WO2010119956A1 - Agent d'amélioration du taux de survie de lactobacillus et procédé de conservation utilisant celui-ci - Google Patents

Agent d'amélioration du taux de survie de lactobacillus et procédé de conservation utilisant celui-ci Download PDF

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WO2010119956A1
WO2010119956A1 PCT/JP2010/056856 JP2010056856W WO2010119956A1 WO 2010119956 A1 WO2010119956 A1 WO 2010119956A1 JP 2010056856 W JP2010056856 W JP 2010056856W WO 2010119956 A1 WO2010119956 A1 WO 2010119956A1
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lactic acid
acid bacteria
survival
galactomannan
molecular weight
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PCT/JP2010/056856
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English (en)
Japanese (ja)
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雅之 鈴木
達哉 裏地
聡 槇島
望美 近藤
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物産フードサイエンス株式会社
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Publication of WO2010119956A1 publication Critical patent/WO2010119956A1/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
    • A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
    • A23C9/13—Fermented milk preparations; Treatment using microorganisms or enzymes using additives
    • A23C9/1307—Milk products or derivatives; Fruit or vegetable juices; Sugars, sugar alcohols, sweeteners; Oligosaccharides; Organic acids or salts thereof or acidifying agents; Flavours, dyes or pigments; Inert or aerosol gases; Carbonation methods
    • 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
    • A23C3/00—Preservation of milk or milk preparations
    • A23C3/08—Preservation of milk or milk preparations by addition of preservatives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
    • C08B37/0087—Glucomannans or galactomannans; Tara or tara gum, i.e. D-mannose and D-galactose units, e.g. from Cesalpinia spinosa; Tamarind gum, i.e. D-galactose, D-glucose and D-xylose units, e.g. from Tamarindus indica; Gum Arabic, i.e. L-arabinose, L-rhamnose, D-galactose and D-glucuronic acid units, e.g. from Acacia Senegal or Acacia Seyal; Derivatives thereof
    • 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
    • C12N1/04—Preserving or maintaining viable microorganisms
    • 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
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

