WO2016104642A1 - Agent de stabilisation d'émulsion, et procédé de stabilisation d'émulsion mettant en œuvre celui-ci - Google Patents

Agent de stabilisation d'émulsion, et procédé de stabilisation d'émulsion mettant en œuvre celui-ci Download PDF

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WO2016104642A1
WO2016104642A1 PCT/JP2015/086095 JP2015086095W WO2016104642A1 WO 2016104642 A1 WO2016104642 A1 WO 2016104642A1 JP 2015086095 W JP2015086095 W JP 2015086095W WO 2016104642 A1 WO2016104642 A1 WO 2016104642A1
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milk
mfg
emulsion
oil
water emulsion
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PCT/JP2015/086095
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English (en)
Japanese (ja)
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幹 松田
武彦 安枝
健司 大島
淑立 市原
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一般財団法人糧食研究会
国立大学法人名古屋大学
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Priority to JP2016566470A priority Critical patent/JP6656172B2/ja
Publication of WO2016104642A1 publication Critical patent/WO2016104642A1/fr

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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

Definitions

  • the present invention relates to an emulsion stabilizer containing MFG-E8 protein as an active ingredient and an emulsion stabilization method using the same.
  • oil-in-water emulsions that are mixed in the form of fat globules in which oil is emulsified, such as milk
  • these fats are adjusted by adjusting the diameter of the fat globules mixed in water to an appropriate size.
  • the spheres are stably dispersed in water, and the emulsion stability of the oil-in-water emulsion is increased.
  • the fat globules mixed in the oil-in-water emulsion generally become associated with each other as the retention time of the oil-in-water emulsion increases, and changes to an aggregate of fat globules. Therefore, even if the fat sphere diameter mixed in the oil-in-water emulsion is initially small, it tends to gradually increase as the holding time becomes longer. Therefore, it is difficult to stabilize the emulsification of the oil-in-water emulsion over a long period of time.
  • fat globule mixed in an oil-in-water emulsion is mechanically sheared with a homogenizer etc. to reduce the fat globule diameter (homogenize), and an emulsifier is added to the oil-in-water emulsion.
  • a method for stabilizing the emulsification of an oil-in-water emulsion is known.
  • the emulsifier added to the oil-in-water emulsion includes, for example, an emulsifier as a chemically synthesized product such as glycerin fatty acid ester that is easily compatible with water and oil, and an emulsifier derived from food such as an emulsifier derived from soybean or an emulsifier derived from egg It is known.
  • an emulsifier as a chemically synthesized product such as glycerin fatty acid ester that is easily compatible with water and oil
  • an emulsifier derived from food such as an emulsifier derived from soybean or an emulsifier derived from egg It is known.
  • emulsifiers as chemically synthesized products are contrary to the trend of evaluating natural materials of consumers in recent years because they recall an artificial image.
  • the food-derived emulsifier is a consumer who is concerned about food allergies when the origin of the oil-in-water emulsion is different, for example, when a soybean-derived emulsifier is added to a milk beverage derived from a milk component. Not good for.
  • an oil-in-water emulsion is orally ingested (administered) as a food or pharmaceutical
  • an emulsifier derived from the same oil-in-water emulsion For example, when the oil-in-water emulsion is milk, it is known to use casein and / or sodium caseinate as an emulsifier derived from milk. It is also known that a milk fat globule membrane fraction of milk can be used as an emulsifier (Patent Document 1).
  • MFG-E8 milk fat globule-EGF factor 8
  • MFG-E8 milk fat globule-EGF factor 8
  • phospholipids phosphatidylserine
  • MFG-E8 gene-deficient mice have been prepared for the purpose of finding out the new functionality (Non-patent Document 2).
  • MFG-E8 has the ability to remove apoptotic cells in vivo and to interact between milk fat globules and epithelial cells in mammary cells. It is known to have functions such as regulation of fat globules in breast milk after breastfeeding and weaning by regulation (Non-patent Document 3).
