JP5565625B2 - Worcester sauce manufacturing method and Worcester sauce obtained thereby - Google Patents

Description

  The present invention relates to a method for producing Worcester sauces and Worcester sauces obtained thereby.

At present, seasonings generally referred to as “sauce” in Japan are classified as Worcester sauces. Worcester sauces are those listed in the following (1) or (2), as defined by Japanese Agricultural Standards, and refer to liquid seasonings that are brown or brown.
(1) Vegetables or fruit juices, boiled juices, purees, or concentrates thereof, prepared by adding sugars, vinegar, salt and spices.
(2) Prepared by adding starch, seasoning, etc. to (1) above.

  Of the Worcester sauces, those having a viscosity of less than 0.2 Pa · s are “Worcester sauce”, those having a viscosity of 0.2 Pa · s or more and less than 2.0 Pa · s are “medium concentration sauce”, and the viscosity is 2.0 Pa.・ Things over s are called “rich sauces”. Worcester sauces are also used as sauces suitable for various uses such as tonkatsu, okonomiyaki, fried noodles, and takoyaki due to differences in material ratios and types of materials.

  Usually, Worcester sauces are processed by pressing or filtering after heating the materials for Worcester sauce, which are mixed with vegetables and fruits (including shredded products and ground products), and their juices, boiled juices and purees. Can be produced by removing the insoluble matter (so-called “sauce cake”) and storing and aging as appropriate. In addition, most of the Worcester sauces currently marketed do not require a fermentation process in the production process.

  However, in recent years, various kinds of Worcester sauces have been reported in which raw vegetables, fruits, and the like (materials for Worcester sauce) are fermented with microorganisms such as yeast, lactic acid bacteria, and acetic acid bacteria to add a new flavor (for example, Patent Document 1). To 6).

JP-A-8-9936 JP-A-6-189723 JP-A-6-189722 JP-A-6-169732 JP-A-6-125745 JP-A 63-68062

  The present inventors also sought to develop Worcester sauces with a new flavor and taste by using microorganisms other than those already reported. However, microorganisms other than those disclosed in the above-mentioned patent documents have problems that they do not grow in the material for Worcester sauce, and the taste of Worcester sauce is deteriorated.

By the way, in recent years, an increase in cancer and lifestyle-related diseases has become a major social problem. One of the causes, excess generated active oxygen in the body ^{_{(1 O 2, O 2 ·}} -, H 2 O 2, the active oxygen species such as · OH) and the like. Active oxygen generated excessively by stress, ultraviolet rays, smoking, etc. causes oxidative damage to DNA, cell membranes, etc. in the living body, and is one of the causes of aging, carcinogenesis, lifestyle-related diseases and the like. In order to protect the body from such excessive active oxygen, in addition to the active oxygen scavenging system in the body, such as active oxygen removal enzyme (SOD), catalase, glutathione peroxidase, the action of antioxidants taken as food is important. The present inventors have repeatedly studied whether or not a function capable of enhancing physiological activity such as antioxidant activity and preventing or ameliorating these diseases can be added also in Worcester sauces by fermentation. We sought to develop useful Worcester sauces.

  The present invention has been made in view of such circumstances, and provides a completely new method for producing Worcester sauces having excellent taste and rich physiological activity such as antioxidant activity, and provision of Worcester sauces obtained thereby. Is the purpose.

  In order to achieve the above object, the present invention provides a Worcester sauce comprising a step of inoculating a basidiomycete having cellulase activity into a mixed solution of Worcester sauce materials mainly composed of vegetables and fruits and fermenting the mixed solution. The manufacturing method of this kind is the first gist.

  Further, Worcester sauces obtained by the production method of the first gist are set as the second gist.

  That is, the present inventors have intensively studied to solve the above problems. In the process, we found that there are some basidiomycetes with cellulase activity. And as a result of fermenting the mixed solution of the material for Worcester sauce which uses vegetables and fruits as the main material using such basidiomycetes, cellulose which is the main component of the cell walls of vegetables and fruits is decomposed, It has been determined that the umami and flavor in the cells can be easily extracted, and that a unique fragrance from basidiomycete is added, resulting in a unique flavor not found in existing Worcester sauces and fermented Worcester sauces. It was. Furthermore, fermentation by the above basidiomycetes makes it rich in physiological activities such as antioxidant activity, and its antioxidant activity is hardly affected even after heat treatment, and has excellent physiological activity that has never existed before. As a result, the present inventors have reached the present invention.

