WO2019065649A1 - 発酵乳及び発酵乳の製造方法 - Google Patents

発酵乳及び発酵乳の製造方法 Download PDF

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Publication number
WO2019065649A1
WO2019065649A1 PCT/JP2018/035502 JP2018035502W WO2019065649A1 WO 2019065649 A1 WO2019065649 A1 WO 2019065649A1 JP 2018035502 W JP2018035502 W JP 2018035502W WO 2019065649 A1 WO2019065649 A1 WO 2019065649A1
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Prior art keywords
milk
raw material
fermented milk
lactose
fermented
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PCT/JP2018/035502
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English (en)
French (fr)
Japanese (ja)
Inventor
堀内　啓史
武文 市村
暢子 井上
奈緒 高木
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Meiji Co Ltd
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Meiji Co Ltd
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Priority to US16/652,068 priority Critical patent/US11344040B2/en
Priority to EP18860859.0A priority patent/EP3753415A4/en
Priority to CN201880076619.7A priority patent/CN111417311A/zh
Publication of WO2019065649A1 publication Critical patent/WO2019065649A1/ja
Anticipated expiration legal-status Critical
Priority to US16/839,256 priority patent/US20200352187A1/en
Priority to US17/730,669 priority patent/US12219969B2/en
Ceased legal-status Critical Current

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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 OR TREATMENT 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
    • A23C9/1238—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt using specific L. bulgaricus or S. thermophilus microorganisms; using entrapped or encapsulated yoghurt bacteria; Physical or chemical treatment of L. bulgaricus or S. thermophilus cultures; Fermentation only with L. bulgaricus or only with S. thermophilus
    • 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 OR TREATMENT 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
    • A23C9/1275—Fermented milk preparations; Treatment using microorganisms or enzymes using microorganisms of the genus lactobacteriaceae and other microorganisms or enzymes, e.g. kefir, koumiss using only lactobacteriaceae for fermentation in combination with enzyme treatment of the milk product; using enzyme treated milk products for fermentation with lactobacteriaceae
    • 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
    • A23V2400/123—Bulgaricus
    • 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
    • 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 fermented milk and a method of producing fermented milk, and more particularly to fermented milk and a method of producing fermented milk in which the amount of polysaccharide produced by lactic acid bacteria is controlled.
  • Fermented milk is produced by fermenting raw material milk to which a lactic acid bacteria starter is added.
  • Lactic acid bacteria such as Bulgarian bacteria and Thermophilus bacteria are used as lactic acid bacteria starter.
  • lactic acid bacteria there are many strains that produce exopolysaccharide (EPS).
  • EPS contributes to the stability of fermented milk
  • OLL 1073 R-1 strain Lactobacillus delbruechi subsp. Bulgaricus OLL 1073 R-1
  • fermented milk produced using the OLL1073 R-1 strain has effects such as activation of NK cells and reduction of common cold.
  • the functional food which contributes to health can be provided by using the lactic acid bacteria which produce EPS, and the EPS produced by lactic acid bacteria.
  • the lactic acid bacteria which produce EPS and the EPS produced by lactic acid bacteria.
  • Patent Document 1 discloses a method of producing fermented milk capable of enhancing the production amount of EPS by fermenting raw material milk to which a phosphate is added.
  • Phosphate functions as a pH buffer in fermented milk. Since the phosphate can extend the period of the pH range in which the lactic acid bacteria can grow during the fermentation of the raw material milk, it can increase the production amount of EPS derived from the lactic acid bacteria.
  • An object of the present disclosure is to provide fermented milk containing a large amount of EPS, and a method of producing fermented milk capable of enhancing the production amount of EPS.
  • Fermented milk according to the present disclosure is produced by fermenting raw material milk. Before the fermentation of the raw material milk is started, the lactose concentration in the raw material milk is 2.5% by mass or less based on the total amount of the raw material milk.
  • the lactose concentration of the fermented milk may be 1.25 mass% or less with respect to the total amount of the fermented milk.
  • the amount of EPS is not less than 1.05 times and not more than 4.2 times the amount of EPS contained in fermented milk obtained by fermenting raw milk without decomposing lactose contained in raw milk. It may be.
  • the fermented milk according to the present disclosure may contain Bulgarian bacteria.
  • the number of Bulgarian bacteria contained in fermented milk is not less than 1.08 times and not more than 4.7 times the number of Bulgarian bacteria contained in fermented milk obtained by fermenting raw milk without decomposing lactose contained in raw milk May be
  • the fermented milk according to the present disclosure may contain the OLL 1073R-1 strain, which is a Bulgarian bacterium, and an EPS of 25 (mg / kg) or more and 100 (mg / kg) or less.
