Koji starter for fermentation, koji for fermentation, and seasoning

Abstract

A seed koji (Aspergillus starter) for brewing is provided. The seed koji is obtained by inoculating koji mold(s) belonging to an Aspergillus genus into a seed koji raw material, where the seed koji raw material is a pea.

Claims

1. Seed koji (Aspergillus starter) for brewing obtained by inoculating koji mold(s) belonging to an Aspergillus genus into a seed koji raw material, wherein the seed koji raw material is a pea.

2. The seed koji for brewing according to claim 1, wherein the pea is a cracked pea.

3. The seed koji for brewing according to claim 2, wherein a grain size of the cracked pea is in a range of 500 to 4000 m.

4. The seed koji for brewing according to claim 1, further including a seed coat of pea as the seed koji raw material.

5. The seed koji for brewing according to claim 4, wherein an amount of addition of the seed coat of pea is in a range of 12.5 to 75.0 wt. %.

6. Koji for brewing obtained by inoculating the seed koji for brewing according to claim 1 into peas and performing cultivation.

7. A seasoning obtained by producing the koji for brewing according to claim 6 with a salt solution, and then performing fermentation and maturation by lactic acid bacteria and yeast(s).

8. The seasoning according to claim 7, wherein a crudely refined protein derived from a food item that does not contain an allergen is added during preparation.

9. The seasoning according to claim 7, containing one or a plurality of components selected from a group consisting of arabitol, mannitol, erythritol, sorbitol, and galactitol, wherein the arabitol is in an amount of 2 mg/mL or more, the mannitol is in an amount of 2 mg/mL or more, the erythritol is in an amount of 1 mg/mL or more, the sorbitol is in an amount of 0.3 mg/mL or more, and the galactitol is in an amount of 0.03 mg/mL or more.

Description

DESCRIPTION OF THE EMBODIMENT

(1) 1. Seed Koji for Brewing and Method of Producing the Same

(2) The seed koji for brewing according to the present embodiment is a seed koji for brewing that is obtained by inoculating koji mold belonging to the Aspergillus genus into a seed koji raw material, where the seed koji raw material is peas.

(3) The peas used in the present embodiment may be any type of peas classified into the pea family, including green peas, yellow peas, red peas, podded peas, and snap peas, for example. In order to obtain a liquid seasoning having a quality that is close to that of soy sauce, it is preferable to use green peas and yellow peas, etc. of Pisum sativum ssp. arvense (L.) Poir.

(4) Before producing seed koji for brewing, the raw material must be processed with the purpose of sterilization and protein denaturation of seed koji raw material. As for the method of processing of the raw material, it is preferable to crack the raw material to an appropriate grain size, thereafter add water, and then perform pressurized steaming and puff-cooking treatment. The processing conditions such as pressure and temperature could conform to those for soybean.

(5) The most important quality index required for seed koji for brewing is the number of spores (also called the number of conidium). If the production of spores is excellent and the number of spores per seed koji weight is large, the growth of the koji mold becomes vigorous during koji-making, and the amount of production of enzymes in koji increases. Therefore, the sugar and amino acid content of the prepared moromi increases, and the growth of the lactic acid bacteria and yeast becomes excellent, and at the same time, excellent results are obtained even from the viewpoint of the flavor and aroma of the final product such as soy sauce and fermented soybean paste. Further, if the number of spores of seed koji is large, it is possible to reduce the weight of seed koji added to koji, and thus it is also possible to reduce the production cost.

(6) The peas used in seed koji for brewing according to the present invention are preferably cracked. As for the method of cracking, industrially available cracking machines, such as crushers like roll crushers and milling machines like hammer mill and roll mill, etc. may be used.

(7) From the viewpoint of obtaining a larger number of spores, the grain size of the cracked peas is preferably in the range of 100 to 6000 m, and more preferably in the range of 500 to 4000 m. If the grain size is too large, the surface area reduces, because of which the number of spores per weight of peas, which are the seed koji raw material, also reduces and is therefore not preferable. On the other hand, if the grain size is too small, the grains come in close contact with each other, the growth of mycelia on koji mold becomes difficult, and the number of spores becomes less, and is therefore not preferable. It is noted that the grain size may be directly measured and checked by using a ruler or micrometer, and in addition to that, the grains may be sorted by size, by using a sieve having appropriate openings, into the grains that pass through the sieve and those that do not. As for the sieve opening, the mesh number and opening (m) are defined according to the Japanese Industrial Standards, for example, 500 m corresponds to #32 mesh and 4000 m corresponds to #5 mesh.

