JPH08280369A - Preservative for food - Google Patents

Abstract

PURPOSE: To obtain a preservative for food having high preserving effect and safety without affecting quality of food by mixing an antiviral material reuterin produced by a lacto bacillus strain, Lactobacillus reuteri with bacteriocin derived produced by a lacto bacillus strain. CONSTITUTION: Reuterin obtain by culturing a lacto bacillus strain, Lactobacillus reuteris [e.g. Lactobacillus reuteri strain DSM20016 (ATCC53609), etc.] is mixed with bacteriocin produced by a lacto bacilluss strain such as Lactococcus lactis, genera Pediococcus, Lactobacillus, Leconostoc and Propionibacterium to obtain the objective preservative for food having high preserving effect and safety without affecting quality of food.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a preservative for foods.

[0002]

2. Description of the Related Art Many conventional preservatives for foods have an antibacterial effect against bacteria, and the deterioration of foods due to the growth of yeasts and molds poses a problem. Especially in the field of food industry such as prepared foods, it tends to be low salt and low sugar due to health-consciousness, so that the storage stability is poor, and microorganisms derived from raw materials of foods,
Alternatively, there is a large problem of alteration of foods due to propagation of fungi mixed during the manufacturing process. Therefore, there is a need for a food preservative that suppresses the growth of these microorganisms.

Under such circumstances, Lactobacillus reuter, which is a type of intestinal bacterium,
Reuterin, which is an antibacterial substance i) produced in a medium under anaerobic conditions, has an antimicrobial effect on some bacteria, yeasts and molds, and therefore, uses reuterin as a food preservative. Was proposed (Tokuheihei 2-503
385 publication).

[0004]

However, since reuterin alone has a narrow antibacterial spectrum and needs to be used in a large amount in order to improve the storability of foods, reuterin alone is used as a food preservative. It was practically difficult to use.

Therefore, the present invention increases the antimicrobial action of reuterin in foods, thereby reducing the amount of reuterin added to foods as much as possible, covering a broad antibacterial spectrum, and providing safe and highly preservable foods. It is an object to provide a food preservative for manufacturing.

[0006]

Means for Solving the Problems The inventors of the present invention have conducted extensive studies to achieve the above object, and as a result, by incorporating bacteriocin produced by lactic acid bacteria into food together with reuterin, antibacterial activity of reuterin in food has been improved. The inventors have found that the activity can be increased, the antibacterial spectrum can be broadened, and the storability of the added food can be dramatically improved, and the present invention has been completed.

That is, the present invention provides a food preservative containing reuterin produced by the lactic acid bacterium Lactobacillus reuteri and bacteriocin produced by the lactic acid bacterium.

Lactobacillus Lactobacillus reuteri
acillus reuteri) is a type of animal intestinal bacterium, is a heterolactic acid bacterium that grows in a medium in the intestine or under anaerobic conditions, and its strain has been deposited with the ATCC in two strains (Accession No. 53608 and 53609).

Reuterin is an antibacterial substance produced by Lactobacillus reuteri in a medium containing glycerin under an anaerobic atmosphere. In the culture supernatant, β-hydroxypropionaldehyde, which is a fermentation product of glycerin, is detected, and this β-hydroxypropionaldehyde is a monomer, a hydrate and a dimer in an aqueous solution. It is presumed to exist in the form and is called reuterin. Reuterin exhibits antibacterial properties against Gram-positive bacteria, Gram-negative bacteria, yeasts and molds.

In the present invention, as reuterin, Lactobacillus reuteri (L. reuteri) is cultured in a medium containing glycerin, and then, for example, Japanese Patent Publication No. 2-5033.
As described in Japanese Patent Publication No. 85, a culture supernatant separated and purified by HPLC or the like is used. Further, a concentrate of the culture supernatant can also be used.

On the other hand, bacteriocin produced by lactic acid bacteria means a proteinaceous antibacterial substance produced by lactic acid bacteria. In the present invention, the bacteriocin produced by lactic acid bacteria includes Lactococcus
s lactis), genus Pediococcus, genus Lactobacillus, Leuconostoc (Le
uconostoc) and Propionibacterium (Propioni
Bacteria) can be mentioned.

Lactococcus
The bacteriocin produced by lactis)
Lactococcus lactis subsp. Nisin produced by lactis (Nisin; Jarvis, B. et al., Biochim. Biophys. Act
a, 168, 153 (1968)], lacticin
481 [Piard, JC et al., Appl. Environ. Microb
iol., 58, 279 (1992)] and Lactostrepcis; Kozak, W. et al., J. Diary Res.,
45, 247 (1978)], Lactococcus lactissubsp.c.
Diplococcin produced by remoris [Davlococcin; Dav
ey, GP etal., Appl. Environ. Microbiol., 41, 84
-89 (1981)], Lactococcus lactis subsp. Diacet.
Bacteriocin S50 produced by ilactis [Bacterioci
n S50 ； Kojic, M. et al., Appl. Environ. Microbio
l., 57, 1835 (1991)].

