JP3435461B2 - Seaweed pickle floor and method for producing pickles using it - Google Patents

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a pickled bed made of seaweed as a raw material, which can easily produce pickled vegetables having a good scent and excellent water retention, and a method for producing pickled vegetables using the same.

[0002]

[Prior Art] Pickles have been an indispensable ingredient on the Japanese table since ancient times. There are various kinds of pickles such as pickled pickles, pickled pickles, pickled nara, etc. Above all, pickled pickles were common people, and each time there was a pickled floor in each family, and pickled vegetables were familiar. However, the peculiar smell of Nuka floor tends to be shunned by young women, it takes a lot of time to manage the floor, such as frequent stirring, and the Nuka floor loosens when the water content is adjusted incorrectly, and good pickles The habit of making pickles at home is gradually being lost in modern homes because there are difficult factors that require experience, such as being unable to make them. On the other hand, the inventors of the present invention have developed a technique for producing finely divided seaweed detritus by decomposing seaweed with an enzyme into single cells and then fermenting it with a microorganism. Regarding the technique of fermenting this seaweed while unicellularizing, the present inventors have invented the invention uniquely in Japanese Patent Application No. 2000-300399 "Production Method of Seaweed Detritus Fermented Feed", but have completed it, but in this invention, use It was an invention whose purpose was limited to the use as an initial feed for aquaculture. However, this fermentation product has a fine particle-like and smooth physical property, has a fruity aromatic odor, is excellent in water retention, is excellent in preservability, and is lactic acid-fermented. As a result of pursuing the potential of the above, as a result of earnest efforts, the inventors have invented a completely new food material called a pickled bed made of seaweed as a raw material, which has characteristics that overcome the problems of the above-mentioned floor.

[0003]

That is, the present invention is
It is possible to produce pickled vegetables that are comparable in flavor to conventional pickled vegetables, have a low salt content, do not loosen the floor even when used again, and do not care about their management. The challenge is to provide. Further, it is an object of the present invention to provide a method for producing a pickle which has a flavor similar to that of conventional pickles using the pickle floor.

[0004]

SUMMARY OF THE INVENTION The present invention is directed to such a section.
It was done to solve the problem,
Seaweed pickled seaweed floor and method of manufacturing pickles using the same
Is. Seaweed by glycolytic enzymes including cellulase
Decomposes and transforms into unicellular particles, as well as lactic acid bacteria
Fermented by a microbial consortium of yeast and yeast
A seaweed pickle bed consisting of the obtained seaweed fermentation product. The seaweed
Manufacturing pickles using a seaweed pickling floor consisting of fermentation products
how to. Microbiota consisting of lactic acid bacteria and yeast in the present invention
As a lactic acid bacterium,Lactobacillus
Genus,StreptococcusGenus,LeuconostocGenus,Pediococcus
Genus,Tet ragenococcusGenus,BifidobacteriumBelong to the genus
Lactic acid bacteria or yeastDebaryomycesGenus,CandidaGenus
andSaccharomycesDesirable to use yeast belonging to the genus
Good In particularLactobacillusFor genus lactic acid bacteria,Lactobacil
lusbrevis NRIFS B5201 strain (FERM BP-7301)De
baryomycesFor genus yeast,Debaryomyces  hanseniiNRIFS
Y5201 strain (FERM BP-7302)Candida For genus yeast,Can
dida  zeylanoides NRIFS Y5206 strain (FERM BP-7303)
It is desirable to use. In addition, the microbial consortium
Means a combination of multiple microbial species.
Also called a group of organisms or complex microorganisms. Not alone
By using multiple microorganisms in combination,
Form stable microflora in the field and simultaneously perform various functions
Since it is possible to demonstrate it, composting of garbage and stone
When decomposing oil, etc., the microorganisms are treated as a consortium.
It is getting more and more attention to throw it in.

[0005]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention will be described in detail below.
The seaweeds used in the present invention include wakame, mackerel,
Examples include brown algae such as Hijiki, red algae such as Amanori, green algae such as Ulva, and flowering plants such as eelgrass, and pickled beds can be produced from all seaweeds. These seaweeds can be used as raw materials in the form of raw or dry powder. Among the above-mentioned seaweeds, wakame seaweed is particularly preferable from the viewpoints of production efficiency of unicellular seaweed particles (hereinafter referred to as SCD), ease of mass procurement of raw materials, and empirically known foods. It is preferable to use kelp, kelp, hijiki or laver.