(8) It is preferable to further include the seed coat of peas as the seed koji raw material. While it depends on the type of seed, the seed coat of peas indicates a thin skin that is approximately 0.1 mm in thickness and covers the cotyledon, radicle, and plumule. By using the seed coat of peas together with the peas as the seed koji raw material, it is possible to secure an appropriate space between the grains of the raw material and to form an appropriate environment for the growth of koji mold from the viewpoint of nutritional balance too, and thus the effect of an increase in the number of spores of koji mold is achieved. It is noted that the seed coat of peas may be obtained by segregating the seed coat that is produced when peas are cracked, with the help of a sieve, etc., or the commercially available seed coat may be used. The commercially available seed coats include, for example, Exafine 2000 (manufactured by Cosucra), etc.

(9) From the viewpoint of obtaining a larger number of spores, the amount of addition of the seed coat of peas is preferably 12.5 to 75.0 wt. %, more preferably 12.5 to 67.5 wt. %, and yet more preferably 12.5 to 32.5 wt. %.

(10) The method of producing seed koji for brewing according to the present invention is as described below. As a culture medium of seed koji, water in an amount of 30 to 120 wt. % based on the weight of the seed koji raw material is mixed with the above-described seed koji raw material (peas and/or seed coat of peas), which is then passed through the steam sterilization process in an autoclave or a well-known seed koji producing machine, etc., and is then cooled to yield a culture medium of seed koji for producing seed koji that does not contain therein any component derived from wheat. If the mixed amount of water is too less, the protein denaturation becomes insufficient, and the growth of koji mold becomes poor. On the other hand, if the mixed amount of water is too much, seed koji solidifies in the form of paste, and the dispersibility during use as seed koji is lost, and is therefore not preferable. The appropriate water content of seed koji after the sterilization process and cooling is approximately 30 to 50 wt. %. Further, as for the sterilization condition, it is possible to use the general sterilization conditions for seed koji for brewing, such as 5 to 30 minutes at 115 to 130 C., as long as the raw material protein is denatured to the appropriate extent, and the saprophytic bacteria in the raw material are killed.

(11) The koji mold used in the present embodiment includes Aspergillus oryzae, Aspergillus sojae, Aspergillus niger, Aspergillus tamarii, Aspergillus kawachii, Aspergillus awamori, Aspergillus saitoi, etc.; however, Aspergillus oryzae or Aspergillus sojae is preferable since they are safe microorganisms that have been used in brewing of soy sauce and fermented soybean paste since the past, and are molds having a high usage frequency in Japanese industries.

(12) In view of growth of the koji mold, sporulation rate, and cost, the amount of addition of source seed koji in seed koji culture medium is preferably 0.01 to 1% of the amount of solid components in the culture medium, and particularly around 0.1%. Further, it is also possible to inoculate an appropriate amount of glycerol stock of koji mold spores and freeze-dried powder.

(13) The seed koji is cultured in the solid seed koji culture medium in a warming condition. The culture temperature is preferably 20 to 40 C., and the culture time could be an optional value, preferably about 72 to 192 hours. Since a temperature of the resulting seed koji will change along with a development of producing processes of the seed koji, the temperature; pH; growth condition, concentration and enzyme activities of the koji mold and other microorganisms are measured and observed with time so that the seed koji may be homogenized (In Japanese teire, which means care), if necessary, by replacing its surface part with its inner part.

(14) The method of checking the number of spores of seed koji includes sampling an appropriate amount of the seed koji thus obtained, suspending the sampled amount in sterile water containing approximately 0.01% of a surface active agent such as Tween 80, and preparing a spore suspension. When the spore suspension is dropped on a Thoma hemocytometer and the number of spores is counted with the regular method using a microscope, then the number of spores per weight of seed koji is known. In addition, it is also possible to use an automatic grain counting device that is used for a similar purpose.