  • the method for producing fermented milk according to the present disclosure includes a preparation step, a lactose decomposition step, and a fermentation step.
  • a preparation process prepares raw material milk.
  • the lactose decomposition step at least a part of lactose contained in the prepared raw material milk is decomposed using lactose degrading enzyme.
  • lactose degrading enzyme In the fermentation step, lactic acid bacteria are added to the raw material milk in which at least a part of lactose has been decomposed, and the raw material milk to which the lactic acid bacteria are added is fermented.
  • the lactose concentration in the raw material milk from which lactose has been degraded is 2.5% by mass or less based on the total amount of the raw material milk.
  • the fermented milk according to the present invention can contain more EPS than conventional fermented milk. Moreover, the manufacturing method of fermented milk which concerns on this invention can raise the production amount of EPS by the lactic-acid-bacteria starter added to raw material milk.
  • FIG. 1 is a table showing the EPS content, the number of Bulgarian bacteria, and the number of Thermophilus bacteria in fermented milk according to Examples 1 to 6 of the present invention.
  • Fermented milk according to the present embodiment is produced by fermenting raw material milk.
  • the fermented milk which concerns on this Embodiment is raw material milk which finished fermentation.
  • raw material milk in which lactose is decomposed before the start of fermentation is used.
  • the lactose concentration of raw material milk before the start of fermentation is preferably 2.5% by mass or less based on the total amount of raw material milk.
  • EPS exopolysaccharide
  • the lactose concentration of raw material milk before the start of fermentation is more preferably 1% by mass or less.
  • Fermented milk obtained by fermenting raw material milk having a lactose concentration of 1% by mass or less before the start of fermentation has more EPS and Bulgarian bacteria than conventional fermented milk.
  • the lactose concentration of raw material milk before the start of fermentation is 0% by mass.
  • Fermented milk obtained by fermenting raw material milk having a lactose concentration of 0% by mass before the start of fermentation has more EPS and Bulgarian bacteria than conventional fermented milk.
  • lactose concentration being 0 mass% shows that lactose is not detected from raw material milk or fermented milk.
  • the method for detecting lactose contained in raw material milk or fermented milk is not particularly limited, and any conventionally known method can be used.
  • the fermented milk which concerns on this Embodiment is fermented milk and a lactic-acid-bacteria drink which were defined by the ministry of milk etc. (December 27, 1951 Ministry of Health ordinance 52nd issue).
  • Fermented milk in the Ministry of Milk, etc. is milk or the like containing non-fat milk solids equal to or more than this fermented with lactic acid bacteria or yeast and made paste-like or liquid, or those frozen.
  • the lactic acid bacteria beverage in the Ministry of Milk, etc. is a beverage obtained by processing milk or the like fermented with lactic acid bacteria or yeast, or using it as a main raw material.
  • the fermented milk according to the present embodiment contains at least Bulgarian bacteria and Thermophilus bacteria as lactic acid bacteria.
  • Thermophilus bacteria According to the United Nations Food and Agriculture Organization (FAO) and the World Health Organization (WHO), yogurt is made from milk and lactic acid bacteria, and lactic acid fermentation by both Bulgaria and Thermophilus bacteria causes dairy products such as milk and skimmed milk It is because it is defined as being made.
  • FAO Food and Agriculture Organization
  • WHO World Health Organization
  • Bactus bacteria refers to lactic acid bacteria of Lactobacillus delbruechii subsp. Bulgaricus species.
  • Thermophilus bacteria refers to lactic acid bacteria of the Streptococcus thermophilus type.
  • the fermented milk according to the present embodiment may include lactic acid bacteria other than Bulgarian bacteria and Thermophilus bacteria.
  • the fermented milk according to the present embodiment may contain gassic bacteria, bifidobacteria and the like.
  • Gasseri bacteria are lactic acid bacteria of the Lactobacillus gasseri (Lactobacillus gasseri) type.
  • Bifidobacterium is a lactic acid bacteria of the Bifidobacterium bifidum type.
  • fermented milk according to the present embodiment (fermented milk produced from raw material milk having a lactose concentration of 2.5% by mass or less before the start of fermentation) is simply referred to as “fermented It describes simply as "milk”.
  • Fermented milk is obtained by fermenting raw material milk having a lactose concentration of 2.5% by mass or less before the start of fermentation. During a period in which the raw material milk is fermented, lactose contained in the raw material milk is consumed by the Bulgarian bacteria and the thermophilus bacteria. Therefore, the lactose concentration of fermented milk is lower than the lactose concentration of raw material milk, and is preferably 1.25% by mass or less with respect to the total amount of fermented milk. More preferably, the lactose concentration of fermented milk is 0% by mass with respect to the total amount of fermented milk.