  In the present invention, basidiomycetes refer to those whose fruiting bodies are said to be “mushrooms”, and in addition to basidiomycetes in the microbe taxonomy (Agaricus, Hidanashitake, Abdominal fungi, and Jellyfish), fruiting bodies If it is said to be a “mushroom”, it is used in a concept that includes a part of the baby fungi. Therefore, even if it is a camphor fungus, when the fruiting body is not what is said to be "mushroom" (for example, fungi such as blue mold, Aspergillus, red mold, Trichoderma fungi, yeast, etc.) It does not fall under “bacteria”.

  In the present invention, a desired Worcester sauce is produced by inoculating a mixed solution of Worcester sauce materials mainly composed of vegetables and fruits with a basidiomycete having cellulase activity and fermenting the mixed solution. This makes it possible to obtain completely new Worcester sauces that have excellent taste, have a flavor unique to fermentation, and are rich in physiological activities such as antioxidant activity.

  In particular, when the mixed solution of Worcester sauce material after fermentation is subjected to heat treatment, it is excellent in processability such that fermentation is controlled and the quality is stabilized, and the aging step can be shortened or omitted. Moreover, even if it heat-processes in this way, the antioxidant activity obtained by the said fermentation is hardly influenced, and Worcester sauce excellent in physiological activity can be obtained.

In addition, when the mixed solution of the above-mentioned Worcester sauce material is one in which at least one of salt and vinegar is removed and seasoned by adding the removed material after fermentation, fermentation by basidiomycetes is better As a result, the taste and fermentation-specific flavor of the obtained Worcester sauces become higher, and the physiological activities such as antioxidant activity are further improved.

  In addition to cellulase activity, the above basidiomycetes can also more easily extract the umami and flavor in the cells of vegetables and fruits if they exhibit pectinase activity, protease activity, amylase activity, and the like. Thus, Worcester sauces having better taste and the like can be obtained.

  The basidiomycetes are basidiomycetes such as mushrooms, mushrooms, oyster mushrooms, maitake mushrooms, and kawatake mushrooms because of their high cellulase activity and excellent ability to add physiological activities such as antioxidant activity by fermentation. Worcester sauces excellent in taste and physiological activity can be obtained.

  Moreover, smoother Worcester sauces can be obtained when the mixed liquid of the said material for Worcester sauce after fermentation passes through processes, such as a pressing process and a filtration process, and an insoluble matter is removed.

  Furthermore, after diluting the insoluble matter (including the insoluble matter produced during the production of conventional Worcester sauce) until the salinity is 0.85% by weight or less, and fermenting it with basidiomycetes having cellulase activity, After passing through the process of reusing in addition to the mixed liquid of the Worcester sauce material, the insoluble matter that has conventionally only been disposed of as a “source cake” can be reused for the production of Worcester sauces.

  In addition, when the basidiomycete having cellulase activity used for recycling the insoluble matter (source koji) is a basidiomycete such as mussel, shirohirotake, oyster mushroom, kawaratake, Himalayan oyster mushroom, muscaria, Napa, the reuse is It will be done more efficiently.

  And, as described above, the Worcester sauces obtained by the production method of the present invention are completely novel Worcester sauces that have excellent taste, have a fermentation-specific flavor, and are rich in physiological activities such as antioxidant activity. Yes, due to its physiological activity, it can be used as a functional / health food. In addition, in the production method of the present invention, the non-heated Worcester sauce has, for example, high activity of a fibrinolytic enzyme having an action of dissolving a thrombus, and also high activity of an antithrombin active substance that makes it difficult to form a thrombus. Therefore, it can exhibit more excellent efficacy in preventing thrombosis such as myocardial infarction and cerebral thrombus.

  Next, embodiments of the present invention will be described in detail.

As described above, the Worcester sauces of the present invention are inoculated with basidiomycetes having cellulase activity in a mixed solution of Worcester sauce materials mainly composed of vegetables and fruits, and subjected to a step of fermenting the mixed solution. Manufactured. The “main material” usually means that it occupies a majority of the whole, and includes the case where the whole consists only of the main material. Here, the “mixed liquid of Worcester sauce material” in the present invention is mainly composed of vegetables and fruits as described above, and vegetables and fruits (the shredded products thereof) as well as general Worcester sauce materials. And a mixture of these squeezed juices, boiled juices, purees, etc., and further concentrates them, and water is also added as appropriate. In addition to the above main ingredients, the Worcester sauce material includes seasonings and additives such as sugars, vinegar, salt, spices, starch, seasonings (amino acids, etc.), honey, liquor, etc. In regard to the above, it may be blended as a material for Worcester sauce before the fermentation step, or may be blended as a material for Worcester sauce after the fermentation step. Examples of the vegetables include tomatoes, onions, carrots, celery, garlic, lettuce, cabbage and the like, and examples of the fruits include apples and tangerines. In addition, before the fermentation step, a protein source such as soybean or a yeast extract may be added to the Worcester sauce material, if necessary, in order to increase the reproductive properties of basidiomycetes.

