WO2022151578A1 - 一种同时改善酱油风味和提高酱油硒含量的方法 - Google Patents

一种同时改善酱油风味和提高酱油硒含量的方法 Download PDF

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WO2022151578A1
WO2022151578A1 PCT/CN2021/081108 CN2021081108W WO2022151578A1 WO 2022151578 A1 WO2022151578 A1 WO 2022151578A1 CN 2021081108 W CN2021081108 W CN 2021081108W WO 2022151578 A1 WO2022151578 A1 WO 2022151578A1
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soy sauce
selenium
content
flavor
improving
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English (en)
French (fr)
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高献礼
冯拓
单培
张雅琼
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江苏大学
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
    • A23L27/50—Soya sauce
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/015—Inorganic compounds
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/03—Organic compounds
    • A23L29/035—Organic compounds containing oxygen as heteroatom
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/03—Organic compounds
    • A23L29/045—Organic compounds containing nitrogen as heteroatom
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/03—Organic compounds
    • A23L29/055—Organic compounds containing sulfur as heteroatom
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/90—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in food processing or handling, e.g. food conservation

Definitions

  • the invention relates to a method for simultaneously improving the flavor of soy sauce and increasing the selenium content of soy sauce, and belongs to the technical field of food processing.
  • Soy sauce originated in my country and is an indispensable condiment in people's daily cooking, catering industry and food industry.
  • the annual output of soy sauce in my country exceeds 10 million tons, accounting for more than 50% of the world's total output.
  • the high-end market of domestic and foreign soy sauce is almost occupied by Japanese soy sauce, and the average price of domestic soy sauce is less than 1/12 of that of Japanese soy sauce.
  • the crude oil of Japanese soy sauce is delicious, mellow, and harmonious in taste, with rich mellow, ester and smoky aromas; domestic soy sauce crude oil has a prominent salty and mellow taste, mainly sauce and ester aroma.
  • its umami, mellow and smoky aroma are insufficient compared with Japanese soy sauce crude oil.
  • domestic soy sauce has a small number of yeast and lactic acid bacteria and a short survival time during the fermentation stage of mash
  • Japanese soy sauce has a large number of yeast and lactic acid bacteria added during the fermentation of mash and has a long survival time
  • these two types of microorganisms are ethanol ( mellow aroma), 4-ethylguaiacol (smoky incense), 4-vinylguaiacol (smoky incense) key production strains. Therefore, improving the protease activity of Aspergillus oryzae (especially the acid protease activity) and the number and survival time of yeast and lactic acid bacteria in the mash is the key to improving the quality of domestic soy sauce crude oil.
  • Mutagenesis and directional screening of Aspergillus oryzae are effective methods to improve the enzyme activity of Aspergillus oryzae, and have achieved remarkable results, but the application of the mutagenized microorganisms in fermented foods is strictly limited.
  • Moromi fermentation is carried out in a high-salt environment (8-22%, w/v), therefore, the growth, reproduction and metabolism of lactic acid bacteria and yeast in the moromi are inhibited by the high-salt environment.
  • the number of lactic acid bacteria and yeast in the high-salt dilute mash reached a peak in the early stage of fermentation, and then decreased rapidly. At about 30 days, the remaining lactic acid bacteria and yeast were only about 1/10 of their peak. Therefore, improving the protease activity of Aspergillus oryzae and the number and survival time of lactic acid bacteria and yeast in the mash is the key technology to improve the taste and aroma of domestic soy sauce.
  • Keshan disease plays an important role in maintaining human health.
  • the occurrence of Keshan disease, Kashin-Beck disease, and cardiovascular disease are all highly related to selenium deficiency. More than one third of people in my country are in a state of selenium deficiency or severe selenium deficiency. Therefore, it is of great significance to increase the selenium intake of Chinese residents.