The glycolytic enzyme used in the present invention includes:
Cellulase must be used because glucose must be produced that can be used as a substrate for fermentation after decomposition. In addition, alginate-degrading enzymes, agarases, xylanases, various hemicellulases derived from microorganisms, pectinases, or from the viewpoint of promoting unicellularization of seaweeds and promoting the production of active nutrients such as oligosaccharides that can be expected to have health functionality, or One or more can be selected from the group of enzymes capable of degrading polysaccharides contained in seaweed such as abalone acetone powder and used in combination with cellulase.

[0007] The higher the concentration of the enzyme used in the process of converting the seaweed into single cells, the higher the efficiency, but from the viewpoint of cost, it is practical to use it at a concentration of 3.0% by weight or less (all% indicate% by weight). Target. When using cellulase alone, S
CD particles (means unicellularized seaweed particles. Sing
le cell detritus (abbreviation of le cell detritus) and the microflora are used as indicators, if it is used at a concentration of 1% or more, it may be possible to singularize and ferment seaweed without adding a starter. But by using the starter
Even with the use of cellulase at a low concentration of 0.1-0.5%,
It is economically advantageous because the seaweed can be efficiently transformed into single cells, the growth of various bacteria can be suppressed, and the fermentation can be reliably carried out. Regarding the temperature for producing the seaweed pickled bed, there is no significant difference in the efficiency of the seaweed to become single cells within the temperature range of 5 to 50 ° C, so strict temperature control is not required. Therefore, it is possible to easily make a pickled bed in a normal home without temperature control equipment in a room temperature environment of 20 to 30 ° C.

The concentration of seaweed when producing a seaweed pickle bed can be 1 to 15% dry weight concentration. In particular, in order to make a uniform normal product without separation of water and solid contents, that is, without loosening the floor, if it is made at a concentration of 5-10%, it becomes a paste-like product suitable for pickling pickles. Most preferred. In general, the water content is
55% is said to be good (New edition: Food Industry General Dictionary: Social Incorporated Society of Food Industry, published by Korin Co., Ltd., p. 964,199
3) However, if you pickle a lot of vegetables, the water content of the vegetables will seep out, the floor of the rice bran will loosen, the salt concentration will decrease, and it will take time and effort to restore it to its original state. On the other hand, the seaweed pickled floor has a moisture content of around 90%, but the moisture in the floor hardly separates, so that anyone can easily manage the floor moisture content. You can make delicious pickles. It is considered that this is because the seaweed-derived polysaccharides typified by alginic acid have a high water-retaining power and thus have a wide allowable range for the water content. In addition, since the pickled bed made from seaweed has a high water content, it has good fluidity,
It also has the advantage that picking and picking vegetables is very easy.

The salt concentration when producing a seaweed pickled bed can be selected in the range of 0 to 10%, but in view of avoiding the risk of spoilage and the saltiness of the pickled product, the salt concentration is 2.5-. 5% is the most preferable. In general,
In the case of Nuka floor, it is said that the salt content around 8% is the best (New edition, Food Industry General Dictionary: Social Incorporated Association of Food Industry, edited by Korin Co., Ltd., p.964, 1993) Can produce delicious pickles with a low salt content of 2.5 to 5%, which matches the recent trend toward health.

