WO2011046194A1 - 乳酸菌増殖促進剤 - Google Patents
乳酸菌増殖促進剤 Download PDFInfo
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- WO2011046194A1 WO2011046194A1 PCT/JP2010/068119 JP2010068119W WO2011046194A1 WO 2011046194 A1 WO2011046194 A1 WO 2011046194A1 JP 2010068119 W JP2010068119 W JP 2010068119W WO 2011046194 A1 WO2011046194 A1 WO 2011046194A1
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- lactic acid
- pineapple
- acid bacteria
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- juice
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- 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
- C12N1/38—Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factors; Stimulation of growth by removal of a chemical compound
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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
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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
- A23C9/133—Fruit or vegetables
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- 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
- C12N2500/00—Specific components of cell culture medium
- C12N2500/70—Undefined extracts
- C12N2500/76—Undefined extracts from plants
Definitions
- the present invention relates to a lactic acid bacteria growth promoter and a method for producing fermented milk using the same.
- Lactic acid bacteria are a general term for microorganisms that generally produce lactic acid by fermentation, and play an important role in improving the flavor, texture, nutritional value of foods, or imparting preservability.
- fermented foods using lactic acid bacteria for example, fermented milk products made from milk of mammals typified by yogurt and cheese, and plants such as vegetables typified by various pickles and ryakuyo are used as raw materials. Pickled vegetables, as well as miso and soy sauce are known.
- various lactic acid bacteria such as the improvement of the intestinal flora by ingesting live lactic acid bacteria or the action of regulating the intestines have been clarified. It has been researched and developed.
- sake lees sake lees extract or enzymatic degradation product of sake lees (see Patent Documents 1 to 3), distillation residue of barley shochu (see Patent Document 4), cocoa mass (Patent Document 5), calcium salt (Patent Document 6), Buttermilk (patent document 7), cytidine and thymidine, fermented wort (patent document 8), banana extract (patent document 9) and the like have been reported to have an effect of promoting the growth of lactic acid bacteria.
- sake lees have a peculiar smell of sake and may adversely affect the flavor of the fermented food, so that there are problems such as limiting the amount of use or limiting the fermented food to be used.
- pineapple fruits are used as flavors of processed foods and foods, or as fermentation raw materials for fermented fruit lactic acid (Patent Document 10). It is not done.
- the present invention relates to providing a lactic acid bacteria growth promoter that does not impair the flavor of food, and a method for producing fermented milk using the same.
- the present inventor surprisingly has an excellent growth promoting effect on lactic acid bacteria, particularly plant lactic acid bacteria, in the fruit of a pineapple genus plant or an extract thereof. And found useful as a lactic acid bacteria growth promoter.
- this invention provides the lactic acid bacteria growth promoter which uses the fruit of a pineapple genus plant, or its extract as an active ingredient.
- the present invention also provides a method for promoting the growth of lactic acid bacteria, comprising culturing lactic acid bacteria in the presence of a fruit of a pineapple genus plant or an extract thereof.
- this invention provides the manufacturing method of fermented milk characterized by fermenting milk using lactic acid bacteria in presence of the fruit of a pineapple genus plant, or its extract.
- this invention provides the solid fermented milk obtained with the manufacturing method of the said fermented milk.
- lactic acid bacterium growth promoter of the present invention can promote the growth of lactic acid bacteria, particularly plant lactic acid bacteria.
- fermented milk with good flavor, especially solid type fermented milk which is difficult to produce by fermentation using plant lactic acid bacteria can be produced efficiently in a short time.
- the lactic acid bacteria growth promotion effect by each fruit juice or each fruit juice and sake lees mixture is shown.
- the solidified state of each fermented milk after 36 hours of lactic acid fermentation is shown.
- -Containing milk medium [No. 1 and 2, no. 6 and 7], milk medium containing pineapple puree (final concentrations 5 and 10% (w / v)) [No. 3 and 4, no. 8 and 9], or milk medium [No. 5 and 10].
- the term “pineapple genus plant” refers to a plant belonging to the genus Bromeliaceae, Ananas, or a related plant thereof (for example, Bromelia pinguin (English name pinguin) of the genus Bromelica). Means.
- a pineapple (scientific name: Ananas comosus English: pineapple) is preferable.
- the pineapple is preferably an edible variety of pineapple classified into Cayenne, Red Spanish, Blanco and Queen.
- fruit juice refers to a juice obtained by squeezing a fruit, preferably a pulp (100% fruit juice), a concentrated fruit juice, a concentrated reduced fruit juice, and a diluted product thereof.
- pasted refers to fruit (preferably, pulp) that is crushed or ground (pure), and further pureed (pure).
- the “residue” means a product obtained by subjecting a discarded part (by-product) of the manufacturing process such as fruit juice, paste or pulp slice to the above physical treatment, and the discarded part includes fruit skin and pulp.
- the fruit processed product can be appropriately diluted, adjusted for pH, sterilized, concentrated, filtered, dried or powdered.
- a pH adjusting agent such as sodium citrate is added to adjust the pH to 4 to 8, preferably pH 5 to 6, or sterilization (at 60 to 120 ° C. for 30 seconds to 20 minutes, And the like subjected to 1 to 10 minutes at 90 to 105 ° C. can be used.
