WO2006106673A1 - Growth inhibitor for thermotolerant acidophilic bacterium, method for inhibition of growth of the bacterium, and method for production of acidic beverage - Google Patents

Growth inhibitor for thermotolerant acidophilic bacterium, method for inhibition of growth of the bacterium, and method for production of acidic beverage Download PDF

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Publication number
WO2006106673A1
WO2006106673A1 PCT/JP2006/306299 JP2006306299W WO2006106673A1 WO 2006106673 A1 WO2006106673 A1 WO 2006106673A1 JP 2006306299 W JP2006306299 W JP 2006306299W WO 2006106673 A1 WO2006106673 A1 WO 2006106673A1
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Prior art keywords
growth
heat
lactic acid
acidophilic bacteria
bacterium
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PCT/JP2006/306299
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French (fr)
Japanese (ja)
Inventor
Tomohiko Yokogawa
Daigo Ide
Kiyoshi Oshima
Takatoshi Shimizu
Shingo Nishimura
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Calpis Co., Ltd.
Nishimura, Hideo
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Priority to JP2007512751A priority Critical patent/JP4988560B2/en
Publication of WO2006106673A1 publication Critical patent/WO2006106673A1/en

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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
    • A01N37/36—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a singly bound oxygen or sulfur atom attached to the same carbon skeleton, this oxygen or sulfur atom not being a member of a carboxylic group or of a thio analogue, or of a derivative thereof, e.g. hydroxy-carboxylic acids
    • 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
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/42—Preservation of non-alcoholic beverages
    • A23L2/44—Preservation of non-alcoholic beverages by adding 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
    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
    • A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
    • A23L3/3463—Organic compounds; Microorganisms; Enzymes
    • 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
    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
    • A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
    • A23L3/3463—Organic compounds; Microorganisms; Enzymes
    • A23L3/3481—Organic compounds containing oxygen
    • A23L3/3508—Organic compounds containing oxygen containing carboxyl groups

Definitions

  • Growth inhibitor for heat-resistant acidophilic bacteria for heat-resistant acidophilic bacteria, method for inhibiting growth, and method for producing acidic beverage
  • the present invention relates to a growth inhibitor of heat-resistant acidophilic bacteria, a method for suppressing the growth of heat-resistant acidophilic bacteria using the same, a method for producing an acidic beverage in which the growth of heat-resistant acidophilic bacteria is suppressed, and the method It relates to the acidic beverage obtained by the above.
  • the heat-resistant acidophilic bacterium belongs to the genus Alicyclobacillus and is a soil bacterium isolated from apple juice in 1984, and has been confirmed to grow in juice so far.
  • thermostable eosinophilic bacterium is an aerobic or facultative anaerobic bacterium, a spore-forming gonococci showing gram-positive or irregular shape, with a growth temperature range of 40-70 ° C and a growth pH range of 2-6. It has the characteristics.
  • heat-resistant acidophilic bacteria form heat-resistant spores, they cannot be sterilized under the general sterilization conditions of soft drink water, and further grow in acidic beverages. Heat resistance Although acidophilic bacteria and their metabolites are not toxic, they cause damage to the appearance and flavor of acidic beverages.
  • Some fungi have also been reported to produce off-flavor components such as guaiacol and 2,6-dibromophenol, which greatly impairs the commercial value.
  • Patent Document 1 describes a bacteriostatic method that suppresses the growth of heat-resistant acidophilic bacteria without impairing the flavor of the beverage.
  • Sodium acetate has been reported as a substance having an effect.
  • Patent Document 2 reports a heat-resistant acidophilic bacterium inhibitor containing diglycerin myristic acid ester and Z or a salt thereof as an active ingredient, and an acid beverage containing the heat-resistant acidophilic bacterium inhibitor.
  • Non-Patent Document 1 describes that lysozyme, polylysine, protamine, acetic acid, fatty acid ester and nisin have a heat-resistant acidophilic bacterial growth inhibitory effect.
  • Patent Documents 1 and 2 describe comparative examples showing that lactic acid has no effect of inhibiting the growth of heat-resistant acidophilic bacteria.
  • Patent Document 1 Japanese Patent Laid-Open No. 2002-315546
  • Patent Document 2 JP 2003-160411 A
  • Non-Patent Document 1 Food stress environment and microorganisms pi 14- 115 (2004) SCIENCE FORUM Invention disclosure
  • Another object of the present invention is to provide an acidic beverage that can effectively suppress the growth of heat-resistant acidophilic bacteria and has a good flavor and appearance, and a method for producing the acidic beverage.
  • a growth inhibitor of heat-resistant acidophilic bacteria comprising as an active ingredient an effective amount of lactic acid that suppresses the growth of heat-resistant acidophilic bacteria.
  • the growth inhibitor of heat-resistant acidophilic bacteria of the present invention contains an effective amount of lactic acid as an active ingredient to suppress the growth of heat-resistant acidophilic bacteria, it has an excellent effect of inhibiting the growth of heat-resistant acidophilic bacteria. Indicates.
  • the planting inhibitor is added to an object for suppressing the growth of heat-resistant acidophilic bacteria, so that the number of heat-resistant acidophilic bacteria can be effectively increased. Breeding can be suppressed.
  • the growth inhibitor of the present invention is added at any point in the production process, so that the growth of heat-resistant acidophilic bacteria is suppressed and the original flavor and appearance of the beverage are impaired. It is possible to efficiently produce an acidic beverage without any problems.
  • FIG. 1 is a graph showing the results of evaluation of the inhibition of the growth of heat-resistant acidophilic bacteria in a milky acidic beverage performed in Example 1.
  • FIG. 2 is a graph showing the results of evaluation of growth inhibition of heat-resistant acidophilic bacteria in an acidic soft drink containing citrate performed in Comparative Example 1.
  • FIG. 3 is a graph showing the results of a growth inhibition evaluation of thermotolerant acidophilic bacteria in an acidic microorganism growth medium performed in Example 2.
  • the growth inhibitor of the present invention contains an effective amount of lactic acid as an active ingredient that suppresses the growth of heat-resistant acidophilic bacteria.
  • Thermostable acidophilic bacteria are cells of the genus Alicyclobacillus, which are Alicyclobacillus acidoterrestris, Alicyclobacillus Examples include Alicyclobacillus acidocaldarius, Alicyclobacillus hesperidum, Alicyclobacillus cycloheptanicus and the like.
  • lactic acid that is an active ingredient may be lactic acid that is usually used in food additives such as commercially available lactic acid manufactured by PUREAC, or chemically synthesized lactic acid.