  In particular, the mixed solution of the material for Worcester sauce before the fermentation step is one in which at least one of salt and vinegar (salt and / or vinegar) is removed, and after the fermentation, the removed material is added and seasoned. As a result, fermentation by basidiomycetes is made better, and the taste and fermentation-specific flavor of the obtained sterol sauces are higher, and the physiological activities such as antioxidant activity are further improved. ,preferable. That is, salt and vinegar can be a factor that inhibits or suppresses fermentation by basidiomycetes, and as described above, problems such as fermentation inhibition can be solved by adding them after fermentation.

  By the way, the cellulase activity of the basidiomycete is roughly determined as to whether or not mycelial growth is observed in the mixed liquid of the above-mentioned Worcester sauce material mainly composed of vegetables and fruits (the Worcester sauce material is used). In detail, the crude enzyme solution collected from the basidiomycete is shaken at 35 ° C. with cellulose (water-insoluble) as a substrate. After the reaction, the reaction can be stopped immediately by heating at 100 ° C. for 10 minutes after the reaction, and the free reducing sugar in the reaction solution can be quantified by measuring by the Somogy-Nelson method.

  In addition to cellulase activity, the above basidiomycetes can also more easily extract the umami and flavor in the cells of vegetables and fruits if they exhibit pectinase activity, protease activity, amylase activity, and the like. It is preferable because Worcester sauces having better taste and the like can be obtained. In addition to the basidiomycetes having the above cellulase activity, the same effects as described above can be obtained even when basidiomycetes having no cellulase activity but exhibiting activities such as pectinase activity, protease activity, and amylase activity are used in combination. Therefore, it is preferable. The pectinase activity is obtained by, for example, reacting a crude enzyme solution collected from the basidiomycete with the substrate pectin while shaking at 35 ° C., and immediately reacting by heating at 100 ° C. for 10 minutes. The free reducing sugar in the reaction solution can be confirmed by a technique such as stopping and measuring by the Somogy-Nelson method.

  The specific basidiomycete used for fermenting the mixture of the material for Worcester sauce is a basidiomycete having cellulase activity or the like as described above, specifically, in forests or the like in various places. Collected, Nichous Bamboo, Suehirotake, Kawaratake, Matsuouji, Kaigaratake, Sanagitake, Mustache, Oystertake, Enokitake, Giberlakin, Oiwaraitake, Murasakimeji, Tsuriganetake, Kitataketake, Samurai bamboo, Samurai bamboo Sujichadaigoke, hanaburi kawatake, yakeatotsumutake, karakasatake, kuritake, numerisugitake, tsutaketake, wahitake, tsukudokopukutake, salmon sabatake, fumizuki kitake, mukitake, bukuryu, nisekin kakukin, oo Awantake, Saskehi Toyotake, Nigakuritake, Hanasanagitake, Tsutsugitaketake, Hitokuchitake, Hariganeotaketake, Eseori Miki, Haratake, Amanitatake, Tamotaketake, Buna Shigeji, Eringi, Aoitake, Bamboo lucidum Basidiomycetes such as maitake, Yamabushitake, Himalayan matsutake, Shirotamogitake, Hoonentake, Napa (a kind of Sarnochoke family, domestically produced in Thailand) are used. These may be used alone or in combination of two or more. The specific basidiomycetes are preferably selected and used only by appropriately screening and the like, showing only cellulase activity, and capable of producing Worcester sauces having better taste and the like.

In addition, among the above-mentioned specific basidiomycetes, when using basidiomycetes such as mushrooms, mushrooms, oyster mushrooms, maitake, kawatake, etc., the cellulase activity is high and the ability to add physiological activity such as antioxidant activity by fermentation is excellent. From these, it is possible to obtain Worcester sauces excellent in taste and physiological activity, etc., which is preferable.

