available starters. Although production is shown, the production amount of 0.4 g / L has not been reached. This is considered to be due to the absence of Streptococcus thermophilus in the tests shown in FIGS.
- Fructose is supplemented to the unadjusted soymilk membrane permeate, but in order to increase the production of D-lactic acid by activating the metabolism of Lactobacillus delbruecki, subspice sheath bulgaricus, simply supply fructose
- Streptococcus thermophilus is considered to have other roles.
- soymilk fermentation undiluted solution commercially available unadjusted soymilk (manufactured by Kikkoman Soy Foods) was used.
- This soymilk fermentation stock solution was inoculated with 1.0% (v / v) of the suspension prepared above and fermented at 42 ° C. for 24 hours. After fermentation, the centrifugal supernatant was collected, and the amount of residual sugar in fructose was measured using F kit D-glucose / fructose (manufactured by JK International). The results are shown in FIG.
- Streptococcus thermophilus strains include St. 496 strain, St. 499 strain, St. It was found that there are strains that accumulate 0.4 g / L or more of fructose in fermented soymilk and strains that do not accumulate, such as 885 strain.
- FIG. 8 shows the measurement results of the amount of D-lactic acid produced in the fermented soymilk.
- a Streptococcus thermophilus strain that accumulates fructose
- 0.4 g / L or more of D-lactic acid is produced, and even when no auxiliary material is added, Lactobacillus delbruecki subspecies Bulgari It was confirmed that a symbiotic relationship with Kas was also established in soy milk.
- Streptococcus thermophilus strain It is considered indispensable to supply a sugar source to the Lactobacillus delbruecki subspecies bulgaricus strain by degrading certain sucrose and accumulating fructose.
- the amount of residual sugars of sucrose and fructose when the Streptococcus thermophilus K1580 strain bred alone was fermented alone, or when the Streptococcus thermophilus K1580 strain and Lactobacillus delbruxii subspecies bulgaricus K1581 strain were co-fermented. was measured over time. The fermentation temperature was set at 42 ° C. The time course of sucrose is shown in FIG. 9, and the time course of fructose is shown in FIG.
- FIG. 11 is a graph showing the transition of fructose concentration over time when soy milk was co-fermented at 42 ° C. with Streptococcus thermophilus K1580 strain and Lactobacillus delbruecki subspecies bulgaricus K1581 strain.
- fructose is metabolized by the K1581 strain, and as a result, contributes to the production of D-lactic acid and other aromatic components.
- Streptococcus thermophilus is divided into a strain that actively assimilate fructose and a strain that does not so, and it is characterized in that it is accumulated in a fermented product without assimilating fructose like the selected strain and breeding strain in this example.
- the amount of D-lactic acid in the fermented product was prepared by appropriately mixing the fermented soymilk product and the co-fermented product that had been fermented separately. That is, 100% co-fermented product (sample 1), 40% co-fermented product (sample 2), 30% co-fermented product (sample 3), 20% co-fermented product (sample 4), 15% co-fermented product (sample 5) ) 10% co-fermented product (sample 6) and 100% single fermented product (sample 7) were prepared.
- Table 2 shows the estimated D-lactic acid and the estimated D / L ratio of each sample.
- Streptococcus thermophilus strains (St. 885 strain, St. 496 strain) derived from commercially available mixed starters YO-MIX885 and YO-MIX496 were isolated.
- Lactobacillus delbruxii subspecies bulgaricus strains (Lb.305 strain, Lb.492 strain, Lb.601 strain) derived from commercially available mixed starters YO-MIX305, YO-MIX492, YO-MIX601 were isolated, Each was stored as a glycerol stock bacterial solution and used for testing.
- Streptococcus thermophilus K1580 and K1584 strains, and Lactobacillus delbruxii subspecies Bulgaricus strains K1581 and K1585 were used as breeding strains.
- the produced Streptococcus thermophilus strain (St. strain) and Lactobacillus delbruecki subspecies bulgaricus strain (Lb. strain) were mixed at a ratio of 1: 1 (for example, K1580 strain + K1581 strain or St. 885 strain + Lb). .305 strain) was fermented with 10 mL of non-adjusted soymilk (manufactured by Kikkoman Soy Foods) to obtain a pre-fermented product. 1.0% (v / v) of this pre-fermented product was inoculated into unadjusted soymilk (manufactured by Kikkoman Soy Foods) and fermented at 42 ° C. for 24 hours to obtain a fermented soymilk product.
- the lactic acid analysis value of each soymilk fermented product is shown below.