  • the fermented milk according to the present embodiment contains a larger amount of EPS than conventional fermented milk produced from raw material milk having a lactose concentration before the start of fermentation of more than 2.5% by mass.
  • conventional fermented milk manufactured from raw material milk whose lactose concentration before fermentation start is higher than 2.5 mass% is described as "conventional fermented milk”.
  • the fermented milk according to the present embodiment contains EPS in an amount of 1.06 times to 4.2 times that of conventional fermented milk. More preferably, the fermented milk which concerns on this Embodiment contains EPS of the quantity of 1.27 times or more and 4.2 times or less compared with the conventional fermented milk. More preferably, the fermented milk according to the present embodiment contains EPS in an amount of 1.48 times to 4.2 times that of conventional fermented milk.
  • the unit of the EPS content in fermented milk is "mg / kg".
  • the fermented milk according to the present embodiment contains more Bulgarian bacteria than conventional fermented milk.
  • the fermented milk according to the present embodiment contains the number of Bulgarian bacteria of 1.08 to 4.7 times the number of conventional fermented milk. More preferably, the fermented milk according to the present embodiment contains 1.18 to 4.7 times as many Bulgarian bacteria as conventional fermented milk. More preferably, the fermented milk according to the present embodiment contains 1.65 times to 4.7 times less Bulgarian bacteria than conventional fermented milk.
  • the unit of the number of Bulgarian bacteria is ( ⁇ 10 7 cfu / g).
  • EPS content and number of Bulgarian bacteria according to production conditions The EPS content and the number of Bulgarian bacteria in the fermented milk according to the present embodiment vary depending on the production conditions.
  • the first to sixth conditions are mentioned as production conditions, and the EPS content and the number of Bulgarian bacteria in fermented milk produced under each condition will be described in more detail.
  • the first to third conditions use Bulgarian bacteria isolated from Meiji Probio Yogurt R-1 (Meiji Co., Ltd.) as a lactic acid bacteria starter.
  • the fourth to sixth conditions use Bulgarian bacteria isolated from Meiji Bulgaria Yogurt LB81 (Meiji Co., Ltd.) as a lactic acid bacteria starter.
  • the Bulgarian fungus isolated from Meiji Probio Yogurt R-1 is Lactobacillus delbruechii subsp. Bulgaricus OLL 1073 R-1 and is deposited under Accession No. FERM BP-10741 at the National Institute of Advanced Industrial Science and Technology Patent Organism Depositary ing.
  • the Bulgarian fungus isolated from Meiji Probio Yogurt R-1 is referred to as "OLL 1037 R-1 strain”.
  • the Bulgarian fungus isolated from Meiji Bulgaria Yogurt LB 81 is described as "Bulgarian fungus MB strain”.
  • the first condition is to produce fermented milk by adding the OLL 1037R-1 strain to raw material milk having a lactose concentration of 2.5% by mass or less before the start of fermentation.
  • EPS content of fermented milk manufactured on the 1st condition is 25 (mg / kg) or more and 100 (mg / kg) or less.
  • the fermented milk produced under the first condition contains EPS in an amount of 1.06 to 4.2 times that of conventional fermented milk using the OLL 1037R-1 strain.
  • the upper limit "100 (mg / kg)" of the EPS content is a value assumed to be produced when the lactic acid bacteria consume all the sugar contained in fermented milk.
  • the number of Bulgarian bacteria of fermented milk manufactured under the first condition is 23 ( ⁇ 10 7 cfu / g) or more and 100 ( ⁇ 10 7 cfu / g) or less.
  • the fermented milk produced under the first condition contains the OLL1037R-1 strain at least 1.08 times and at most 4.7 times as much Bulgarian bacteria as a conventional fermented milk using the OLL1037R-1 strain.
  • the second condition is to produce fermented milk by adding the OLL 1037R-1 strain to raw material milk having a lactose concentration of 1% by mass or less before the start of fermentation. That is, the lactose concentration of the raw material milk under the second condition is lower than the lactose concentration of the raw material milk under the first condition.
  • the fermented milk produced under the second condition contains EPS in an amount of 30 (mg / kg) or more and 100 (mg / kg) or less.
  • the fermented milk produced under the second condition contains EPS in an amount of 1.27 to 4.2 times that of conventional fermented milk using the OLL 1037R-1 strain.