  Fermentation of the material mixture for Worcester sauce by the basidiomycete is usually performed at a temperature of 20 to 30 ° C. for 20 to 100 days. Note that this fermentation treatment may be performed under anaerobic conditions or aerobic conditions. That is, when it is performed under anaerobic conditions, there is an advantage that propagation of various bacteria can be suppressed, and when it is performed under an aerobic condition, there is an advantage that fermentation is promoted. Further, as the basidiomycete, it is preferable to use one that has been purely cultured and grown as a mycelium by liquid culture. Moreover, it is preferable to adjust suitably the pH at the time of fermentation so that it may become optimal conditions with the basidiomycete to be used (normally pH = 4.0-6.0). And after this fermentation stage passes, it can heat-process, refrigerate, etc., and can store and age | cure | ripen suitably, The Worcester sauce of this invention can be manufactured.

  When the heat treatment is not performed after the fermentation treatment, the non-heated Worcester sauces obtained thereby have, for example, a high fibrinolytic enzyme activity (fibrinolytic activity) that dissolves the thrombus, and further creates a thrombus. Since the activity (antithrombin activity) of the antithrombin active substance that makes it difficult is high, it is possible to exert more excellent effects in the prevention of thrombosis such as myocardial infarction and cerebral thrombus. Furthermore, it also has excellent antioxidant activity.

  However, the above Worcester sauce material mixed solution is usually subjected to a heat treatment after the fermentation treatment, for reasons of processing. Thereby, although the fibrinolytic activity and antithrombin activity obtained by the said fermentation fall, antioxidant activity is hardly influenced and the Worcester sauce excellent in physiological activity can be obtained. In addition, heat treatment as described above also has advantages such as fermentation control and the like, stabilizing the quality, and shortening or omitting the aging step.

  And by the manufacturing method of the present invention as described above, it is possible to obtain completely new Worcester sauces that are excellent in taste, have a flavor unique to fermentation, and are rich in physiological activities such as antioxidant activity. Moreover, since the oxidation inhibitory rate based on the chemiluminescence method of the said Worcester sauce is 70% or more, since it is excellent in antioxidant activity, it can be used more preferentially as a functional and health food, and is preferable.

  Among the above Worcester sauces, those having a viscosity of less than 0.2 Pa · s are “Worcester sauce”, those having a viscosity of 0.2 Pa · s or more and less than 2.0 Pa · s are “medium-concentrated sauce”, and a viscosity of 2. Those with 0 Pa · s or higher can be considered as “rich sauce”. Moreover, it can be used as a source suitable for various uses such as for tonkatsu, okonomiyaki, yakisoba, and takoyaki due to the difference in the material ratio of the above Worcester sauces, the difference in the type of material, and the like.

  In addition, in the manufacturing method of the Worcester sauces of this invention, it is also possible to manufacture the seasoning (for example, dressing etc.) which is not included in the definition of Worcester sauces by the difference in the material ratio, the difference in the kind of material, etc. . The seasoning thus obtained is also excellent in taste, has a flavor unique to fermentation, and is rich in physiological activities such as antioxidant activity, like the Worcester sauce of the present invention.

In addition, when the mixed liquid of the above-mentioned fermented Worcester sauce material is subjected to a treatment such as a pressing treatment or a filtration treatment to remove insoluble matters (vegetables and fruit fibers, etc.), smoother Worcester sauces are obtained. Can do. The filtering process includes a backside process. Moreover, after adding water to the said insoluble matter and diluting until the salt concentration will be 0.85 weight% or less, fermenting with the basidiomycete which has cellulase activity, this is added to the liquid mixture of the material for Worcester sauce. Through the process of reusing, the insoluble matter, which has been conventionally disposed of as “source soot”, can be reused for the production of Worcester sauces. In addition, fermentation by a basidiomycete comes to be made favorable by reducing salt concentration to 0.85 weight% or less as mentioned above.

  In addition, in the manufacturing method of the Worcester sauces of this invention, since the fermentation by the basidiomycete which has cellulase activity is performed from the beginning, the production amount of the said insoluble matter is not so much. However, for example, in the conventional process of producing Worcester sauce, such insoluble matter is produced in large quantities as “source cake”, and therefore, this insoluble matter is recycled in the production method of Worcester sauce of the present invention according to the above procedure. When used, the above-mentioned insoluble matter that has only been disposed of as a “source soot” can be effectively used, and further, there is an environmental advantage as the amount of waste is reduced.