- lactic acid bacteria growth promoter of the present invention in addition to using the fruit of a pineapple genus plant, various solvent extracts of the fruit (including processed fruits) may be used as long as the lactic acid bacteria growth promoting action can be exerted. it can.
- the extraction means is not particularly limited, and examples thereof include solid-liquid extraction, liquid-liquid extraction, immersion, decoction, leaching, reflux extraction, ultrasonic extraction, microwave extraction, and stirring.
- the extraction solvent may be either a polar solvent or a nonpolar solvent.
- a polar solvent for example, water; alcohols such as methanol, ethanol and butanol (preferably alcohols having 1 to 4 carbon atoms); propylene glycol, butylene glycol and the like Polyhydric alcohols; Esters such as ethyl acetate; Hydrocarbons such as n-hexane; Halogenated hydrocarbons such as chloroform; Supercritical carbon dioxide, etc. These may be used alone or in combination of two or more. can do.
- the extraction conditions are not particularly limited, and can be carried out, for example, at 4 to 120 ° C., preferably 80 to 105 ° C. for 1 minute to several days.
- the pH may be 5 to 6.
- separation and purification techniques such as activated carbon treatment, liquid chromatography, liquid-liquid separation, gel filtration, and distillation may be used as appropriate.
- Lactic acid bacteria to which the fruit of the pineapple genus plant or an extract thereof can be applied as a lactic acid bacteria growth promoter are not particularly limited.
- lactobacillus bacteria, streptococcus bacteria, leuconostoccus bacteria, pediococcus bacteria, bacillus Genus bacteria, lactococcus bacteria, enterococcus bacteria, terragenococcus bacteria, bifidobacteria bacteria and the like are not particularly limited.
- plant lactic acid bacteria isolated from plant sources such as cereals, vegetables, fruits, flowers, and medicinal plants are generally less proliferative than animal lactic acid bacteria isolated from animal sources such as livestock meat and milk. It is difficult to produce a solid type yogurt. Therefore, the lactic acid bacteria growth promoter of the present invention can be more effectively applied to plant lactic acid bacteria.
- examples of the plant lactic acid bacteria include Lactobacillus plantarum, Lactobacillus hilgardi, Lactobacillus brevis, Lactobacillus lactis, and Lactobacillus lactatum.
- Lactobacillus spp . Leuconostoc mesenteroides and other Leuconostoc genus bacteria; Enterococcus munditi, Enterococcus abium and other Enterococcus genus bacteria; Tetragenococ Tetragenococcus genus bacteria such as Tetragenococcus halophilus; Pediococcus pentoaceus, Pediococcus assidiolacty genus B. Can be mentioned.
- the plant lactic acid bacteria include Lactobacillus plantarum SN13T strain (NITE P-7), Lactobacillus plantarum SN26T strain (NITE P-8), Lactobacillus plantarum SN35N strain (NITE P-6), Lactobacillus Plantarum SN35M strain (NITE P-5), Lactococcus lactis subspecies Lactis SN26N strain (NITE P-9), Enterococcus species SN21I strain (NITE P-11), Enterococcus munti SN29N strain (NITE P-10) and Enterococcus abium (NITE BP-142), Lactobacillus plantarum JCM 1149 strain, Lactobacillus plantarum JCM 8348 strain, Lactobacillus plantarum IFO307 Ltd.
- Lactobacillus plantarum is preferable, and Lactobacillus plantarum SN13T strain (NITE P-7) and Lactobacillus plantarum SN35N strain (NITE P-6) are particularly preferable.
- lactic acid fermentation proceeds rapidly by growing lactic acid bacteria in the presence of a fruit of a pineapple genus plant or an extract thereof. Therefore, the fruit of a pineapple genus plant or an extract thereof can be used as a lactic acid bacteria growth promoter when lactic acid fermentation is performed, thereby exhibiting the effect of promoting lactic acid bacteria growth and efficiently producing various fermented products in a short time. It becomes possible. Fermentation may be performed by inoculating a fermentation raw material with lactic acid bacteria according to a known method in the presence of a fruit of a pineapple genus plant or an extract thereof.
- fermented raw materials such as milk (preferably containing 0.1 to 20% (w / v) of milk (in terms of dry matter) in 1 L of a medium containing the raw materials) the fruit of a pineapple genus plant or an extract thereof, In 1 L of medium, it is preferably 0.1 to 20% (w / v), more preferably 0.1 to 10% (w / v), still more preferably 0.5 to 2 (w / v) in terms of dry matter. %), Inoculated with lactic acid bacteria, and cultured at 25 to 45 ° C. for 10 to 80 hours.
- the case of producing fermented milk will be described as an example.
- milk medium a liquid medium containing milk
- the fruit of the pineapple genus plant of the present invention or an extract thereof in 1 L of the medium is added so as to be 0.1 to 5% (w / v) in terms of dry matter, and the fruit of the pineapple genus plant or A milk medium containing the extract is prepared.