(15) 2. Koji for Brewing and Method for Producing the Same

(16) The koji for brewing according to the present embodiment is obtained by inoculating the above-described seed koji for brewing into peas and performing cultivation.

(17) Although normally, before producing soy sauce koji (koji-making), the raw material is processed with the purpose of denaturing the protein of soybean, it is also possible to process the raw material for the peas used in the present embodiment by a method similar to that for soybean. The widely used industrial methods include performing pressurized steaming and puff-cooking treatment after cracking the raw material to an appropriate grain size. The processing conditions such as pressure and temperature could conform to those for soybean.

(18) In the koji-making process of koji for brewing according to the present embodiment, except the fact that the seed koji for brewing according to the present embodiment is used for the koji mold, it is possible to perform the process in a similar way to that for regular soy sauce koji, and it is also possible to employ methods such as koji-making by using a typical cultivation box (koji-buta) and a method that makes use of a ventilating koji-making device that is widely used industrially. It is possible to obtain koji by adjusting the water content and temperature of the peas for which the raw material has been processed, inoculating the spores (conidia) of the seed koji for brewing according to the present embodiment, and then culturing for 12 to 240 hours at 25 to 45 C. at a humidity level of 85 to 95%. Homogenization (teire) is performed during the process of koji-making in order to regulate the temperature. Further, it is also possible to produce koji by adding whole grains such as rice, which do not contain allergens, and then mixing with powders.

(19) The seed koji for brewing that is obtained based on the present embodiment may also be used as seed koji, etc. for producing enzymes for reagents and medical drugs, in addition to seed koji for brewing for the production of soy sauce, fermented soybean paste, refined sake, etc.

(20) 3. Soy Sauce-Like Seasoning and Method of Producing the Same

(21) The soy sauce-like seasoning according to the present embodiment is obtained by preparing the above-described koji for brewing with a salt solution, and then performing fermentation and maturation by lactic acid bacteria and yeasts.

(22) The soy sauce-like seasoning according to the present embodiment is a liquid seasoning used for a similar purpose to that of the soy sauce (shoyu) stipulated by the Japanese Agricultural Standards. However, on the basis of the purpose of the present invention, the soy sauce-like seasoning refers to an item that either does not include any food item containing an allergenic substance (allergen) specified as allergens for which labeling is either mandatory or recommended on the basis of the Food Sanitation Act as well as any food item specified as an item to be labeled according to the CODEX specified by the Joint FAO/WHO Food Standards Programme (Codex Alimentarius Commission), or includes only up to an amount that does not induce an allergy, in the main raw material and auxiliary raw material. The soy sauce-like seasoning according to the present embodiment may include auxiliary raw materials such as fruit juice, vegetable soup, various extracts, stocks, saccharides, seasonings, alcoholic beverages, fermented seasonings, acidifiers, and flavoring agents, as long as there is no deviation from the purpose of the present invention.

(23) Here, food items containing allergenic substances (allergens) indicates 27 food items specified in Japan as of March 2014 (eggs, milk, wheat, buckwheat, peanuts, abalone, squid, salmon roe, shrimp/prawns, oranges, crab, kiwifruit, beef, walnuts, salmon, mackerel, soybeans, chicken, pork, matsutake mushrooms, peaches, bananas, yams, apples, gelatin, sesame, and cashew nuts), and eight kinds of food items specified by CODEX (cereals containing gluten, i.e., wheat, rye, barley, oats, spelt or their hybridized strains and products of these; crustacea and products of these; eggs and egg products; fish and fish products; peanuts, soybeans and products of these; milk and milk products (lactose included); tree nuts and nut products; and food products made of sulfite, or containing 10 mg/kg or more sulfite.)

(24) The method of producing the soy sauce-like seasoning according to the present embodiment is as described below. The above-described koji for brewing is prepared together with a salt solution having an appropriate concentration in the regular preparation proportion in soy sauce brewing, and moromi is obtained. While the concentration of salt in the moromi juice may differ depending on the preparation temperature, it is preferably around 0.5 to 19%, as long as it is possible to sufficiently inhibit the growth of harmful microorganisms during fermentation. If necessary, Tetragenococcus halophilus and Zygosaccharomyces rouxii or Candida versatilis are added to moromi, and lactic-acid fermentation and yeast fermentation are performed. Fermentation and maturation is performed for 3 to 6 months while appropriately stirring at 15 to 35 C. according to the regular soy sauce brewing method, or for 1 day to 2 months while appropriately stirring at 35 to 55 C. according to a well-known low-salt high-temperature decomposition method, and matured moromi having the flavor of soy sauce is obtained.