  As a specific basidiomycete used for fermenting the “source koji”, a basidiomycete that exhibits cellulase activity and can grow even in the presence of some salt (salt concentration of 0.85 wt% or less). More specifically, fungi are used. Chawantake, Nigakuritake, Bunashimeji, Coral Haritake, Shiitake, Nameko, Naratakemodoki, Numerisugitake, Himalayan oyster mushroom, Bunaharitake, Maitake, Nabushitake, Himalayan matsutake, Shirotamotake, Hanatake These may be used alone or in combination of two or more. The specific basidiomycetes can be appropriately screened, and can produce Worcester sauces that exhibit more cellulase activity, can grow in the presence of salt, and have superior taste and the like. It is preferable to select and use only bacteria.

  Further, among the above-mentioned specific basidiomycetes, when using basidiomycetes such as mussel, shirohirotake, oyster mushroom, kawaratake, himalaya oyster mushroom, mukitake, and napa, the above-mentioned “sauce koji” is excellent. Can be more efficiently performed, which is preferable. Furthermore, the basidiomycete is also excellent in ability to add physiological activity such as antioxidant activity due to its fermentation, and from this viewpoint, it is excellent in the reuse of the above-mentioned “source koji”.

  By the way, since the “source koji” after the fermentation treatment can be decomposed products of cellulose and pectin (oligosaccharides), bioethanol can also be produced by adding yeast to this and performing alcoholic fermentation. Is possible.

  Next, examples will be described. However, the present invention is not limited to these examples.

[Availability of basidiomycete source material fermentation ability]
First, according to the blending ratio shown in Table 1 below, a Worcester sauce material was prepared by mixing a vegetable or fruit puree with an auxiliary material. Next, in order to select basidiomycetes suitable for fermentation of the said Worcester sauce material, the fermentation was performed using 96 types of 216 strains of basidiomycetes. That is, 30 ml each of the above-prepared Worcester sauce material was dispensed into a 50 ml Erlenmeyer flask and autoclaved (120 ° C., 1.2 atm, 20 min) to prevent contamination with other microorganisms. Mycelia of various basidiomycetes (5 mm square sections, 5 sides) cultured on potato dextrose agar medium at 25 ° C. for 14 days are aseptically inoculated and cultured at room temperature for 42 days with rotary shaking (100 rpm) It was. And the growth state of the mycelium in the test strain (basidiomycete) and the material for Worcester sauce was as shown in Tables 2 and 3 below. In addition, strain No. shown in Table 2 and Table 3 is used. “NBRC + number” indicates a strain purchased from NBRC, “alphabet 2 letter-number” indicates a commercially available basidiomycete, and “alphabet 1 letter-number” indicates a wild-type basidiomycete. In addition, in the evaluation of the growth state, “−” indicates that the mycelia did not grow, “+” indicates that the mycelium slightly grew, “++” indicates that the mycelia grew so much as to cover the surface of the Worcester sauce material, and the triangle. A sample in which mycelia grew along the side of the flask was evaluated as “++++”.

From the above results, the presence or absence of cellulase activity was confirmed for basidiomycetes in which mycelial growth was observed (evaluation other than-). That is, the crude enzyme solution collected from the basidiomycete was reacted with the substrate cellulose (water-insoluble) while shaking at 35 ° C., and immediately after the reaction, the reaction was stopped by heating at 100 ° C. for 10 minutes. The free reducing sugar in the reaction solution was measured by the Somogy Nelson method. As a result, cellulase activity was observed in all basidiomycetes.

  Next, the material for Worcester sauce in which the growth of mycelia was recognized as described above was further subjected to stationary culture for 50 days. As a result, the viscosity of the Worcester sauce material decreased, the transparency increased, and the basidiomycete hyphae covered the upper part of the liquid surface. To 150 g of this fermented liquid, 300 g of sugar, 50 g of salt, 290 g of vinegar, 3.6 g of spices, 30 g of paste, and 290 g of water are added, and heat treatment is performed at 100 ° C. for 15 minutes. The insoluble matter was removed by scouring, and concentrated sauces (Example products) as Worcester sauces were prepared. The thick sauce obtained in this way is completely new compared to the thick sauce prepared without performing fermentation for comparison, and has an increased umami, excellent taste, and a flavor peculiar to fermentation. It became a thick sauce. This sensory evaluation was performed by 10 expert panelists.

  In addition, according to the above-mentioned paneler evaluation, for umster sauces produced by fermentation of mutake and maitake mushrooms, the amount of free amino acids in the material for fermented wooster sauce (before seasoning with salt, etc.) was measured according to the following procedure. Went. The result is combined with the measurement result regarding the material for unfermented Worcester sauce, and is shown in Table 4 below.

