JPS61128867A - Fish meat paste product - Google Patents

Abstract

PURPOSE:The title paste food such as KAMABOKO (boiled fish-meat paste) that contains ethanol, alcohol oxidase and catalase, thus enabling the product to be stored for a prolonged period by inhibiting contamination with microorganisms and their propagation without any adverse effect on its flavor, taste color and viscoelasticity. CONSTITUTION:The objective paste product is produced by adding, to fish meat, (A) ethanol, preferably 0.1-2wt%, (B) alcohol oxidase [EC1.1,3,13], preferably 0.1-5 U/g and (C) catalase [EC1,11,1.6], preferably in an amount 2-20 times the amount of component B added. Component B is preferably obtained together wtih component C by culturing a strain of Lipomyces in a medium containing only methanol as a carbon source, separating the culture mixture from cell bodies and purifying it.

Description

[Detailed Description of the Invention] [Object of the Invention] The present invention relates to a fish paste product that can be stored for a long period of time, and more specifically to a fish paste product containing a small amount of ethyl alcohol and specific amounts of alcohol oxidase and catalase. It is related to. The aim is to create nine color tones of flavor and flavor for fish paste products.

To provide a fish paste product that can be stored for a long period of time by suppressing contamination and proliferation of microorganisms that cause spoilage without impairing physical properties such as viscoelasticity.

[Industrial application field]

The present invention relates to fish paste products such as kamahoko and chikuwa.

Manufacture of hanpen, fish sauce, fish cakes, datemaki, fish ham, fish noodles, fish paste, etc.

It is used in the food manufacturing industry for the purpose of improving food preservation during the distribution stage.

[Conventional technology]

When fish paste products are wrapped in cellophane or the like and kept in high humidity, a colony of slimy bacteria called Neto will form, so it is usually used as a preservative.
Nrupinate salts such as potassium nrubate and sodium nrubate and hydrogen peroxide have been used, but since the Ministry of Health and Welfare pointed out the carcinogenicity of hydrogen peroxide in January 1981, hydrogen peroxide has been used. Instead, alcohols, amino acids, organic acids, enzymes such as suzozyme, etc. are used alone or in combination.

[Problems that Hamoe is trying to solve] ``Of the preservatives mentioned above, sorbic acids have excellent antiseptic effects, but their bactericidal and bacteriostatic effects on spore-forming bacteria are weak. Among alcohols, ethyl alcohol has a bactericidal effect by itself, but in order to achieve this effect, 3
A concentration of 0% or higher is required. However, as the amount added increases, the unique alcohol odor of ethyl alcohol will stick to the food, impairing the flavor and taste of the fish paste product. Furthermore, ethyl alcohol has a protein denaturing effect, and adding it in high concentrations will change the physical properties of the fish paste product. For this reason, the amount used is limited, and the concentration of ethyl alcohol added to food is limited to about 3q6.

Therefore, such low concentrations are sufficient for sterilization.

Difficult to obtain bacteriostatic effect.

[Means to solve the problem]

The present inventors have lowered the concentration of ethyl alcohol to improve the quality of fish paste products without affecting the flavor, taste, physical properties, etc., and when using sorbic acid alone or adding other existing preservatives to sorbic acid. As a result of searching for a substance that enhances the preservative effect, we found that by coexisting alcohol oxidase [Ecl, 1.3.13] and catalase [EC1, 11, 1, 6], which are enzymes that oxidize ethyl alcohol, this The inventors have discovered that the object can be achieved and have developed the present invention.
That is, the present invention uses ethyl alcohol, preferably 0.1
~2 fk% alcohol and alcohol oxidase, preferably 0. IU/g~5. The present invention provides a fish paste product containing U/'g of alcohol oxidase and camerase, preferably catalase in an amount of 2 to 20 times the added amount of alcohol oxidase.

To obtain the monoalcohol oxidase used in the present invention, a strain that grows by assimilating methanol as the only carbon source is preferred, but any strain that grows alcohol oxidase may be used. .