- this medium is inoculated with 0.1 to 6% (w / v) of lactic acid bacteria prepared as a starter and having a high concentration. After inoculation, fermentation is carried out at a temperature of 30 to 45 ° C. for 3 to 72 hours until the amount of lactic acid bacteria reaches 10 million cells / mL or more.
- milk containing 1% (w / v) or more in an aqueous solution
- an aqueous solution prepared by mixing other raw materials with it at a temperature of 65 to 130 ° C. for 1 second to 30 minutes Heat sterilize for hours and then cool to 30-45 ° C.
- 0.1 to 6% (w / v) of lactic acid bacteria are inoculated as a starter.
- fermentation is performed at a temperature of 30 to 45 ° C. for 12 to 72 hours in the presence of the fruit of the pineapple genus plant of the present invention or an extract thereof.
- the cooled product is used as a fermented milk beverage.
- the fermented milk drink may be used as a drink as it is, or may be further diluted or sterilized.
- the content of the fruit of the pineapple genus plant of the present invention or the extract thereof in 1 L of an aqueous solution prepared with raw materials such as milk is 0.1 to 2% (w / v) in terms of dry matter. Is preferred.
- fermented milk in addition to milk as a fermentation raw material, gelatin, agar, sugar, fragrance, pulp, nitrogen source, pH adjuster, and other lactic acid bacteria growth promoting components are usually used for the production of fermented milk.
- Raw materials can also be used.
- sugars such as sucrose, glucose, fructose, palatinose and trehalose; sugar alcohols such as sorbitol, xylitol, erythritol and reduced starch syrup; high-intensity sweeteners such as aspartame and acesulfame potassium; Emulsifiers such as carrageenan, xanthan gum, guar gum, etc .; acidulants such as citric acid, lactic acid, malic acid; fruit juices such as lemon juice, orange juice, cuwi juice, pear juice; vitamins; calcium, iron, Minerals such as manganese and zinc; herbal medicines such as licorice, katsura, and ginger; or food additives such as herbs, sodium glutamate, gardenia pigments, silicone, and phosphates can be used.
- sugar alcohols such as sorbitol, xylitol, erythritol and reduced starch syrup
- high-intensity sweeteners such as aspartam
- milk that is a fermentation raw material
- animal milk such as cow's milk, goat milk, and sheep milk
- milk is particularly preferable.
- Such milk may be either non-sterilized milk or pasteurized milk, and concentrated milk or condensed milk prepared from these milks, skim milk, partially skimmed milk or powdered milk obtained by drying these powders. Etc.
- the culture of lactic acid bacteria used as a starter is not particularly limited, and the culture medium, culture conditions, etc. may be adopted using known methods, but the pineapple genus of the present invention described above is used. It is preferable to use a plant fruit or an extract thereof as it is, or a solution obtained by adjusting pH or sterilizing it, and further adding a sake lees thereto.
- the content of the pineapple plant fruit or extract thereof in 1 L of the starter medium is not particularly limited, but is 0.1 to 20% (w / v), more preferably 10 to 20% (w / v) in terms of dry matter.
- the mass ratio of the fruit of a pineapple genus plant or its extract: liquor: 1: 0.01 to 0.1, in terms of dry matter, : 0.05 to 0.09 is preferable.
- a lactic acid bacterium cultured under such conditions as a starter the fermentation promoting effect can be further improved.
- the “sake lees” commercially available products or freeze-dried and spray-dried liquors can be used, but the remaining residue after squeezing the sake from “Moromi”, that is, to the liquor mother cultivating a large amount of yeast. Steamed rice and koji are added, and alcohol fermentation is performed for about 3 weeks to obtain “aged moromi”, which is a fermentation product.
- rice shochu lees or rice shochu may be freeze-dried or spray-dried.
- the fermented product (fermented milk) produced using the lactic acid bacteria growth promoter of the present invention thus obtained has a rapid progress of lactic acid fermentation as shown in the examples described later, that is, usually has a very low growth ability. Even when using a plant lactic acid bacterium that is said to be difficult to produce solid type fermented milk, it quickly becomes solid (solid fermented milk). Moreover, the fermented milk is imparted with an excellent flavor and a fragrance specific to pineapple, and is also excellent in texture, appearance and color of the fermented milk.
- the ability to grow lactic acid bacteria can be adjusted as appropriate, and fermented milk of any form such as plain type, soft type, drink type, solid (hard) type, frozen type, etc. can be produced. Is possible.
- Example 1 Fermented milk solidification test using each fruit juice ⁇ Bacteria used> Lactobacillus plantarum plantarum SN13T strain or L. plantarum SN35N strain was used. These are to the National Institute for Product Evaluation Technology Patent Microorganisms Depositary (Address: 2-8-8 Kazusa-Kamashita, Kisarazu City, Chiba Prefecture), Lactobacillus plantarum SN13T strain (NITE P-7) and Lactobacillus plantarum It has been deposited as the SN35N strain (NITE P-6) (Deposit date: July 6, 2004).