[Free amino acid content]
First, 4 ml of 99.5% ethanol was added to 1 ml of fermented Worcester sauce material, followed by extraction at 4 ° C. overnight, followed by centrifugation (8000 rpm, 10 min, 4 ° C.). The obtained supernatant was dried under reduced pressure and then dissolved in 10 ml of distilled water. Thereafter, the solution is applied to an ion exchange resin (Amberlite IR-120), the non-adsorbed substance is washed away with water, eluted with 3N ammonia, dried under reduced pressure, dissolved in 10 ml of 0.02N hydrochloric acid, and released. A sample for amino acid analysis was used. The free amino acid analysis sample thus prepared was appropriately diluted with 0.02N hydrochloric acid, and then the free amino acid concentration (composition) was determined using a high-speed amino acid analyzer (L-8800 type amino acid analysis system: Hitachi High-Technologies). It was measured.

  From the above results, the free amino acid concentration of the Worcester sauce material fermented with Mukitake was greatly increased compared to the unfermented material, and in particular, the concentrations of glutamic acid and aspartic acid exhibiting umami increased greatly. In addition, the material for Worcester sauce fermented with maitake also had more amino acids than the unfermented material.

  By the way, about the material for Worcester sauce in which the mycelial growth was recognized (those other than evaluation-) shown in Table 2 and Table 3 were not subjected to heat treatment (non-heated), and heated at 100 ° C. for 10 minutes. The treatment (heating) was measured for fibrinolytic activity, antithrombin activity, and antioxidant activity according to the following criteria. The results are shown in Tables 5 to 8 below.

[Fibrinolytic activity]
To a flat petri dish, 4 ml of a fibrinogen solution and 2 ml of a thrombin solution were added, immediately mixed without unevenness, and allowed to stand until solidified. On the fibrin plate thus obtained, 30 μl of the fermented liquid (sample) of the material for Worcester sauce was placed, and the dissolution area after standing for 24 hours was measured.

[Antithrombin activity]
50 μl of fermented liquor (sample) of Worcester sauce material and 50 μl of 12.5 U / ml thrombin (produced by Ito Ham Co., Ltd., derived from cow plasma) were preincubated at 37 ° C. for 5 minutes, and further 200 μl of 0.33% fibrinogen solution Was added, and the thrombin clotting time (the time from the change from fibrinogen to fibrin and clotting) was measured using a core gromometer (Henrich Amlang). In addition, the thing exceeding 300 seconds was shown as ">300".

[Antioxidant activity]
0.1 U xanthine oxidase dissolved in 10 μl of 300 μM MPEC (luminescent reagent) and 0.1 M KH ₂ PO ₄ / NaOH buffer solution (pH 7.5, containing 0.05 mM EDTA) per 10 μl of fermented liquid of Worcester sauce material 60 μl (manufactured by Sigma) and 170 μl of 0.1 M KH ₂ PO ₄ / NaOH buffer solution (pH 7.5, containing 0.05 mM EDTA) were mixed. This mixed solution is set in a luminescence sensor (luminescence sensor PSN AB-2200, manufactured by ATTO Corporation), and 0.72 mM hypoxanthine (manufactured by Wako) is dispensed with a built-in pump for 30 seconds. The luminescence integrated value of was measured. Based on the result, the light emission inhibition rate (oxidation inhibition rate) (%) was calculated based on the chemiluminescence method.

  From the above results, about half or more of the materials for Worcester sauce fermented by basidiomycetes exhibiting cellulase activity showed high antioxidant activity regardless of heating or non-heating. In particular, the material for fermented Worcester sauce with mussels (especially NBRC6432), mukitake (especially NBRC30526), oyster mushrooms (especially Hr-17), maitake (especially Mi-3), and kawatake (especially W18) has antioxidant activity. Etc. showed high activity. Therefore, Worcester sauces prepared by using the same basidiomycete fermented Worcester sauce material and seasoned with salt, vinegar, spices, etc. also exhibit high antioxidant activity regardless of heating or non-heating. From the above results, some test strains showed high activity in fibrinolytic activity and antithrombin activity. In particular, the fibrinolytic activity was higher in the non-heated one.

  Next, an experiment was conducted as follows with respect to the reuse of the soot sauce (insoluble matter removed by squeezing or filtration) generated in large quantities during the production of conventional Worcester sauces.