  • a material containing lactic acid can also be used, and examples of such a material include fermented lactic acid bacteria containing lactic acid and lactate such as calcium lactate and sodium lactate. Can be used as
  • the content ratio of lactic acid, which is an active ingredient is such that the heat-resistant acidophilic bacteria can easily grow depending on the type of the object to be suppressed in the growth of the heat-resistant acidophilic bacteria. Therefore, the effective amount can be appropriately selected and determined according to the type of the object.
  • the total amount of lactic acid relative to the total amount of the acidic beverage can be 0.075% by weight or more, preferably 0.075 to 3.0% by weight.
  • the total amount of lactic acid with respect to the total amount of the microorganism culture medium can be adjusted to an amount of 0.15% by weight or more.
  • the lactic acid content ratio of the active ingredient in the growth inhibitor of the present invention can be the amount obtained by subtracting the amount of lactic acid contained in advance. .
  • the growth inhibitor of the present invention may contain other components in addition to the above-mentioned active ingredients as long as the desired effects of the present invention are not impaired.
  • Other components include, for example, oxidizing agents such as citrate, phosphoric acid, adipic acid, malic acid, darconic acid, tartaric acid, acetic acid, fumaric acid and the like.
  • Additives usually used in preparations such as corn starch and inorganic salts.
  • the form of the growth inhibitor of the present invention is not particularly limited as long as it can be easily contained in an object such as food, and it may be liquid, powder, solid, or the like.
  • the method for inhibiting the growth of heat-resistant acidophilic bacteria according to the present invention is applied to a target for suppressing the growth of heat-resistant acidophilic bacteria such as foods such as acidic beverages, pharmaceuticals or their raw materials.
  • a target for suppressing the growth of heat-resistant acidophilic bacteria such as foods such as acidic beverages, pharmaceuticals or their raw materials.
  • the present invention Adding a growth inhibitor.
  • the growth inhibitor according to the present invention comprises a growth inhibitory agent according to the present invention containing an effective amount of lactic acid to inhibit the growth of the above-mentioned heat-resistant acidophilic bacteria according to the type of the object. It can be carried out by a method of adding an agent.
  • the method for producing an acidic beverage of the present invention is characterized in that the growth inhibitor of the present invention is added at the time of!
  • the acidic beverage is usually a soft drink having a pH of less than 4.6, and usually includes a fruit beverage, a vegetable beverage, a sports beverage, a lactic acid bacteria beverage, a lactic acid beverage, and a carbonated beverage.
  • Sports drinks, lactic acid bacteria drinks, lactic acid drinks and carbonated drinks may or may not contain fruit juice and Z or vegetable juice.
  • the step of producing the acidic beverage to which the growth inhibitor of the present invention is added Usually, it can be added before the final sterilization step.
  • the timing of addition may be divided into once or multiple times.
  • the amount of the growth inhibitor of the present invention to be added is an amount that gives a total lactic acid amount power of 0.075% by weight or more with respect to the total amount of the obtained acidic beverage, preferably 0.075 to 5%.
  • the amount can be 3.0% by weight.
  • the acidic beverage is a fruit beverage
  • the heat-resistant acidophilic bacteria are likely to grow, so that the total amount of lactic acid relative to the total amount of the obtained acidic beverage is preferably 0.15% by weight or more. If the content is small, the bacteriostatic action of sufficient heat-resistant acidophilic bacteria cannot be expected.
  • the acidity of the acidic beverage may be impaired due to the acidity derived from lactic acid.
  • an acidic beverage having a low fruit juice ratio for example, a fruit juice ratio of 3% by weight or less, a sufficient inhibitory effect can be obtained when the total amount of lactic acid relative to the total amount of the acidic beverage is 0.075 to 0.3% by weight.
  • the resulting acidic beverage contains 0.075 to 0.3% by weight of lactic acid, and further 0.15 to 0.3% by weight of lactic acid, and can effectively suppress the growth of thermotolerant acidophilic bacteria, and further does not impair the flavor and appearance. It is a beverage.
  • the container filled with the acidic beverage of the present invention is not particularly limited, and examples thereof include PET bottles, steel cans, aluminum cans, and paper packs.
  • % in an example means weight%.
  • YSG agar media shown in Table 1 Three YSG agar media shown in Table 1 were prepared, and the pH of each medium was adjusted to 3.70 using sulfuric acid or sodium hydroxide. Subsequently, each medium was separately inoculated with Alicyclobacillus acid telestris (ATCC4902 5), Alicyclobacillus acidocardarius (ATCC27009) or Alicyclobacillus cicum mouthpiece (ATCC49029), The cells were cultured at 45 ° C for 3 days. Next, the bacteria were scraped from each medium and suspended in 5 ml of 0.9 wt% physiological saline.
  • the number of bacteria in each suspension was measured with a microscope, and diluted with 0.9% by weight physiological saline so that the number of bacteria became 1.0 ⁇ 10 6 Zml.
  • a spore bacterial solution was prepared by maintaining at 80 ° C. for 10 minutes.
  • Lactic acid (Pureac) is added to the dairy drinking water shown in Table 2 so that the lactic acid concentration shown in Table 3 is reached, and the pH is adjusted to 3.70 using 10% or 10% sodium quenate. 200 ml of adjusted dairy acidic drinking water was prepared. Next, each of the milky acidic drinking waters of levels 1 to 6 was sterilized at 96 ° C. for 30 seconds, and 200 1 each of the spore bacteria solution prepared above was inoculated.
  • Each inoculated dairy acidic drinking water was statically cultured in an incubator at a constant temperature of 45 ° C for 0 days, 3 days, 7 days, and 14 days. After each culture, 50 1 was applied to each agar medium shown in Table 1, and the number of bacteria per ml was measured from the number of colonies formed. The results are shown in Figure 1.
  • thermotolerant acidophilic bacteria in acidic soft drinks containing citrate As shown in Table 2 prepared in Example 1 so that the concentration of citrate instead of lactic acid is as shown in Table 4 200 ml of acidic soft drinks, each of which was added to dairy drinking water having a composition and adjusted to pH 3.70 using 10% citrate or 10% sodium quenate, were prepared. Next, each beverage of level 1 to 3 was sterilized at 96 ° C for 30 seconds, and 200 1 each of the spore bacteria solution prepared in Example 1 was inoculated.
  • Each inoculated beverage was statically cultured in an incubator at a constant temperature of 45 ° C for 0 days, 3 days, 7 days, and 14 days. After each culture, 50 1 each was applied to the agar medium shown in Table 1 prepared in Example 1, and the number of bacteria per ml was measured from the number of colonies formed. The result is shown in figure 2.
  • the acidic soft drink containing citrate instead of lactic acid confirmed the growth of thermostable acidophilic bacteria, and the inhibitory effect was strong.