(25) While an excellent liquid seasoning having the aroma and umami of soy sauce is obtained even from moromi that is obtained from the above-described preparations, when a crudely refined protein obtained by performing extraction, crude refining, and concentration of the protein component from a raw material that does not contain allergens is added during preparation, the soluble total nitrogen (TN), free amino acid, and peptides of the liquid seasoning obtained after fermentation and maturation increases, resulting in an excellent liquid seasoning comparable to high-quality soy sauce. There are no particular restrictions with regard to the crudely refined protein derived from a raw material that does not contain allergens, and for example, a crudely refined protein derived from peas, potato, corn, and rice, etc. may be used. Among these, protein derived from peas is preferable. As the protein derived from peas, for example, Emvital E7 (product name, manufactured by Emsland), PP-CS (product name, manufactured by Organo Food Tech Corporation), Nutralys F85F (product name, manufactured by Roquette), etc. are generally available. From the viewpoint of flavor and protein decomposition ratio, the amount of addition of protein derived from peas is preferably 5 to 100 wt. % of the above-described koji for brewing.

(26) As a method of obtaining a clear soy sauce-like fermented liquid by removing the solid content from matured moromi, it is possible to use a well-known method such as the filter press method by which moromi is wrapped in a filter cloth made of synthetic fibers of nylon, etc. and then folded up and pressurized, or the filter press method by which moromi is placed inside a filter cloth pasted on a filtration plate and a compression plate, and then pressurized by compressed air, etc.

(27) Clarification may also be performed for the soy sauce-like seasoning thus obtained. There are no restrictions regarding the clarification method, and a method that is conventionally known may be used, such as the membrane treatment, diatomaceous earth filtration, centrifugal separation, coagulation method, and sedimentation method.

(28) When adding a taste component to the liquid seasoning of the present embodiment in order to adjust the flavor of the soy sauce-like seasoning, there are no particular restrictions regarding food items as long as it does not contain an allergen, and amino acid, yeast extract, nucleic acid, organic acid, protein hydrolysate, saccharides, beet sugar, vegetable extracts, meat extracts, fish sauce, alcoholic beverages, mirin, alcohol, thickeners, emulsifiers, inorganic salts, etc. may be added. It is possible to add these taste components either alone or in combination.

(29) Sterilization or bacteria elimination may also be performed for the soy sauce-like seasoning thus obtained. In the case of sterilization, the soy sauce-like seasoning goes through a heat sterilization process called pasteurization. During pasteurization, the heating conditions performed in the well-known soy sauce making process may be used. It is preferable to perform heating for 20 to 60 minutes at 80 to 85 C., or for 5 to 20 seconds at 110 to 120 C., and then perform cooling thereafter. Since sediment may be produced due to heating, the soy sauce-like seasoning is obtained after allowing to stand still for a few days, and then separating the supernatant from the sediment. Further, in the case of bacteria elimination, filtration and bacteria elimination, etc. are performed through a well-known MF membrane filter, and the filtrate is obtained as the soy sauce-like seasoning.

(30) The soy sauce-like seasoning according to the present invention may be used in a similar way to that of the soy sauce (shoyu) stipulated by the Japanese Agricultural Standards, and may be blended with any food and beverage. For example, the soy sauce-like seasoning may be used by adding to food items such as broths, gravies, ponzu sauce, dressings, soups, sauces, and elements of daily dish.

(31) The sugar alcohols included in the soy sauce-like seasoning according to the present invention for adding umami and mellowness include, for example, arabitol, mannitol, erythritol, sorbitol, galactitol, threitol, xylitol, ribitol, iditol, volemitol, perseitol, inositol, quercitol, etc., and particularly, arabitol, mannitol, erythritol, sorbitol, and galactitol are preferable. Moreover, the amount of addition of these sugar alcohols is preferably 0.01 mg/mL or more. More specifically, as for the amount of addition of each sugar alcohol, 2 mg/mL or more of arabitol, 2 mg/mL or more of mannitol, 1 mg/mL or more of erythritol, 0.3 mg/mL or more of sorbitol, and 0.03 mg/mL or more of galactitol are preferable. Each sugar alcohol may be added either individually, or as a combination of 2 or more sugar alcohols.