  • An object of the present invention is to achieve emulsification in which fat globules mixed in an oil-in-water emulsion are stably dispersed in water and the emulsion stability of the oil-in-water emulsion is increased even when the content (addition) is relatively small. It is to provide a stabilizer and an emulsion stabilization method using the emulsion stabilizer. Moreover, the subject of the present invention is a milky food containing milk and / or dairy products. Even if the amount (addition) is relatively small, fat globules mixed in the milky food are stably dispersed in water. Another object of the present invention is to provide an emulsion stabilizer that improves the emulsion stability of milky foods, and an emulsion stabilization method using the emulsion stabilizer.
  • the present inventors have found that the emulsification stability of an oil-in-water emulsion is increased by containing MFG-E8, which is a protein, as an active ingredient. It was. Specifically, the fat globule diameter of the breast milk of MFG-E8 gene-deficient mice is particularly long compared to the fat globule diameter of breast milk of normal mice not deficient in the MFG-E8 gene. As it became, it was confirmed that the fat globule diameter was increased by associating the fat globules with other fat globules in the breast milk.
  • the emulsion stabilizer of the present invention is, for example, an oil-in-water solution even if the MFG-E8 is contained (added) in a relatively small amount of 0.001 to 1 part by mass with respect to 100 parts by mass of the oil-in-water emulsion.
  • the present inventors have found that fat globules mixed in a mold emulsion are stably dispersed in water, and the emulsion stability of the oil-in-water emulsion is increased.
  • emulsion stabilizer of the present invention by adding the emulsion stabilizer of the present invention to, for example, milk and / or dairy foods containing dairy products among oil-in-water emulsions, fat globules mixed in dairy foods are stable in water. It was found that the emulsification stability of dairy foods is enhanced.
  • the emulsion stabilizer of the present invention is derived from milk and the oil-in-water emulsion is a dairy food consisting of milk and / or milk-derived dairy products
  • the emulsion stabilizer and the oil-in-water emulsion are All are derived from milk and can be made into a product with high added value such as “processed milk” standard or “component-adjusted milk” standard instead of “milk beverage” standard.
  • the present invention is as follows.
  • MFG-E8 milk fat globule-EGF factor 8
  • MFG-E8 is derived from skim milk.
  • a method for stabilizing the emulsion of an oil-in-water emulsion comprising the step of adding MFG-E8, which is a protein, to the oil-in-water emulsion.
  • MFG-E8 which is a protein
  • the presence of MFG-E8, which is a protein allows fat globules mixed in an oil-in-water emulsion to be stably dispersed in water even when contained (added) in a relatively small amount. It has been found that the emulsification stability of the mold emulsion is enhanced.
  • the present invention relates to an emulsion stabilizer for an oil-in-water emulsion containing MFG-E8 (milk fat globule-EGF factor 8), which is a protein, as an active ingredient, and an oil-in-water type comprising a step of adding the emulsion stabilizer.
  • An emulsion stabilization method for an emulsion can be provided.
  • FIG. 1A is a photomicrograph of fat globules (MFG) in each milk sample.
  • FIG. 1B is a graph showing the number of fat globules (MFG) for each particle diameter in the microscopic field of each milk sample.
  • FIG. 2A is a view of fat globules (MFG) in each milk sample observed with a microscope.
  • FIG. 2B is a graph showing the number of fat globules (MFG) having a diameter of 15 ⁇ m or more in the microscopic field of each milk sample.
  • FIG. 3A is a view of fat globules (MFG) in each milk sample observed with a microscope.
  • FIG. 1A is a photomicrograph of fat globules (MFG) in each milk sample.
  • FIG. 1B is a graph showing the number of fat globules (MFG) for each particle diameter in the microscopic field of each milk sample.
  • FIG. 2A is a view of fat glob
  • FIG. 3B is a graph showing the number of fat globules (MFG) having a diameter of 15 ⁇ m or more in the microscopic field of each milk sample.
  • FIG. 3C is a diagram showing immunostaining of fat globules (MFG) in each milk sample with anti-MFG-E8 antibody.
  • FIG. 4 is a diagram of observation of milk fat globules (bovine fat globules) that have not been homogenized after milking under a phase contrast microscope.
  • FIG. 5 is a diagram showing the inhibitory effect on the association of fat globules (bovine fat globules) in milk that has not been homogenized after milking during storage by MFG-E8 derived from bovine milk.
  • FIG. 6 is a diagram showing the inhibitory effect of MFG-E8 derived from mouse milk on the association between milk fat globules (bovine fat globules) that have not been homogenized after milking during storage.