The microorganism used in the present invention has the precondition that it can grow in a system in which only seaweed is present as a nutrient source, and that it causes lactic acid fermentation using glucose as a substrate and imparts aromaticity. A combination of microorganisms satisfying the two conditions can be selected and used.
That is, lactic acid bacteria as microorganisms that perform lactic acid fermentation and yeast as microorganisms that produce aroma may be added at the same time. Specifically, the former microorganisms include Lactobacillus , Streptococcus , and Leuconos.
One or more lactic acid bacteria selected from the genus toc, the genus Pediococcus, and the genus Tetragenococcus . In addition, it is possible to use bacteria of the genus Bifidobacterium which are considered to be lactic acid bacteria in a broad sense. Also, as the latter microorganism, Debaryomyc
One or more yeasts selected from the genera es , Candida and Saccharomyces can be selected and used.
In particular, it has been experimentally confirmed that various bacteria derived from seaweed are well suppressed, and the present inventors have isolated a microbial consortium from a fermented sample of sea lettuce, that is, Lactobacillus.
brevis NRIFS 5201 strain, Debaryomyces hansenii NRIFS
It is effective to add three strains of Y5201 strain and Candid a zeylanoides NRIFS Y5206 strain at the same time.

For details of the method for separating this microbial consortium and the taxonomical properties of each microorganism, see Japanese Patent Application No.
It is as described in 000-300399. That is,
In the laboratory of the present inventors, a sample left at 2 ° C. for 1 year and 5 months under the condition that sea urchin treated with cellulase and converted into SCD in a plastic bottle was sealed in the laboratory of the present invention, fermented to give a fermentation odor. I found that I was in a state. It was observed that when this fermented Ulva sample was subcultured to a new Ulva sample together with cellulase in an amount of about 1%, the Ulva sample could be newly introduced into a fermented state. After about 1 year after that, detailed analysis was carried out, and the salt concentration was 5
%, Cellulase concentration 1%, inoculate 1% of fermented sample, and incubate at 20 ℃ for 1 week, the sample can be maintained with the ability as a starter for fermenting seaweed even when subcultured every several months. I found that I could maintain it.
Therefore, it was decided to obtain a group of microorganisms capable of fermenting Ulva from the fermented Ulva sample. Direct analysis of the microflora in the transferred sea urchin sample, detailed analysis by the agar plate method, and the results from the direct counting method using a fluorescence microscope also showed that in this sample,
One type of lactic acid bacterium, two types of yeast, and three types of filamentous fungi were dominant, and it was found that the microflora was stably maintained before and after transplanting.

Of these, filamentous fungi generally have a strong activity for a glycolytic enzyme and are useful in the step of producing sugar as a substrate for fermentation. In the present invention, a commercially available glycolytic enzyme is used. Therefore, it is judged that it is not necessary to use filamentous fungi, and one type of lactic acid bacterium (ie, NRIFS B5201 strain) and two types of yeast (ie, NRIFS Y5201 strain and NRIFS Y5206 strain) are used.
Strains) were separated and used as fermentation starters. When a small amount of a mixture of these three types of microorganisms was inoculated on a new seaweed sample together with cellulase, it was found that it can be used as a starter for inducing a wide variety of seaweeds such as wakame, mackerel, etc. into a fermented state. did. The term "fermented state" as used herein means that, even if the seaweed sample is suspended in an aqueous solution and left at 20 ° C for 1 week or longer, not only does it not have a rotten odor, but also a fruity ester-like odor. Means being induced to the state in which it occurred. Further, at this time, it is characterized in that lactic acid or ethyl alcohol is produced in the supernatant fraction of the sample.

The microorganism obtained here is used in the Burgey's Manual
of Saystematic Bacteriology (NR krieg and JG
Holt, Wiliams & Wilkins 1984) and The Yeast 4th e
dition (Kurtzman, CPand Fell, JW, Elsevier Scie
nce BV, 1998).
201 strains are Lactobacillus brevis as shown in Table 1, and yeast NRIFS Y5201 strain and yeast NRIFS Y5206 strains are Debaryomyces hansenii and Cand as shown in Tables 2 and 3.
id a zeylanoides .