(32) Hereinafter, the present invention will be further specifically described with reference to Examples. However, the technical scope of the present invention will not be limited in any way by these Examples.

EXAMPLES

(33) 1. Preparation of Seed Koji Using Peas Having Different Cracked-Grain Size

(34) (1) Cracking of Peas

(35) Green peas (produced in Canada) were cracked using MILTON Special A type mill (manufactured by Maru-roku Milling Machines). The cracked peas were then sorted by grain size by using a stainless sieve (manufactured by Tokyo Screen Co., Ltd).

(36) (2) Preparation of Seed Koji

(37) 4 g of seed koji raw material was put into an erlenmeyer flask having a capacity of 150 mL, to which 3.2 mL of tap water was added and mixed thoroughly. The opening was plugged with cotton, and sterilization was performed for 30 minutes at 121 C. in an autoclave. After cooling, 0.1 g of source seed koji of the koji mold was inoculated and stirred uniformly. The inoculated mixture was stationarily cultured for four days at 30 C.

(38) (3) Measurement of the Number of Spores

(39) 2 g of the seed koji thus obtained was sampled, water containing 20 mL of 0.01% Tween80 was added, and the mixture was stirred and suspended vigorously. The suspension was taken up in a dropper and dropped on a Thoma hemocytometer, and the number of spores was measured by using a microscope. It is noted that if the number of spores was too large, the suspension was diluted appropriately before performing microscopy. The results are shown in Table 1. Here, Not cracked, not sieved indicates round-shaped peas for which these processes are not performed. Cracked, but not sieved indicates that the entire quantity of the cracked peas was used without sorting by sieving. #5 or above indicates the grains that were sorted by a #5-mesh sieve, and did not pass through (the grains are larger than the sieve openings). #8 or above, #5 or below indicates the grains that did not pass through an #8 mesh from among the grains that had passed through a #5 mesh.

(40) TABLE-US-00001 TABLE 1 No. of spores of seed koji Cracked-grain size of peas (10.sup.7 spores/g) Not cracked, not sieve 3 Cracked, but not sieved 290 #5 (4.00-mm opening) or above 110 #8 (2.36-mm opening) or above 800 #10 (1.70-mm opening) or above 1000 #16 (1.00-mm opening) or above 720 #32 (500-m opening) or above 340 #32 (500-m opening) or less 96 #8 or above, #5 or below 1100 #10 or above, #8 or below 1000 #16 or above, #10 or below 820 #32 or above, #16 or below 510 Wheat bran (comparison) 1100

(41) As shown in Table 1, an excellent spore formation was observed even when peas were used as the seed koji raw material. Since the number of spores is approximately 110010.sup.7 spores/g when wheat bran, which is the general seed koji raw material, is used, it can be said that the peas are suitable as seed koji raw material. Particularly, the number of spores increases significantly when peas are cracked as compared to the case when peas are not cracked, and moreover, by sorting the grains of an appropriate grain size by a sieve, the number of spores further increases, and an excellent seed koji is obtained in much the same case as that of the existing wheat bran.

(42) 2. Examination of the Amount of Addition of Seed Coat of Peas

(43) When preparing the seed koji, the effect of the amount of addition of the seed coat of peas on the number of spores of seed koji was examined according to the procedures described below. That is, the cracked peas (Cracked, but not sieved) obtained from the above 1 and the seed coat of peas (product name Exafine 2000, manufactured by Cosucra) were mixed in appropriate amounts, and then the mixture was put into an erlenmeyer flask so that the total weight became 4 g. This mixture was used to prepare seed koji by using a similar manner to that of Example 1, and the number of spores was measured.