  • the emulsion stabilizer for oil-in-water emulsions of the present invention is characterized by containing MFG-E8 (milk fat globule-EGF factor 8) as an active ingredient.
  • MFG-E8 of the present invention is a membrane-bound protein that binds to phospholipids (phosphatidylserine) in the membrane of fat globules mixed in breast milk, but is originally required as a component of milk fat globule membrane. It is said that it is not a membrane-bound protein.
  • the presence of MFG-E8 is known, for example, from human origin, mouse origin, rat origin, pig origin, and bovine origin.
  • MFG-E8 is contained in a supernatant fraction (so-called whey fraction) obtained by centrifuging milk or skim milk (for example, reduced skim milk powder) under a predetermined condition to remove casein.
  • whey fraction a supernatant fraction obtained by centrifuging milk or skim milk (for example, reduced skim milk powder) under a predetermined condition to remove casein.
  • the whey fraction is purified by known protein purification techniques such as various chromatographies, salting out, dialysis, membrane separation, etc., to thereby purify MFG-E8 or partially (for example, MFG-E8 purified to any stage (such as phosphatidylserine) can be obtained.
  • the MFG-E8 of the present invention is obtained by, for example, centrifuging purified MFG-E8, partially purified MFG-E8, and milk or skim milk (for example, reduced skim milk powder)
  • the supernatant fraction from which casein has been removed can be used.
  • the shape of the MFG-E8 of the present invention is not limited, and examples thereof include liquid, paste, powder, and frozen.
  • the origin of MFG-E8 of the present invention is not limited, and for example, derived from milk (derived from human milk, derived from mouse milk, derived from rat milk, derived from pig milk, derived from bovine milk, etc.), derived from skim milk (human Derived from milk, derived from mouse milk, derived from rat milk, derived from pig milk, derived from bovine milk, and the like.
  • milk derived from human milk, derived from mouse milk, derived from rat milk, derived from pig milk, derived from bovine milk, etc.
  • skim milk human Derived from milk, derived from mouse milk, derived from rat milk, derived from pig milk, derived from bovine milk, and the like.
  • a supernatant fraction (so-called whey fraction) from which milk or skim milk (for example, reduced skim milk powder), which is an embodiment of MFG-E8 of the present invention, is centrifuged under predetermined conditions to remove casein. ) Is a so-called industrial waste fraction in the production process of casein, which is important as a milk protein raw material.
  • the use of these whey fractions as MFG-E8 of the present invention is preferable from the viewpoint of effective use of resources, environmental protection or cost reduction.
  • the whey fraction preferably contains a large amount of MFG-E8 of the present invention.
  • the whey fraction used as the MFG-E8 of the present invention is milk or skim milk (for example, reduced skim milk powder), and the centrifugal conditions are such that MFG-E8 can be obtained. There are no restrictions on the number of revolutions, the time required for centrifugation, the temperature at which centrifugation is performed, and the like.
  • Centrifugal force is, for example, 5000-100000 ⁇ g, 6000-80000 ⁇ g, 7000-60000 ⁇ g, 8000-40000 ⁇ g, 9000-30000 ⁇ g, 10000-20000 ⁇ g, 12000-18000 ⁇ g, and 14000. ⁇ 16000 ⁇ g.
  • a centrifugal force of 5000 ⁇ g or more is preferable because the purity of MFG-E8 can be increased.
  • the centrifugal force is 100000 ⁇ g or less, it is not excessive centrifugal force and can be efficiently centrifuged, which is preferable.
  • the time required for centrifugation is, for example, 2 to 60 minutes, 4 to 50 minutes, 6 to 40 minutes, 8 to 30 minutes, 10 to 20 minutes, 12 to 18 minutes, and 14 to 16 minutes. If the time required for centrifugation is 2 minutes or more, the purity of MFG-E8 can be increased, which is preferable. In addition, if the time required for the centrifugation is 60 minutes or less, it is not a time required for the excessive centrifugation, and it can be efficiently centrifuged.