[0014]

[Table 1]               Properties of NRIFS B 5201 strain of lactic acid bacteria ────────────────────────────     Test item Test result ────────────────────────────     Gram stain +     Form bacillus                                      1.0 × 1.5 to 2.0 μm     Spore     Motility −     O / F test F     Catalase-     Oxidase     Village color cream     Lactic acid production from glucose (efficiency) + (50%)     Ethanol production from glucose +     Gas production from glucose +     Lactic acid fermentation type Hetero type     Growth temperature       5 ° C-       15 ℃ +       30 ° C +       45 ° C-     Acid production from carbohydrates       Glucose +       Fructose +       Galactose +       D-xylose +       L-xylose-       Mannose-       Glycerol-       Erythritol −       D-arabinose-       L-arabinose +       Ribose +       Adonitol-       β-methyl-D-xylose-       Sorbose −       Rhamnose-       Dulcitol-       Inositol-       Mannitol-       Sorbitol       α-methyl-D-mannose-       α-methyl-D-glucose ±       N-acetylglucosamine ±       Amygdalin −       Arbutin-       Esculin +       Salicin −       Cellobiose −       Lactose-       Meribiose-       Sucrose-       Trehalose-       Inulin −       Melettitose-       Raffinose-       Starch-       Glycogen −       Xylitol-       Gentiobiose-       D-Tranose-       D-Liquisose-       D-Tagatose-       D-Fucose −       L-Fucose −       D-arabitol −       L-arabitol −       Gluconate +       2-ketogluconic acid −       5-ketogluconic acid ±     GC content (mol%) 45     16S rRNA gene (E. coli No. 41-     (No. 1507)                                                                              Lavtobacillus brevis (Accession 99.9%      No.dbj D37785) homology   ───────────────────────────────

[0015]

[Table 2] Properties of yeast NRIFS Y5201 strain ───────────────────────────── Test item Test results (Y5201 strain) ── ──────────────────────────── Vegetative cell morphology Globular-oval growth format Multipolar budding Pseudohyphal non-formation Ascospore formation ( Malt medium, 15 ° C, 3 weeks) Spherical shape, surface not smooth Growth test 35 ° C-In the presence of 0.01% cycloheximide-Acetic acid production-Vitamin deficient medium-Sugar fermentation test D-Glucose weak D-Galactose-Sucrose weak Maltose-Lactose-Raffinose-Carbon compound assimilation test D-glucose + D-galactose + L-sorbose + D-glucosamine + D-ribose-D-xylose + L-arabinose + L-rhamnose + sucrose + maltose + α , Α-train Hallose + methyl α-D-glucoside + cellobiose + melibiose-lactose-raffinose + meletitose + glycerol + meso-erythritol + D-sorbitol + D-mannitol + myo-inositol-2-keto-D-gluconic acid + D-gluconic acid. + D-Glucuronic acid + DL- Lactic acid + Nitrogen compound assimilation test Nitrate-ethylamine + L-lysine + gadaverine + D-glucosamine-18S rRNA gene ( Saccharomyces cerevisiae 22nd to 1771th) nucleotide sequence Comparison of Debaryomyces hansenii (Accessien No.dbj AB013555) and 99.9% homology ────────────────────────────

[0016]

[Table 3] Properties of yeast NRIFS Y5206 strain ────────────────────────────── Test item Test results ─────── ──────────────────────── morphology of vegetative cells oval to oblong growth form budding pseudohyphae forming ascospores not forming (malt And acetic acid medium) Growth test Under 35 ° C-In the presence of 0.01% cycloheximide + Production of acetic acid-Sugar fermentation test D-glucose-D-galactose-sucrose-maltose-lactose-raffinose-Carbon compound utilization test D-glucose + D-galactose-L-sorbose + D-glucosamine Weak D-ribose-D-xylose-L-arabinose-L-rhamnose-sucrose-maltose-α, α-trehalose + methyl α-D-glucoside-cellobiose-medium Biose-Lactose-Raffinose-Meletetose-Glycerol + meso-erythritol-D-sorbitol + D-mannitol + myo-inositol-2-keto-D-gluconic acid + D-gluconic acid-D-glucuronic acid-DL-lactic acid-nitrogen Compound assimilation test Nitrate-ethylamine + L-lysine + cadaverine + D-glucosamine-18S rRNA gene ( Saccharomyces cerevisiae Nos. 22-1771) Comparison of nucleotide sequences Candid a zeylanoides (Accessien No.dbj AB013509 ) And 99.5% homology ───────────────────────────

These microorganisms are respectively deposited under the consignment number FERM BP-730 at the Institute of Biotechnology, Institute of Biotechnology, Ministry of International Trade and Industry (now: Institute of Biotechnology, Institute of Biotechnology).
1, FERM BP-7302, FERM BP-7303 have been deposited.