[With or without basidiomycete source fermentability]
First, in order to lower the salinity concentration of the recovered sauce cake to some extent, after mixing at a sauce sauce / water ratio of 1/9 (w / w) and diluting to a salt concentration of 0.56% by weight, NaOH is used to adjust the pH to 5. Adjusted to 6. 5 ml of this was dispensed into a test tube, capped with a silicon stopper, and autoclaved (120 ° C., 1.2 atm, 20 min). Next, in order to select basidiomycetes suitable for fermentation of the sauce koji medium prepared in this way, 96 types of 216 strains of basidiomycetes were used for fermentation. That is, mycelia of various basidiomycetes (three 5 mm square sections) cultured on a potato dextrose agar medium at 25 ° C. for 14 days were aseptically inoculated into the above-mentioned source koji medium and rotated at room temperature for 100 days. Endoculture (100 rpm) was performed. And the growth state of the mycelium in the test strain (basidiomycetes) and the source koji medium was as shown in Table 9 and Table 10 below. In addition, strain No. shown in Table 9 and Table 10. “NBRC + number” indicates a strain purchased from NBRC, “alphabet 2 letter-number” indicates a commercially available basidiomycete, and “alphabet 1 letter-number” indicates a wild-type basidiomycete. In addition, in the evaluation of the growth state, “−” indicates that the mycelia did not grow, “+” indicates that the mycelium slightly grew, “++” indicates that the mycelia grew so as to cover the surface of the source koji medium, and “triangle” A sample in which mycelia grew along the side of the flask was evaluated as “++++”.

From the above results, the test strains in which mycelium growth was observed overlap with the test strains in which mycelial growth was observed in Tables 2 and 3. Therefore, these test strains have cellulase activity. However, the test strains in which mycelial growth was observed from the above results were less in kind than those in Table 2 and Table 3 where mycelial growth was observed. This indicates that the test strains that can grow even in the presence of salt are limited. It turns out that it is suitable. That is, when the salmon was fermented with the above-mentioned test strain with the salt concentration lowered to some extent as described above, the taste of the source salmon increased due to the cellulase activity of the strain, as in the above Worcester sauces. A special flavor of fermentation is added. Therefore, when this fermented sauce cake is added to the previous mixed solution of Worcester sauce materials and reused in the production method of Worcester sauces of the present invention, resources can be effectively utilized.

  By the way, as shown in Table 9 and Table 10, for the source koji culture medium in which mycelial growth was observed (evaluation other than −), heat treatment was not performed (non-heated), 100 ° C. × 10 minutes heating The fibrinolytic activity, antithrombin activity, and antioxidant activity were measured according to the above-mentioned standard for the treated product (heating). The results are shown in Tables 11 and 12 below.

  From the above results, the source koji culture medium that showed cellulase activity and was fermented by basidiomycetes that can grow even in the presence of salt, whether heated or non-heated, most of them are from those before fermentation Also showed high antioxidant activity. In particular, the fermented sauce koji medium with mussel, Suehirotake, oyster mushroom, kawatake, Himalayan oyster mushroom, mukitake, and Napa showed high activity in antioxidant activity and the like. Therefore, when the same fermentation basidiomycete by basidiomycete is added to the mixture of the above-mentioned materials for Worcester sauce and reused in the method for producing the Worcester sauces of the present invention, it has high antioxidant activity regardless of heating or non-heating. The following Worcester sauces will be obtained. From the above results, some test strains showed high activity in fibrinolytic activity and antithrombin activity. In particular, the fibrinolytic activity was higher in the non-heated one.

  By the way, 1 ml of the fermentation source sputum medium with mussel mushroom (NBRC6432), Suehirotake (NBRC6504), oyster mushroom (Hr-15), Kawaratake (W17), Himalaya oyster mushroom (Hm-1), Mukitake (W2), Napa was observed with a microscope. As a result, compared with the unfermented sauce cake, the decomposition of the sauce cake by basidiomycetes was clearly observed. Although few were completely decomposed, it was confirmed that the inside of the cell wall became scarce and a part of the cell wall was decomposed. In addition, basidiomycetes were wrapped around the cell walls in the source basket, and mycelia were entangled. Therefore, using a hematometer, the average value of changes in the amount of solids (the volume of the cocoon and the mycelium volume) and the content ratio of the solids in 1 ml of the fermentation source koji medium was measured. The results are shown in Table 13 below.

  From the above results, the ratio of cocoons and mycelia in the solids of the sauce mash after fermentation was higher for all mycelia. Moreover, the amount of solids in 1 ml (mycelium + cocoon) decreased in mushrooms, shirohirotake, oyster mushrooms, mukitake, and napa. For this reason, waste is reduced by fermentation of the sauce cake with the cells.