- the soymilk fermented product produced using the lactic acid bacteria mixed starter according to the present invention (lactic acid bacteria mixed starter comprising a selected or bred strain) has a superior flavor. And found to have a taste.
- YO-MIX300, YO-MIX305 both are product numbers
- Fermented soymilk fermented product obtained by fermentation, and fermented soymilk fermented by selected or bred strains K1580 strain + K1581 strain, St.496 + Lb.601, St.885 + Lb.305) are uniformly stirred, and laser The particle size distribution was measured using a diffraction type particle size distribution measuring device (SALD-2200 (Shimadzu Corporation)).
- the measurement results are shown in FIG. In the graph, the integrated results are shown together with the appearance frequency of the particle diameter with the same symbol for each sample.
- the proportion of particles having a particle size of 100 micrometers or more is 10% or less, It was confirmed that it was a fermented soymilk with a smooth touch.
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Abstract
Description
本発明の豆乳発酵物は、豆乳に酵素処理など特別な加工処理を加えずに、ストレプトコッカス・サーモフィラスとラクトバチルス・デルブルッキー・サブスピーシス・ブルガリカスの2種類の乳酸菌のみで発酵することにより得られる。
ヨーグルト様の良好な豆乳発酵物を得るためには、D-乳酸だけでなく、ストレプトコッカス・サーモフィラスによって生成されるL-乳酸も含まれている必要がある。
(1)乳酸菌混合スターター
乳酸菌混合スターターとして、市販のヨーグルト製造用の乳酸菌混合スターター(ダニスコ社製、ストレプトコッカス・サーモフィラス株及びラクトバチルス・デルブルッキー・サブスピーシース・ブルガリカス株の両方を含む凍結乾燥品。以下、「市販混合スターター」ということがある)を使用した。この市販混合スターターは、ストレプトコッカス・サーモフィラス株及びラクトバチルス・デルブルッキー・サブスピーシース・ブルガリカス株の性質に応じて、種々のものが存在する。具体的には、YO-MIX300、YO-MIX305、YO-MIX496、YO-MIX499、YO-MIX505、YO-MIX511、YO-MIX863、YO-MIX883、YO-MIX885、YO-MIX401、YO-MIX421、YO-MIX495、YO-MIX601である(いずれも製品番号)。
豆乳発酵原液には、市販の無調整豆乳(キッコーマンソイフーズ社製)を用いた。一方、各乳酸菌混合スターターの凍結乾燥物を10mLのLM17培地(Terzaghi及びSandine,Appl.Microbiol.29,807~813,1975)で培養し、培養液を0.9%NaCl溶液で遠心・洗浄し、10mLの0.9%NaCl溶液に再懸濁した。この調製した懸濁液を上記豆乳発酵原液に1.0%(v/v)接種し、42℃、24時間発酵した。そして、24時間経過後に発酵を終了し、豆乳発酵物中のD-乳酸量を、酵素電極法(王子計測社製、バイオセンサーBF-5)により測定した。
結果を図4に示す。24間発酵させて得られた豆乳発酵物中のD-乳酸量を測定したところ、D-乳酸はほとんど生成されず、所望のヨーグルト様の風味を持つ豆乳発酵物が得られないことを確認した(図4)。
(1)ストレプトコッカス・サーモフィラス