The amount of inoculum of the microorganism used is 1 to 0.1% of the cell suspension prepared by cultivating the cultured cells of each cell at a concentration of turbidity OD _{(660 nm)} = 1 with respect to the seaweed suspension. It is the simplest to directly inoculate a part of the fermented seaweed in an amount corresponding to 1 to 10% and use it as a bed seed. When adding the seaweed fermenting microorganisms,
Although it may be added after conversion to D, it is preferable to add them simultaneously in the first step of adding the glycolytic enzyme, as this saves time.

With the seaweed pickle bed thus prepared,
To make pickles, you can make pickles basically in the same way as traditional pickled pickles. That is, at room temperature, 1
Just leave it for about two weeks from the day, for an appropriate period depending on the type of vegetables. However, it is not necessary to agitate the flooring that is necessary for pickling in soup, and even if it is agitated, the bed has a high fluidity, so it is easy to shake the container without soiling the hands. Will be achieved.

As described above, according to the present invention, the problems of the peculiar odor and the looseness of the floor, which were the drawbacks of the conventional floor pad, are solved, and the pickled floor with a good fragrance and excellent water retention and easy management is provided. It can be easily made at home, and if it is used to make pickles, it is expected that it will be accepted by many people as a new pickle with a good aroma and a healthy image.

[0021]

The present invention will be described in more detail with reference to the following examples, but the present invention is not limited to these examples.

[Example 1] Dry powder of red seaweed, brown seaweed, green seaweed and various seaweeds belonging to flowering plants shown in Table 4 (particle size 1 mm
0.5g to 0.1g of Cellulase Onozuka R-10 and 9m
Place the solution in a test tube with a lid together with a 3.5% NaCl aqueous solution that has been sterilized by autoclaving, and then use 3 strains ( Lactobacillus brevis NRIFS B5201 strain, as a starter).
Debaryomyces hansenii NRIFS Y5201 strain and Candid a
zeylanoides NRIFSY5206 strain) was inoculated with a seaweed fermentation type microbial consortium. The method of inoculation was to inoculate 0.05 ml of each suspension of the cultured bacterial cells of each strain in physiological saline at a concentration of OD _{660 nm} = 1. The culture is
In a tightly closed state, the test tube was slowly rotated (5 rpm) at 20 ° C. for 7 days, and after culturing, the amounts of lactic acid and ethanol produced by microbial fermentation were measured and summarized in Table 4.

[0022]

[Table 4]

From Table 4, although some brown algae produced a small amount of lactic acid, production of lactic acid or ethanol was observed in all seaweeds. The sample had an ester-like fruity scent. From this result, it is understood that all types of seaweed can be lactic acid-fermented or ethanol-fermented by adding the above-mentioned seaweed-fermenting microbial consortium developed by the present inventor together with cellulase.

[0024]

[Example 2] 10.0 g of a dry powder of wakame seaweed (trade name, Wakamidori, Riken Vitamin Co., Ltd.) was dissolved in 180 ml of sterilized saline having a concentration of 3.5%, and cellulase (Onozuka R-10, Yakult) was dissolved. The sample to which 1% of (Head office) was added was allowed to stand under various conditions of reaction temperature from 5 ° C to 50 ° C to prepare a seaweed pickled bed. 5.8 μm diameter for these samples
The seaweed particles having a size of ˜11.5 μm were regarded as unicellular particles (SCD particles), and the state in which the weight percentage of unicellular seaweed particles produced in the pickled bed was increased was investigated and shown in FIG.

From FIG. 1, it can be seen that the preparation (reaction) time of the seaweed pickled bed is in the range of 5 ° C. to 20 ° C. for 24 hours or 6 days, and the higher the temperature, the higher the SCD production rate. It can be seen that if the period is as long as 14 days, there is no difference in the SCD production rate between 5 ° C and 50 ° C. From this result, it can be seen that the seaweed pickled bed can be easily prepared at home at room temperature.