  • the purpose of this invention is to develop a kind of by adding substances such as selenium, gum arabic, VC and VB to improve the taste and aroma substances of soy sauce, while improving the method for selenium content in soy sauce.
  • the present invention provides a kind of using soybean or soybean meal, flour or pulverized fried wheat as the main raw material, using Aspergillus oryzae (Aspergillus oryzae), Aspergillus sojae (Aspergillus sojae), lactic acid bacteria and yeast as fermentation strains.
  • Selenium, gum arabic, VC and VB and other substances improve the taste and aroma of soy sauce, and at the same time increase the selenium content of soy sauce.
  • (1) Preparation of selenium-gum arabic compound using water as a solvent, prepare a mixed solution of gum arabic and sodium selenite or sodium selenate (that is, selenium-gum arabic solution); The mass concentration is 0.1%-30%; the concentration of sodium selenite or sodium selenate in the mixed solution is 1-1000mM/L; then VC (vitamin C), VB1 (vitamin B1), methionine are added to the mixed solution , and stir evenly to obtain a selenium-Gum Arabic compound; the molar ratio of the sodium selenite or sodium selenate to VC, VB1 and methionine is 1:(2-10):(0-5):(0- 5);
  • saline mixture containing glycerol, VB (vitamin B), choline, inositol, and saline first prepare saline with a certain mass concentration, and then add glycerol, VB5 (vitamin B5), and VB7 (vitamin B7) to the saline , choline and inositol, mix well to obtain a saline mixture;
  • the mass ratio of saline, glycerol, VB5, VB7, choline and inositol in the saline mixture is 1000000:(1000-15000):(0-100):(0 -1):(50-5000):(10-1000);
  • Sauce mash preparation, fermentation and soy sauce preparation add Tetragenococcus halophilus to the brine mixture obtained in step (5) to make its content in the brine mixture reach 1 ⁇ 10 4 -1 ⁇ 10 7 CFU/ml, mix evenly, then add the Daqu prepared in step (4) and mix to make mash, and after fermenting for a few days, a mash is obtained; then add Zygosaccharomyces rouxii to make its content in the mash reach 5. ⁇ 10 4 -1 ⁇ 10 7 CFU/ml, fermented at room temperature for 30-360 days, and after fermentation, the soy sauce is obtained by pressing or pouring oil, filtering, sterilizing and filling.
  • step (1) the mass concentration of gum arabic in the mixed solution is 0.2%-10%; the concentration of sodium selenite or sodium selenate in the mixed solution is 10-100mM/L.
  • the molar ratio of sodium selenite or sodium selenate to VC, VB1 and methionine described in step (1) is 1:(3-7):(0.5-3):(0.5-3).
  • the selenium content in the selenium-acacia compound aqueous solution is 0.1-10 mg/L; preferably, the selenium content is 0.5-8 mg/L.
  • the mass ratio of soybean or soybean meal to flour or pulverized fried wheat described in step (3) is 100:(15-100).
  • the soaking time of soybean or soybean meal described in step (3) is 0.5h-24h
  • the cooking temperature in step (3) is 110-130° C.
  • the cooking time is 30-60 min.
  • step (4) the mass ratio of Aspergillus oryzae or Aspergillus sojae spores to flour or pulverized fried wheat is 1:(100-10000).
  • the mass ratio of soybean or soybean meal and flour or pulverized fried wheat in the premix described in step (4) is 100:(15-500); the preferred mass ratio is 100:(15-40).
  • the mass concentration of saline in step (5) is 5%-25%; the mass ratio of saline, glycerol, VB5, VB7, choline and inositol in the saline mixture is 1000000:(2000-10000):( 5-50):(0.05-0.5):(50-1000):(50-500).
  • the content of Tetraphylococcus halophila in the brine mixture is 5 ⁇ 10 4 -5 ⁇ 10 6 CFU/ml; the content of the rumeni in the mash is 1 ⁇ 10 5-5 x 106 CFU/ml.