Definitions

  • the present invention relates to an agent for improving the survival of lactic acid bacteria during storage such as refrigeration or freezing storage using galactomannan whose molecular weight is controlled by molecular weight reduction treatment, and a storage method using the same.
  • a lactic acid bacterium refers to a bacterium that uses saccharides as a raw material and does not produce a spoilage substance by fermentation and uses lactic acid as a main product, and as described in Non-Patent Document 1, a homolactic lactic acid bacterium that produces only lactic acid as a final product And heterolactic acid bacteria that produce alcohol and acetic acid in parallel.
  • lactic acid bacteria are known for industrial use. According to the definition using lactic acid production efficiency as an index, there is an interpretation that the genus Bifidobacterium is excluded from lactic acid bacteria, but since it produces lactic acid, it is generally considered as a lactic acid bacterium in a broad sense.
  • Representative bacteria that are often referred to as lactic acid bacteria, including the genus include the genus Lactobacillus, the genus Enterococcus, the genus Lactococcus, and the genus Pediococcus.
  • Leuconostoc There are six genera of Leuconostoc, and the former three are used for yogurt and intestinal regulating agents. In both cases, it is thought that by producing a large amount of lactic acid by fermentation and changing its own living environment to acidic, it inhibits the growth of other microorganisms and thereby suppresses contamination.
  • lactic acid bacteria inhibit the growth of other microorganisms, but are resistant to lactic acid produced by themselves. Low. Therefore, the number of viable bacteria decreases with time in the storage process before ingestion of the product added with lactic acid bacteria. Therefore, by adding an excessive amount of lactic acid bacteria at the time of production, it is possible to secure a sufficient number of living lactic acid bacteria at the expiration date and surpass the decrease in the number of living bacteria.
  • Patent Document 1 proposes a method for finding a strain of the genus Lactobacillus having a low killing rate even in a highly acidic environment at the end of the culture by a screening operation and improving survival.
  • Patent Document 2 describes the survival of Bifidobacterium sp. By culturing propionic acid or naphthoquinone ring compounds that promote the growth of Bifidobacterium sp. A method for improving the performance has been proposed. Further, in Patent Document 3, when Lactobacillus helveticus is used, co-fermentation with Lactobacillus acidophilus suppresses an increase in lactic acid acidity during storage and improves survival. A method has been proposed.
  • Patent Document 4 a food or drink containing Bifidobacterium is treated with a specific resin to selectively remove an acid that inhibits the survival of Bifidobacterium, Methods for improving the survival of bacteria belonging to the genus Bacteria have been proposed.
  • Patent Documents 5 to 12 propose methods for improving survival by adding a physiologically active compound to a medium or culture of lactic acid bacteria.
  • a physiologically active compound include sorbitol, bovine lactoferrin, bovine apolactoferrin, bovine lactoferrin iron, malic acid or a salt thereof, lactitol, phospholipid, dead lactic acid bacteria, manganese, etc.
  • the method of improving the survival using the improved survival strain found by making full use of the screening operation sacrifices versatility in exchange for high specificity, and is widely used in industrial applications.
  • the desired lactic acid bacteria intended by the practitioner cannot be survived.
  • Techniques that secrete growth-promoting substances of the target lactic acid bacteria by co-fermentation or suppress the production of survival inhibitory substances are also effective for improving survival, but have high specificity for the combination of strains, Lactic acid bacteria intended by the practitioner cannot survive.
  • Technology that improves survival by adsorbing / removing acidic substances with resin can achieve the purpose of improving survival, but the acid adsorption / removal process is complicated, and it is actually a high-value-added product.
  • the application mode is extremely limited because it is not applicable to solid lactic acid bacteria foods.
  • the method of improving the survival by adding a physiologically active compound to the culture medium or culture of lactic acid bacteria has proven its effectiveness, although there are unclear parts in its mechanism of action. While having versatility and simplicity, high concentrations of physiologically active compounds are required to improve the survival of lactic acid bacteria depending on the type of the compound. In this case, the food and drink containing the lactic acid bacteria has an influence on the taste.
  • Orla-Jensen, S.M. The Lactic Acid Bacteria, Host and Son, Copenhagen (1919) Sir Erie Metnikov, “The Longevity of Life”, University of Japan Civilization Association Office, p236 (1912) Honmichi: History of bifidobacteria research, bifidobacteria research, Tomokazu Kitaoka, Japan bifidobacteria center, Tokyo p1 (1994) Japanese Patent Laid-Open No. 2003-219861 JP 7-227207 A Japanese Patent Laid-Open No. 10-099018 Japanese Patent Laid-Open No.
  • the present invention has been made to solve the above-described problems, and is prepared from a medium or culture by adding an appropriate physiologically active compound to a medium or culture of lactic acid bacteria in a trace amount and at an optimum concentration.
  • An object of the present invention is to provide an agent for improving the survival of lactic acid bacteria in storage of food and drink, and a method using the same.
  • the lactic acid bacteria survival improver of the present invention which has been made to achieve the above object, has a molecular weight controlled to 5 to 310 kDa, a main chain of D-mannose units bonded with ⁇ -1,4, and ⁇ -1,6.
  • a low molecular weight-treated galactomannan having a side chain of bound D-galactose units and having a composition ratio of D-mannose to D-galactose of 2.5: 1 to 15: 1 is contained as an active ingredient.
  • Such a low molecular weight treatment galactomannan can be obtained by subjecting at least one kind of galactomannan selected from a group of different galactomannans to a low molecular weight treatment.
  • the lactic acid bacteria survival improver is added to the culture medium or culture of the lactic acid bacteria, so that the culture medium or the culture and the food and drink containing it This improves the survival of the lactic acid bacteria during storage.
  • the storage may be low-temperature storage selected from refrigerated storage and frozen storage.