[0026]

[Example 3] 10.0 g of a commercially available dry powder of wakame seaweed (the same product as in Example 2) was dissolved in 180 ml of sterile saline having a concentration of 3.5%,
Different amounts of cellulase ranging from 0% to 3% (Example 2
(Same product as above) and reacting at 20 ℃,
Seaweed pickled floor samples were prepared. When a microbial consortium was added at the start of preparation (see Example 1 for the addition method and amount)
The same as the above) and the case where no addition was made, and the addition amount of cellulase required for fermenting seaweed was examined. Judgment whether the seaweed pickle bed was fermented or rotted was 20 ° C for 12 days.
It was determined whether or not the sensory odor and the lactic acid bacteria formed a dominant microflora at the time of static culture in the culture. The confirmation that lactic acid bacteria were dominant was judged by whether colonies considered to be lactic acid bacteria that turned yellow were dominant when the number of microorganisms in the sample was counted on a BCP-added count agar plate medium. The results are shown in Table 5.

[0027]

[Table 5]

From Table 5, it can be seen that when the microbial consortium is not added, the fermentation cannot be reliably performed unless the cellulase concentration is 1% or more, whereas when the microbial consortium is added, the cellulase concentration is 0.1%. %
However, it turns out that fermentation is possible and economical. As a result of examining the microorganisms separated from both fermented samples at the gene level, those who added the microbial consortium
The three types of added microorganisms were dominant, but those who did not add the consortium had different types of microorganisms,
There was a safety and quality control problem.

[0029]

Example 4 A seaweed pickled bed was prepared from wakame as a raw material in the same manner as in Example 3 except that the concentration of cellulase added was 0.1% and the concentration of wakame was 10%. Add two bottles (500m each) of this pickle floor to 300ml each.
l volume, polycarbonate material, manufactured by Nalgen),
We pickled three 50g cucumbers, both ends of which were cut into one side, and another 5g of conus, about 20g, which had been dropped. When left at room temperature with the bottle cap closed,
The cucumber was pickled after 2 days and the conus after 5 days.
10 experts (ages 20 to 60, male 5) engaged in research and development of seafood to evaluate the quality of these pickles.
We asked the people (five women) to actually eat this as panelists, and asked them about the overall quality including flavor (taste and fragrance) and appearance, compared to the commercially available pickled cucumber and Konas. It was The results are shown in Table 6.

[0030]

[Table 6]

From the results shown in Table 6, it can be seen that by using a pickle floor made of wakame seaweed, a pickle which is comparable in flavor to commercial nuka pickles can be obtained. I repeated making pickled vegetables three times using the same floor, but I was able to easily make delicious pickled vegetables without taking care to manage the floor without loosening the floor.

[Brief description of drawings]

FIG. 1 is a comparison of the weight percentage of SCD fraction particles produced in a seaweed pickle bed when the reaction temperature is changed in Example 2.

─────────────────────────────────────────────────── --Continued from the front page (56) References JP-A-6-225721 (JP, A) JP-A 52-120157 (JP, A) JP-A 63-39542 (JP, A) JP-A 55- 118342 (JP, A) JP 2002-101826 (JP, A) (58) Fields surveyed (Int.Cl. ⁷ , DB name) A23B 7/10 JISST file (JOIS)

Claims (5)

(57) [Claims] 1. A seaweed pickling bed comprising a seaweed fermentation product obtained by decomposing seaweed with a saccharide-degrading enzyme containing cellulase to convert it into unicellular particles and fermenting it with a microbial consortium composed of lactic acid bacteria and yeast. . 2. The seaweed pickled bed according to claim 1, wherein the seaweed is seaweed or kelp. 3. The microbial consortium is Lactobacillu.
Lactobacillus s and Debaryomyces , Candida and Sacchar
The seaweed pickled bed according to claim 1 or 2, comprising one or more yeasts selected from the genus omyces . 4. The microbial consortium is Lactobacillu.
s brevis NRIFS B5201 strain (FERM BP-7301) and Debaryomyc
es hansenii NRIFS Y5201 strain (FERM BP-7302), and
Candid a zeylanoide s NRIFS Y5206 strain (FERM BP-7303)
The seaweed pickled bed according to claim 3, comprising: 5. A method for producing a pickle, which comprises producing a pickle using the seaweed pickle floor according to any one of claims 1 to 4.