  • fermented milk produced under the second condition has Bulgarian bacteria of 25 ( ⁇ 10 7 cfu / g) or more and 100 ( ⁇ 10 7 cfu / g) or less.
  • the fermented milk produced under the second condition contains 1.18 to 4.7 times less than the conventional fermented milk using the OLL 1037R-1 strain.
  • the third condition is to produce fermented milk by adding the OLL 1037R-1 strain to raw material milk having a lactose concentration of 0% by mass before the start of fermentation. That is, the lactose concentration of the raw material milk under the third condition is lower than the lactose concentration of the raw material milk under the first condition and the second condition.
  • the fermented milk produced under the third condition contains an EPS of 35 (mg / kg) or more and 100 (mg / kg) or less.
  • the fermented milk produced under the third condition contains EPS in an amount of 1.48 times to 4.2 times that of conventional fermented milk using the OLL 1073R-1 strain.
  • fermented milk produced under the third condition has Bulgarian bacteria of 35 ( ⁇ 10 7 cfu / g) or more and 100 ( ⁇ 10 7 cfu / g) or less.
  • the fermented milk produced under the third condition contains 1.65 to 4.7 times less Bulgarian bacteria as compared to the conventional fermented milk using the OLL 1037R-1 strain.
  • the fourth condition is to produce fermented milk by adding the Bulgarian fungus MB strain to raw material milk having a lactose concentration of 2.5% by mass or less before the start of fermentation.
  • the fermented milk produced under the fourth condition contains EPS in an amount of 50 (mg / kg) or more and 100 (mg / kg) or less.
  • the fermented milk produced under the fourth condition contains EPS in an amount of 1.18 times to 2.36 times that of conventional fermented milk using the Bulgarian fungus MB strain.
  • the fermented milk manufactured on the 4th condition has 15 (* 10 ⁇ 7 > cfu / g) or more and 100 (* 10 ⁇ 7 > cfu / g) or less Bulgarian bacteria.
  • the fermented milk produced under the fourth condition contains 1.2 to 8.0 times as much Bulgarian bacteria as fermented milk using the Bulgarian strain MB strain.
  • the fifth condition is to produce fermented milk by adding the Bulgarian fungus MB strain to raw material milk having a lactose concentration of 1% by mass or less before the start of fermentation. That is, the lactose concentration of the raw material milk under the fifth condition is lower than the lactose concentration of the raw material milk under the fourth condition.
  • the fermented milk produced under the fifth condition contains EPS in an amount of 55 (mg / kg) or more and 100 (mg / kg) or less.
  • the fermented milk produced under the fifth condition contains EPS in an amount of 1.3 times or more and 2.36 times or less as compared with conventional fermented milk using the Bulgarian fungus MB strain.
  • the fermented milk produced under the fifth condition has Bulgarian bacteria of 25 ( ⁇ 10 7 cfu / g) or more and 100 ( ⁇ 10 7 cfu / g) or less.
  • the fermented milk produced under the fifth condition is compared with the conventional fermented milk produced by adding the Bulgarian fungus MB strain to raw material milk having a lactose concentration before the start of fermentation higher than 2.5% by mass. It contains 0 to 8.0 times less Bulgarian bacteria.
  • the sixth condition is to produce fermented milk by adding the Bulgarian fungus MB strain to raw material milk having a lactose concentration of 0% by mass before the start of fermentation. That is, the lactose concentration of the raw material milk under the sixth condition is lower than the lactose concentration of the raw material milk under the fourth and fifth conditions.
  • the fermented milk produced under the sixth condition contains EPS in an amount of 65 (mg / kg) or more and 100 (mg / kg) or less.
  • the fermented milk produced under the sixth condition contains EPS in an amount of 1.53 times to 2.36 times that of conventional fermented milk using the Bulgaria strain MB strain.
  • fermented milk produced under the sixth condition has 40 ( ⁇ 10 7 cfu / g) or more and 100 ( ⁇ 10 7 cfu / g) or less of Bulgarian bacteria.
  • the fermented milk produced under the sixth condition contains at least 3.2 times and at most 8 times less than the conventional fermented milk using the Bulgarian fungus MB strain.
  • raw material milk is prepared.
  • raw materials used for preparation of raw material milk for example, water, raw milk, skimmed milk powder, whole milk powder, buttermilk, butter, cream, whey protein concentrate (WPC), whey protein isolate (WPI), ⁇ -lactalbumin , ⁇ -lactoglobulin and the like.
  • Raw material milk should just contain the milk component for performing lactic acid fermentation by lactic acid bacteria. For this reason, raw material milk may not contain all the raw materials listed above, and may use raw materials other than the raw materials listed above.