  • Lactic acid (manufactured by Pulac Co., Ltd.) was added to the agar medium having the composition shown in Table 1 prepared in Example 1 so that the lactic acid concentration shown in Table 5 was obtained, and 10% citrate or 10% sodium quenate was used. 200 ml of each microbial growth medium adjusted to pH 3.70 was prepared. Next, each medium of levels 1 to 6 was sterilized at 96 ° C. for 30 seconds, and 200 ⁇ l of each of the spore bacteria solution prepared in Example 1 was inoculated.
  • Each inoculated medium was statically cultured in an incubator at a constant temperature of 45 ° C for 0, 3, and 10 days. After each culture, 50 1 each was applied to the agar medium shown in Table 1 prepared in Example 1, and the number of bacteria per ml was measured from the number of colonies formed. The results are shown in Figure 3.

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Abstract

Disclosed are a growth inhibitor which is applied to a product intended to inhibit the growth of a thermotolerant acidophilic bacterium and has an excellent growth inhibitory effect against a thermotolerant acidophilic bacterium; a method for inhibiting the growth of the bacterium; an acidic beverage which can inhibit the growth of a thermotolerant acidophilic bacterium effectively and has good flavor and appearance; and a process for producing the acidic beverage. The growth inhibitor comprises, as the active ingredient, lactic acid in such an amount that is effective in inhibiting the growth of a thermotolerant acidophilic bacterium. The method for inhibiting the growth of the bacterium and the process for producing the acidic beverage utilize the growth inhibitor.

Description

明 細 書  Specification
耐熱性好酸性菌の増殖抑制剤、増殖抑制方法及び酸性飲料の製造方 法  Growth inhibitor for heat-resistant acidophilic bacteria, method for inhibiting growth, and method for producing acidic beverage
技術分野  Technical field
[0001] 本発明は、耐熱性好酸性菌の増殖抑制剤、これを用いた耐熱性好酸性菌の増殖 抑制方法、耐熱性好酸性菌の増殖が抑制された酸性飲料の製造方法及び該方法 により得られた酸性飲料に関する。  [0001] The present invention relates to a growth inhibitor of heat-resistant acidophilic bacteria, a method for suppressing the growth of heat-resistant acidophilic bacteria using the same, a method for producing an acidic beverage in which the growth of heat-resistant acidophilic bacteria is suppressed, and the method It relates to the acidic beverage obtained by the above.
背景技術  Background art
[0002] 酸性飲料の開発において、外観や風味を損ねないまま、長期保存することが望ま れている。酸性飲料において、外観や風味を損なう一つの要因としては、微生物の 増殖による影響が挙げられる。特に耐熱性好酸性菌の増殖による影響が問題となる 。耐熱性好酸性菌は、アリシクロバチルス (Alicyclobacillus)属に属し、 1984年にリンゴ 果汁から単離された土壌菌であって、これまでに果汁中での増殖が確認されている。 耐熱性好酸性菌は、好気性又は通性嫌気性の細菌で、グラム陽性あるいは不定形 を示す芽胞形成桿菌であり、発育温度域が 40〜70°C及び発育 pH域が 2〜6であると いう特徴を有する。また、耐熱性好酸性菌は、耐熱性芽胞を形成するため、清涼飲 料水の一般的な殺菌条件では殺菌できず、更に酸性の飲料中で増殖する。耐熱性 好酸性菌自体やその代謝物には毒性は無いが、酸性飲料の外観や風味を損なう要 因となる。また一部の菌で、グアイャコールや、 2,6-ジブロモフエノールといった異臭 成分を生成することも報告されており、商品価値が大きく損なわれる。  [0002] In the development of acidic beverages, it is desired to store them for a long time without impairing the appearance and flavor. One of the factors that impair the appearance and flavor of acidic beverages is the effect of microbial growth. In particular, the effect of the growth of heat-resistant acidophilic bacteria becomes a problem. The heat-resistant acidophilic bacterium belongs to the genus Alicyclobacillus and is a soil bacterium isolated from apple juice in 1984, and has been confirmed to grow in juice so far. A thermostable eosinophilic bacterium is an aerobic or facultative anaerobic bacterium, a spore-forming gonococci showing gram-positive or irregular shape, with a growth temperature range of 40-70 ° C and a growth pH range of 2-6. It has the characteristics. In addition, since heat-resistant acidophilic bacteria form heat-resistant spores, they cannot be sterilized under the general sterilization conditions of soft drink water, and further grow in acidic beverages. Heat resistance Although acidophilic bacteria and their metabolites are not toxic, they cause damage to the appearance and flavor of acidic beverages. Some fungi have also been reported to produce off-flavor components such as guaiacol and 2,6-dibromophenol, which greatly impairs the commercial value.
[0003] 上記問題を防止するために、製品中での耐熱性好酸性菌の増殖を抑制することが 必要となる。耐熱性好酸性菌の増殖抑制には、 UHT殺菌の実施、静菌剤の使用等 が挙げられる。しかし、 UHT殺菌は、通常の殺菌条件よりも過酷な条件であるため飲 料の風味や外観が損なわれる。また静菌剤の使用は、その有効量添カ卩により飲料の 風味を損なう恐れがある。従って、酸性飲料の風味を損なわない方法での耐熱性好 酸性菌の増殖抑制方法及び耐熱性好酸性菌の増殖抑制剤の開発が望まれている。  [0003] In order to prevent the above problems, it is necessary to suppress the growth of heat-resistant acidophilic bacteria in the product. Inhibiting the growth of heat-resistant acidophilic bacteria includes the implementation of UHT sterilization and the use of bacteriostatic agents. However, UHT sterilization is more severe than normal sterilization conditions, so the flavor and appearance of the drink are impaired. In addition, the use of bacteriostatic agents may impair the flavor of the beverage due to its effective amount. Therefore, development of a method for suppressing the growth of heat-resistant acidophilic bacteria and a growth-inhibiting agent for heat-resistant acidophilic bacteria by methods that do not impair the flavor of acidic beverages is desired.
[0004] 特許文献 1には、飲料の風味を損なわずに、耐熱性好酸性菌の増殖を抑える静菌 効果を有する物質として、酢酸ナトリウムが報告されている。また、特許文献 2には、 ジグリセリンミリスチン酸エステル及び Z又はその塩を有効成分として含む耐熱性好 酸性菌抑制剤、並びに該耐熱性好酸性菌抑制剤を含む酸性飲料が報告されて ヽる 。更に、非特許文献 1には、リゾチーム、ポリリジン、プロタミン、酢酸、脂肪酸エステル 及びナイシンに耐熱性好酸性菌増殖抑制効果があることが記載されている。 [0004] Patent Document 1 describes a bacteriostatic method that suppresses the growth of heat-resistant acidophilic bacteria without impairing the flavor of the beverage. Sodium acetate has been reported as a substance having an effect. Patent Document 2 reports a heat-resistant acidophilic bacterium inhibitor containing diglycerin myristic acid ester and Z or a salt thereof as an active ingredient, and an acid beverage containing the heat-resistant acidophilic bacterium inhibitor. . Furthermore, Non-Patent Document 1 describes that lysozyme, polylysine, protamine, acetic acid, fatty acid ester and nisin have a heat-resistant acidophilic bacterial growth inhibitory effect.
