WO1994024890A1

WO1994024890A1 - Food preservative and food preservation

Info

Publication number: WO1994024890A1
Application number: PCT/JP1993/000537
Authority: WO
Inventors: Yoshio Hidaka; Kentaro Furube; Yasuaki Matsuda
Original assignee: Eisai Co., Ltd.
Priority date: 1993-04-26
Filing date: 1993-04-26
Publication date: 1994-11-10
Also published as: KR950701194A

Abstract

A food preservative containing lysozyme and alanine, another food preservative containing lysozyme, alanine, L-cystine and/or fumaric acid, and a method of preserving food by adding thereto the food preservative(s).

Description

Specification

Food preservatives and preservation methods

Industrial applications

The present invention relates to a food preservative having an excellent bacteriostatic effect on food and a method for preserving food.

Conventional technology

Conventionally, food preservatives and bacteriostatic agents, lysozyme, or lysozyme combined with lower fatty acid monoglyceride, glycine, gallic acid, e-polylysine, cystine, etc., enhance food preservation effects It has been reported that

The present inventors have already invented a method for preserving a food or drink, to which glycine, cystine, cystine, invertase, vitamin, and the like, and a microbial cell wall lysing enzyme represented by lysozyme are added. (Japanese Patent Publication No. 547-1780). In addition, for example, a food preservative containing lysozyme and a fatty acid glyceride and glycine (Japanese Patent Application Laid-Open No. 2-316666), and lysozyme, gallic acid, phytin or vegetarian Food preservatives that combine glycerin and ε-polylysine (Japanese Patent Publication No. 2-2831).

However, glycine has a peculiar off-flavor called “gourmet taste”, and there is a problem that adding glycine makes food taste worse. Also, glycine had the disadvantage that browning progressed quickly because the browning reaction occurred quickly.

Other food preservatives have the disadvantage that they have little preservative effect on foods in which lactic acid bacteria or Bacillus subtilis cause deterioration in quality. Problems to be solved by the invention

Therefore, the present invention relates to foods in which lactic acid bacteria or Bacillus subtilis cause deterioration in quality, for example, food products such as processed meat products, meat dishes, fishery products, pickles, vegetables, custard dough, soft cream, etc. A preservative that has an excellent preservative effect on food and does not adversely affect the taste of food And to develop preservation methods.

Means for solving the problem

The present inventors have proposed a food effective against the lactic acid bacteria Leuconostoc Lactis, Leuconostoc mesenteroides, and general viable bacteria which cause the above foods to spoil. After repeated research on preservatives and preservation methods, it was found that the combination of lysozyme and alanine exhibited a strong bacteriostatic effect.

Furthermore, the present inventors have found that the addition of L-cystine or fumaric acid to lysozyme and alanine enhances the preservative effect.

Furthermore, the present inventors have also found C in which the preservative effect is enhanced by combining L-cystine and fumaric acid with lysozyme and alanine.

The foods that are the subject of the present invention are optional, but include, among others, processed meats, meat dishes, fishery products, pickles, fish, custard cream, soft cream, etc. it can.

In practicing the present invention, lysozyme and alanine are blended by an ordinary method. Or mix lysozyme, alanine and L-cystine, or mix lysozyme, alanine and fumaric acid. Alternatively, combine lysozyme, alanine, L-cystine and fumaric acid.

Lysozyme, alanine, L-cystine and fumaric acid were added in the following amounts: lysozyme 0.0005 to 0.05% by weight, alanine 0.1 to 1.0% by weight, L-cystine 0.002 to 2% by weight, fumaric acid 0.01 to 0.15% Combinations in the range of weight% are preferred.

The minimum doses of lysozyme 0.0005% by weight, alanine 0.1% by weight, L-cystine 0.002% by weight, and fumaric acid 0.01% by weight are the minimum concentrations showing the antibacterial effect. Lysozyme 0.05% by weight as the maximum dose is an amount that does not increase the effect even if the dose is increased further, and 1.0% by weight of alanine gives a stronger taste when administered more than that. I do not like it.