- pineapple juice fruit juice derived from Okinawa pineapple fruit (fruit juice 100%: manufactured by Pineapple Winery Co., Ltd.) (hereinafter referred to as “pineapple fruit juice 1”), or fruit juice derived from Costa Rica pineapple fruit (fruit juice 100%) [hereinafter, "Pineapple juice 2" was used. These are squeezed pineapple fruits of the pineapple family pineapple genus plant. All juices were found to be turbid, but the Costa Rica pineapple juice was more turbid than the Okinawa pineapple juice. At this time, when pineapple juice 1 or 2 (50 mL) was freeze-dried, the average was 8 g.
- Pineapple fruit juice 1 or 2 with sodium citrate added to adjust the pH to 5-6 is sterilized by heating at 100-104 ° C. for 5-10 minutes and sterilized pineapple fruit juice 1 or 2 (pineapple fruit in the total amount)
- the content of was 16% (w / v) in terms of dry matter.
- kiwi or pear pulp sterilized orange juice, sterilized kiwi juice, or sterilized pear juice was prepared in accordance with the pineapple fruit.
- a starter (Lactobacillus plantarum SN13T strain) was prepared according to the above ⁇ Preparation of starter>.
- the final concentration of pineapple juice is 0% or 1.0% (w / v) in 1 L of the medium under aseptic conditions.
- a pineapple fruit is added to a milk medium so that it may become 0.16% (w / v) in terms of dry matter, and a pineapple fruit juice-free milk medium or a 1.0% (w / v) pineapple fruit milk medium was prepared as the main culture medium. 1 mL of each prepared main culture medium was put into each 1.5 mL polyethylene microtube.
- pineapple juice 1 When pineapple juice 1 was used, the fermented milk solidified about 70%, and when pineapple juice 2 was used, the fermented milk was completely solidified after 40 hours of culture. On the other hand, fermented milk solidified to about 50% when cucumber juice or pear juice was used, and fermented milk only solidified to about 20% with orange juice. That is, it was confirmed that the use of pineapple fruit juice could promote the growth of lactic acid bacteria and solidify the fermented milk.
- Example 2 Fermented milk solidification test using pineapple juice or sake lees ⁇ Preparation of sake lees and sake lees milk medium> As the sake lees, commercially available liquors obtained by spray drying were used. A sake lees spray-dried product (hereinafter also referred to as “Sake lees SD”) is added to the above milk medium to a final concentration of 1.5% (w / v) and contains 1.5% (w / v) sake lees. A main culture medium was prepared.
- each starter was used as an inoculum at 37 ° C. For 48 hours, shaking culture was performed to produce fermented milk, and this solidified state was confirmed. In addition, as a control, fermented milk was produced in the same manner as a main culture medium with nothing added to the milk medium.
- Example 3 Lactic acid bacteria growth promoting effect test using pineapple juice or pineapple juice / sake lees mixture as starter medium ⁇ Preparation of pineapple juice / sake lees mixture>
- the sake lees SD was added to the sterilized pineapple juice 1 to a final concentration of 1.0% (w / v) to prepare a pineapple juice 1 / sake lees mixture.
- the mass ratio of pineapple juice: liquor was 16: 1 in terms of dry matter.
- Example 1 the sterilized pineapple juice 1 described in Example 1 or the pineapple juice 1 / sake lees mixture was used as it was. According to ⁇ Preparation of starter> in Example 1 above, each of the starter media was inoculated with the Lactobacillus plantarum SN13T strain, then the bacteria were cultured, and the number of bacteria at this time was measured.
- Example 4 Lactobacillus growth promoting effect test using each fruit juice and each fruit juice / sake lees mixture as a starter medium ⁇ Preparation of fruit juice, fruit juice / sake lees mixture>
- pineapple juice concentrated reduced juice obtained by diluting concentrated fruit juice (manufactured by Nomura Dairies) to 100% fruit juice was used.
- the concentrated reduced fruit juice which was adjusted to pH 5.7 by adding sodium bicarbonate, was sterilized by heating at 100 ° C. for 8 minutes to obtain a sterilized pineapple fruit juice 3.
- the sake lees SD was added to the sterilized pineapple juice 3 to a final concentration of 1.0% (w / v) to prepare a pineapple juice 3 and sake lees mixture.
- the mass ratio of pineapple juice: liquor was 16: 1 in terms of dry matter.
- sterilized fruit juice and a mixture of each fruit juice and sake lees were prepared from the concentrated fruit juice in accordance with Pineapple Juice 3.
- starter medium each of the above sterilized fruit juices or each fruit juice / sake lees mixture was used as it was.
- a culture solution of Lactobacillus plantarum strain SN13T (Lp.13T) previously cultured in an MRS medium (referred to as a pre-preculture medium) is used for each starter medium.
- a pre-preculture medium a culture solution of Lactobacillus plantarum strain SN13T (Lp.13T) previously cultured in an MRS medium (referred to as a pre-preculture medium) is used for each starter medium.
- a pre-preculture medium referred to as a pre-preculture medium
- the bacteria were cultured at 37 ° C. for 24 hours.
- pineapple fruit has an effect of promoting the growth of Lactobacillus plantarum.
- carrot juice, apple juice, and banana juice had a weak growth promoting effect on Lactobacillus plantarum.