(44) TABLE-US-00002 TABLE 2 Cracked peas Seed coat of No. of spores (wt. %) peas (wt. %) (10.sup.7 spores/g) 100 0 700 87.5 12.5 2000 75.0 25.0 2100 67.5 32.5 2900 50 50 1500 32.5 67.5 1300 25.0 75.0 1000 12.5 87.5 690 0 100 300

(45) As shown in Table 2, when the seed coat of peas was mixed with cracked peas, the number of spores increased depending on the amount of the addition, and the quality of seed koji improved. However, when the amount of addition of the seed coat of peas exceeded the amount of addition of cracked peas, the number of spores gradually reduced, and when only the seed coat of peas was present, the number of spores reduced. From these results, it became clear that it is possible to obtain excellent seed koji by adding an appropriate amount of the seed coat of peas to the cracked peas.

(46) 3. Preparation of Fermented Seasoning of Peas

(47) (1) Method of Preparation

(48) 1.5 kg of peas (yellow peas, produced in Canada) were cracked by using MILTON Special A type mill (manufactured by Maru-roku Milling Machines), 1 L of water was added and mixed, and the mixture was steamed for three minutes at 121 C. in an autoclave. After allowing the mixture to stand and cool down to 40 C. or below, 2 g of seed koji of peas produced by using the cracked peas (Cracked, but not sieved) obtained from the above 1 was mixed, and the koji-making process was carried out for three days in a constant temperature/humidity machine (temperature 30 C. and humidity 95%) on a cultivation box (Prototype example 1). Moreover, a test section was also prepared in which the seed koji of peas produced by using the cracked peas (Cracked, but not sieved) obtained from the above 1 was inoculated after mixing pregelatinized rice flour in an amount of 20 wt. % of the peas (Prototype example 2). During the koji-making process, teire was performed depending on the growth of the koji mold. 2 L of salt solution was added to the koji thus obtained, which was then stirred well to yield moromi. At this time, the concentration of salt in moromi was 17% w/v. Similar preparation management to that of the regular Koikuchi soy sauce was performed for moromi, which was then matured through lactic-acid fermentation and yeast fermentation. After 6 months from preparation, moromi was placed in a nylon filter cloth, and compression was performed by placing a weight. In addition, diatomaceous earth filtration, pasteurization, and supernatant separating were performed according to the usual method of soy sauce producing, and fermented seasoning of peas (Prototype examples 1, 2) was obtained.

(49) (2) General Component Analysis

(50) The general components of soy sauce were analyzed according to the method specified in the Soy Sauce Test Method (Japan Soy Sauce Research Institute Foundation (ed.), issued on Mar. 1, 1985). The aroma components, such as HEMF (sweet, caramel-like aroma) were analyzed quantitatively according to the gas chromatography method (see Journal of Agricultural and Food Chemistry Vol. 39, 934 (1991)). The results are shown in Table 3. It is noted that as a reference, a commercially available regular Koikuchi soy sauce (manufactured by Kikkoman Corporation) using soybean and wheat as the raw material, was analyzed.

(51) TABLE-US-00003 TABLE 3 Total Glutamic Lactic HEMF Sample nitrogen (TN) Salt acid acid Ethanol Color (ppm name (% w/v) (% w/v) (% w/v) (% w/v) (% w/v) pH No. w/v) Prototype 1.40 16.3 0.9 0.9 3.2 4.7 17 14 example 1 Prototype 1.21 15.9 0.8 0.7 3.8 4.6 25 13 example 2 Regular 1.68 15.8 1.0 0.9 2.9 4.8 9 29 Koikuchi soy sauce

(52) The fermented seasoning of peas thus obtained had an appearance and flavor like the regular Koikuchi soy sauce, and was a preferable soy sauce-like seasoning even from the viewpoint of taste. From the analysis values of Table 3, it was understood that the components contributing to the taste were included in almost the same proportion as the regular Koikuchi soy sauce. Moreover, it became clear that HEMF, which is known as the characteristic flavor component of the flavor of soy sauce, was also contained in a sufficient amount.

(53) 4. Preparation of a Fermented Seasoning of Peas to which Pea Protein is Added

(54) (1) Method of Preparation

(55) Similar to the Prototype example 1 described above, the preparation of seed koji and koji producing were performed. The pea koji thus obtained and pea protein (manufactured by Emsland) were mixed in the weight ratio of 90:10 (Prototype example 3), 80:20 (Prototype example 4), and 50:50 (Prototype example 5), and similar to the above 3 (1), after carrying out preparation and fermentation and maturation, filter press and pasteurization were performed, and a fermented seasoning of peas (Prototype examples 3 to 5) was obtained.