  • the dosage ratio of Daqu and brine mixture described in step (6) is 1g:(1-4)ml.
  • the fermentation days in step (6) are specifically 3-15 days; the normal temperature fermentation is 30-180 days.
  • the soy sauce produced by the inventive method has the following advantages and beneficial effects:
  • the method of the present invention can simultaneously significantly improve the enzyme activity of Daqu protease, and increase the number and survival time of lactic acid bacteria and yeast in the mash;
  • the soy sauce taste, aroma substance and selenium content produced by the present invention are significantly improved, and the umami, sweet taste, mellow feeling, mellow aroma and smoky aroma are significantly improved, and have both health care functions ;
  • the present invention has important economic value and social significance.
  • Fig. 1 is a graph showing the changes in the quantity of yeast luteria and Tetraphylococcus halophila in the fermentation process of Example 1 and its control;
  • Fig. 2 is embodiment 1 and its contrast taste (A) and aroma (B) evaluation result graph;
  • Fig. 3 is a graph showing the change in the quantity of yeast luteria and Tetraphylococcus halophila in the fermentation process of Example 2 and its control;
  • Fig. 4 is embodiment 2 and its comparative taste (A) and aroma (B) evaluation result graph;
  • Fig. 5 is a graph of changes in the number of yeast lutella and Tetraphylococcus halophila in the fermentation process of Example 3 and its control;
  • Fig. 6 is embodiment 3 and its contrast taste (A) and aroma (B) evaluation result graph;
  • T.halophilus refers to Tetraphylococcus halophilus in the experimental group
  • Z.rouxii refers to Saccharomyces rubella in the experimental group
  • D-T.halophilus refers to Tetraphylococcus halophilus in the control group
  • D-Z.rouxii refers to Saccharomyces rutheli in the control group.
  • the protease enzyme activity, amino acid nitrogen content, flavor substance content in soy sauce and the viable count of lactic acid bacteria and yeast involved in the present invention refer to methods such as literature SB/T 10317-1999 and Gao Xianli (Accelerating aroma formation of raw soy sauce using low intensity sonication[J]. Food Chemistry, 2020, 329, 127118).
  • the determination methods of total selenium, organic selenium and inorganic selenium content refer to GB 5009.93-2017 and Jing Dawei and other methods (Selenium enrichment, fruit quality and yield of winter jujube as affected by addition of sodium selenite [J]. Scientia Horticulturae, 2017, 225 , 1-5).
  • Embodiment 1 take soybean as protein raw material and adopt the inventive method to produce soy sauce
  • An aqueous solution of arabic was prepared, and sodium selenite was added to the aqueous solution of gum arabic to obtain a mixed solution; the mass concentration of gum arabic in the mixed solution was 0.1%, and the concentration of sodium selenite solution was 1 mM/L; VC was slowly added to the above mixed solution to adjust The molar ratio of sodium selenite and VC is 1:2, and stirring evenly is the selenium-acacia compound; the selenium-acacia compound is used as the raw material to prepare an aqueous solution of the selenium-acacia compound, so that the selenium content reaches 0.1 mg /L.
  • soybean and flour as raw materials, the ratio of soybean and flour is 100:15 (w/w), the soybean is washed with water first, and then soaked in the aqueous solution of selenium-Gum Arabic compound until the soybean fully absorbs water and swells, and the soybean is dried. Cook at 100°C for 180min. Pre-mix Aspergillus oryzae with flour, and then mix with soybeans cooled to room temperature. The ratio of soybeans and flour is 100:15 (w/w), and cultivated at 20°C until the surface of the mixture is pale yellow-green, that is, Daqu is obtained. .
  • Tetragenococcus halophilus to the saline containing glycerol, VB, choline and inositol to make its content in saline reach 1 ⁇ 10 4 CFU/ml, mix well, take Daqu to saline ratio as 1:1 (w/v) mixing to make mash, when fermenting to the 5th day, add Zygosaccharomyces rouxii to make the content in the mash reach 5 ⁇ 10 4 CFU/ml, ferment at room temperature for 30 days, ferment Soy sauce is obtained by pressing, filtering, sterilizing and filling at the end.