しかし、乳酸が、耐熱性好酸性菌の増殖抑制作用を示すことについては従来提案 が無い。尚、特許文献 1及び 2には、乳酸に、耐熱性好酸性菌の増殖抑制効果がな Vヽことを示す比較例が記載されて!、る。  However, there has been no proposal for lactic acid to show the effect of inhibiting the growth of heat-resistant acidophilic bacteria. Patent Documents 1 and 2 describe comparative examples showing that lactic acid has no effect of inhibiting the growth of heat-resistant acidophilic bacteria.
特許文献 1：特開 2002— 315546号公報  Patent Document 1: Japanese Patent Laid-Open No. 2002-315546
特許文献 2：特開 2003— 160411号公報  Patent Document 2: JP 2003-160411 A
非特許文献 1 :食品のストレス環境と微生物 pi 14- 115 (2004) SCIENCE FORUM 発明の開示  Non-Patent Document 1: Food stress environment and microorganisms pi 14- 115 (2004) SCIENCE FORUM Invention disclosure
発明が解決しょうとする課題  Problems to be solved by the invention
[0005] 本発明の課題は、酸性飲料や酸性の微生物増殖培地等の耐熱性好酸性菌の増 殖を抑制しょうとする対象物に使用し、優れた耐熱性好酸性菌の増殖抑制作用を示 す増殖抑制剤、該増殖抑制剤を利用した耐熱性好酸性菌の増殖抑制方法を提供 することにある。 [0005] An object of the present invention is to use an excellent anti-growth effect of thermotolerant acidophilic bacteria by using it for an object intended to inhibit the growth of thermotolerant acidophilic bacteria such as an acidic beverage or an acidic microorganism growth medium. It is an object of the present invention to provide a growth inhibitory agent and a method for inhibiting the growth of thermostable acidophilic bacteria using the growth inhibitor.
本発明の別の課題は、耐熱性好酸性菌の増殖を有効に抑制でき、風味や外観も 良好な酸性飲料及び該酸性飲料の製造方法を提供することにある。  Another object of the present invention is to provide an acidic beverage that can effectively suppress the growth of heat-resistant acidophilic bacteria and has a good flavor and appearance, and a method for producing the acidic beverage.
課題を解決するための手段  Means for solving the problem
[0006] 本発明者らは上記課題を解決するために鋭意研究を行った結果、耐熱性好酸性 菌の増殖を抑制しょうとする対象物が、ある一定量以上の乳酸を含有することで耐熱 性好酸性菌の増殖抑制効果を示すことを見出し、本発明を完成するに至った。 [0006] As a result of diligent research to solve the above problems, the present inventors have found that an object to suppress the growth of heat-resistant acidophilic bacteria contains a certain amount or more of lactic acid. The present invention has been completed by finding that it exhibits the growth-inhibiting effect of the acidophilic bacterium.
[0007] 本発明によれば、耐熱性好酸性菌の増殖を抑制する有効量の乳酸を有効成分とし て含む耐熱性好酸性菌の増殖抑制剤が提供される。 [0007] According to the present invention, there is provided a growth inhibitor of heat-resistant acidophilic bacteria comprising as an active ingredient an effective amount of lactic acid that suppresses the growth of heat-resistant acidophilic bacteria.
また本発明によれば、耐熱性好酸性菌の増殖を抑制しょうとする対象物に、上記増 殖抑制剤を添加する工程を含む耐熱性好酸性菌の増殖抑制方法が提供される。 更に本発明によれば、酸性飲料の製造工程のいずれかの時点で、上記増殖抑制 剤を添加する耐熱性好酸性菌の増殖を抑制した酸性飲料の製造方法が提供される 更にまた本発明によれば、上記製造方法により得られた、耐熱性好酸性菌の増殖 を抑制した酸性飲料が提供される。 In addition, according to the present invention, there is provided a method for inhibiting the growth of heat-resistant acidophilic bacteria, which comprises the step of adding the growth inhibitor to an object for which the growth of heat-resistant acidophilic bacteria is to be inhibited. Furthermore, according to the present invention, at any point in the production process of acidic beverage, A method for producing an acidic beverage that suppresses the growth of heat-resistant acidophilic bacteria to which an agent is added is provided. Furthermore, according to the present invention, an acid that suppresses the growth of heat-resistant acidophilic bacteria obtained by the above production method is provided. Beverages are provided.
また本発明によれば、耐熱性好酸性菌の増殖抑制剤を製造するための、乳酸の使 用が提供される。  Further, according to the present invention, use of lactic acid for producing a growth inhibitor of heat-resistant acidophilic bacteria is provided.
発明の効果  The invention's effect
[0008] 本発明の耐熱性好酸性菌の増殖抑制剤は、耐熱性好酸性菌の増殖を抑制する有 効量の乳酸を有効成分として含むので、優れた耐熱性好酸性菌の増殖抑制作用を 示す。本発明の耐熱性好酸性菌の増殖抑制方法は、該増植抑制剤を、耐熱性好酸 性菌の増殖を抑制しょうとする対象物に添加するので、有効に耐熱性好酸性菌の増 殖を抑制することができる。  [0008] Since the growth inhibitor of heat-resistant acidophilic bacteria of the present invention contains an effective amount of lactic acid as an active ingredient to suppress the growth of heat-resistant acidophilic bacteria, it has an excellent effect of inhibiting the growth of heat-resistant acidophilic bacteria. Indicates. In the method for inhibiting the growth of heat-resistant acidophilic bacteria according to the present invention, the planting inhibitor is added to an object for suppressing the growth of heat-resistant acidophilic bacteria, so that the number of heat-resistant acidophilic bacteria can be effectively increased. Breeding can be suppressed.
本発明の酸性飲料の製造方法は、製造工程のいずれかの時点で、本発明の増殖 抑制剤を添加するので、耐熱性好酸性菌の増殖が抑制され、飲料本来の風味や外 観を損なうことのない酸性飲料を効率良く製造することができる。  In the method for producing an acidic beverage of the present invention, the growth inhibitor of the present invention is added at any point in the production process, so that the growth of heat-resistant acidophilic bacteria is suppressed and the original flavor and appearance of the beverage are impaired. It is possible to efficiently produce an acidic beverage without any problems.
図面の簡単な説明  Brief Description of Drawings
[0009] [図 1]実施例 1で行った、乳性酸性飲料での耐熱性好酸性菌の増殖抑制評価の結果 を示すグラフである。  [0009] FIG. 1 is a graph showing the results of evaluation of the inhibition of the growth of heat-resistant acidophilic bacteria in a milky acidic beverage performed in Example 1.