  • the temperature for centrifugation is, for example, 0-20 ° C, 0-15 ° C, 0-12 ° C, 0-10 ° C, 0-8 ° C, 0-7 ° C, 0-6 ° C, 0-5 ° C, 1 -5 ° C, 2-5 ° C, and 3-5 ° C. If the temperature at which the centrifugation is performed is 0 ° C. or higher, it is preferable that the milk or skim milk (for example, a product obtained by reducing skim milk powder) can be centrifuged without freezing. In addition, if the temperature at which the centrifugation is performed is 20 ° C. or less, it is particularly preferable because milk or skimmed milk that has not been heat-sterilized in advance (for example, a product obtained by reducing skimmed milk powder) can be hygienically centrifuged.
  • the whey fraction used as MFG-E8 of the present invention can qualitatively and quantitatively confirm the presence of MFG-E8 in these whey fractions using an antigen-antibody reaction.
  • the antigen-antibody reaction is carried out by reacting with MFG-E8 in the whey fraction using, for example, a reaction with an anti-MFG-E8 monoclonal antibody and a fluorescently labeled secondary antibody. The presence and concentration of MFG-E8 can be confirmed.
  • the whey fraction used as MFG-E8 of the present invention can appropriately set the amount added to the oil-in-water emulsion according to the stability of emulsification in the actual oil-in-water emulsion. .
  • the emulsion stabilizer of the present invention is not particularly limited with respect to the oil-in-water emulsion as long as the emulsion of the oil-in-water emulsion can be stabilized.
  • the emulsion stabilizer of the present invention derived from milk of mammals is unlike an emulsifier of a chemically synthesized product, has little artificial image, and has no worries about side effects due to ingestion due to its eating habits. Addition to the emulsion is preferred.
  • the oil-in-water emulsion that can be taken orally as referred to herein is, for example, a food that is an oil-in-water emulsion or a pharmaceutical that is an oil-in-water emulsion.
  • a food that is an oil-in-water emulsion or a pharmaceutical that is an oil-in-water emulsion.
  • the association of fat globules mixed in the oil-in-water emulsion tends to occur in a highly fluid system. Therefore, the emulsion stabilizer of the present invention can be expected to have an effect of stabilizing emulsification with respect to an oil-in-water emulsion having a fluid or paste-like fluid shape.
  • Examples of the food that is an oil-in-water emulsion in the present invention include milk (vegetable milk such as cow's milk, goat milk, buffalo milk, sheep milk, and sheep milk, soy milk, vegetable milk such as coconut milk, coffee, etc. Artificial milk such as whitener), dairy products, milk and / or dairy foods containing dairy products, and mayonnaise. Of these, dairy foods containing milk and / or dairy products are preferred.
  • dairy products refer to all processed milk, such as concentrated milk, whole milk powder, defatted concentrated milk, defatted milk powder, partially defatted concentrated milk, partially defatted milk powder, condensed milk, sweetened condensed milk, cream, butter , Fermented milk, cheese, whey, whey powder, milk protein concentrate, and whey protein concentrate.
  • the dairy food containing milk and / or dairy products is, for example, cow's milk, processed milk, ingredient-adjusted milk, low-fat milk, milk drink, coffee-containing milk drink, fruit-containing milk drink, mineral-enriched milk drink, Contains vitamin-enriched milk drinks, functional ingredient-enriched milk drinks, yogurt, lactic acid bacteria drinks, whey drinks, acidic milk drinks, cheese, condensed milk, sweetened condensed milk, fresh cream, compound cream, vegetable fat cream, and coffee whitener .
  • the pharmaceutical that is an oil-in-water emulsion in the present invention is not particularly limited as long as it is a known pharmaceutical.
  • MFG-E8 derived from the same food or MFG-E8 derived from the same animal is used for the purpose of preventing contamination with allergens. It is preferable to use it.
  • the emulsion stabilizer of the present invention is MFG-E8 derived from milk (derived from cow).
  • milk derived from cow
  • the emulsion stabilizer and the oil-in-water emulsion are all derived from milk, and can be made into high value-added products such as “processed milk” standards or “component-adjusted milk” standards instead of “milk beverage” standards. .
  • the emulsion stabilization method of the present invention is characterized by including the step of adding MFG-E8, which is a protein, to an oil-in-water emulsion.
  • MFG-E8 of the present invention before being added to the oil-in-water emulsion is, for example, purified MFG-E8, partially purified MFG-E8, and milk or skim milk (for example, A supernatant fraction (so-called whey fraction) from which the casein has been removed by centrifuging the reduced skim milk powder) can be used.