  • Control group 1 In control group 1, soybeans were not soaked in an aqueous solution of selenium-gum arabic compound, and glycerol, choline and inositol were not added to the saline.
  • the neutral and alkaline protease activity of Daqu produced by the method of the present invention is significantly higher than the Daqu enzyme activity of the control sample, and its amino acid nitrogen, total sugar and reducing sugar content of the soy sauce produced by the method of the present invention Significantly higher than the corresponding value of the control soy sauce.
  • the sensory evaluation results show ( FIG. 2A ) that, compared with the control soy sauce, the soy sauce produced by the method of the present invention has significantly improved umami, sweetness and mellow taste.
  • the adoption of the patented method of the present invention can significantly increase the number of lactic acid bacteria and yeast in the mash in the fermentation process, and significantly increase the content of key flavor compounds in the final product (see Table 3).
  • the sensory evaluation results showed (Fig. 2B) that the mellow and smoky aroma of soy sauce was significantly improved by the patented method of the present invention.
  • the method of the present invention significantly improves the taste and aroma of soy sauce.
  • the total selenium and organic selenium content of the soy sauce produced by the method of the invention are 0.049mg/L and 0.034mg/L respectively, and the total selenium and organic selenium content of the control are 0.021mg/L and 0.016mg/L respectively, indicating that the method of the invention is adopted. It can significantly increase the total selenium and organic selenium content of soy sauce.
  • Table 2 embodiment 1 and its control group 1 prepare the taste substance content of soy sauce
  • a,b,c compound identification method (a) by comparing the retention index (RI) and GC-MS chromatogram with standard compounds under the same conditions; (b) by comparing with the NIST05 database; (c) by comparing with the NIST05 database The aroma characteristics of standard compounds smelled by GC-O were compared.
  • d RI was calculated on DB-wax and TR-5MS columns, respectively.
  • Embodiment 2 take soybean meal as protein raw material and adopt the inventive method to produce soy sauce
  • the ratio of soybean meal to pulverized fried wheat is 100:100 (w/w). 5min. Aspergillus sojae spores (Aspergillus sojae) were premixed with crushed fried wheat, and then mixed with soybean meal cooled to room temperature. The ratio of soybean meal to crushed fried wheat was 100:100 (w/w), and incubated at 35°C to the surface of the mixture Showing light yellow-green, that is, get the big song.
  • Tetragenococcus halophilus to the saline containing glycerol, VB, choline and inositol to make its content in saline reach 1 ⁇ 10 7 CFU/ml, mix well, take Daqu to saline ratio as 1:4 (w/v) mixing to make mash, when fermented to the 5th day, add Zygosaccharomyces rouxii to make the content in the mash reach 1 ⁇ 10 7 CFU/ml, ferment at room temperature for 360 days, ferment The soy sauce is obtained by pouring oil, filtering, sterilizing and filling at the end.
  • Control group 2 In control group 2, soybeans were not soaked in an aqueous solution of selenium-acacia gum complex, and glycerol, choline and inositol were not added to the saline, and the raw material ratio and production process were the same as those in Example 2.
  • the acidity, neutrality and alkaline protease activity of the Daqu produced by the method of the present invention is significantly higher than that of the control sample Daqu enzyme, and the soy sauce produced by the method of the present invention is its total nitrogen amino acid nitrogen, total
  • the sugar and reducing sugar contents were significantly higher than the corresponding values of the control soy sauce.
  • the sensory evaluation results show ( FIG. 4A ) that, compared with the control soy sauce, the soy sauce produced by the method of the present invention has significantly improved umami, sweetness and mellow feeling.