[図 2]比較例 1で行った、クェン酸を含有した酸性の清涼飲料水での耐熱性好酸性 菌の増殖抑制評価の結果を示すグラフである。  FIG. 2 is a graph showing the results of evaluation of growth inhibition of heat-resistant acidophilic bacteria in an acidic soft drink containing citrate performed in Comparative Example 1.
[図 3]実施例 2で行った、酸性の微生物増殖培地での耐熱性好酸性菌の増殖抑制評 価の結果を示すグラフである。  FIG. 3 is a graph showing the results of a growth inhibition evaluation of thermotolerant acidophilic bacteria in an acidic microorganism growth medium performed in Example 2.
発明を実施するための最良の形態  BEST MODE FOR CARRYING OUT THE INVENTION
[0010] 以下本発明を更に詳細に説明する。 [0010] Hereinafter, the present invention will be described in more detail.
本発明の増殖抑制剤は、耐熱性好酸性菌の増殖を抑制する有効量の乳酸を有効 成分として含有する。  The growth inhibitor of the present invention contains an effective amount of lactic acid as an active ingredient that suppresses the growth of heat-resistant acidophilic bacteria.
耐熱性好酸性菌は、アリシクロバチルス (Alicyclobacillus)属の菌体であって、アリシ クロバチルス ·ァシドテレストリス (Alicyclobacillus acidoterrestris)、アリシクロバチルス · ァシドカルダリウス (Alicyclobacillus acidocaldarius)、アリシクロバチルス ·ヘスペリダム ( Alicyclobacillus hesperidum)、アリシクロバチルス ·シクロへプタ-カス (Alicyclobacillus cycloheptanicus)等が挙げられる。 Thermostable acidophilic bacteria are cells of the genus Alicyclobacillus, which are Alicyclobacillus acidoterrestris, Alicyclobacillus Examples include Alicyclobacillus acidocaldarius, Alicyclobacillus hesperidum, Alicyclobacillus cycloheptanicus and the like.
[0011] 本発明の増殖抑制剤において、有効成分である乳酸は、例えば、ピューラック社製 等の市販の乳酸、化学合成した乳酸等の通常食品添加剤に使用する乳酸を用いる ことができる。また、乳酸を含む材料を用いることもでき、このような材料としては、例 えば、乳酸を含む乳酸菌の発酵物や、乳酸カルシウム、乳酸ナトリウム等の乳酸塩が 挙げられ、使用に際しては単独若しくは混合物として用いることができる。  [0011] In the growth inhibitor of the present invention, lactic acid that is an active ingredient may be lactic acid that is usually used in food additives such as commercially available lactic acid manufactured by PUREAC, or chemically synthesized lactic acid. A material containing lactic acid can also be used, and examples of such a material include fermented lactic acid bacteria containing lactic acid and lactate such as calcium lactate and sodium lactate. Can be used as
[0012] 本発明の増殖抑制剤において、有効成分である乳酸の含有割合は、耐熱性好酸 性菌の増殖を抑制しょうとする対象物の種類によって、耐熱性好酸性菌の増殖し易 さが異なるため、該対象物の種類に応じて、その有効量を適宜選択して決定すること ができる。  [0012] In the growth inhibitor of the present invention, the content ratio of lactic acid, which is an active ingredient, is such that the heat-resistant acidophilic bacteria can easily grow depending on the type of the object to be suppressed in the growth of the heat-resistant acidophilic bacteria. Therefore, the effective amount can be appropriately selected and determined according to the type of the object.
例えば、対象物が酸性飲料の場合、該酸性飲料の全量に対する全乳酸量が 0.075 重量％以上となる量、好ましくは 0.075〜3.0重量％となる量とすることができる。また、 酸性の微生物培養培地の場合、微生物培養培地の全量に対する全乳酸量が、 0.15 重量％以上となる量とすることができる。  For example, when the object is an acidic beverage, the total amount of lactic acid relative to the total amount of the acidic beverage can be 0.075% by weight or more, preferably 0.075 to 3.0% by weight. In the case of an acidic microorganism culture medium, the total amount of lactic acid with respect to the total amount of the microorganism culture medium can be adjusted to an amount of 0.15% by weight or more.
従って、前記対象物が予め乳酸を含有する場合には、本発明の増殖抑制剤にお ける有効成分の乳酸含有割合は、該予め含有される乳酸量を差し引いた量とするこ とがでさる。  Therefore, when the object contains lactic acid in advance, the lactic acid content ratio of the active ingredient in the growth inhibitor of the present invention can be the amount obtained by subtracting the amount of lactic acid contained in advance. .
[0013] 本発明の増殖抑制剤は、前記有効成分の他に、本発明の所望の効果を損なわな い範囲で、他の成分を含んでも良い。他の成分としては、例えば、クェン酸、リン酸、 アジピン酸、リンゴ酸、ダルコン酸、酒石酸、酢酸、フマル酸等の酸化剤、更に、例え ば、デンプン、乳糖、白糖、マンニット、カルボキシメチルセルロース、コーンスターチ 、無機塩類等の製剤に通常用いられる添加剤等が挙げられる。  [0013] The growth inhibitor of the present invention may contain other components in addition to the above-mentioned active ingredients as long as the desired effects of the present invention are not impaired. Other components include, for example, oxidizing agents such as citrate, phosphoric acid, adipic acid, malic acid, darconic acid, tartaric acid, acetic acid, fumaric acid and the like. Additives usually used in preparations such as corn starch and inorganic salts.
本発明の増殖抑制剤の形態は、食品等の対象物に含有せしめ易い形態であれば 特に限定されず、液状でも、粉末又は固形の剤等の何れであっても良い。  The form of the growth inhibitor of the present invention is not particularly limited as long as it can be easily contained in an object such as food, and it may be liquid, powder, solid, or the like.
[0014] 本発明の耐熱性好酸性菌の増殖抑制方法は、酸性飲料等の食品、医薬品又はそ れらの原料等の、耐熱性好酸性菌の増殖を抑制しょうとする対象物に、上記本発明 の増殖抑制剤を添加する工程を含む。 [0014] The method for inhibiting the growth of heat-resistant acidophilic bacteria according to the present invention is applied to a target for suppressing the growth of heat-resistant acidophilic bacteria such as foods such as acidic beverages, pharmaceuticals or their raw materials. The present invention Adding a growth inhibitor.
前記本発明の増殖抑制剤の添カ卩は、対象物に、該対象物の種類に応じて、上述し た耐熱性好酸性菌の増殖を抑制する有効量の乳酸を含む本発明の増殖抑制剤を 添加する方法等により行なうことができる。  The growth inhibitor according to the present invention comprises a growth inhibitory agent according to the present invention containing an effective amount of lactic acid to inhibit the growth of the above-mentioned heat-resistant acidophilic bacteria according to the type of the object. It can be carried out by a method of adding an agent.