When L-cystine is administered at 0.2% by weight or more, sufficient solubility cannot be obtained. In addition, when fumaric acid is administered in an amount of 0.15% by weight or more, the pH (approximately 5.8) decreases, and it does not coagulate sufficiently in food that is coagulated by heating. _(If the range of the dose of lysozyme and alanine is indicated by the mixing ratio, Zozyme, alanine = 1: 2 to 2000. The ratio of lysozyme, alanine and L-cystine is lysozyme: alanine: L-cystine = 1: 2 to 200: 0.04. In the case of lysozyme, alanine and fumaric acid, the ratio is as follows: lysozyme: alanine: fumaric acid-1: 2-2000: 0.2 to 300. In addition, lysozyme, aranine, L The ratio of cystine and fumaric acid is preferably 1: 2 to 2000: 0.04 to 400: 0.2 to 300.

In practicing the present invention, lactose, dextrin, salt and the like may be added to the food together with the preservative composition shown in the present invention.

Example

Next, the present invention will be specifically described with reference to Examples, Comparative Examples, and Experimental Examples. However, the present invention is not limited to only these examples. Example 1

Knead 0.0006% by weight of lysozyme and 0.15% by weight of alanine into the following ingredients to make four sausages of 20 to 25 g per one. After boiled for 30 minutes at a center temperature of 71 ° C or more, they were individually packed and stored at 10 ° C. Sausage combination

Pork lean 6 500 g

Fat 3 50 g

250 g of water

1 7 g

Sodium nitrite 0.13 g

The number of initial bacteria and the number of bacteria on the 5th, 10th, and 15th day of storage were examined. The number of initial bacteria: <10 ² , the number of 5th day of storage: <10 ² , the 10th day of storage number: Ku 1 0 ^2, Save 1 day 5 number of bacteria: 4.4x 1 0 ^3, and the no change in the number of bacteria to store 1 day 0, _c example 2 was found to exhibit good preservative effect

A sausage was prepared by kneading 0.0006% by weight of lysozyme, 0.15% by weight of alanine and 0.003% by weight of L-cystine into the same sausage material and formulation as in Example 1. Boyle in the same manner as in Example 1, save days and, as a result of examining the number of bacteria, number of initial bacteria: <1 0 ^2, stored for 5 days Number of bacteria: <1 0 ^2, Save 1 Day 0 Number of bacteria: <1 0 ^2, stored for 1 day 5 the number of bacteria: became a 2.6X 1 0 ^3. There was no change in the bacterial count up to the 10th day of storage, indicating a good storage effect.

Example 3

A sausage was prepared by kneading 0.0006% by weight of lysozyme, 0.15% by weight of alanine and 0.03% by weight of fumaric acid into the same sausage material and formulation as in Example 1. The boil, the number of storage days, and the number of bacteria were examined in the same manner as in Example 1. As a result, the number of initial bacteria: 10 ² , the number of 5 days of storage: <10 ² , the number of 10 days of storage : <1 0 ^2, stored for 1 day 5 the number of bacteria: became 2.2x 1 0 ^3. There was no change in the bacterial count up to the 10th day of storage, indicating a good storage effect.

Example 4

To the same sausage material and formulation as in Example 1, 0.0006% by weight of lysozyme, 0.15% by weight of alanine, 0.003% by weight of L-cystine and A sausage was prepared by kneading in 0.03% by weight of malic acid. And Boyle real Example 1 and the same method, storage days and the results of examining the number of bacteria, number of initial bacteria: <1 0 ^2, stored for 5 days Number of bacteria: <1 0 ^2, Save 1 Day 0 bacteria Number: 10 ² , Storage 15 days after bacterial count: 3.8 × 10 ³ There was no change in the bacterial count up to the 10th day of storage, indicating a good storage effect.