□Specific examples of these strains include Candida (Co).
ndlda), genus Ha'nm'enula
), Plchlm and Torrogsis (T
orulopalg), Kloeckella
＠ra), Lipomyces (Llpomycea), and more specifically, Hansenula polymorph 7 (Hansenula polymorph 7).
), Candida? Idi = (Candlda b)
oJdjn), Vichia pastoris (PIchia
pastorla), Behear Contigure (Ple
hla eontlgua), L'ipomyces methanosylviensis
-sllvlenslm) and other methanol-assimilating bacteria. Among these, particularly preferred in the present invention are strains of the genus Ribomyces, specifically strains of the genus Ribomyces.
Methanosylviensis reeds, this natural.

Even artificial mutant strains can be used if they produce alcohol oxidase using methanol as a carbon source.

In addition, the above-mentioned IJf Myces methanosylviensis was already registered in April 1980 as Microorganism Accession No. 5489.
It was deposited with the Agency of Industrial Science and Technology's Institute of Microbial Technology (Rζ) on April 15th, and its mycological properties were published in the
This is as described in Japanese Patent No. 7-28551.

The medium used for alcohol oxidase production by the yeast used in the present invention contains methanol or ethanol, insolopanol, n-propanol, etc. as the main carbon source, and appropriate amounts of nitrogen sources, inorganic substances, vitamins, and other growth-promoting substances. Although either a synthetic medium or a natural medium can be used, a medium containing methanol as the sole carbon source is preferable.

The yeast used in the present invention can grow up to a concentration of 4% by weight of methanol, which is the main carbon source in the medium, but the culture method is as follows:
In order to reduce the evaporation loss of methanol due to aeration during culture, it is recommended to keep the initial concentration of methanol in the culture medium as low as possible and add methanol as methanol is consumed to keep the concentration of methanol in the culture medium at a low level. preferable.

As a nitrogen source, a nitrogen compound such as ammonium salt, urea, cornstarch liquor, yeast extract, or pegtone is used. In addition, examples of other inorganic groups include magnesium salts, phosphates, potassium salts, calcium salts, iron salts, etc., and as necessary substances essential for growth or growth promoting substances such as vitamins and amino acids. It is also preferable to add.

The yeast used in the present invention can be grown at a culture temperature of 20 to 42°C, but a temperature of 34 to 39°C is particularly preferable from the viewpoint of production of alcohol oxidase. Also, P) 13
It is possible to grow at a pH of ~7 to 7, but a pH of 3.5 to 5.5 is preferable, and furthermore, in order to produce alcohol oxidase by conducting continuous culture for a long period of time without contamination with bacteria other than the target yeast. It is particularly preferable to culture at pH 4 to 5. Furthermore, the culture method may be either batch culture or continuous culture.

Alcohol oxidase and catalase in the culture obtained by smelling are mainly produced within the bacterial cells. Therefore, in order to isolate the alcohol oxidase produced in the culture solution, collect the bacterial cells by centrifugation, then crush the cells using a Dynomyl cell crusher, Braun cell crusher, etc., and then add them to a phosphate buffer. Crude cell extract,
Furthermore, salting out method, solvent precipitation method2. It is isolated as a partially purified product or purified product obtained by purification methods such as column chromatography, but usually ammonium sulfate is added to this crude extract to a saturation of about 40%, and the resulting precipitate is removed using a centrifuge. , Add ammonium sulfate to the supernatant from which the precipitate was removed and add 50
% saturation. The resulting precipitate was suspended in sodium phosphate buffer and then dialyzed against alcohol oxidase. -ze ratio is 1:10~20
(U ratio of activity) is obtained. In order to further purify alcohol oxidase, it is adsorbed onto a DEAE-Sephacel column equilibrated with the same buffer. Then, elute with a continuous sodium chloride concentration gradient of 0.25M to 0.4M and collect both fractions containing alcohol oxidase activity. Concentrate the active fraction and use Sephadex, G-=20
Purified enzyme can be obtained by performing filtration and collecting the active fraction.