- Example 5 Lactobacillus growth-promoting effect test using pineapple juice, pineapple puree or pineapple juice / sake lees-containing milk medium ⁇ Preparation of puree> A pineapple fruit ground with a mixer (pure form) was obtained. At this time, when purey 50mL was dried, it was 8 mg. Pineapple puree, pineapple puree with sodium citrate added to adjust pH to 5-6, sterilized by heat sterilization at 100-104 ° C for 5-10 minutes (contains pineapple fruit in the total amount) The amount was 16% (w / v) in terms of dry matter.
- each starter was inoculated into each main culture medium for 36 hours to produce fermented milk, and this solidified state was confirmed.
- a main culture medium containing only a milk medium was used.
- a starter prepared using pineapple juice as a starter medium was inoculated into a main culture medium containing 10% (w / v) of pineapple juice 1 and cultured for 16 hours.
- the solidification rate of fermented milk was 100%.
- the solidified state of fermented milk after 36-hour culture is shown in FIG.
- the solidification rate of the fermented milk was 100% after culturing for 16 hours.
- the content of sake lees in the main culture medium is as small as 0.01% (w / v)
- solidification of the fermented milk is considered to be due to the pineapple fruit contained in the main culture medium.
- the fermented milk produced by this starter had no smell peculiar to sake, and was given a scent and flavor peculiar to pineapple. Furthermore, when the puree was added to the milk medium so as to be 5% (w / v), when cultured for 16, the solidification rate of the fermented milk became 100%, whereas pineapple juice 1 was 5% (w / V), the solidification rate of the fermented milk was 100% in 24 to 36 hours (see FIG. 2). From this, it was considered that a puree containing more dietary fiber than fruit juice enhances the growth of lactic acid bacteria and also improves the solidification of fermented milk. In addition, plant lactic acid bacteria grew slowly and it was difficult to solidify fermented milk. However, using pineapple juice or puree promotes lactic acid fermentation by plant lactic acid bacteria, and fermented milk can be obtained in 16 to 24 hours. Since it was solidified, it was thought that the fermentation time could be shortened.
- pineapple fruit has the effect of promoting the growth of lactic acid bacteria, especially plant lactic acid bacteria, so that pineapple fruit is added to milk medium and lactic fermentation is performed to produce solid type fermented milk with good flavor and aroma. Became possible.