Comparative example

Using only 0.0006% by weight of lysozyme, a sausage was prepared in the same manner as in Example 1 and the number of bacteria was examined. As a result, no increase in the number of initial bacteria was observed by the 5th day of storage, but the storage was continued. 1 day 0 1.3X 1 0 ³ is stored for 1 day 5 was seen an increase of 3.9x 1 0 ⁸ and the number of bacteria. This demonstrates that the addition of alanine, L-cystine and / or fumaric acid to lysozyme extends the shelf life.

Next, the effects of the present invention will be shown by experimental examples.

Experimental example 1 Minimum inhibitory concentration for lactic acid bacteria

Measurement of the minimum growth inhibitory concentration of lysozyme, lysozyme + alanine, lysozyme + alanine + L-cystine, lysozyme + alanine + fumaric acid, lysozyme + alanine + L-cystine + fumaric acid against lactic acid bacteria Was done.

Test method

The test was performed by a two-fold dilution method using a conventional liquid medium.

The proportions in the formulation were lysozyme 0.2%, alanine 50.0%, fumaric acid 10.0%, L-cystine 1.0%, and the remainder was dextrin, lysozyme, lysozyme + alanine, lysozyme + alanine. Addition tests were performed on the blends of + L-cystine, lysozyme + aranine + fumaric acid, and lysozyme + aranan + L-cystine + fumaric acid. The drug concentration was determined in five steps of 0.5%, 0.25%, 0.125%, 0.063% and 0% for each formulation.

The strain used is Leuconostoc Lactis IF 0 1 2 4 5 5.

As a test medium, glucose, magnesium sulfate, and peptide medium (pH 7.0) were used. Incubated at C for 48 hours. As a medium for measuring the number of bacteria, a plate count medium supplemented with BCP was used and incubated at 37 ° C. for 48 hours, and the number of bacteria was counted.

Test results

Table 1 shows the test results. The numbers in the table indicate the number of bacteria.

table 1

Fruit

Lysozyme alone had no bacteriostatic effect at a concentration of 0.5%.

The minimum growth inhibitory concentration when lysozyme and alanine are combined is as low as 0.25%, indicating high antibacterial activity against lactic acid bacteria. In addition, the minimum growth inhibitory concentration when lysozyme, alanine and L-cystine are combined, and when lysozyme, alanine and fumaric acid are combined are 0.125%, respectively, and the antibacterial activity is further enhanced. Is shown. Furthermore, when lysozyme, alanine, L-cystine and fumaric acid were added, the minimum growth inhibitory concentration was 0.063%, indicating a stronger antibacterial activity.

Experimental example 2 Minimum growth inhibitory concentration against Bacillus subtilis

For the combination of lysozyme, lysozyme + alanine, lysozyme + alanine + L-cystine, lysozyme + alanine + fumaric acid, and lysozyme + alanine + L-cystine + fumaric acid, the minimum growth inhibitory concentration against lactic acid bacteria was A measurement was made.

The strain used is Bacillus subtilis T0A303 3.

The test was performed in the same manner as in Experimental Example 1, using the formulation mixture ratio and medium.

Test results

Table 2 shows the test results. The numbers in the table indicate the number of bacteria. Table 2

Lysozyme alone had no bacteriostatic effect at a concentration of 0.5%. The minimum growth inhibitory concentration when lysozyme and alanine are combined is 0.5%, indicating antibacterial activity against Bacillus subtilis.

The minimum anti-proliferative concentration when lysozyme, alanine and L-cystine are combined, and when lysozyme, alanine and fumaric acid are combined are 0.25%, respectively, to further enhance the antibacterial activity. Is shown.

Furthermore, when lysozyme, alanine, L-cystine and fumaric acid were added, the minimum growth inhibitory concentration was 0.125%, indicating a stronger antibacterial activity.

Next, the effect of glycine and alanine on the taste of food was subjected to a sensory test using a power star cream and sausage. These sensory tests were conducted by 10 panelists of the Japan Association of Food and Fat Testing.