[0015] 本発明の酸性飲料の製造方法は、酸性飲料の製造工程の!/、ずれかの時点で、本 発明の増殖抑制剤を添加することを特徴とする。  [0015] The method for producing an acidic beverage of the present invention is characterized in that the growth inhibitor of the present invention is added at the time of!
前記酸性飲料は、通常、 pH4.6未満の清涼飲料水であって、通常、果実飲料、野 菜飲料、スポーツ飲料、乳酸菌飲料、乳酸飲料、炭酸飲料が挙げられる。スポーツ 飲料、乳酸菌飲料、乳酸飲料、炭酸飲料は、果汁及び Z又は野菜汁を含有してもし なくても良い。  The acidic beverage is usually a soft drink having a pH of less than 4.6, and usually includes a fruit beverage, a vegetable beverage, a sports beverage, a lactic acid bacteria beverage, a lactic acid beverage, and a carbonated beverage. Sports drinks, lactic acid bacteria drinks, lactic acid drinks and carbonated drinks may or may not contain fruit juice and Z or vegetable juice.
[0016] 本発明の酸性飲料の製造方法において、本発明の増殖抑制剤を添加する、酸性 飲料の製造工程の!/、ずれかの時点とは、上記各種酸性飲料の製造工程の!/、ずれの 時点でも良ぐ通常、最終の殺菌工程の以前に添加することができる。添加時期は、 1回でも複数回に分けても良 、。  [0016] In the method for producing an acidic beverage of the present invention, the step of producing the acidic beverage to which the growth inhibitor of the present invention is added! Usually, it can be added before the final sterilization step. The timing of addition may be divided into once or multiple times.
[0017] 本発明の酸性飲料の製造方法において、本発明の増殖抑制剤の添加量は、得ら れる酸性飲料の全量に対する全乳酸量力 S0.075重量％以上となる量、好ましくは 0.07 5〜3.0重量％となる量とすることができる。また、酸性飲料が、果実飲料の場合には、 耐熱性好酸性菌が増殖し易 ヽため、得られる酸性飲料の全量に対する全乳酸量が 0 .15重量％以上となる量が好ましい。少ない含有量では、十分な耐熱性好酸性菌の 静菌作用は期待できず、 0.3重量％より多い含有量では乳酸由来の酸味のために、 酸性飲料本来の風味を損なう恐れがある。但し、果汁の割合が低い、例えば、果汁 率が 3重量％以下の酸性飲料の場合には、酸性飲料の全量に対する全乳酸量が 0.0 75〜0.3重量％で十分な抑制効果が得られる。  [0017] In the method for producing an acidic beverage of the present invention, the amount of the growth inhibitor of the present invention to be added is an amount that gives a total lactic acid amount power of 0.075% by weight or more with respect to the total amount of the obtained acidic beverage, preferably 0.075 to 5%. The amount can be 3.0% by weight. Further, when the acidic beverage is a fruit beverage, the heat-resistant acidophilic bacteria are likely to grow, so that the total amount of lactic acid relative to the total amount of the obtained acidic beverage is preferably 0.15% by weight or more. If the content is small, the bacteriostatic action of sufficient heat-resistant acidophilic bacteria cannot be expected. If the content is more than 0.3% by weight, the acidity of the acidic beverage may be impaired due to the acidity derived from lactic acid. However, in the case of an acidic beverage having a low fruit juice ratio, for example, a fruit juice ratio of 3% by weight or less, a sufficient inhibitory effect can be obtained when the total amount of lactic acid relative to the total amount of the acidic beverage is 0.075 to 0.3% by weight.
従って、得られる酸性飲料は、 0.075〜0.3重量％の乳酸、更には 0.15〜0.3重量％ の乳酸を含み、有効に耐熱性好酸性菌の増殖を抑制でき、更に風味及び外観を損 なわない酸性飲料である。  Therefore, the resulting acidic beverage contains 0.075 to 0.3% by weight of lactic acid, and further 0.15 to 0.3% by weight of lactic acid, and can effectively suppress the growth of thermotolerant acidophilic bacteria, and further does not impair the flavor and appearance. It is a beverage.
[0018] 本発明の酸性飲料の製造方法においては、 pH4.0以下の清涼飲料水における 65 °Cで 10分間の加熱又は同等以上の加熱、 pH4.0以上 4.6未満の清涼飲料水における 85°Cで 30分間の加熱又は同等以上の加熱による殺菌工程を行うことができる。 [0018] In the method for producing an acidic beverage of the present invention, in soft drinks having a pH of 4.0 or less, heating at 65 ° C for 10 minutes or equivalent or higher, in soft drinks having a pH of 4.0 or more and less than 4.6 It can be sterilized by heating at 85 ° C for 30 minutes or equivalent or higher.
本発明の酸性飲料を充填する容器は特に限定されず、例えば、 PETボトル、スチー ル缶、アルミ缶、紙パック等が挙げられる。  The container filled with the acidic beverage of the present invention is not particularly limited, and examples thereof include PET bottles, steel cans, aluminum cans, and paper packs.
実施例  Example
[0019] 以下、本発明を実施例により更に詳細に説明するが、本発明はこれに限定されな い。尚、例中の％は、重量％を意味する。  Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited thereto. In addition,% in an example means weight%.
実施例 1〜3及び比較例 1〜3  Examples 1-3 and Comparative Examples 1-3
(1)芽胞菌液の調製  (1) Preparation of spore bacteria solution
表 1に示す YSG寒天培地を 3枚用意し、硫酸又は水酸ィ匕ナトリウムを用いて、各培 地の pHを 3.70に調整した。続いて、アリシクロバチルス ·ァシドテレストリス (ATCC4902 5)、アリシクロバチルス ·ァシドカルダリウス (ATCC27009)又はアリシクロバチルス ·シク 口へプタ-カス (ATCC49029)を各培地に別々に接種し、 45°Cで 3日間培養した。 次に、各培地から菌を搔きとり、 0.9重量％生理食塩水 5mlに懸濁させた。各懸濁液 の菌数を顕微鏡で測定し、菌数が 1.0 X 106個 Zmlとなるように、 0.9重量％生理食塩 水で希釈した。得られた各希釈液を等量ずつ混合し、芽胞の発芽効率を高めるため に、 80°C、 10分間保持して芽胞菌液を調製した。 Three YSG agar media shown in Table 1 were prepared, and the pH of each medium was adjusted to 3.70 using sulfuric acid or sodium hydroxide. Subsequently, each medium was separately inoculated with Alicyclobacillus acid telestris (ATCC4902 5), Alicyclobacillus acidocardarius (ATCC27009) or Alicyclobacillus cicum mouthpiece (ATCC49029), The cells were cultured at 45 ° C for 3 days. Next, the bacteria were scraped from each medium and suspended in 5 ml of 0.9 wt% physiological saline. The number of bacteria in each suspension was measured with a microscope, and diluted with 0.9% by weight physiological saline so that the number of bacteria became 1.0 × 10 6 Zml. In order to increase the germination efficiency of the spores by mixing equal amounts of each of the obtained dilutions, a spore bacterial solution was prepared by maintaining at 80 ° C. for 10 minutes.