Bacteriocin produced by a bacterium belonging to the genus Pediococcus is generally pediocine (Pedioc).
in), for example Pediococcus acidilacti
ciH produces pediocin AcH [Pediocin AcH; Bh
unia, AK et al., J. Appl. Bacteriol., 65, 261 (1
988)], pediocin PA-1 produced by Pediococcus acidilactici PAC1.0 [Pediocin PA-1; Gonzalez,
CF and Kunka, BS, Appl. Environ. Microbiol.,
53, 2534 (1987)] and Pediococcus pentosace.
Pediocin A; D produced by ous FBB 61
aeschel, MAet al., Appl. Environ. Microbiol., 5
0, 1538 (1985)].

The bacteriocin produced by a bacterium belonging to the genus Lactobacillus is, for example, Lactobacillus.
s helveticus LP27 produces lactocin 27 [Lactoc
in 27 ； Upreti, GC, Antimicrob. Agents Chemoth
er., 4,487 (1973)], Lactobacillus acidophilu
s TK8912 produced by Acidocin 891
2), Lactobacillus plantarum C-11 produced plantaricin A [Plantaricin A; Daeschel, MA et a
L., Food Microbiol., 7, 91 (1990)] and Lact.
A bacteriocin produced by obacillus piscicola LV17 can be mentioned.

Examples of bacteriocins produced by Leuconostoc spp. Include Leuconostoc.
Leuconosine S [Leucon produced by paramesenteriodes
ocinS ； Lewus, CB, Appl. Environ. Microbiol., 5
8, 143 (1992)], Leuconostoc gelidum UAL187-produced leucosine A-UAL187 [Leucocin A-UAL
187; Hastings, JW et al., J. Bacteriol., 173,
7491 (1991)] and Leuconostoc mesenteroides.
5 [Mesenterocin 5; Daba,
H. et al., Appl. Environ. Microbiol., 57, 3450 (1
991).] Can be mentioned.

[0016] Further, Propionibacterium (Propioni
Examples of the bacteriocin produced by a bacterium belonging to the genus (bacterium) include, for example, Jenseniin G; Grinstead, DA et a produced by Propionibacterium jensenii P126.
L., Appl. Environ. Microbiol., 58, 215 (1992)] and Propionibacterium thoenii P127 produced Propionicin PLG-1 [Lyo].
n, W. et al., Appl. Environ. Microbiol., 57, 701
(See (1991)]. Two or more of these bacteriocins can be used in combination.

The preservative for food of the present invention is added so that reuterin is contained in the food in an amount of preferably 0.003 to 0.5% by weight, more preferably 0.01 to 0.2% by weight. To use. If the food is a seasoning liquid or solution,
It is recommended to use within this range even when storing in an aqueous solution.

In addition, when the above-mentioned reuterin and bacteriocin are separately added to the food or its material,
It is included in the range of the food preservative of the present invention.

[0019]

The reuterin in the food preservative of the present invention is DN depending on the ribonucleotide reductase activity of the microorganism.
It is known to exhibit antibacterial properties by inhibiting the synthesis of A. The mechanism by which co-existing bacteriocin produced by lactic acid bacteria enhances the antibacterial action of reuterin against bacteria, yeasts and molds (fungi) is not clear, but these bacteriocins coexist with reuterin in microbial cells. Therefore, it is presumed that DNA synthesis is synergistically inhibited.

[0020]

The present invention will be described in more detail with reference to the following examples. In the examples,% is% by weight unless otherwise specified. The production methods of reuterin and bacteriocin used in the examples are as follows.

[Production Method of Reuterin] Lactobacillus Lactobacillus reuteri Strain DSM20016 (ATCC5360)
9) the medium (peptone 1%, meat extract 1%, yeast extract 1%, glucose 1%, ammonium citrate 0.2)
%, Sodium acetate 0.5%, magnesium sulfate 0.0
1%, manganese sulphate 0.005%, dipotassium phosphate 0.2%, pH 7.0) 50 ml of platinum after inoculating one platinum loop, 3
50 ml of the culture solution that had been statically cultured at 7 ° C overnight was inoculated into 1 liter of a medium (production medium) in which 4.6% of glycerin was added to the same medium, and statically cultured at 37 ° C overnight. This culture solution was centrifuged at 4,000 rpm for 10 minutes, and the resulting supernatant was further filtered through a membrane filter having a pore size of 0.45 μm to remove bacteria. Approximately 1 liter of the sterilized culture solution was heated to 100 at 40 ° C using a rotary evaporator.
It was concentrated under reduced pressure to g. This concentrate contains about 1% reuterin.
It was contained, and this concentrated solution was used as reuterin in the examples.