  • MFG-E8 of the present invention is not limited, and examples thereof include liquid, paste, powder, and frozen.
  • the origin of MFG-E8 of the present invention is not limited. For example, it is derived from milk (from human milk, from mouse milk, from rat milk, from porcine milk, from bovine milk, etc.) and from skim milk ( Derived from human milk, derived from mouse milk, derived from rat milk, derived from pig milk, derived from bovine milk, and the like.
  • milk or skimmed milk for example, reduced skim milk powder
  • the supernatant fraction (so-called whey fraction) that has been centrifuged under conditions to remove casein is a fraction that is so-called industrial waste in the casein production process that is important as a milk protein raw material.
  • whey fraction preferably contains a large amount of MFG-E8 of the present invention.
  • the whey fraction used as the MFG-E8 of the present invention before being added to the oil-in-water emulsion is milk or skim milk (for example, reduced skim milk powder). Obtained by centrifugation.
  • the centrifugation conditions are not limited to the centrifugal force (number of rotations), the time required for the centrifugation, the temperature at which the centrifugation is performed, and the like.
  • Centrifugal force is, for example, 5000-100000 ⁇ g, 6000-80000 ⁇ g, 7000-60000 ⁇ g, 8000-40000 ⁇ g, 9000-30000 ⁇ g, 10000-20000 ⁇ g, 12000-18000 ⁇ g, and 14000. ⁇ 16000 ⁇ g.
  • a centrifugal force of 5000 ⁇ g or more is preferable because the purity of MFG-E8 can be increased.
  • the centrifugal force is 100000 ⁇ g or less, it is not excessive centrifugal force and can be efficiently centrifuged, which is preferable.
  • the time required for centrifugation is, for example, 2 to 60 minutes, 4 to 50 minutes, 6 to 40 minutes, 8 to 30 minutes, 10 to 20 minutes, 12 to 18 minutes, and 14 to 16 minutes. If the time required for centrifugation is 2 minutes or more, the purity of MFG-E8 can be increased, which is preferable. In addition, if the time required for the centrifugation is 60 minutes or less, it is not a time required for the excessive centrifugation, and it can be efficiently centrifuged.
  • the temperature for centrifugation is, for example, 0-20 ° C, 0-15 ° C, 0-12 ° C, 0-10 ° C, 0-8 ° C, 0-7 ° C, 0-6 ° C, 0-5 ° C, 1 -5 ° C, 2-5 ° C, and 3-5 ° C. If the temperature at which the centrifugation is performed is 0 ° C. or higher, it is preferable that the milk or skim milk (for example, a product obtained by reducing skim milk powder) can be centrifuged without freezing. In addition, if the temperature at which the centrifugation is performed is 20 ° C. or less, it is particularly preferable because milk or skimmed milk that has not been heat-sterilized in advance (for example, a product obtained by reducing skimmed milk powder) can be hygienically centrifuged.
  • the whey fraction used as the MFG-E8 of the present invention before being added to the oil-in-water emulsion is the presence of MFG-E8 in these whey fractions. It can be confirmed qualitatively and quantitatively using antigen-antibody reaction.
  • the antigen-antibody reaction is carried out by reacting with MFG-E8 in the whey fraction using, for example, a reaction with an anti-MFG-E8 monoclonal antibody and a fluorescently labeled secondary antibody. The presence and concentration of MFG-E8 can be confirmed.
  • the whey fraction used as MFG-E8 of the present invention can appropriately set the amount added to the oil-in-water emulsion according to the stability of emulsification in the actual oil-in-water emulsion. .
  • the time when the emulsion stabilizer containing MFG-E8 of the present invention as an active ingredient is added to the oil-in-water emulsion is determined in advance in the production process of the oil-in-water emulsion.
  • a known method for stabilizing the emulsion of an oil-in-water emulsion is used.
  • the known stabilization method of the emulsification of the oil-in-water emulsion is, for example, the fat globule diameter by mechanically shearing the fat globules mixed in the oil-in-water emulsion with a homogenizer or the like.
  • the fat mixed in the oil-in-water emulsion can be reduced (homogenized) by mechanically shearing the sphere with a homogenizer or the like.