The method for measuring alcohol oxidase activity is to use a methanol/peroxidase/chromadon reagent solution (7 Btt water).
r1. Methanol 1ml So, 5M phosphate buffer PH8
,71omp, 0.4% peroxy/--h" f8
Liquid (100 units/m9) 10Mg, 196 o
-Dianisidine solution 1 ml) 4. ml was kept warm at 25°C for 10 minutes, 1 ml of enzyme solution was added and reacted for 5 minutes, then 0.2N of 4M hydrochloric acid was added to stop the reaction, and 410 n
Measure the absorbance at m. 1 μm per minute under the above conditions
The amount of enzyme that generates hydrogen peroxide in ole was defined as 1 unit (U>).

Alcohol oxidase produced and accumulated in the bacterial cells by culturing a particularly preferred strain of the present invention, Myces methanosylvini/cis, in a liquid medium has (A) activity, temperature range: 25-4
5°C, optimal temperature = 40°C, - range: 6-9, optimal pH:
7.5, (B) Molecular weight 740,000, Subunit molecular weight: 89,000, (C) km value for ethanol: 50 mM, Q)) Stability, temperature:
Below 40°C, it is an alcohol oxidase with the characteristics of pl(: 6 to 7.5).

The catalase of the present invention is produced together with alcohol oxidase in the methanol-assimilating OX cells, and crude extracts obtained from these cells by the method described above, as well as partially purified products or purified products, are used. In addition, 9 catalases derived from animals such as commercially available liver catalase, Mleroecus spp., Aspergillus spp., Penicillium (P@nla1)
Catalase derived from other microorganisms such as the genus 111um and the genus Blcllua can also be used.

To measure the activity of catalase, dilute the enzyme to about 40 U/rul with 0.05 M salt/acid buffer (pH 7,0) and use it as a sample.
ml and 9.0.05M phosphate buffer (pH 7,0
) to a volume of 5 ome and use it as a substrate solution. 2.9 tnl of the substrate solution was put into a 10-sized quartz cell, and after leaving it in a constant temperature water bath at 25°C for 10 minutes, the enzyme solution 01. ITIL
e and measure the time required for the absorbance at 240 nm to decrease from 0.45 to 0.40 using a spectrophotometer.

What is the unit of Rad activity? 1 minute per minute at 25℃
The enzyme activity for oxidizing hydrogen peroxide in μmot is assumed to be about 1 year (U).

The method for incorporating the preservative composition consisting of ethyl alcohol, alcohol oxidase, and catalase into fish paste products is not particularly limited, such as dipping, spraying, and rolling, but preferably rolling. The concentration of ethyl alcohol added is preferably 0.1 to 2% by weight, and the amount of alcohol oxidase added is 0.1 to 2% by weight. IU
, #-Food food Kushi et al. 5U/II-Food per unit is preferable,
The amount of catalase to be added is preferably 2 to 20 times that of alcohol oxidase in terms of U. The concentration of ethyl alcohol is 0
．． If it is less than 1%, there will be no preservative effect, and if it is more than 2%, it will change the flavor of the fish paste product, which is not desirable.

Also, the amount of alcohol oxidase added is Q, I U/
g-If the food content is too low, the preservative effect will be lost, 5U/I
! - If there is too much per food item, it is not preferable because it impairs economic efficiency. Furthermore, if the amount of catalase added is more than 20 times (in terms of U) the amount of alcohol oxidase, the preservative effect will decrease, and if it is less than 2 times, it is undesirable that peroxide accumulation will remain.

Examples of the fish paste products of the present invention include kamahoko, chikuwa, hanpen, tsumire, fish cake, datemaki, fish ham, fish meat sage, and minced fish.

Any method for manufacturing such a fish paste product may be any commonly used manufacturing method. That is, the process is to wash the fish meat with water, mash it, add additives such as reinforcing agents and seasonings, mash it, shape it, and heat it (steaming, baking), preferably at 80 to 100°C. It becomes more. Any type of fish meat may be used as the sushi meat, but white fish with low oil content, such as sea bream, flounder, flying fish, shark, yellowtail, kisu, cod, croaker, and conger conger, are preferably used. As the reinforcing agent, one that imparts viscosity, such as alginic acid, gelatin, and starch, is used. In addition, as seasonings, cilantro, sugar, salt, starch syrup, monosodium glutamate, spices (pepper, ginger), fats and oils, etc. are used.