  • Raw material milk is prepared by a conventionally known method. For example, raw milk can be prepared by producing a mixture by mixing the raw materials listed above and homogenizing the produced mixture. Raw milk prepared in this manner contains lactose. Lactose is contained in raw materials derived from milk such as raw milk, skimmed milk powder, whole milk powder and the like.
  • lactase decomposition process In the lactose decomposition step, lactase is added to the prepared raw material milk, and at least a part of lactose contained in the prepared raw material milk is decomposed with lactase. Lactase degrades lactose to produce glucose and galactose.
  • the type of lactase added is not particularly limited as long as the optimum pH of the added lactase is in the neutral region or the acidic region. For example, commercially available lactase can be added to raw material milk.
  • lactose contained in the raw material milk is decomposed with lactase until the lactose concentration in the raw material milk becomes 2.5% by mass or less.
  • lactose concentration in the raw material milk prepared by the preparation step is, for example, 5% by mass, lactose is decomposed by lactase until the lactose decomposition rate reaches 50% or more.
  • lactose contained in the raw material milk is decomposed by lactase until the lactose concentration in the raw material milk becomes 1% by mass or less.
  • lactose concentration of the raw material milk prepared by the preparation step is, for example, 5% by mass
  • lactose is decomposed by lactase until the lactose decomposition rate reaches 80% or more.
  • lactose contained in the raw material is decomposed with lactase until the lactose concentration in the raw material milk becomes 0% by mass.
  • lactose is decomposed by lactase until the lactose decomposition rate reaches 100%.
  • Decomposition of lactose by lactase is performed until fermentation of raw milk by Bulgarian bacteria and Thermophilus bacteria is started.
  • the start timing of the fermentation is, for example, the timing when the lactic acid bacteria starter (Bulgarian bacteria and Thermophilus bacteria) is added to the raw material milk.
  • the sterilization process may be performed before the lactose decomposition process. In this case, since lactase can continue to decompose lactose in a fermentation step described later, the lactose concentration of fermented milk can be further lowered.
  • a lactic acid bacteria starter is added to the pasteurized raw material milk, and the raw material milk is fermented under predetermined fermentation conditions.
  • the raw material milk after the fermentation is refrigerated as fermented milk according to the present embodiment.
  • Fermentation conditions such as fermentation temperature and fermentation time may be appropriately adjusted in consideration of the type of lactic acid bacteria starter added to the raw material milk, the flavor of the desired fermented milk, and the like. For example, by placing the raw material milk in an environment of 30 ° C. or more and 50 ° C. or less, fermentation by lactic acid bacteria can be promoted. The fermentation time is appropriately adjusted according to the fermentation temperature, the type of lactic acid bacteria starter added to the raw material milk, the desired lactic acid level in fermented milk, and the like.
  • the method for producing fermented milk according to the present embodiment decomposes lactose contained in raw material milk so that the lactose concentration of raw material milk before starting fermentation becomes 2.5% by mass or less, Ferment fermented raw milk.
  • Raw milk produced in this manner can contain more EPS than conventional fermented milk.
  • the timing of the fermentation start in raw material milk was defined as a timing in which a lactic-acid-bacteria starter is added to raw material milk.
  • the number of lactic acid bacteria starters added to the raw material milk does not increase in the induction phase (period until the logarithmic growth phase starts), lactose is hardly consumed in the induction phase. Therefore, it is possible to define the timing of the initiation of fermentation as the timing at which the logarithmic growth phase of lactic acid bacteria is initiated.
  • the sterilization step is performed prior to the lactose degradation step. That is, lactase is added to the heat-killed raw material milk.
  • the start timing of the logarithmic growth phase is the start timing of fermentation
  • the decomposition of lactose by lactase continues even after adding the lactic acid bacteria starter to the raw material milk.
  • the lactose concentration in the raw material milk may be 1.5 mass% or less.
  • Example 1 The fermented milk which concerns on Example 1 respond
  • Raw material milk was prepared by mixing 500.0 g of raw milk, 53.2 g of skimmed milk powder, 23.0 g of fresh cream and 403.6 g of tap water.
  • the lactose concentration of the prepared raw material milk was 5% by mass.
  • 0.2 g of lactase (GODO-YNL, manufactured by Kyoshu Shusei Co., Ltd.) was added to the raw material milk to decompose lactose contained in the raw material milk. Specifically, the decomposition of lactose was continued until the lactose decomposition rate of the raw material milk was 50%. The method of measuring the lactose decomposition rate will be described later.