Additives were prepared as in Tables 3, 4, 5, and 6 below. Control additives A-1, A-2, A-3 and additives B-11, B-2, B-3 are added to the custard cream and control additives C-11 are added to the sausage. , C-12, and additives D-1, D-2.

The control additives A-1, A-2, A-3 and the control additives C-11, C-12 consist of the same components of lysozyme, glycine, cystine and lactose, but with different doses (addition rates) It is an additive. The additive ratios of the control additives A-1, A-2, and A-3 were 0.5, 1.0, and 2.0% with respect to the castor cream to which no additives were added, and the additives were added. The additive ratios of the control additives C-11 and C-2 to the sausages without the addition were 0.5 and 1.0%. Additives B-1, B-2 and B-3 and additives D-1 and D-2 consist of the same components of lysozyme, alanine, cystine and lactose, but the dose (addition rate) Different additives. Additives B-II, II-2 and III-3 were added to Custard Dream to which no additives were added at 0.5, 1.0, and 2.0%, and additives were added. The additive ratio of additives D-1 and D-2 to the sausage was 0.5 and 1.0%, respectively. Table 3

Additives to be added to the caster dough

*): The addition ratio indicates the weight ratio of the additive to the weight of the custard cream without the additive. Table 4

Additives to be added to the castor lyme Lysozyme alanine cystine lactose Π a 1 Addition rate (¾) 0.002 0.45 0.0075 0.0405 0.5

Dose (mg) 7.0 1579.5 26.3 142.2 1755.0 Addition rate (¾) 0.004 0.90 0.015 0.081 1.0 Appendix B-2

Dose (mg) 14.0 3159.0 52.7 284.3 3510.0 Addition rate (%) 0.008 1.80 0.03 0.162 2.0 Addendum B-3

Dose (mg) 28.1 6318.0 105.3 568.6 7020.0 Table 5

Control additive added to sausage

*): Addition ratio indicates the weight ratio of additive to the weight of sausage without additive.

Table 6

Additives added to sausages

Test 1 Sensory test using Custard Dream

Test method

① 2 yolks (60 g), granulated sugar 502, control in a bowl Add Excipient A-1 and mix with a whisk until white. Separately, put the milk in a pan over medium heat and boil until just before the film is formed. ② Combine 13 g of flour and 13 g of corn starch with ① and mix with a sieve and whisk. Mix until powdery.

③ Add boiled milk 200 cc to 2-3 in 2 or 3 times.

After mixing well, put it back in the pot containing milk, put it on medium heat, gently mix the surface with a whisk, and when it becomes thick, mix quickly like rubbing the bottom of the pot.

たら When the glue appeared on the surface and the surface became shiny and shiny, it was removed from the fire, added with margarine (15 g, salt-free), and quickly mixed with a whisk to form a custard cream.

Instead of Control Additive A-1, make a caster dream containing Control Additive A-2, Control Additive A-3, Additive B-1 and Additive B-2 and Additive B-3, respectively. The taste was compared. In addition, we made a Custard Dream without additives and used it as a blank control. [Result]

Custard cream without additives had a good taste. Control Additive A-1 gives a slightly harsh taste, Control Additive A-12 adds a little harsh taste, and Control Additive A-3 adds a strong harsh taste. Taste was exhibited. Since the control additives A-1, A-2 and A-3 contain glycine that is not contained in additives B-1, B-2 and B-3, this taste is due to glycine. it is conceivable that. On the other hand, the addition of additives B-1, B-2 and B-3 containing alanine gave the custard cream a good taste. Table 7 shows the results. Table 7 Performance evaluation of custom cream

Test 2 Sensory test using sausage

Test method

① Crush 10 g of ice with a mixer, 70 g of pork, 20 g of lard, 1.5 g of salt, 0.1 lg of sodium nitrite, 0.3 g of sodium glutamate, Koshi Pour 0.25 g, nutmeg 0.1 g, allspice 0.05 g, cinnamon 0.03 g, and control additive C-11. Add stickiness with a mixer and mix until air enters.