- the use of a starter medium in which about 0.5 to 1.5% of sake lees are added to pineapple fruit has a higher effect of promoting the growth of lactic acid bacteria. It has become possible to produce solid type fermented milk that has no odor and has a pineapple flavor and aroma.
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Abstract
Description
従来から、乳酸菌を用いた発酵食品としては、例えばヨーグルト、チーズに代表される哺乳動物の乳を原料としたはっ酵乳製品を始め、各種の漬物・らっきょうに代表される野菜等の植物を原料とした漬物類、さらには味噌、醤油類が知られている。
また、近年、生きた乳酸菌の摂取による腸内菌叢の改善効果又は整腸作用等の様々な乳酸菌の生理的効果が明らかとなり、当該乳酸菌の生理作用を利用した食品、健康食品、医薬品等が研究開発されてきている。
しかしながら、例えば酒粕には酒独特の臭いがあり、発酵食品の風味に悪影響を及ぼす可能性があるので、使用量を制限するか使用する発酵食品が限定される等の問題があった。
また、本発明は、パイナップル属植物の果実又はその抽出物の存在下、乳酸菌を培養することを特徴とする乳酸菌増殖促進方法を提供するものである。
また、本発明は、パイナップル属植物の果実又はその抽出物の存在下、乳酸菌を用いて乳を発酵することを特徴とするはっ酵乳の製造方法を提供するものである。
また、本発明は、前記のはっ酵乳の製造方法にて得られた固形はっ酵乳を提供するものである。
当該パイナップルとしては、カイエン系、レッドスパニッッシュ系、ブランコ系及びクイーン系等の系統に分類される食用品種のパイナップルが好ましい。
ここで、「果汁」とは、果実、好ましくは果肉を搾汁して得られる液汁(100%果汁)、濃縮果汁、濃縮還元果汁、及びこれらを希釈したものを云う。
また、「ペースト状」とは、果実、好ましくは果肉を破砕や摩砕したもの(ピューレ状)、及び更にピューレ状のものを裏ごししたもの(ピューレ)を云う。
また、「残渣」とは、果汁、ペースト状又は果肉スライス等の製造過程の廃棄部分(副産物)に上記物理的処理を施したものを云い、廃棄部分には果実表皮や果肉等が含まれる。
当該果実処理物は、適宜希釈、pH調整、殺菌処理、濃縮、濾過、乾燥又は粉末化したものを用いることができる。
例えば、果実処理物に、クエン酸ナトリウム等のpH調整剤を添加して、pH4~8、好ましくはpH5~6に調整したもの、また、殺菌処理(60~120℃で30秒~20分間、90~105℃で1~10分間等)を施したものを用いることができる。
抽出手段としては、特に限定されないが、例えば、固液抽出、液液抽出、浸漬、煎出、浸出、還流抽出、超音波抽出、マイクロ波抽出、攪拌等が挙げられる。
パイナップル属植物の果実の抽出物は、不純物や夾雑物等を除去するため、適宜活性炭処理、液体クロマトグラフィー、液液分液、ゲルろ過、蒸留等の分離精製技術を用いてもよい。
このうち、穀物、野菜、果物、花、薬用植物等の植物源から分離された植物乳酸菌は、一般的に、畜肉・乳等の動物源から分離された動物乳酸菌に比べて増殖能が低いため、固形タイプのヨーグルトを製造することが困難である。従って、本発明の乳酸菌増殖促進剤は、植物乳酸菌に対して、より有効的に適用できる。
このうち、ラクトバチルス プランタルムが好ましく、特にラクトバチルス プランタルムSN13T株(NITE P-7)及びラクトバチルス プランタルムSN35N株(NITE P-6)が好ましい。
発酵は、パイナップル属植物の果実又はその抽出物の存在下、公知の方法に従って、発酵原料に乳酸菌を接種して行えばよい。例えば、乳等の発酵原料(好ましくは、原料を含む培地1L中、乳(乾燥物換算)0.1~20%(w/v)含有)に、パイナップル属植物の果実又はその抽出物を、培地1L中、乾燥物換算で、好ましくは0.1~20%(w/v)、より好ましくは0.1~10%(w/v)、更に好ましくは0.5~2(w/v)%になるように添加し、これに乳酸菌を接種して、25~45℃で10~80時間培養することにより行うことができる。以下に、はっ酵乳を製造する場合を例に挙げて、説明する。
スターター培地1L中のパイナップル属植物の果実又はその抽出物の含有量は、特に限定されないが、乾燥物換算で、0.1~20%(w/v)、より10~20%(w/v)であるのが好ましく、これに酒粕を併用する場合には、パイナップル属植物の果実又はその抽出物:酒粕の質量比が、乾燥物換算で、1:0.01~0.1、より1:0.05~0.09であるのが好ましい。斯かる条件で培養した乳酸菌をスターターとして用いることにより、発酵促進効果をより向上させることができる。
ここで、「酒粕」として、市販品や酒粕の凍結乾燥品や噴霧乾燥品を用いればよいが、「もろみ」から清酒を搾り取った後の残りかす、すなわち、酵母を大量に培養した酒母に蒸米と麹を加えて、約3週間アルコール発酵を行って、発酵産物である「熟成もろみ」とし、これより清酒を搾った残りかすを用いてもよい。また、酒粕の代わりに米焼酎の粕もしくは米焼酎粕を凍結乾燥や噴霧乾燥処理したものでもよい。
<使用する菌>
ラクトバチルス プランタルムの一種L. plantarum SN13T株又はL. plantarum SN35N株を用いた。
これらは、独立行政法人製品評価技術基盤機構特許微生物寄託センター(住所:千葉県木更津市かずさ鎌足2丁目5番8号)へ、ラクトバチルスプランタルムSN13T株(NITE P-7)及びラクトバチルス プランタルムSN35N株(NITE P-6)として寄託されているものである(寄託日:平成16(2004)年7月6日)。