市販スターター(いずれもストレプトコッカス・サーモフィラス及びラクトバチルス・デルブルッキー・サブスピーシス・ブルガリカスの2菌種を含む)から、ストレプトコッカス・サーモフィラス株のみを単離した。単離した株は、市販スターターの製品番号に基づき、ストレプトコッカス・サーモフィラス(Streptococcus thermophilus:St.)45株、St.40株、St.505株、St.511株、St.863株、St.883株、St.885株、St.300株、St.305株、St.495株、St.496株、St.499株、St.187株、St.205株、St.207株、St.208株、St.211株、St.401株、St.421株、St.492株、St.601株とした。
上記市販スターターから、ラクトバチルス・デルブルッキー・サブスピーシース・ブルガリカス株のみを単離した。単離した株は、市販スターターの製品番号に基づき、ラクトバチルス・デルブルッキー・サブスピーシース・ブルガリカス(Lactobacillus delbrueckii subsp.bulgaricus:Lb.)300株、Lb.305株、Lb.401株、Lb.Lb.421株、Lb.492株、Lb.495株、Lb.496株、Lb.499株、Lb.505株、Lb.511株、Lb.601株、Lb.863株、Lb.883株、Lb.885株とした。
(1)供試菌株
市販混合スターターから単離した乳酸菌及び選抜・育種した乳酸菌(上記2参照)を用いて豆乳を発酵させ、得られた豆乳発酵物中のD-乳酸量を測定・比較した。
発酵原料は、成分無調整豆乳(キッコーマンソイフーズ社製)を用いた。この発酵原料に、ストレプトコッカス・サーモフィラス株とラクトバチルス・デルブルッキー・サブスピーシース・ブルガリカス株からなる乳酸菌混合スターターを構成したものを添加し、42℃、24時間発酵した。発酵終了後、得られた豆乳発酵物中のD-乳酸量を酵素電極法(王子計測社製、バイオセンサーBF-5)で測定した。
図1は、各種ストレプトコッカス・サーモフィラス株と、育種株であるラクトバチルス・デルブルッキー・サブスピーシース・ブルガリカスK1581株とで乳酸菌混合スターターを構成し、発酵して得られた豆乳発酵物中のD-乳酸量を測定した結果を示す図である。
(1)上述のように、ストレプトコッカス・サーモフィラス株の特徴によってラクトバチルス・デルブルッキー・サブスピーシース・ブルガリカス株によるD-乳酸生成量が大きく変化したことが分かった。そこで、選抜・育種したラクトバチルス・デルブルッキー・サブスピーシース・ブルガリカス株と市販スターターから単離したラクトバチルス・デルブルッキー・サブスピーシース・ブルガリカス株の豆乳膜透過中での代謝能の違いを確認するため、予めフラクトースを補糖した豆乳膜透過液中でのD-乳酸生成量を比較した。豆乳膜透過液は豆乳から高分子の不溶性タンパク質や脂質が取り除かれたものであり、豆乳中の遊離アミノ酸、可溶性のペプチド、糖分やミネラルなど大きな違いはないものと考えられる。
市販の無調整豆乳(キッコーマンソイフーズ社製)を中空糸状の限外ろ過モジュール(型式:USP-143(旭化成社製))を用い、真空ポンプで吸引し(-60kPa)、膜を透過した液を成分無調整の豆乳膜透過液とした。この液に対し、終濃度1.0%(v/v)となるようにフラクトースを無菌的に添加した(無調整豆乳膜透過液)。更に、この豆乳膜透過液に塩酸を添加し、pH5.0に調整した酸性豆乳膜透過液を作製した。
市販のヨーグルト製造用粉末混合スターター(ダニスコ社製:ストレプトコッカス・サーモフィラス株およびラクトバチルス・ブルガリカス株を含む凍結乾燥品)から単離したラクトバチルス・デルブルッキー・サブスピーシース・ブルガリカス株、育種したラクトバチルス・デルブルッキー・サブスピーシース・ブルガリカス株3株(K1581株、K1583株、K1585株)、野生株(ATCC11842株)をそれぞれMRS培地で培養した。得られた菌液を0.85%滅菌食塩水で2度遠心・洗浄し、得られたペレットを無調整豆乳膜透過液に懸濁し、スターター菌液とした。
市販の乳酸菌混合スターター(ダニスコ社製)YO-MIX300、305、496、499、501、885からストレプトコッカス・サーモフィラス株(St.300株、St.305株、St.496株、St.499株、St.501株、St.885株)のみを単離した。単離した各菌株を10mLのLM17培地で培養し、培養液を0.9%NaCl溶液で遠心・洗浄し、10mLの0.9%NaCl溶液に再懸濁した。豆乳発酵原液には、市販の無調整豆乳(キッコーマンソイフーズ社製)を用いた。この豆乳発酵原液に上述で調製した懸濁液を1.0%(v/v)接種し、42℃、24時間発酵した。発酵後、遠心上清を回収し、フラクトースの残糖量をFキット D-グルコース/果糖(ジェイ・ケイ・インターナショナル社製)により測定した。結果を図7に示す。