  • the adoption of the patented method of the present invention can significantly increase the number of lactic acid bacteria and yeast in the mash in the fermentation process, and significantly increase the content of key flavor compounds in the final product (see Table 6).
  • the sensory evaluation results showed (Fig. 4B) that the mellow and smoky aroma of soy sauce was significantly improved by the patented method of the present invention.
  • the method of the present invention significantly improves the taste and aroma of soy sauce.
  • the total selenium and organic selenium content of the soy sauce produced by the method of the invention are 0.95mg/L and 0.60mg/L respectively, and the total selenium and organic selenium content of the control are 0.011mg/L and 0.07mg/L respectively, indicating that the method of the invention is adopted. It can significantly increase the total selenium and organic selenium content of soy sauce.
  • Table 5 embodiment 2 and control group 2 prepare the taste substance content of soy sauce
  • a,b,c compound identification method (a) by comparing the retention index (RI) and GC-MS chromatogram with standard compounds under the same conditions; (b) by comparing with the NIST05 database; (c) by comparing with the NIST05 database The aroma characteristics of standard compounds smelled by GC-O were compared.
  • d RI was calculated on DB-wax and TR-5MS columns, respectively.
  • Embodiment 3 take soybean as protein raw material and adopt the inventive method to produce soy sauce
  • an aqueous solution of arabic To prepare an aqueous solution of arabic, add sodium selenite to the aqueous solution of gum arabic to obtain a mixed solution; the mass concentration of gum arabic in the mixed solution is 1%, and the concentration of sodium selenite is 25 mM/L; slowly add VC and VB1 to the above mixed solution , methionine, adjust the molar ratio of sodium selenite to VC, VB1, and methionine to 1:5:2.5:2.5, and stir evenly to obtain a selenium-arabic gum complex.
  • VC and VB1 To prepare an aqueous solution of arabic compound, add sodium selenite to the aqueous solution of gum arabic to obtain a mixed solution; the mass concentration of gum arabic in the mixed solution is 1%, and the concentration of sodium selenite is 25 mM/L; slowly add VC and VB1 to the above mixed solution , methionine, adjust the molar ratio of sodium sele
  • soybean and crushed fried wheat as raw materials, the ratio of soybean and crushed fried wheat is 100:500 (w/w), the soybean is washed with water first, and then soaked with selenium-acacia compound aqueous solution until the soybean fully absorbs water After swelling, the soybean meal was fully infiltrated with an aqueous solution of selenium-gum arabic complex, and the drained soybeans were cooked at 130 °C for 20 min. Aspergillus oryzae and crushed fried wheat were pre-mixed, and then mixed with soybeans cooled to room temperature. The ratio of soybeans and crushed fried wheat was 100:500 (w/w), and cultivated at 30°C until the surface of the mixture appeared. Light yellow-green, that is to get Daqu.
  • Tetragenococcus halophilus to the saline containing glycerol, VB, choline and inositol to make its content in saline reach 5 ⁇ 10 5 CFU/ml, mix well, take Daqu to saline ratio as 1:2.5 (w/v) mixed to make mash, when the fermentation to the 5th day, Zygosaccharomyces rouxii was added to make the content in the mash reached 1 ⁇ 10 6 CFU/ml, fermented at room temperature for 180 days, fermented
  • the soy sauce is obtained by pouring oil, filtering, sterilizing and filling at the end.
  • Control group 3 is the soy sauce with the same raw material ratio and production process as in Example 3 without soaking soybeans in the aqueous solution of selenium-acacia gum complex, without adding glycerol, choline and inositol in the brine.
  • the acidity, neutrality and alkaline protease activity of the Daqu produced by the method of the present invention is significantly higher than that of the control sample Daqu, and the soy sauce produced by the method of the present invention is its total nitrogen amino acid nitrogen, total
  • the sugar and reducing sugar contents were significantly higher than the corresponding values of the control soy sauce.