[Production method of bacteriocin] Two 1 liter Erlenmeyer flasks were prepared, 500 ml of a medium consisting of 5% corn syrup and 0.5% yeast extract was put in each, and after sterilized by heating at 98 ° C for 30 minutes, Lactococcu
s lactis subsp. diacetilactis and Pediococcus ac
Inoculate idilactici PAC1.0 one by one, 3
It was cultured at 7 ° C. for about 20 hours. This culture solution is at 80 ° C for 10
After heating for 1 minute, the mixture was concentrated and spray-dried to obtain powders containing bacteriocin S50 and pediocin PA1, respectively.

In another 3 Erlenmeyer flasks, 500 ml of MRS medium having the following composition was placed, and after similarly heat sterilized, La
ctobacillus helveticus LP27, Leuconostoc gelidum
Cultivation was performed in the same manner as above except that UAL187 and Propionibacterium jensenii P126 were inoculated, and the culture solution was heated, concentrated, and lyophilized to obtain powders containing lactocin 27, leucosine A-UAL 187, and jensenin G, respectively. It was These powders were used as bacteriocin.

Composition of MRS medium (g / l) Polypeptone [manufactured by Daigo Nutrition Co., Ltd.] 10; meat extract [manufactured by Wako Co., Ltd.] 10; yeast extract [manufactured by Oriental Yeast Co., Ltd.] 5; glucose 20; Tween 80 1; K ₂ H
PO ₄ 2; anhydrous sodium acetate 5; ammonium citrate 2; MgSO ₄ .H ₂ O 0.1; MnSO ₄ .4 ~
5H ₂ O 0.5

Example 1 Alaska pollack frozen surimi 2.5 kg, salt 75 g, mirin 50 g, sodium glutamate 25 g, sugar 25 g,
To the basic composition of 175 g of potato starch and 1 kg of ice water, the preservatives shown in Table 1 were added in the proportions shown in Table 1, and after crushing for 30 minutes, the resulting meat paste was mixed with vinylidene chloride film (folded). About 100 g was packed in a diameter of 48 mm), both ends were ligated, heated in hot water of 90 ° C. for 30 minutes, and then cooled in running water for 30 minutes to obtain a kamaboko. The obtained kamaboko was used as a sample for a storage test together with the kamaboko obtained in the same manner without adding a preservative. In the preservation test, 10 kamaboko were stored in a thermostat at 25 ° C. per test section, and the appearance was visually observed to determine the antiseptic effect. That is,

0 point: No change. 0.5 point: An extremely small spot appears. 1 point: 1 colony-like spot or 1 partial expansion, syneresis and slightly cloudy. 2 points: 2 or more colony-like spots or 2 partial expansions
Individually, the water is a little cloudy. 3 points: Many colony-like spots or many small partial expansions. 4 points: Partially expanded many or partially softened. 5 points: The whole is softened and expanded.

The number of days until the evaluation reached 1 point was obtained for each of the 10 test specimens, and the average was taken as the number of effective storage days. Table 2 shows the results. In addition, as a result of the sensory test, the test section to which the preservative of the present invention was added showed no difference in taste, color, odor, etc., compared with the control section to which the control product was added, and the adverse effect on the quality due to addition was observed. Was not recognized.

[0028]

[Table 1]

[0029]

[Table 2]

It is clear from Table 2 that the preservative of the present invention has a far longer effective preservation period than the control group.

Example 2 A preservative having the composition shown in Table 1 was added to a basic composition containing 500 g of strong flour, 60 g of water and 5 g of flour, and the mixture was sufficiently mixed to prepare noodle strings with a small noodle making machine. It was boiled in boiling water for 4 minutes and cooled with water. After draining, 25 g of this was put in a polyethylene bag and sealed, and 10 bags per test section were stored in a thermostat at 25 ° C., and the change in appearance was observed. The number of days until the evaluation reached 1 point was obtained for each of the test specimens of 1 and the average was taken as the number of effective storage days. The results are shown in Table 3.