  • the addition of the oil-in-water emulsion of the emulsion stabilizer containing MFG-E8 of the present invention as an active ingredient can be used as a method to replace the preliminary emulsification before homogenizing the oil-in-water emulsion. .
  • the fat sphere diameter is reduced (homogenized).
  • the emulsion stabilizer containing MFG-E8 of the present invention as an active ingredient can also be added to the oil-in-water emulsion.
  • an oil-in-water emulsion of an emulsion stabilizer containing MFG-E8 of the present invention increases the stability of emulsification of the oil-in-water emulsion, and makes MFG-E8 of the present invention an active ingredient
  • the oil-in-water emulsion can be stored for a longer period of time.
  • the oil-in-water emulsion is not particularly limited as long as the emulsion of the oil-in-water emulsion can be stabilized.
  • the emulsion stabilizer of the present invention derived from milk of mammals is unlike an emulsifier of a chemically synthesized product, has little artificial image, and has no worries about side effects due to ingestion due to its eating habits. Addition to the emulsion is preferred.
  • the oil-in-water emulsion that can be taken orally as referred to herein is, for example, a food that is an oil-in-water emulsion or a pharmaceutical that is an oil-in-water emulsion.
  • a food that is an oil-in-water emulsion or a pharmaceutical that is an oil-in-water emulsion.
  • the association of fat globules mixed in the oil-in-water emulsion tends to occur in a highly fluid system. Therefore, the emulsion stabilizer of the present invention can be expected to stabilize the emulsification with respect to an oil-in-water emulsion having a fluid shape such as liquid or paste.
  • examples of the food that is an oil-in-water emulsion include milk (animal milk such as milk, goat milk, buffalo milk, sheep milk, and sheep milk, soy milk, coconut milk, etc. Plant milk, artificial milk such as coffee whitener), dairy products, milk and / or dairy foods containing dairy products, and mayonnaise, among others, milk and / or dairy foods containing dairy products preferable.
  • milk animal milk such as milk, goat milk, buffalo milk, sheep milk, and sheep milk, soy milk, coconut milk, etc.
  • Plant milk artificial milk such as coffee whitener
  • dairy products milk and / or dairy foods containing dairy products
  • mayonnaise among others, milk and / or dairy foods containing dairy products preferable.
  • dairy products refer to all processed milk, such as concentrated milk, whole milk powder, defatted concentrated milk, defatted milk powder, partially defatted concentrated milk, partially defatted milk powder, condensed milk, sweetened condensed milk, cream, butter , Fermented milk, cheese, whey, whey powder, milk protein concentrate, and whey protein concentrate.
  • the dairy food containing milk and / or dairy products is, for example, cow's milk, processed milk, ingredient-adjusted milk, low-fat milk, milk drink, coffee-containing milk drink, fruit-containing milk drink, mineral-enriched milk drink, Contains vitamin-enriched milk drinks, functional ingredient-enriched milk drinks, yogurt, lactic acid bacteria drinks, whey drinks, acidic milk drinks, cheese, condensed milk, sweetened condensed milk, fresh cream, compound cream, vegetable fat cream, and coffee whitener .
  • the drug that is an oil-in-water emulsion is not particularly limited as long as it is a known drug.
  • the emulsion stabilizer containing the MFG-E8 of the present invention as an active ingredient is used for foods that are oil-in-water emulsions, the same is used for the purpose of preventing contamination of allergens. It is preferable to use MFG-E8 derived from food or MFG-E8 derived from the same animal.
  • the emulsion stabilizer containing MFG-E8 of the present invention as an active ingredient is derived from milk.
  • MFG-E8 derived from bovine
  • the emulsion stabilizer and the oil-in-water emulsion are all derived from milk, and can be made into high value-added products such as “processed milk” standards or “component-adjusted milk” standards instead of “milk beverage” standards. .
  • the amount added is 100 parts by mass of the oil-in-water emulsion.
  • purified product for example, 0.001 to 1 part by mass, 0.005 to 1 part by mass, 0.01 to 0.1 part by mass, 0.01 to 0.05 part by mass, 0.01 to 0 0.03 parts by mass and 0.02 to 0.03 parts by mass.
  • the amount added is 100 parts by mass of the oil-in-water emulsion.
  • the amount is 0.001 part by mass or more in terms of purified product, the effect of stabilizing the emulsion of the present invention is obtained, which is preferable.