〔effect〕

The fish meat paste product of the present invention obtained in this way has excellent storage stability, and is also excellent in physical properties such as flavor, taste/color tone, and viscoelasticity.

〔Example〕

A detailed explanation will be given below with examples.
"H" shall be based on weight.

Example 1. Preservation effect of C-casing stuffed kamaboko) Table-1
According to the blending ratio shown in , these were mixed with 7-dokatsuta squid, partially dissolved in cold water, and mixed with ethyl alcohol and alcohol oxidase.

Catalase was added to the concentration shown in Table 2. For comparison purposes, a sample was prepared in which a commercially available sorbic acid preparation was added in place of ethyl alcohol, alcohol oxidase, and catalase. It's easy to slide up after the lift. wo M diameter 4 am
Filled with 1209 into a vinylidene chloride film, left to sit at 20°C for 16 hours, boiled in hot water at 90 to 95°C for 40 minutes, cooled with running water for 20 minutes, and stored in a thermostat at 20°C.
Sensory observations were made by Neto et al. The evaluation method, which involves sensory observation of the growth and reproduction status of bacterial colonies such as Neto over time, is based on the method of Shohin, Matsuda et al. (Nippon Suisan Jitsu Vow, 31, A2, 1965). , −(
0), ±(0,5).

10'(1,0') , -)+ (2,0)+++ (
A seven-point scoring system of 3,0) i (4,0> and 1+14) (5,0) was used. +(1,0) was used as the basic value for the number of effective days of storage.

The number of samples in each ward shown in Table 2 is 8, and the basic daily calculation value is + (1, 0), so the number of samples x rating = 8
Those with the following ratings were judged to be effective, and the effective storage days were calculated as shown in Table 3. The scores for each day in Figure 1 were expressed as the sum of scores for all samples in each district.

The above dates t'+point' are shown in Figure 1. From this, the effective storage days are shown as shown in Table 3, and the addition of ethyl alcohol, alcohol oxidase, and catalase has a remarkable ability to extend the storage period compared to those without preservatives or those without sorbic acid; (1A'* Example 2 (Steamed Kamabako with a plate) According to the blending ratio shown in Table 1, the fish fillets were diced using a food cutter, and the minced fish was molded onto a veneer with a plate attached.
Alcohol oxidase, adjusted to 15 °C after sitting at room temperature for 30 minutes and evaporating at 85-90 °C for 30 minutes;
After immersing it in a catalase and ethyl alcohol solution for 20 seconds and wrapping it in moisture-proof cellophane, it was stored at 20° C. in a thermostatic chamber, and a storage test was conducted in the same manner as in Example 1.

The composition of the dipping solution is alcohol oxidase 1000
U/A, catalase 10.0 OOU/l, and ethyl alcohol 511/l'ljx were used. The oral scores of the above tests are shown in Figure 2. From this, the effective storage days are shown as shown in Table 4, and a remarkable preservation effect was observed by immersion in a solution of alcohol oxidase, catalase, and ethyl alcohol.

:15)

[Brief explanation of drawings]

Figure 1 shows the present invention (A
-1 to 3 sections), the additive-free section, and the sorbic acid preparation (SOA section) as a comparative example, and the sensory observation scores and effective storage days. Figure 2 shows the sensory observation scores and effective storage days of the present invention and the additive-free group using the immersion method for Shikamaboko of the Itatsuki type.

Claims (1)

[Claims] 1. Ethyl alcohol and alcohol oxidase [EC
1,1,3,13] and catalase [EC1,11,1
, 6]. 2. The fish paste product according to claim 1, wherein the concentration of ethyl alcohol is 0.1 to 2% by weight. 3. The amount of alcohol oxidase added is 0.1 U/g ~
The fish paste product according to claim 1, characterized in that the amount of fish meat is 5 U/g. 4. The fish paste product according to claim 1, wherein the amount of catalase added is 2 to 20 times the amount of alcohol oxidase added.