フラクトースを蓄積する特徴のあるストレプトコッカス・サーモフィラス株(St.499株)と蓄積しない株(St.305株)を市販混合スターターYO-MIX505、511、863から単離したラクトバチルス・デルブルッキー・サブスピーシーズ・ブルガリカス株(Lb.505株、Lb.511株、Lb.863株)と1:1で組合せた乳酸菌混合スターターを構成した。これを市販の無調整豆乳(キッコーマンソイフーズ社製)に添加し42℃で24時間、発酵して豆乳発酵物を得た。そして、発酵終了後、得られた豆乳発酵物中のD-乳酸量を酵素電極法(王子計測社製、バイオセンサーBF-5)で測定した。
ストレプトコッカス・サーモフィラス株とラクトバチルス・デルブルッキー・サブスピーシーズ・ブルガリカス株が豆乳中で共生関係を構築するためには、ストレプトコッカス・サーモフィラス株によって豆乳の主要糖分であるスクロースを分解し、フラクトースを蓄積することで、ラクトバチルス・デルブルッキー・サブスピーシーズ・ブルガリカス株に糖源を供給することが不可欠であると考えられる。今回、育種したストレプトコッカス・サーモフィラスK1580株を単独で発酵した時、また、ストレプトコッカス・サーモフィラスK1580株とラクトバチルス・デルブルッキー・サブスピーシーズ・ブルガリカスK1581株を共発酵した時のスクロース、フラクトースの残糖量を経時的に測定した。発酵温度は42℃に設定した。スクロースの経時変化を図9に、フラクトースの経時変化を図10に示す。
育種株であるストレプトコッカス・サーモフィラスK1580株とラクトバチルス・デルブルッキー・サブスピーシーズ・ブルガリカスK1581株からなる乳酸菌混合スターターを調製し、これを用いて、無調整豆乳(キッコーマンソイフーズ社製)を42℃、24時間発酵させて豆乳発酵物を得た。一方、ストレプトコッカス・サーモフィラスK1580株を単独で無調整豆乳に添加し、42℃、24時間発酵させ、豆乳発酵物を得た。それぞれの成分分析値は以下のとおりである。
◎:非常に風味が良い
○:風味が良い
△:風味が少し悪い
×:風味が悪い
市販のヨーグルト製造用粉末混合スターター(ダニスコ社製:ストレプトコッカス・サーモフィラス株およびラクトバチルス・ブルガリカス株を含む凍結乾燥品)YO-MIX300、YO-MIX305、YO-MIX511、YO-MIX863(いずれも製品番号)の粉末を滅菌水で溶解し、同量の懸濁液を無調整豆乳(キッコーマンソイフーズ社製)に接種し、42℃、24時間発酵させ、豆乳発酵物を得た。
豆乳発酵物の物性を測定し、評価を実施した。物性の評価に関しては、豆乳発酵物中にどのような粒子径の粒子が、どのような割合で含まれているかを示す指標である粒度分布を用いた。粒度分布を測定することで、発酵物を口に含んだときの舌触りや滑らかさを評価することができる。
Claims (6)
- 豆乳に接種し培養した際に、発酵物中に0.4g/L以上のフラクトースを蓄積可能であるストレプトコッカス・サーモフィラスと、
豆乳に接種して培養した際に、発酵物中に0.4g/L以上のD-乳酸を蓄積可能であるラクトバチルス・デルブルッキー・サブスピーシーズ・ブルガリカスと、
を含む乳酸菌混合スターターを使用して得られた豆乳発酵物。 - 前記ストレプトコッカス・サーモフィラス及び前記ラクトバチルス・デルブルッキー・サブスピーシーズ・ブルガリカスが、豆乳を主成分とする培地で育種されたものである、請求項1に記載の豆乳発酵物。
- 前記ストレプトコッカス・サーモフィラス及び前記ラクトバチルス・デルブルッキー・サブスピーシーズ・ブルガリカスが、豆乳を主成分とする培地に同時に接種して育種されたものである、請求項1に記載の豆乳発酵物。
- 請求項1に記載した豆乳発酵物であって、前記豆乳発酵物の粒度分布において粒子径100マイクロメートル以上の粒子の全体に占める比率が10%以下である、豆乳発酵物。
- 豆乳に接種し培養した際に、発酵物中に0.4g/L以上のフラクトースを蓄積可能であるストレプトコッカス・サーモフィラスと、
豆乳に接種して培養した際に、発酵物中に0.4g/L以上のD-乳酸を蓄積可能であるラクトバチルス・デルブルッキー・サブスピーシーズ・ブルガリカスと、
を含む乳酸菌混合スターター。 - 前記乳酸菌混合スターターが、豆乳発酵物の製造に使用される、請求項6に記載の乳酸菌混合スターター。
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JP2018064521A (ja) * | 2016-10-20 | 2018-04-26 | 京都府 | 乳酸発酵豆乳の製造方法 |
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CN116508836B (zh) * | 2023-02-22 | 2024-05-10 | 洽洽食品股份有限公司 | 一种直投式酸奶发酵剂及其制备方法 |
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