  • the lactose concentration in the raw material milk in which the decomposition of lactose was completed was 2.5% by mass. Then, the raw material milk from which lactose was decomposed was heat-sterilized at a temperature of 95 ° C., and the heat-sterilized raw material milk was cooled to a temperature of 43 ° C.
  • the measuring method of the lactose decomposition rate in raw material milk is demonstrated. First, the lactose content per solid content in raw milk before lactase is added is measured. Next, the glucose content per solid content in the raw material milk is measured from the glucose concentration in the raw material milk in which lactose has been decomposed.
  • the lactose content can be measured using arginine fluorescence method by high performance liquid chromatography (BUNSEKI KAGAKU, vol. 32, p. E207, published by The Japan Society of Analytical Chemistry, 1983).
  • the glucose content can be measured, for example, using Medisafe Mini (manufactured by Terumo Corporation).
  • Lactose concentration can be calculated from solid content concentration and lactose content in raw material milk.
  • lactic acid bacteria isolated from Meiji Probio Yogurt R-1 (Meiji Co., Ltd.) were added to the raw material milk after heat sterilization as a lactic acid bacteria starter.
  • the lactic acid bacteria isolated from Meiji Probio Yogurt R-1 include Thermophilus bacteria and the OLL 1073R-1 strain which is a Bulgarian bacteria.
  • the addition amount of the lactic acid bacteria starter is 20 g.
  • the raw material milk to which the lactic acid bacteria starter was added was filled into a cup container (volume: 100 mL, made of plastic).
  • the raw material milk filled in the cup container was subjected to stationary fermentation in a fermentation chamber at a temperature of 43 ° C. until the lactic acid acidity became 0.7%.
  • the cupped raw material milk which stationary fermentation finished was cooled in 10 degreeC refrigerator as fermented milk which concerns on Example 1. As shown in FIG. The lactose concentration of the fermented milk which concerns on Example 1 immediately after the completion of fermentation was 1.25 mass%.
  • the number of Bulgarian bacteria and the number of thermophilus bacteria in fermented milk can be measured by a conventionally known method. For example, a predetermined amount of fermented milk is appropriately diluted, and a diluted solution of fermented milk is smeared on a BL medium.
  • the viable bacteria smeared on the BL medium can be anaerobically cultured for 72 hours in a temperature environment of 37 ° C., and the number of colonies on the BL medium after culture can be determined.
  • the measuring method of the EPS content of fermented milk is demonstrated.
  • 10 g of fermented milk according to Example 1 was placed in a 50 mL tube, and 1 mL of 100% trichloroacetic acid was added to the tube into which fermented milk according to Example 1 was placed.
  • the contents of the tube were stirred and the stirred contents centrifuged at 12000 G relative centrifugal force.
  • the supernatant obtained by centrifugation was transferred to a new tube of 50 mL in volume.
  • twice the amount of ethanol of the supernatant was added to the supernatant in a fresh tube.
  • the mixture of supernatant and ethanol was allowed to stand overnight at a temperature of 4 ° C.
  • the stationary mixture was centrifuged at a relative centrifugal force of 12000 G, and the supernatant was discarded from the centrifuged mixture.
  • 10 mL of purified water was added to the precipitate in the centrifuged mixture to completely dissolve the precipitate in purified water.
  • the purified water in which the precipitate was dissolved was injected into high performance liquid chromatography (HPLC) using a 0.45 ⁇ m-filtered syringe. Then, the ratio of “the peak area of a single peak detected by the RI detector in the vicinity of the time when 16 minutes pass from the start of injection” to the “total peak area” is the content of EPS included in Example 1 And
  • HPLC system ACQUITY UPLC H-Class (Waters) Column: OHpak SB-806HQ, SB-G (Shodex) Column temperature: 40 ° C Solvent: 0.2 M NaCl aqueous solution flow rate: 0.5 mL / min Detector: RI detector 2414 (Waters) Detection temperature: 40 ° C Sample injection: 150 ⁇ L Analysis time: 50 min Example 2 The fermented milk which concerns on Example 2 respond
  • the fermented milk which concerns on Example 2 is the same as the said Example 1 except the point which decomposes
  • the lactose concentration in the raw material milk according to Example 2 was 1% by mass before the start of fermentation.
  • the lactose concentration in the fermented milk according to Example 2 was 0% by mass immediately after the completion of the fermentation. That is, immediately after the completion of the fermentation, lactose was not detected from the fermented milk according to Example 2.