  In addition, it was confirmed that Napa, in which the decomposition of the source rice cake was particularly remarkable, has pectinase activity in addition to cellulase activity. The pectinase activity was determined by reacting the crude enzyme solution collected from Napa with the substrate pectin while shaking at 35 ° C., immediately after the reaction, heating at 100 ° C. for 10 minutes to stop the reaction, The free reducing sugar was confirmed by measuring by the somology-Nelson method.

  In addition, it is suggested that polysaccharides such as cellulose, which is the main component of the cell wall, are decomposed by the cellulase activity and the like, and it is suggested that oligosaccharides are produced. Therefore, the above-mentioned mushrooms, Shirohirotake, oyster mushrooms, kawatake, Himalayan oyster mushrooms, It was investigated whether yeast could perform alcoholic fermentation using these oligosaccharides by adding yeast to the fermentation source koji medium by Mukitake and Napa.

That is, the fermentation source koji medium was filtered with gauze and divided into a liquid part and a solid part, and the following three conditions were used. For yeast for alcohol fermentation, Saccharomyces sake (Kyokai No. 6) cultured at 25 ° C. for 2 days in a 2% malto medium was used, and 5% of the culture solution was aseptically added to the following three conditions. Then, it was statically cultured for 2 days in an artificial meteorological apparatus at 30 ° C. and fermented. Moreover, also in the sauce koji culture medium which is not fermenting by basidiomycete, the alcoholic fermentation by yeast was performed like the above (blank).
(1) Stock solution: The liquid part was used without centrifugation.
(2) Concentrated liquid: The liquid part was concentrated to 1/10 with a centrifugal evaporator.
(3) 粕: 2.5 g of the solid part was suspended in 2.5 ml of distilled water.

  In each of the above conditions, the alcohol concentration was determined by HPLC [column: TSKgel Oapak-A (manufactured by Tosoh Corporation), column size: Φ7.8 mm D.I. × 30.0 cm, eluent: ultradistilled water, flow rate: 1 ml. / Min, detector: RI-8020, injection volume: 20 μl, measurement temperature: room temperature]. The results are shown in Table 14 below.

  From the above results, the stock solution and koji of the fermentation source koji medium by basidiomycetes had a lower alcohol concentration than the blank, but the concentrated solution of the fermentation source koji by oyster mushroom, kawaratake, Himalayan oyster mushroom, and Napa was clearer than the blank. The alcohol concentration was high. Therefore, it can be seen that the concentrated solution of fermentation source cake with oyster mushrooms, oyster mushrooms, Himalaya oyster mushrooms, and Napa is useful for producing alcohol (bioethanol) by yeast fermentation.

Claims (9)

  A method for producing Worcester sauces comprising a step of inoculating a mixed solution of Worcester sauce materials mainly comprising vegetables and fruits with a basidiomycete having cellulase activity and fermenting the mixed solution.   The manufacturing method of the Worcester sauces of Claim 1 provided with the process of heat-processing the liquid mixture of the said material for Worcester sauce after the said fermentation.   The Worcester sauce according to claim 1 or 2, wherein the mixture of the material for Worcester sauce is obtained by removing at least one of salt and vinegar, and adding the seasoned product after the fermentation. The manufacturing method of the kind.   The Worcester sauce according to any one of claims 1 to 3, wherein the basidiomycete exhibits at least one activity selected from the group consisting of pectinase activity, protease activity and amylase activity in addition to cellulase activity. The manufacturing method.   The method for producing Worcester sauces according to any one of claims 1 to 4, wherein the basidiomycete is at least one basidiomycete selected from the group consisting of mussel, mushroom, oyster mushroom, maitake and kawatake.   The manufacturing method of the Worcester sauces as described in any one of Claims 1-5 provided with the process of removing the insoluble matter by subjecting the mixed liquid of the said Worcester sauce material after fermentation to at least one process of a pressing process and a filtration process. .   The insoluble matter produced during the compression treatment and filtration treatment during the production of Worcester sauce is diluted until the salt concentration becomes 0.85% by weight or less, and this is fermented with a basidiomycete having cellulase activity. The manufacturing method of the Worcester sauces as described in any one of Claims 1-6 provided with the process of reusing in addition to the liquid mixture of the material for use.   8. The method for producing Worcester sauces according to claim 7, wherein the basidiomycete is at least one basidiomycete selected from the group consisting of mussel, Suehirotake, oyster mushrooms, Kawaratake, Himalayan oyster mushrooms, Mukitake and Napa.   Worcester sauces obtained by the production method according to any one of claims 1 to 8.