<果汁の調製>
パイナップル果汁として、沖縄産パイナップル果実由来の果汁(果汁100%:株式会社パイナップルワイナリー製)〔以下、「パイナップル果汁1」と云う〕、又はコスタリカ産パイナップル果実由来の果汁(果汁100%)〔以下、「パイナップル果汁2」と云う〕を用いた。これらは、パイナップル科パイナップル属植物のパイナップルの果実を搾汁した液汁である。何れの果汁も果汁由来の混濁が認められたが、コスタリカ産パイナップル果汁の方が沖縄産パイナップル果汁より、濁っていた。
このとき、パイナップル果汁1又は2(50mL)を凍結乾燥すると、平均8gであった。
パイナップル果汁1又は2にクエン酸Naを添加してpHを5~6に調整したものを、100~104℃で5~10分加熱殺菌して殺菌済パイナップル果汁1又は2(全量中のパイナップル果実の含有量は、乾燥物換算で16%(w/v))とした。
また、オレンジ、キウイ又はナシの果肉についてもパイナップルの果実に準じて、殺菌済オレンジ果汁、殺菌済キウイ果汁、又は殺菌済ナシ果汁を調製した。
無菌下で、スターター培地5mLに、乳酸菌を植菌し、37℃で24時間種培養を行い、スターターを調製した。スターター培地として、上記殺菌済パイナップル果汁1をそのまま用いた。
脱脂粉乳(PM)13%(w/v)水溶液(pHをクエン酸ナトリウム溶液で5.0~5.5に調整したもの)を121℃、15分間高圧滅菌し、殺菌済乳培地を調製した。
上記<スターターの調製>に従って、スターター(ラクトバチルス プランタルムSN13T株)を調製した。
上記<果汁の調製>に記載の殺菌済パイナップル果汁1又は2を用い、無菌下で、パイナップル果汁の最終濃度が、培地1L中、0%又は1.0%(w/v)(全量中、パイナップル果実が、乾燥物換算で、0.16%(w/v))になるように乳培地に添加し、パイナップル果汁無添加の乳培地又は1.0%(w/v)パイナップル果汁乳培地を本培養培地として調製した。
調製した各本培養培地1mLを各1.5mL容ポリエチレン製マイクロチューブに入れた。更に、上述の調製したスターター(種菌)が1.0%(w/v)となるように、各マイクロチューブに入れ、マイクロチューブを密閉後、37℃、40時間振とう培養し、はっ酵乳を製造した。
また、殺菌済パイナップル果汁を上記の殺菌済オレンジ果汁、殺菌済キウイ果汁、又は殺菌済ナシ果汁に代えた以外は、上記と同様にして、振とう培養を行い、はっ酵乳を製造した。
各はっ酵乳の固化状況を確認した。
<酒粕及び酒粕乳培地の調製>
酒粕として、市販品の酒粕を噴霧乾燥したものを用いた。
酒粕噴霧乾燥品(以下「酒粕SD」とも云う)を最終濃度1.5%(w/v)になるように、上記の乳培地に添加し、酒粕1.5%(w/v)含有の本培養培地を調製した。
スターター培地として上記<果汁の調製>に記載の殺菌済パイナップル果汁1を用い、乳酸菌として上記ラクトバチルス プランタルムSN13T株又はラクトバチルス プランタルムSN35N株を用いて、上記<スターターの調製>に従って、2種のスターターをそれぞれ調製した。
また、コントロールとして、乳培地に何も添加しないものを本培養培地として同様にはっ酵乳を製造した。
従来、植物乳酸菌は、動物乳酸菌に比べて増殖能が低く、植物乳酸菌で固形タイプはっ酵乳を得ることは非常に困難で、ドリンクタイプのはっ酵乳が主流であった。この結果から、植物乳酸菌のように増殖能が低いものであっても、パイナップル果汁を用いれば非常に乳酸菌の増殖能が高まると共に、はっ酵乳が固化できることが確認された。
また、ポジティブコントロールとして、酒粕を乳培地中0.15%(w/v)で用いたが、酒独特の臭いがあり、はっ酵乳の匂いや風味が損なわれていたのに対し、パイナップル果汁を用いた場合には、はっ酵乳に優れた風味やパイナップル特有の香気が付与されるので、商品的価値の高いはっ酵乳を製造することができた。
<パイナップル果汁・酒粕混合物の調製>
上記酒粕SDを最終濃度1.0%(w/v)になるように、上記殺菌済パイナップル果汁1に添加し、パイナップル果汁1・酒粕混合物を調製した。このとき、パイナップル果汁:酒粕の質量比は、乾燥物換算で16:1であった。
スターター培地として、上記実施例1に記載の殺菌済パイナップル果汁1又は上記パイナップル果汁1・酒粕混合物をそのままの状態で使用した。
上記実施例1の<スターターの調製>に従って、上記各スターター培地に上記ラクトバチルス プランタルムSN13T株を接種した後、菌を培養し、このときの菌数を測定した。
<果汁、果汁・酒粕混合物の調製>
パイナップル果汁として、濃縮果汁(野村乳業製)を100%果汁に希釈した濃縮還元果汁を用いた。濃縮還元果汁に、炭酸水素ナトリウムを添加してpHを5.7に調整したものを、100℃で8分加熱殺菌して殺菌済パイナップル果汁3とした。
上記酒粕SDを最終濃度1.0%(w/v)になるように、上記殺菌済パイナップル果汁3に添加し、パイナップル果汁3・酒粕混合物を調製した。このとき、パイナップル果汁:酒粕の質量比は、乾燥物換算で16:1であった。
また、人参果汁、リンゴ果汁及びバナナ果汁についてもパイナップル果汁3に準じて、濃縮果汁からそれぞれ殺菌済果汁と、各果汁・酒粕混合物を調製した。
スターター培地として、上記それぞれの殺菌済果汁又は各果汁・酒粕混合物をそのままの状態で使用した。
上記実施例1の<スターターの調製>に従って、各スターター培地に、あらかじめMRS培地にて培養したラクトバチルス プランタルムSN13T株(L.p.13T)の培養液(これを前前培養液と呼ぶ)を各スターター培地100mLに対して1mL接種した(最終濃度で1(V/v)%)後、37℃で24時間菌を培養した。各スターター培地から1mLを採取し、0.85%Naclで・10倍から1010倍までの希釈系列を作製して希釈後、BCPアガー培地(日水製薬株式会社)に1mL接種して嫌気的条件下で培養後、菌数を測定した。結果を図1に示す。