  • Example 3 The fermented milk which concerns on Example 3 respond
  • the process of producing fermented milk according to Example 3 is the same as Example 1 except that lactose contained in the raw material milk is degraded by lactase until the lactose decomposition rate becomes 100%.
  • the lactose concentration in the raw material milk according to Example 3 was 0% by mass before the start of fermentation.
  • the lactose concentration in the fermented milk according to Example 3 was 0 mass% immediately after the completion of the fermentation. That is, lactose was not detected from the fermented milk according to Example 3 both before and after the start of fermentation.
  • Comparative Example 1 The fermented milk according to Comparative Example 1 corresponds to conventional fermented milk using the OLL 1037R-1 strain.
  • the lactose decomposition process is omitted from the production method in Example 1.
  • the lactose decomposition rate in the raw material milk according to Comparative Example 1 was 0%, and the lactose concentration in the raw material milk according to Comparative Example 1 was 5% by mass before the start of fermentation.
  • the lactose concentration in the fermented milk according to Comparative Example 1 was 3.75 mass% immediately after the completion of the fermentation.
  • Example 4 The fermented milk which concerns on Example 4 respond
  • the process for producing fermented milk according to Example 4 is the same as the process for producing fermented milk according to Example 1, except that the lactic acid bacteria starter is not the OLL1073R-1 strain but the Bulgarian bacteria MB strain.
  • the lactose concentration in the raw material milk according to Example 4 was 2.5% by mass before the start of fermentation.
  • the lactose concentration in the fermented milk according to Example 4 was 1.25% by mass immediately after the completion of the fermentation.
  • Example 5 The fermented milk which concerns on Example 5 respond
  • the process for producing fermented milk according to Example 5 is the same as the process for producing fermented milk according to Example 2 except that the lactic acid bacteria starter is a Bulgarian bacteria MB strain instead of the OLL 1073R-1 strain.
  • the lactose concentration in the raw material milk according to Example 5 was 1% by mass before the start of fermentation.
  • the lactose concentration in the fermented milk according to Example 5 was 0% by mass immediately after the completion of the fermentation. That is, immediately after the completion of fermentation, lactose was not detected from the fermented milk according to Example 5.
  • Example 6 The fermented milk which concerns on Example 6 respond
  • the process for producing fermented milk according to Example 6 is the same as the process for producing fermented milk according to Example 3, except that the lactic acid bacteria starter is not the OLL 1073 R-1 strain but the MB strain of Bulgarian bacteria.
  • the lactose concentration in the raw material milk according to Example 6 was 0% by mass before the start of fermentation.
  • the lactose concentration in the fermented milk according to Example 6 was 0 mass% immediately after the completion of the fermentation. That is, lactose was not detected from the fermented milk according to Example 6 both before the start of fermentation and immediately after the end of fermentation.
  • the process for producing fermented milk according to Comparative Example 2 is the same as the process for producing fermented milk according to Comparative Example 1 except that the lactic acid bacteria starter is not the OLL 1073R-1 strain but the Bulgarian bacteria MB strain.
  • the lactose decomposition rate in the raw material milk according to Comparative Example 2 was 0%, and the lactose concentration in the raw material milk according to Comparative Example 2 was 5% by mass before the start of fermentation.
  • the lactose concentration in the fermented milk according to Comparative Example 2 was 3.75 mass% immediately after the completion of the fermentation.
  • FIG. 1 is a table showing the EPS content, the number of Bulgarian bacteria, and the number of thermophilus bacteria in fermented milk according to Examples 1 to 6 and Comparative Examples 1 and 2. The relationship between the lactose concentration of raw material milk and the EPS content of fermented milk and the relationship between the lactose concentration of raw material milk and the number of Bulgarian bacteria of fermented milk will be described with reference to FIG.
  • the lactose concentration of raw material milk before the start of fermentation is 5% by mass, and the EPS content of fermented milk is 23.6 (mg / kg).
  • the lactose concentration of raw material milk before fermentation start is 2.5 mass%, and the EPS content of fermented milk is 26.2 (mg / kg).
  • the lactose concentration of raw material milk before the start of fermentation is 1% by mass, and the EPS content of fermented milk is 31.6 (mg / kg).
  • the lactose concentration of raw material milk before the start of fermentation is 0% by mass, and the EPS content of fermented milk is 36.5 (mg / kg).
  • the lactose concentration of raw material milk before the start of fermentation is 5% by mass, and the EPS content of fermented milk is 42.4 (mg / kg).
  • the lactose concentration of raw material milk before fermentation start is 2.5 mass%, and the EPS content of fermented milk is 52.4 (mg / kg).