0 point: No change. 1 point: Discoloration, softening, net and mold occur in one place. 2 points: discoloration, softening, netting, mold occurred in 2 places or 1
The deterioration of the place spreads. 3 points: Discoloration, softening, neto, and mold spread to half of the whole. 4 points: Discoloration, softening, net and mold spread to 3/4 of the whole. 5 points: Discoloration, softening, neto, and mold spread throughout.

[0033]

[Table 3]

It is clear from Table 3 that the noodles to which the preservative of the present invention is added have a longer effective storage period than the noodles to which the control product is added.

Example 3 1,000 g of ground minced meat, 300 g of onion, 10 g of salt,
The preservative shown in Table 1 was added to a basic composition containing 60 g of wheat flour and 50 g of water, and the mixture was sufficiently mixed, then molded into 10 hamburgers, steamed for 25 minutes, and cooled. After that, 10 pieces per test section were stored at 25 ° C., the change in appearance was observed, and the number of effective storage days was determined according to the same criteria as in Example 2 and the results are shown in Table 4. As shown in Table 4, the effective preservative days were longer when the preservative of the present invention was added than when the control product was added. In addition, as a result of the sensory test, the test plots to which the preservative of the present invention was added were compared with the control plots in color, taste, and
No difference was found in odor and morphology, and no adverse effect on quality due to addition was observed.

[0036]

[Table 4]

Example 4 160 g of egg yolk, 1,440 g of milk, 38 g of sugar, 6.5 g of wheat flour and 6.5 g of cornstarch were used as basic compositions, to which the preservative shown in Table 1 was heated under low heat with sufficient stirring. , 10% of the total weight was boiled down. After cooling this custard cream, it was filled in a cup and stored at 25 ° C. to observe the change in appearance. The number of days until the number of general viable bacteria reached 1 × 10 ⁶ cells / g was defined as the number of effective storage days. The results are shown in Table 5. As shown in Table 5, the addition of the preservative of the present invention
The effective storage days were much longer than those to which the control product was added. Further, as a result of the sensory test, the test group to which the preservative of the present invention was added showed no difference in taste, color, odor, morphology, etc., as compared with the control group, and adverse effects on quality due to addition were recognized. There wasn't.

[0038]

[Table 5]

Example 5 40 ml of commercially available soy milk (pH 7.0) was dispensed into a glass bottle and autoclaved. The preservative having the composition shown in Table 1 was added to and mixed with sterilized soymilk so that the amount shown in Table 1 was obtained, and the total amount was 50 ml. Then, a spore suspension of Bacillus subtilis was added to soy milk, and the spores were about 10 ²
The number of cells / ml was inoculated, heated in a water bath at 90 ° C. for 40 minutes, cooled with water, stored at 25 ° C., and the number of bacteria was measured daily. The number of days until the number of bacteria reached 10 ⁶ cells / ml was defined as the number of effective storage days. The results are shown in Table 6.

[0040]

[Table 6]

Example 6 To 6 kg of an equal mixture of minced pork and mutton,
15% of lard, 2.5% of salt, 0.1% of polymerized phosphate, 0.5% of spice, 70 ppm of sodium nitrite and 10% of ice water were added, and cutting was performed with a silent cutter for 10 minutes. About 15 g of the obtained emulsion meat was filled into the sheep intestine using a manual stuffer. This was dried in a smoke house for 40 minutes, then smoked and steamed, and heated so that the center temperature became 75 ° C. to prepare a wiener sausage. After storing this Wiener sausage in a refrigerator overnight, it was immersed in an aqueous solution of a preservative having the composition shown in Table 1 (prepared so that the amount of each component in the aqueous solution would be the amount shown in Table 1) for 2 minutes, and drained and air dried. , Sterile petri dish 1
Ten pieces of one test section each containing two pieces of wiener sausage were prepared and stored at 25 ° C. to observe changes in appearance. The number of days of effective storage was determined according to the same criteria as in Example 2. The results are shown in Table 7.

[0042]

[Table 7]

[0043]

The food preservative of the present invention can remarkably improve the storability of food, and is particularly effective for extending the shelf life of food contaminated with yeast or mold. Moreover, the original taste and color tone of the food are not changed, and the quality of the food is not adversely affected by the addition, which is extremely effective for preservation of various foods.

Claims (2)

[Claims] 1. A lactic acid bacterium Lactobacillus reuteri (Lact)
reuterin produced by obacillus reuteri)
uterin) and a preservative for food containing bacteriocin produced by lactic acid bacteria. 2. The bacteriocin produced by lactic acid bacteria is Lactococcus lacti.
s), genus Pediococcus, genus Lactobacillus, Leuconos
toc) or Propionibacter
The food preservative according to claim 1, which is a bacteriocin produced by the genus (ium).