(56) (2) General Component Analysis

(57) The results of the analysis performed similarly to the above 3 (2) are shown in Table 4.

(58) TABLE-US-00004 TABLE 4 Glutamic HEMF Sample (TN) Salt acid Lactic acid Ethanol Color (ppm name (% w/v) (% w/v) (% w/v) (% w/v) (% w/v) pH No. w/v) Prototype 1.65 16.2 1.2 1.1 3.1 4.7 16 16 example 3 Prototype 1.89 16 1.3 1.1 3.8 4.6 17 15 example 4 Prototype 2.43 15.6 1.5 1.3 2.2 4.7 21 12 example 5

(59) In Prototype example 1 that was prepared only by pea koji, the total nitrogen (TN) was 1.4 which corresponds to the JAS superior grade soy sauce (see Table 3), but from the results of Table 4, in Prototype example 3 to which pea protein was added during preparation, the total nitrogen (TN) became 1.65 which corresponds to the JAS special grade soy sauce. Moreover, glutamic acid that has a large contribution to the umami of the soy sauce-like seasoning also improved from 0.9 to 1.2. In addition, by increasing the amount of addition of pea protein, the TN and glutamic acid improved depending on the increased amount, and a significantly stronger umami could be tasted. On the other hand, in the recent years, when used as a raw material for processing, the demand for soy sauce having a light color has been increasing. When peas are used as the raw material, the color becomes lighter, and it was found that by adding rice (see Table 3) and pea protein (see Table 4), a soy sauce-like seasoning having a further lighter color was obtained. Since pea protein also does not contain specific allergens, the addition of pea protein made it possible to improve the quality of umami, color, etc. while retaining the characteristic of an allergen-free soy sauce-like seasoning.

(60) 5. Sensory Evaluation

(61) The sensory evaluation of each test item was performed by a panel of five trained persons having the discrimination ability, and the taste and aroma of the Prototype example 1 and Prototype example 4 described earlier was evaluated by the semantic differential method in terms of the intensity of umami and soy sauce-like characteristics. Prototype examples 1 and 4 were presented blindly as stock solutions without revealing the contents, and a comparison was made by smelling the aroma and then tasting a quantity of 0.2 mL. The rating scale was based on the criteria described below and the average rating among panelists was calculated.

(62) (Rating Scale)

(63) 1. Quite weak or almost no umami/soy sauce-like characteristics

(64) 2. Somewhat weak umami/soy sauce-like characteristics

(65) 3. Some umami/soy sauce-like characteristics is experienced

(66) 4. Somewhat strong umami/soy sauce-like characteristics

(67) 5. Quit strong umami/soy sauce-like characteristics

(68) TABLE-US-00005 TABLE 5 Soy sauce-like Sample name umami characteristics Prototype example 1 4.6 4.8 Prototype example 4 4.8 4.4

(69) From the sensory evaluation results of Table 5, it was found that both the Prototype examples 1 and 4 have a quite strong umami and soy sauce-like characteristics. The fact that the pH, umami of glutamic acid, organic acid generated by fermentation and maturation, and the color components generated by the Maillard reaction are included in an amount close to that of the regular soybean and wheat soy sauce seems to contribute to the soy sauce-like taste. Further, the fact that the characteristic aroma components of soy sauce generated by fermentation, including the HEMF, are included in an amount close to that of the regular soybean and wheat soy sauce seems to contribute to the soy sauce-like aroma. In addition, it was confirmed that the Prototype example 4 to which pea protein was added had an even stronger umami. It is noted that the comment from the panelists was that the blind tasting of Prototype examples 1 and 4 indicates soy sauce-like characteristics to an extent that is indistinguishable from the regular soy sauce.