[0020] [表 1] [0020] [Table 1]
Figure imgf000007_0001
Figure imgf000007_0001
[0021] (2)乳性酸性飲料水での耐熱性好酸性菌の増殖抑制評価  [0021] (2) Evaluation of growth inhibition of thermotolerant acidophilic bacteria in milky acidic drinking water
表 2に示す乳性飲料水に、表 3に示す乳酸濃度となるように乳酸 (ピューラック社製） を添カ卩し、 10%クェン酸又は 10%クェン酸ナトリウムを用いて pHを 3.70に調整した乳 性酸性飲料水 200mlをそれぞれ調製した。次いで、水準 1〜6の各乳性酸性飲料水を 、 96°C、 30秒間殺菌し、上記で調製した芽胞菌液をそれぞれ 200 1ずつ植菌した。  Lactic acid (Pureac) is added to the dairy drinking water shown in Table 2 so that the lactic acid concentration shown in Table 3 is reached, and the pH is adjusted to 3.70 using 10% or 10% sodium quenate. 200 ml of adjusted dairy acidic drinking water was prepared. Next, each of the milky acidic drinking waters of levels 1 to 6 was sterilized at 96 ° C. for 30 seconds, and 200 1 each of the spore bacteria solution prepared above was inoculated.
[0022] [表 2] 原材料 配合率 果糖ぶどう糖液糖 4.48 [0022] [Table 2] Raw materials Composition ratio Fructose Glucose liquid sugar 4.48
脱脂粉乳 0.300  Nonfat dry milk 0.300
大豆多糖類 0.100  Soy polysaccharide 0.100
水 95.12  Water 95.12
[0023] [表 3] [0023] [Table 3]
Figure imgf000008_0001
Figure imgf000008_0001
[0024] 植菌した各乳性酸性飲料水を、 45°C定温のインキュベータ一中で、 0日、 3日、 7日 、 14日間静置培養した。各培養後、表 1に示す寒天培地に 50 1ずつ塗布し、形成し たコロニー数から lml当たりの菌数を測定した。結果を図 1に示す。  [0024] Each inoculated dairy acidic drinking water was statically cultured in an incubator at a constant temperature of 45 ° C for 0 days, 3 days, 7 days, and 14 days. After each culture, 50 1 was applied to each agar medium shown in Table 1, and the number of bacteria per ml was measured from the number of colonies formed. The results are shown in Figure 1.
図 1から、乳性酸性飲料水中の乳酸濃度が 0.075%以上となる水準 4〜6において 有意に耐熱性好酸性菌の増殖を抑えることができた。  From Fig. 1, it was possible to significantly suppress the growth of heat-resistant acidophilic bacteria at levels 4 to 6 where the lactic acid concentration in the acidic milk water was 0.075% or higher.
[0025] 比較例 1 [0025] Comparative Example 1
(1)クェン酸を含有した酸性清涼飲料水での耐熱性好酸性菌の増殖抑制評価 乳酸の代わりにクェン酸を表 4に示す濃度となるように、実施例 1で調製した表 2に 示す組成の乳性飲料水に添加し、 10%クェン酸又は 10%クェン酸ナトリウムを用いて pHを 3.70に調整した酸性清涼飲料水 200mlをそれぞれ調製した。次いで、水準 1〜3 の各飲料を、 96°C、 30秒間殺菌し、実施例 1で調製した芽胞菌液をそれぞれ 200 1 ずつ植菌した。  (1) Evaluation of growth inhibition of thermotolerant acidophilic bacteria in acidic soft drinks containing citrate As shown in Table 2 prepared in Example 1 so that the concentration of citrate instead of lactic acid is as shown in Table 4 200 ml of acidic soft drinks, each of which was added to dairy drinking water having a composition and adjusted to pH 3.70 using 10% citrate or 10% sodium quenate, were prepared. Next, each beverage of level 1 to 3 was sterilized at 96 ° C for 30 seconds, and 200 1 each of the spore bacteria solution prepared in Example 1 was inoculated.
[0026] [表 4] クェン酸含有量 H [0026] [Table 4] Chenic acid content H
(重量％)  (% By weight)
水準 1 0.095 3.70  Level 1 0.095 3.70
水準 2 0.24 3.70  Level 2 0.24 3.70
水準 3 0.395 3.70 [0027] 植菌した各飲料を、 45°C定温のインキュベータ一中で、 0日、 3日、 7日、 14日間静 置培養した。各培養後、実施例 1で調製した表 1に示す寒天培地に 50 1ずつ塗布し 、形成したコロニー数から lml当たりの菌数を測定した。結果を図 2に示す。 Level 3 0.395 3.70 [0027] Each inoculated beverage was statically cultured in an incubator at a constant temperature of 45 ° C for 0 days, 3 days, 7 days, and 14 days. After each culture, 50 1 each was applied to the agar medium shown in Table 1 prepared in Example 1, and the number of bacteria per ml was measured from the number of colonies formed. The result is shown in figure 2.
図 2から、乳酸の代わりにクェン酸を含有させた酸性清涼飲料水では、耐熱性好酸 性菌の増殖が確認され、抑制効果は認められな力つた。  From Fig. 2, the acidic soft drink containing citrate instead of lactic acid confirmed the growth of thermostable acidophilic bacteria, and the inhibitory effect was strong.
[0028] 実施例 2  [0028] Example 2
(1)微生物増殖培地での耐熱性好酸性菌の増殖抑制評価  (1) Evaluation of growth inhibition of thermotolerant acidophilic bacteria in microbial growth medium
実施例 1で調製した表 1に示す組成の寒天培地に、表 5に示す乳酸濃度となるよう に乳酸 (ピューラック社製)を添加し、 10%クェン酸又は 10%クェン酸ナトリウムを用い て pHを 3.70に調整した微生物増殖培地をそれぞれ 200ml調製した。次いで、水準 1 〜6の各培地を、 96°C、 30秒間殺菌し、実施例 1で調製した芽胞菌液をそれぞれ 200 μ 1ずつ植菌した。  Lactic acid (manufactured by Pulac Co., Ltd.) was added to the agar medium having the composition shown in Table 1 prepared in Example 1 so that the lactic acid concentration shown in Table 5 was obtained, and 10% citrate or 10% sodium quenate was used. 200 ml of each microbial growth medium adjusted to pH 3.70 was prepared. Next, each medium of levels 1 to 6 was sterilized at 96 ° C. for 30 seconds, and 200 μl of each of the spore bacteria solution prepared in Example 1 was inoculated.