  • the amount added is 100 mass of the oil-in-water emulsion.
  • Wild-type C57BL / 6 mice (WT) (purchased from Japan SLC) and MFG-E8-deficient C57BL / 6 mice (KO) (provided by Professor Shigekazu Nagata, graduate School of Medicine, Kyoto University) are used as experimental animals. did. These mice were fed with research food (manufactured by SLC, Japan) and adjusted so that the number of litters in the mice was 6-8 when the mother mice gave birth.
  • milk was milked 3 hours (W3h) and 48 hours (W48h) after forced weaning, and this was used as a milk sample.
  • These milk samples are dispensed into capped tubes and penicillin G and streptomycin are added in the presence of 57 ⁇ g / ml penicillin G and 100 ⁇ g / ml streptomycin for 24 hours at 37 ° C. The temperature was kept constant for 48 hours.
  • composition of these milk samples was confirmed by measuring the fat content by Triglyceride E-test kit (Wako Pure Chemical Industries) and the protein content by BCA assay kit (Thermo Fisher Scientific). These milk samples were centrifuged at 15000 ⁇ g for 15 minutes at 4 ° C., and the supernatant fraction was used as the whey fraction.
  • MFG-E8 in fat globules is an antigen in the fat globule (MFG) fraction, anti-mouse MFG-E8 antibody or anti-bovine MFG-E8 monoclonal antibody, and Alexa Fluor 488-labeled secondary antibody (Life Technology).
  • MFG fat globule
  • the antibody was reacted and the fat globules stained with immunofluorescence were observed with a phase-contrast fluorescence microscope (IX71, Olympus).
  • FIG. 1B The result of the distribution of fat globule diameter is shown in FIG. 1B.
  • milk samples extracted from the breast milk of MFG-E8-deficient C57BL / 6 mice (KO) are large (fat globule diameter 6.6 to 9.9 ⁇ m) or extra large (fat globule diameter 10 ⁇ m or more).
  • the ratio of the number of classified fat globules was determined for both the wild-type C57BL / milk sample obtained by milking milk 3 hours after forced weaning (W3h) and the milk sample milked 48 hours after forced weaning (W48h).
  • WT wild-type C57BL / milk sample obtained by milking milk 3 hours after forced weaning
  • W48h the milk sample milked 48 hours after forced weaning
  • MFG-E8 is a milk sample extracted from the breast milk of MFG-E8-deficient C57BL / 6 mice (KO), with the aim of reproducing the fat globule morphology of the milk sample that was milked 48 hours after forced weaning (W48h).
  • a milk sample extracted from the breast milk of a deficient C57BL / 6 mouse (KO) is dispensed into a capped tube and penicillin G and streptomycin are added so that 57 ⁇ g / ml penicillin G and 100 ⁇ g / ml streptomycin are present. After the addition, the temperature was kept constant at 37 ° C.
  • FIG. 2A As shown in FIG. 2A, the ratio of the number of fat globules classified as oversized in the milk samples of MFG-E8-deficient C57BL / 6 mice (KO) with holding for 24 hours (24 h) or 48 hours (48 h) increased.
  • the number of abnormal extra-large fat globules with a fat globule diameter of 15 ⁇ m or more should be maintained at 37 ° C. for 48 hours in milk samples extracted from the breast milk of MFG-E8-deficient C57BL / 6 mice (KO). The number increased significantly.
  • the number of abnormal extra-large fat globules having a fat globule diameter of 15 ⁇ m or more did not change even when kept at 37 ° C. for 48 hours.
  • a whey fraction (50% by mass) of the breast milk of another MFG-E8-deficient C57BL / 6 mouse was added and kept at 37 ° C. for 24 hours or 48 hours to maintain a constant temperature.
  • penicillin G and streptomycin were added to all milk samples so as to be in the presence of 57 ⁇ g / ml penicillin G and 100 ⁇ g / ml streptomycin.
  • the fat globules in these milk samples were observed with a phase contrast microscope. The result is shown in FIG. 3A.
  • WT wild-type C57BL / 6 mice
  • MFG-E8 MFG-E8 purified from milk
  • the ratio of the number of fat globules classified as extra large did not increase during the holding at 37 ° C. for 48 hours.