  • the addition amount of the low molecular weight-treated galactomannan is 0.01 to 1.0% by weight with respect to the weight of the medium or the culture.
  • the lactic acid bacterium-containing food or drink of the present invention is a product to which a lactic acid bacterium survival improver is added.
  • the lactic acid bacteria survival improver of the present invention and the preservation method for improving the survival of lactic acid bacteria using the same will be described in detail later, but the lactic acid bacteria containing a low molecular weight-treated galactomannan as an active ingredient
  • a survival improver to the medium or culture of lactic acid bacteria, it is possible to improve the survival of lactic acid bacteria during storage such as distribution and storage of the culture medium / culture and foods and drinks produced therefrom. Has the function and effect.
  • the storage method of the present invention when the culture medium / culture product or food and drink containing lactic acid bacteria are stored, especially refrigerated or frozen, the decrease in the number of surviving lactic acid bacteria is small even after storage for 3 weeks.
  • the survivability during distribution storage is stable at a high value, and the original purpose of using lactic acid bacteria in a medium / culture or food / drink can be achieved.
  • the storage method of the present invention can be carried out simply, inexpensively and safely by using an appropriate amount of the survival improver of lactic acid bacteria of the present invention in a small amount in a medium / culture or food / drink.
  • a storage method for improving the survival of lactic acid bacteria using a lactic acid bacteria survival improver to which the present invention is applied and a method for storing lactic acid bacteria by the number of times of freezing when the lactic acid bacteria culture medium is frozen and stored in a storage method not applying the present invention It is a figure which shows the comparison of residual property.
  • another storage method for improving the viability of lactic acid bacteria using the lactic acid bacteria survival improver to which the present invention is applied and in the refrigerated storage time when the lactic acid bacteria culture medium is refrigerated and stored in a storage method not applying the present invention. It is a figure which shows the comparison of lactic acid bacteria survivability of.
  • the preservation method for improving the survival of lactic acid bacteria using the lactic acid bacteria survival improver of the present invention is a specific polysaccharide having a high water retention function as an active ingredient, that is, a low molecular weight reduction. Addition of lactic acid bacteria survival improver containing treated galactomannan (lower molecular weight-treated galactomannan) to 0.01 to 1.0% by weight with respect to the medium or culture of lactic acid bacteria To solve this problem.
  • the food and drink to which the survival improver of lactic acid bacteria of the present invention is added solves the above technical problem by using a galactomannan subjected to a low molecular weight treatment as an effective and essential component. is there.
  • the addition amount of the galactomannan subjected to the low molecular weight treatment is less than 0.01% by weight relative to the medium or culture of the lactic acid bacteria, and the survival of the lactic acid bacteria is almost the same as in the non-added group, The addition effect of the said galactomannan is hardly seen.
  • the addition amount exceeds 1.0% by weight the survival of lactic acid bacteria loses the dependency on the addition amount and almost reaches a plateau, so that the survival improvement effect according to the addition amount cannot be obtained. .
  • the medium used in the present invention includes milk such as MRS medium and Briggsliver broth medium in addition to a medium obtained by adding a growth promoting substance to whole milk, skim milk, or reduced milk from these milk powders. It is also possible to apply a medium that does not contain. In addition, fermentation in these culture media is performed under aerobic conditions.
  • the obtained culture may be used for food as it is as a food containing lactic acid bacteria, or may be used for beverages as dairy lactic acid bacteria fermented dairy products after adjusting the taste and texture.
  • Lactic acid bacteria that improve the viability used in the present invention are not particularly limited, and include, for example, the aforementioned Bifidobacterium genus, Lactobacillus genus, Enterococcus genus, Lactococcus genus, Pediococcus genus, Leuconostoc genus, and the like, more specifically Lactobacillus. . casei, L .; acidophilus, L. et al. Helveticus is a preferred example.
  • the galactomannan used as a raw material for the low molecular weight treated galactomannan has a main chain of ⁇ -1,4-linked D-mannose units and a side chain of ⁇ -1,6-linked D-galactose units.
  • the origin of the galactomannan is not limited to the raw material, i.e., the origin, and need not be highly purified.
  • locust bean seed powder is preferable, but as other galactomannan, guar gum, cassia gum, soy bean full derived from soybean seed coat, tamson gum and the like can be mentioned. These are highly safe substances that are used as thickeners in the food industry and as tableting aids in the pharmaceutical industry.
  • galactomannans of different origins differ in the composition ratio of mannose units and galactose units, they may be derived from a single source or a galactomannan group consisting of a mixture of different sources as raw materials for the low molecular weight processed galactomannan. It may also be an aqueous solution or suspension. Furthermore, there is no restriction
  • the molecular structure obtained after the treatment has a molecular weight of 5 to 310 kDa, a main chain of D-1,4-linked D-mannose units and ⁇ -1,6 bonds. It must have a side chain of D-galactose units so that the constituent ratio of D-mannose and D-galactose is 2.5: 1 to 15: 1.
  • a method for adjusting the ratio is as follows: Alternatively, a chemical method using an acid catalyst or the like, or a physical method such as a high-pressure hydrothermal reaction using heat or pressure may be used.
  • Microorganisms used for fermentation may be naturally occurring wild strains or mutant strains obtained using genetic recombination techniques, but in terms of fermentation rate, ease of handling, safety, etc.
  • Yeast is practical. Many food yeasts can be used in the present invention, but are preferably selected from the genus Candida and the genus Saccharomyces.