(70) 6. Extraction of Sugar Alcohol and Derivatization

(71) (1) Test Method

(72) 2 L of each test sample was transferred to a micro tube respectively, 1000 L of a mixed solution of Methanol:Water:Chloroform (5:2:2) and 60 L of 0.2 mg/mL of ribitol (manufactured by Wako Pure Chemical Industries, Ltd.) as an internal standard substance were added, and then stirred. After the stirring, centrifugal separation was performed for 16,000g for three minutes at 4 C., and 900 L of supernatant was collected and then transferred to another micro tube. Thereafter, 400 L of distilled water was added, stirred, and performed centrifugal separation again in the similar manner. 900 L of the supernatant was transferred to yet another micro tube, and was concentrated to become 200 L or less by a centrifugal concentrator. Thereafter, a dry solid was obtained by using a freeze dryer. Next, the dried solid was dissolved in 100 L of anhydrous pyridine (manufactured by Wako Pure Chemical Industries, Ltd.) containing 20 mg/mL of methoxyamine hydrochloride (manufactured by Sigma-Aldrich), and was maintained under stirring for 90 minutes at 30 C. Then, 50 L of N-methyl-N-trimethylsilyl-trifluoro-acetamide (manufactured by GL Sciences Inc.) was added, and trimethylsilylation was performed by allowing the solution to react for 30 minutes at 37 C. to obtain a sample for gas chromatography mass spectrometry (GC/MS).

(73) (2) GC/MS Analysis

(74) 7890A-5975C (manufactured by Agilent Technologies) was used for the GC/MS analysis, and as a standard, arabitol, mannitol, erythritol, sorbitol, and galactitol (all manufactured by Wako Pure Chemical Industries, Ltd.) were derivatized and measured in a similar manner, and quantitative determination was performed from the resulting calibration curve. The device conditions were as follows.

(75) Column: CP-SIL 8CB-MS (30 mm0.25 mm, manufactured by Agilent Technologies)

(76) Temperature-rising conditions: After retaining for 2 minutes at 80 C., 80 to 320 C., 15 C./minute, and retention for 6 minutes at 320 C.

(77) Carrier gas and flow rate: Helium gas, 1 mL/minute

(78) Injection temperature: 230 C.

(79) TABLE-US-00006 TABLE 6 Prototype Prototype Regular Koikuchi Substance example 1 example 4 soy sauce name (mg/mL) (mg/mL) (mg/mL) Arabitol 9.93 5.81 1.64 Mannitol 9.28 8.53 1.34 Erythritol 6.20 3.79 0.63 Sorbitol 0.53 0.46 0.24 Galactitol 0.19 0.18 0.02

(80) From the results of Table 6, it was found that the fermented seasoning of peas contains approximately 2 to 10 times more sugar alcohols as compared to the regular Koikuchi soy sauce. Specifically, the fermented seasoning of peas of the preset invention was found to contain, at least, 2 mg/mL or more of the arabitol, 2 mg/mL or more of the mannitol, 1 mg/mL or more of the erythritol, 0.3 mg/mL or more of the sorbitol, and 0.03 mg/mL or more of the galactitol. Since these sugar alcohols are known to exhibit a unique umami and mellowness, these sugar alcohols are believed to provide preferable characteristics to the taste of the fermented seasoning of peas.

(81) It is possible to blend the sugar alcohols that are not derived from peas, which are the fermented ingredients, as food additives; however, when taking into consideration the recent demand for natural and additive-free food items, sugar alcohols derived from peas, which are the fermented ingredients, are preferable. The fermented seasoning of peas of the present invention is possible to contain sugar alcohols derived from peas in a significant amount, without the use of food additives, by using seed koji produced from a pea raw material and performing fermentation and maturation with natural food materials (particularly, food items like peas that do not contain allergens) as the raw material.

(82) From the above results, by performing fermentation and maturation through the addition of peas, and if necessary, a crudely refined protein derived from food items that do not contain allergens, a soy sauce-like seasoning having the favorable taste and aroma of soy sauce was obtained without the use of any food item containing an allergenic substance. In addition, a seasoning capable of imparting a distinctive umami and richness due to a significant amount of sugar alcohols as compared to the regular soy sauce, was realized. The soy sauce-like seasoning according to the present invention is possible to be used in conventional Japanese cuisine in a similar manner to the soy sauce, and is also possible to be widely used in Chinese and Western cuisines, and thus it is expected to contribute to an improvement in QOL of the eating habits of patients allergic to food items.