  • the whey fraction (50% by mass) of the breast milk of another MFG-E8-deficient C57BL / 6 mouse (KO) was added, the ratio of the number of fat globules classified as extra large increased.
  • MFG-E8-deficient C57BL / 6 mouse breast milk supplemented with whey fraction of wild-type C57BL / 6 mouse (WT) breast milk; MFG-E8 supplemented with MFG-E8 purified from cow's milk Breast milk of deficient C57BL / 6 mice (KO) (white circle); Breast milk of MFG-E8 deficient C57BL / 6 mice (KO) mice supplemented with whey fraction of breast milk of MFG-E8 deficient C57BL / 6 mice (KO) ( Black triangles): The results expressed as mean ⁇ SD for breast milk (black circles) of MFG-E8-deficient C57BL / 6 mice (KO) supplemented with phosphate buffered saline are shown in FIG. 3B.
  • WT wild-type C57BL / 6 mice
  • MFG-E8 MFG-E8 purified from milk
  • the milk sample to which was added the number of abnormal extra-large fat globules having a fat globule diameter of 15 ⁇ m or more did not increase even when maintained at 37 ° C. for 48 hours.
  • the milk sample (50% by mass) of the breast milk of another MFG-E8-deficient C57BL / 6 mouse (KO) and the milk sample added with phosphate buffered saline were kept at 37 ° C. for 48 hours.
  • the number of abnormally large fat globules with fat globule diameters of 15 ⁇ m or more increased.
  • the whey fraction (50% by mass) of the breast milk of another MFG-E8-deficient C57BL / 6 mouse (KO) was added, and at 37 ° C. for 1, 2, 3, 4, or 7 days Hold at constant temperature.
  • sodium azide was added to each so that it might become final concentration 0.1%.
  • the milk fat globule fraction immediately after milking was prepared by centrifuging 1 ml of milk (300 ⁇ g, 10 minutes, 20 ° C.) and removing whey and casein (300 ⁇ g ppt) with an injection needle. 500 ⁇ l of TBS (10 mM Tris-HCl buffer, 150 mM sodium chloride) was added and centrifuged again (300 ⁇ g, 10 minutes, 20 ° C.) to wash the fat globules. Washing with TBS was repeated three times, and this was used as the fat globule fraction.
  • TBS Tris-HCl buffer, 150 mM sodium chloride
  • the emulsion stabilizer of the oil-in-water emulsion of the present invention contains a relatively small amount (addition), fat globules mixed in the oil-in-water emulsion are stably dispersed in water, and the oil-in-water emulsion This can contribute to the stabilization of the emulsification of the product.
  • the milk-derived emulsion stabilizer of the present invention can suppress the use of an emulsion stabilizer in a small amount as compared with conventional dairy foods containing milk and / or dairy products.

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Dairy Products (AREA)
  • General Preparation And Processing Of Foods (AREA)
  • Emulsifying, Dispersing, Foam-Producing Or Wetting Agents (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)

Abstract

Le procédé de l'invention fournit un agent de stabilisation d'émulsion dans lequel sont dispersés dans l'eau de manière stable des globules gras mélangés à une émulsion huile dans l'eau, y compris à raison d'une teneur (addition) de relativement faible quantité, et qui accroît la stabilité de l'émulsion huile dans l'eau. L'invention fournit, en outre, un procédé de stabilisation d'émulsion mettant en œuvre cet agent de stabilisation d'émulsion. L'agent de stabilisation d'émulsion de l'invention est caractéristique en ce qu'il comprend en tant que composant actif un ＭＦＧ－Ｅ８(globule gras de lait – facteur 8 EGF) consistant en une protéine. De plus, le procédé de stabilisation d'émulsion est caractéristique en ce qu'il inclut une étape au cours de laquelle le ＭＦＧ－Ｅ８ est additionné à l'émulsion huile dans l'eau. De préférence, l'émulsion huile dans l'eau consiste en un aliment lactique comprenant du lait et/ou un produit laitier, et le ＭＦＧ－Ｅ８consiste en un dérivé du lait.
PCT/JP2015/086095 2014-12-25 2015-12-24 Agent de stabilisation d'émulsion, et procédé de stabilisation d'émulsion mettant en œuvre celui-ci WO2016104642A1 (fr)

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