  • the genus Phaffia the genus Phodotorula, the genus Zygosaccharomyces, the genus Kluyveromyces, the genus Torulaspora, and the like can also be used. These can be cultured alone or in combination.
  • the above fermentation is performed by a degrading enzyme that yeast inductively produces with galactomannan as a raw material.
  • Degrading enzymes that are inductively produced by yeast contain ⁇ -galactosidase, ⁇ -mannosidase, and ⁇ -mannanase, and the enzyme group catalyzes a low molecular weight reaction of galactomannan as a raw material. Therefore, for production of the low molecular weight-treated galactomannan, not only live yeasts but also the degrading enzyme group, hemicellulase showing substrate specificity for galactomannan and the like can be used.
  • the enzyme may be a naturally occurring wild type or a mutant obtained by subcloning, glycosylation or amino acid modification using gene recombination techniques. Further, it may be a fusion protein with other protein or peptide, or an enzyme fragment.
  • the amount of ⁇ -galactosidase, ⁇ -mannosidase, and ⁇ -mannanase used is not particularly limited, but is preferably 0.0001 unit or more and 1000 units or less per mL of the reaction solution, and more preferably 0.00 per unit of reaction solution. 001 unit or more and 100 unit or less.
  • One unit of ⁇ -galactosidase and ⁇ -mannosidase is defined as the amount of enzyme that produces 1 mmol of p-nitrophenol per minute from a p-nitrophenolated substrate at 37 ° C. and pH 7.5.
  • One unit of ⁇ -mannanase is defined as the amount of enzyme showing a reducing power equivalent to 1 mmol of mannose from mannan at 37 ° C. and pH 7.5.
  • Yeast and enzyme groups that are used as a catalyst or a catalyst for the production of the low molecular weight treatment galactomannan can be subjected to a low molecular weight treatment in a free form or an immobilized form.
  • the immobilized form refers to cross-linking to a support and inclusion / encapsulation in a semipermeable membrane.
  • Candida utilis was inoculated in a liquid medium containing 0.5% by weight locust bean seed powder as a raw material galactomannan, ammonium sulfate and a trace amount of inorganic salts as a nitrogen source, and liquid culture was performed at 30 ° C. for 24 hours. After completion of the culture, the supernatant was concentrated and dialyzed to obtain a crude galactomannan extract with a reduced molecular weight. This was subjected to gel filtration chromatography to obtain a fraction having a molecular weight of about 270 kDa.
  • Lactic acid dehydrogenase is an enzyme with extremely low resistance to freezing denaturation.
  • the residual activity of the enzyme after freezing and thawing is an indicator of the effect of preventing freezing denaturation.
  • Untreated galactomannan and the low molecular weight-treated galactomannan obtained in Example 1 were added to lactate dehydrogenase to a final concentration of 0.005% by weight, cooled at a rate of 1 ° C. per minute, ⁇ It was kept frozen at 20 ° C. for 24 hours.
  • the residual activity in the untreated galactomannan addition group was 45%, whereas in the low-molecular-weight treated galactomannan addition group, 100% remaining Showed activity.
  • the low molecular weight-treated galactomannan was found to have an excellent antifreezing effect at an extremely low concentration of 0.005%.
  • Preparation Example 2 Dissolve 520 g of whole milk powder and 0.75 g of yeast extract in pure water to make 2.1 L. Dispense 300 mL into triangular Kolben and sterilize in a hot water bath at 95 ° C. for 30 minutes. A whole fat medium was used. Here, in the same whole fat medium, L. 1.0 mL of a culture solution obtained by growing caseiJCM1134 strain to 1.0 ⁇ 10 9 cfu / mL was inoculated and statically cultured at 30 ° C. for 18 hours.
  • the appropriate diluted culture solution and a solution of pure water, erythritol, lactitol, xylitol, or low molecular weight-treated galactomannan are 1: 1 was followed, and the time-dependent change in the number of viable bacteria during freezing or refrigerated storage was followed.
  • the number of viable bacteria in the storage test is 100 mL by dissolving 1.0 g glucose, 0.5 g milk casein hydrolyzate, 0.3 g yeast extract, 0.3 g malt extract, and 1.0 g agar in pure water. Of these, 10 mL was dispensed into a petri dish and used as a plate medium for viable cell count measurement. In this flat plate medium, 1.0 mL of the liquid medium after storage was seeded and left to stand overnight at 30 ° C., and the formed colonies were counted.
  • Example 1 L After culturing casei in the above liquid medium, pure water (D.W.), erythritol (ERT), lactitol (LCT), low molecular weight-treated galactomannan (1.0% 10 9 cfu / mL) (SCLBG). The final concentration of each was set to 0.50%. This was frozen at ⁇ 18 ° C. and then thawed repeatedly at room temperature to monitor changes in the number of viable bacteria. The result is shown in FIG.
  • Example 2 After culturing casei in the liquid medium, it was diluted with pure water, erythritol, lactitol, and low molecular weight-treated galactomannan so as to be 2.0 ⁇ 10 10 cfu / mL. At this time, final concentrations of erythritol, lactitol, and xylitol were set to 0.4 mol / L, and final concentrations of low molecular weight-treated galactomannan were set to 0.05 wt%, 0.1 wt%, and 0.5 wt%. . This was followed over 3 weeks for changes in the number of viable bacteria during storage at 4 ° C. The result is shown in FIG.
  • Example 3 10 g of commercially available yogurt starter was cultivated in the above liquid medium in 500 g of skim milk sterilized at 80 ° C. for 10 minutes in a water bath. Casei was added and mixed with 5.0 mL of galactomannan having a reduced molecular weight as a lactic acid bacteria survival improver, and dispensed into a cup by 50 mL. The final concentration of the low molecular weight treatment galactomannan at this time was set to 0.20%. After the preparation, it was fermented at 40 ° C. for 5 hours to produce yogurt. As a control, yogurt was prepared with the same composition except that the low molecular weight-treated galactomannan was removed. Changes in the number of viable bacteria during storage of these prepared yogurts during storage at 4 ° C. were followed over 3 weeks. The results are shown in FIG.
  • the lactic acid bacteria are stable for a long period of time even if the medium or culture of lactic acid bacteria and foods and drinks containing them are refrigerated and frozen using the lactic acid bacteria survival improver of the present invention. Therefore, it was shown that the effective period of foods and drinks containing lactic acid bacteria can be set as long as several days to several weeks.
  • this invention contributes to the improvement of the storage technique in the product group which appealed for the utilization of live lactic acid bacteria, and also contributes to the improvement of the distribution system.