② Each piece is shaped into a disc of 30 g and steamed for 10 minutes with a steamer.

Instead of control additive C-1, sausage containing control additive C-12, additive D-1 and additive D-2 was prepared, and the taste was compared. In addition, sausages without additives were prepared and used as blank controls.

[Result]

The sausage without added additives had a good taste. The one to which control additive C-1 was added exhibited a slightly harsh taste, and the one to which control additive C-2 was added exhibited a strong harsh taste. Glycine not included in Additives D-1 and D-2 was included in Control Additive C-1 and Control Additive C-2. It is considered that this taste is due to glycine. On the other hand, the addition of the additives D-1 and D-2 containing alanine exhibited a good taste. Table 8 shows the results.

Table 8 Evaluation of government functions

Next, for glycine and alanine, the transmittance and the reduction rate of amino acid were examined in order to narrow down the progress rate of browning due to the Maillard reaction.

Test 3 Permeability due to browning

Test method

A solution of 2 g of glycine and 10 g of glucose dissolved in 188 ml of Cortov buffer (pH 6.0) (concentrations of glycine and glucose were 1% (w / v) and 5% (/ V) Put 15 m 1 of the solution into a test tube and spread a cork た with a wrapping paper. The test tube is heated in a thermostat at 120 ° C for 1 hour. After quenching with permanent water, the transmittance (500 nm) was measured using Shimadzu spectrophotometer UV 240.

In the above test, a solution to which 2 g of alanine was added instead of 2 g of glycine was prepared, heated under the same conditions, and its transmittance (500 nm) was measured and compared. The transmittance of glycine and alanine at pH 6.2, pH 6.5, pH 7.0 and pH 8.0, respectively, was also measured and examined. BRIEF DESCRIPTION OF THE FIGURES

Figure 1

It is explanatory drawing which shows the transmittance | permeability by browning.

Figure 2

FIG. 3 is an explanatory diagram showing a time-dependent decrease rate of an amino acid.

[Result]

As shown in Fig. 1, the value of transmittance (500 nm) is higher for alanine than for glycine and less browning due to the Maillard reaction at each pH between 6.0 and 8.0. Is shown.

Test 4 Reduction rate of amino acid

5 g of glycine and 20 g of glucose were dissolved in 100 ml of distilled water, stirred, heated at 100, and heated to 100 to destroy the amino acid to observe the progress of the Maillard reaction. Fraction was measured over 24 hours over time.

[Result]

Compared to glycine, the percentage of alanine decreased over time (0, 4, 7, 13,

2 4.5 hours), it was found that glycine was less likely to cause browning due to the Maillard reaction. The result is shown in figure 2.

The invention's effect

According to the food preservative and preservation method of the present invention, a combination of lysozyme and alanine, a combination of lysozyme, alanine and L-cystine, a combination of lysozyme, alanine and fumaric acid, and The combination of zozyme, alanine, L-cystine and fumaric acid shows remarkable antibacterial activity and achieves a preservative effect.

In addition, since each component is a natural product, it has high safety, does not affect the taste of food, and hardly causes browning. Therefore, it is suitable as a food preservative and method. Therefore, it is possible to increase the amount of additives, so it is excellent in processed meat, meat dish, fishery products, pickles, varieties, custard cream, soft cream, etc. A preservation effect can be obtained.

Claims

The scope of the claims

1. A food preservative containing lysozyme and alanine.

2. The food preservative according to claim 1, wherein the ratio of lysozyme to alanine is 1: 2 to 2000.

3. A preservative for food, characterized by containing lysozyme, alanine, L-cystine and / or fumaric acid.

4. A food preservation method characterized by containing lysozyme and alanine.

5. A method of preserving food, comprising lysozyme, alanine, L-cystine and / or fumaric acid.