スターター培地として、上記殺菌済パイナップル果汁3又はパイナップル果汁3・酒粕混合物をそのままの状態で使用した。各スターター培地にペディオコッカス・ペントサセウスLP28株を接種した以外は、上記乳酸菌増殖促進効果試験(1)と同様にして菌を培養後、菌数を測定した。なお、ペディオコッカス・ペントサセウスLP28株は、独立行政法人製品評価技術基盤機構特許微生物寄託センター(住所:千葉県木更津市かずさ鎌足2丁目5番8号)へ、NITE P-700として寄託されているものである(寄託日:平成21(2009)年1月27日)。
その結果、ペディオコッカス・ペントサセウスLP28株を、パイナップル果汁3で培養すると、培地1mL当たり22億の乳酸菌数となったことから、パイナップル果実そのものにペディオコッカス・ペントサセウスの増殖促進効果があることが確認された。また、パイナップル果汁に酒粕を添加したパイナップル果汁3・酒粕混合物で培養すると、培地1mL当たり24億の乳酸菌数となったことから、更に酒粕を併用することによって乳酸菌増殖促進効果がより高まることも見出した。
<ピューレ状の調製>
パイナップル果実をミキサーで摩砕したもの(ピューレ状)を得た。このとき、ピューレ状50mLを乾燥すると、8mgであった。
パイナップルピューレ状にクエン酸Naを添加してpHを5~6に調整したパイナップルピューレ状を、100~104℃で5~10分加熱殺菌して殺菌済パイナップルピューレ状(全量中のパイナップル果実の含有量は、乾燥物換算で16%(w/v))とした。
上記実施例3記載の2種のスターター(パイナップル果汁1又はパイナップル果汁1・酒粕混合物をスターター培地として使用)を用いた。
上記実施例1に記載の殺菌済パイナップル果汁1、上記実施例3に記載のパイナップル果汁1・酒粕混合物、上記殺菌済パイナップルピューレ状を、それぞれ乳培地に添加して、本培養培地とした。このとき、各本培養培地の調製法は、実施例1の方法に準じて行い、各本培養培地中のパイナップル果汁の含有量は、培地1L中、0%、5%(w/v)(全量中のパイナップル果実の含有量は、乾燥物換算で0.8%(w/v))、10%(w/v)(全量中のパイナップル果実の含有量は、乾燥物換算で1.6%(w/v))とした。
上記実施例1のはっ酵乳の製造法に従って、上記各スターターを各本培養培地に接種後36時間培養し、はっ酵乳を製造し、この固化状態を確認した。なお、コントロールとして、乳培地のみの本培養培地を用いた。
一方、パイナップル果汁1に酒粕を1%(w/v)添加した混合物をスターター培地とした場合、16時間培養するとはっ酵乳の固化率が100%となった。このとき、本培養培地中の酒粕の含有量は0.01%(w/v)と微量になるので、はっ酵乳の固化は本培養培地に含まれるパイナップル果実によるものと考える。また、このスターターで製造されたはっ酵乳に酒独特の臭いはなく、しかもパイナップル特有の香りや風味が付与されていた。
更に、ピューレ状を5%(w/v)になるように乳培地に添加した場合、16培養すると、はっ酵乳の固化率が100%となったのに対し、パイナップル果汁1を5%(w/v)になるように乳培地に添加した場合、24時間~36時間で、はっ酵乳の固化率が100%となった(図2参照)。このことから、果汁よりも、食物繊維質を多く含むようなピューレ状の方が、乳酸菌の増殖性を高めると共に、はっ酵乳の固化も良好になると考えた。
また、植物乳酸菌は増殖が遅く、しかも発酵乳を固化することが困難であったが、パイナップルの果汁又はピューレ状を用いれば、植物乳酸菌による乳酸発酵を促進させて16~24時間で発酵乳を固化させることができたので、発酵時間も短縮することも可能となると考えた。
更に、スターター調製の際、パイナップル果実に酒粕を0.5~1.5%程度添加したスターター培地を用いた方が、乳酸菌の増殖促進効果がより高くなると共に、これを用いても酒独特の臭いがなく、パイナップルの風味や香りのある固形タイプのはっ酵乳を製造することも可能となった。
Claims (10)
- パイナップル属植物の果実又はその抽出物を有効成分とする乳酸菌増殖促進剤。
- 乳発酵用の乳酸菌増殖促進剤である請求項1記載の乳酸菌増殖促進剤。
- パイナップル属植物の果実が、果実処理物である請求項1又は2記載の乳酸菌増殖促進剤。
- 乳酸菌が、植物乳酸菌である請求項1~3の何れか1項記載の乳酸菌増殖促進剤。
- 植物乳酸菌が、ラクトバチルス プランタルムである請求項4記載の乳酸菌増殖促進剤。
- パイナップル属植物の果実又はその抽出物の存在下、乳酸菌を培養することを特徴とする乳酸菌増殖促進方法。
- パイナップル属植物の果実又はその抽出物の存在下、乳酸菌を用いて乳を発酵することを特徴とするはっ酵乳の製造方法。
- パイナップル属植物の果実若しくはその抽出物、又はパイナップル属植物の果実若しくはその抽出物と酒粕の存在下で培養した乳酸菌をスターターとして用いる請求項7記載のはっ酵乳の製造方法。
- 乳酸菌が植物乳酸菌である請求項7又は8記載のはっ酵乳の製造方法。
- 請求項7~9の何れか1項記載のはっ酵乳の製造方法にて得られた固形はっ酵乳。
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JP5750785B2 (ja) | 2015-07-22 |
TW201119584A (en) | 2011-06-16 |
KR101780176B1 (ko) | 2017-09-21 |
TWI520685B (zh) | 2016-02-11 |
KR20120095361A (ko) | 2012-08-28 |
JPWO2011046194A1 (ja) | 2013-03-07 |
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