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Abstract

La présente invention concerne une substance, dont des quantités très faibles sont très efficaces dans l'amélioration du taux de survie de lactobacillus conservés au réfrigérateur ou au congélateur. L'agent d'amélioration du taux de survie de lactobacillus contient, en tant que substance active, un polysaccharide spécifique à forte rétention d'eau, c'est-à-dire une galactomannane traitée pour réduire le poids moléculaire de celle-ci (galactomannane à faible poids moléculaire). En ajoutant l'agent d'amélioration du taux de survie de lactobacillus à une culture ou un milieu de culture de lactobacillus de sorte que la concentration finale soit comprise entre 0,01 % et 1,0 % en poids, le taux de survie des lactobacillus étant conservés au réfrigérateur ou au congélateur peut être amélioré. Cela permet en outre la production d'aliments et de boissons auxquels un agent d'amélioration du taux de survie de lactobacillus, contenant une galactomannane de faible poids moléculaire en tant que substance active, a été ajouté.
PCT/JP2010/056856 2009-04-17 2010-04-16 Agent d'amélioration du taux de survie de lactobacillus et procédé de conservation utilisant celui-ci WO2010119956A1 (fr)

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005110107A1 (fr) * 2004-05-19 2005-11-24 Morinaga Milk Industry Co., Ltd. Lait fermenté
JP2008143986A (ja) * 2006-12-07 2008-06-26 Nikken Kasei Kk 低分子化処理を施したガラクトマンナンおよびその製造法ならびにその使用方法

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005110107A1 (fr) * 2004-05-19 2005-11-24 Morinaga Milk Industry Co., Ltd. Lait fermenté
JP2008143986A (ja) * 2006-12-07 2008-06-26 Nikken Kasei Kk 低分子化処理を施したガラクトマンナンおよびその製造法ならびにその使用方法

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
HIROMI MANABE: "Suiyosei Shokumotsu Sen'i 'San Fiber' no Kino to Yoghurt eno Oyo", NEW FOOD INDUSTRY, vol. 40, no. 10, 1998, pages 27 - 32 *

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