[0029] [表 5]  [0029] [Table 5]
Figure imgf000009_0001
Figure imgf000009_0001
[0030] 植菌した各培地を、 45°C定温のインキュベータ一中で、 0日、 3日、 10日間静置培養 した。各培養後、実施例 1で調製した表 1に示す寒天培地に 50 1ずつ塗布し、形成 したコロニー数から lml当たりの菌数を測定した。結果を図 3に示す。  [0030] Each inoculated medium was statically cultured in an incubator at a constant temperature of 45 ° C for 0, 3, and 10 days. After each culture, 50 1 each was applied to the agar medium shown in Table 1 prepared in Example 1, and the number of bacteria per ml was measured from the number of colonies formed. The results are shown in Figure 3.
図 3から、微生物増殖培地中の乳酸濃度が 0.15%以上となる水準 4〜6において有 意に耐熱性好酸性菌の増殖を抑えることができた。  From Fig. 3, it was possible to significantly suppress the growth of heat-resistant acidophilic bacteria at levels 4 to 6 where the lactic acid concentration in the microorganism growth medium was 0.15% or higher.
[0031] 実施例 3 [0031] Example 3
(1)果実飲料水での耐熱性好酸性菌の増殖抑制及び風味評価  (1) Growth suppression and flavor evaluation of thermotolerant acidophilic bacteria in fruit drinking water
表 6に示す組成の果実飲料水に、表 7に示す乳酸濃度及びクェン酸濃度となるよう に乳酸及びクェン酸を添加し、クェン酸ナトリウムを用いて pHを 3.70に調整した果実 飲料水をそれぞれ 200ml調製した。次いで、水準 1〜9の各飲料水を、 96°C、 30秒間 殺菌し、実施例 1で調製した芽胞菌液をそれぞれ 200 1ずつ植菌した。尚、表 6中の 55° オレンジ果汁の配合率は、 JAS法に基づいて換算すると果汁率は 3%となる。 植菌した各飲料水を、 45°C定温のインキュベータ一中で、 10日間静置培養した。各 培養後、実施例 1で調製した表 1に示す寒天培地に 50 ずつ塗布し、形成したコロ ニー数から lml当たりの菌数を測定した。増殖が抑制されたものを〇、抑制されなか つたものを Xとした。 Fruits with the composition shown in Table 6 added with lactic acid and citrate to the lactic acid concentration and citrate concentration shown in Table 7, and adjusted to pH 3.70 using sodium citrate. 200 ml of drinking water was prepared for each. Subsequently, each level 1-9 drinking water was sterilized at 96 ° C. for 30 seconds, and 200 1 each of the spore bacteria solution prepared in Example 1 was inoculated. The blending ratio of 55 ° orange juice in Table 6 is 3% when converted based on the JAS method. Each inoculated drinking water was statically cultured for 10 days in a 45 ° C constant temperature incubator. After each culture, 50 cells were applied to the agar medium shown in Table 1 prepared in Example 1, and the number of bacteria per ml was measured from the number of colonies formed. The case where growth was inhibited was marked as ◯, and the case where growth was not inhibited was marked as X.
前記 10日間培養後の各飲料水について風味の官能評価を行った。風味が良いも のを〇、酸味が強いが飲むことができるものを△、酸味が著しく飲むことができないも のを Xとした。結果を表 7に示す。  The sensory evaluation of the flavor was performed on each drinking water after culturing for 10 days. ◯ indicates that the taste is good, △ indicates that the sour taste is strong but can be drunk, and X indicates that the taste is not so drastic. The results are shown in Table 7.
[表 6] [Table 6]
Figure imgf000010_0001
Figure imgf000010_0001
[表 7] [Table 7]
Figure imgf000010_0002
Figure imgf000010_0002
表 7から、果汁飲料水中の乳酸濃度が 0.075〜0.3%となる水準 4〜8において有意 に耐熱性好酸性菌の増殖を抑え、かつ風味が良いか、飲用できる果汁飲料水が得 られることが判った。 From Table 7, fruit juice drinking water is obtained that significantly suppresses the growth of thermotolerant acidophilic bacteria at levels 4 to 8 where the lactic acid concentration in the juice drinking water is 0.075 to 0.3% and is tasty or drinkable. It was found that

Claims

請求の範囲 The scope of the claims
[1] 耐熱性好酸性菌の増殖を抑制する有効量の乳酸を有効成分として含む耐熱性好 酸性菌の増殖抑制剤。  [1] A growth inhibitor of heat-resistant acidophilic bacteria comprising an effective amount of lactic acid as an active ingredient to suppress the growth of heat-resistant acidophilic bacteria.
[2] 耐熱性好酸性菌の増殖を抑制しょうとする対象物に、請求項 1記載の増殖抑制剤 を添加する工程を含む耐熱性好酸性菌の増殖抑制方法。  [2] A method for inhibiting the growth of heat-resistant acidophilic bacteria, comprising the step of adding the growth inhibitor according to claim 1 to an object for inhibiting the growth of heat-resistant acidophilic bacteria.
[3] 酸性飲料の製造工程の!/、ずれかの時点で、請求項 1記載の増殖抑制剤を添加す る耐熱性好酸性菌の増殖を抑制した酸性飲料の製造方法。 [3] A method for producing an acidic beverage that suppresses the growth of heat-resistant acidophilic bacteria, to which the growth inhibitor according to claim 1 is added at the time of the production process of the acidic beverage.
[4] 前記増殖抑制剤の添加量が、酸性飲料全量に対する全乳酸量が 0.075重量％以 上となる量である請求項 3記載の製造方法。 [4] The production method according to claim 3, wherein the amount of the growth inhibitor added is such that the total amount of lactic acid relative to the total amount of the acidic beverage is 0.075% by weight or more.
[5] 請求項 3又は 4記載の製造方法により得られた、耐熱性好酸性菌の増殖を抑制し た酸性飲料。 [5] An acidic beverage obtained by the production method according to claim 3 or 4 and suppressing the growth of heat-resistant acidophilic bacteria.
[6] 耐熱性好酸性菌の増殖抑制剤を製造するための、乳酸の使用。  [6] Use of lactic acid to produce a growth inhibitor of heat-resistant acidophilic bacteria.
PCT/JP2006/306299 2005-03-31 2006-03-28 Growth inhibitor for thermotolerant acidophilic bacterium, method for inhibition of growth of the bacterium, and method for production of acidic beverage WO2006106673A1 (en)

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