  • the lactose concentration of raw material milk before the start of fermentation is 1% by mass, and the EPS content of fermented milk is 59.3 (mg / kg).
  • the lactose concentration of raw material milk before the start of fermentation is 0% by mass, and the EPS content of fermented milk is 69.6 (mg / kg).
  • the lactose concentration of raw material milk before the start of fermentation is 5% by mass, and the number of Bulgarian bacteria of fermented milk is 21.2 ( ⁇ 10 7 cfu / kg).
  • the lactose concentration of raw material milk before the start of fermentation is 2.5% by mass, and the number of Bulgarian bacteria in fermented milk is 24.7 ( ⁇ 10 7 cfu / kg).
  • the lactose concentration of raw material milk before the start of fermentation is 1% by mass, and the number of Bulgarian bacteria of fermented milk is 25.3 ( ⁇ 10 7 cfu / kg).
  • the lactose concentration of raw material milk before the start of fermentation is 0% by mass, and the number of Bulgarian bacteria in fermented milk is 37.9 ( ⁇ 10 7 cfu / kg).
  • the OLL1073R-1 strain is a lactic acid bacteria starter
  • the number of Bulgarian bacteria increases as the lactose concentration of the raw material milk before the start of fermentation decreases.
  • the lactose concentration of raw material milk before the start of fermentation is minimum (0% by mass)
  • the number of Bulgarian bacteria in fermented milk is maximum.
  • thermophilus bacteria of Examples 1 to 3 and Comparative Example 1 in FIG. 1 will be referred to.
  • the number of Thermophilus bacteria in the fermented milk according to Examples 1 to 3 is larger than the number of Thermophilus bacteria in the fermented milk according to Comparative Example 1. That is, when the lactose of the raw material milk is decomposed so that the lactose concentration of the raw material milk is 2.5% by mass or less before the start of the fermentation, the number of thermophilus bacteria can be increased.
  • the thermophilus bacteria count is the largest in Example 2 in which the lactose concentration of the raw material milk before the start of fermentation is 1% by mass, the relationship between the lactose concentration and the thermophilus bacteria count is Not as clear as relationship with.
  • the number of Bulgarian bacteria increases as the lactose concentration of the raw material milk decreases before the start of fermentation, it is considered that the Bulgarian bacteria contribute to an increase in the EPS content more than the thermophilus bacteria. .
  • the lactose concentration of raw material milk before the start of fermentation is 5% by mass, and the number of Bulgarian bacteria in fermented milk is 12.5 ( ⁇ 10 7 cfu / kg).
  • the lactose concentration of raw material milk before the start of fermentation is 2.5% by mass, and the number of Bulgarian bacteria in fermented milk is 18.0 ( ⁇ 10 7 cfu / kg).
  • the lactose concentration of raw material milk before the start of fermentation is 1% by mass, and the number of Bulgarian bacteria of fermented milk is 26.5 ( ⁇ 10 7 cfu / kg).
  • the lactose concentration of raw material milk before the start of fermentation is 0% by mass, and the number of Bulgarian bacteria of fermented milk is 41.0 ( ⁇ 10 7 cfu / kg).
  • the number of Bulgarian bacteria increases as the lactose concentration of raw material milk before the start of fermentation decreases.
  • the lactose concentration of raw material milk before the start of fermentation is minimum (0% by mass)
  • the number of Bulgarian bacteria in fermented milk is maximum.
  • the Bulgarian bacteria MB strain contained in the raw material milk is activated in the fermentation process It is thought that the EPS derived from was efficiently produced.
  • thermophilus bacteria of Examples 4 to 6 and Comparative Example 2 The thermophilus bacteria count of the fermented milk which concerns on Example 4-6 is larger than the thermophilus bacteria count of the fermented milk which concerns on the comparative example 2. As shown in FIG. That is, when the lactose of the raw material milk is decomposed so that the lactose concentration of the raw material milk is 2.5% by mass or less before the start of the fermentation, the number of thermophilus bacteria can be increased.
  • thermophilus bacteria count is the largest in Examples 1 and 2 where the lactose concentration of the raw material milk is 2.5 mass% and 1 mass% before the start of fermentation, and the relationship between the lactose concentration and the thermophilus bacteria count is Not as clear as the relationship between lactose concentration and the number of Bulgarian bacteria.
  • the number of Bulgarian bacteria increases as the lactose concentration of the raw material milk decreases before the start of fermentation, it is considered that the Bulgarian bacteria contribute to an increase in the EPS content more than the thermophilus bacteria. .

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JP2022162949A (ja) * 2021-04-13 2022-10-25 株式会社明治 菌体外多糖の検出方法

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