  • the sensory evaluation results show (FIG. 6A) that, compared with the control soy sauce, the soy sauce produced by the method of the present invention has significantly improved umami, sweetness and mellow feeling.
  • the adoption of the patented method of the present invention can significantly increase the number of lactic acid bacteria and yeast in the mash in the fermentation process, and significantly increase the content of key flavor compounds in the final product (see Table 9).
  • the sensory evaluation results showed (Fig. 6B) that the mellow and smoky aroma of soy sauce was significantly improved by the patented method of the present invention.
  • the method of the present invention significantly improves the taste and aroma of soy sauce.
  • the total selenium and organic selenium content of the soy sauce produced by the method of the invention are 0.81mg/L and 0.52mg/L respectively, and the total selenium and organic selenium content of the control are 0.017mg/L and 0.012mg/L respectively, indicating that the method of the invention is adopted. It can significantly increase the total selenium and organic selenium content of soy sauce.
  • Table 8 embodiment 3 and control group 3 prepare the taste substance content in soy sauce
  • a,b,c compound identification method (a) by comparing the retention index (RI) and GC-MS chromatogram with standard compounds under the same conditions; (b) by comparing with the NIST05 database; (c) by comparing with the NIST05 database The aroma characteristics of standard compounds smelled by GC-O were compared.
  • d RI was calculated on DB-wax and TR-5MS columns, respectively.
  • the present invention utilizes selenium, vitamins, choline, inositol and glycerol to significantly increase the activity of soy koji enzymes, the number of lactic acid bacteria and yeast in the soy mash, improve the content of taste and aroma substances, and at the same time increase the total selenium in the soy sauce. and organic selenium content.
  • the soy sauce produced by the method not only improves the flavor of the soy sauce, but also imparts the health care function of the soy sauce, thereby increasing the added value of the soy sauce, and has a good market prospect. Therefore, the present invention has important economic value and social significance.

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Abstract

一种同时改善酱油风味和提高酱油硒含量的方法；首先配制硒-阿拉伯胶溶液，加入VC、VB1和蛋氨酸，搅拌得硒-阿拉伯胶复合物；配制硒-阿拉伯胶复合物水溶液(硒含量为0.1-10mg/L)，加入大豆或豆粕浸泡，浸泡后经蒸煮、冷却后与接种曲霉的面粉或粉碎的炒小麦混合培养得到大曲；在盐水中添加甘油、VB5、VB7、胆碱、肌醇及乳酸菌，加入大曲混合发酵数天后，添加酵母菌继续常温发酵，最后通过压榨或淋油、过滤、杀菌和灌装得到酱油。与传统方法生产的酱油相比，生产得到的酱油滋味、香气物质及硒含量显著提高，鲜味、醇厚感、醇香及烟熏香显著改善，并兼具保健功能，具有重要的经济价值和社会意义。

Description

一种同时改善酱油风味和提高酱油硒含量的方法 技术领域
本发明涉及一种同时改善酱油风味和提高酱油硒含量的方法，属于食品加工技术领域。
背景技术
酱油起源于我国，是人们日常烹饪、餐饮行业和食品工业中必不可少的调味品。我国酱油年产量超过1000万吨，占世界总产量的50％以上。但国内外酱油高端市场几乎被日本酱油占据，国产酱油的平均价格不及日本酱油的1/12。日本酱油的原油滋味鲜美、醇厚、口感协调，具有浓郁的醇香、酯香和烟熏香；国产酱油原油咸味突出、醇厚，以酱香、酯香为主。但与日本酱油原油相比其鲜味、醇香和烟熏香不足。国产酱油发酵采用米曲霉制曲，日本酱油采用酱油曲霉制曲。与酱油曲霉相比，米曲霉产蛋白酶酶活，特别是酸性蛋白酶酶活偏低，这导致其在酱醪发酵阶段对大豆蛋白降解不充分，鲜味氨基酸和小分子量量肽含量低，这是其与日本酱油原油相比鲜味弱的重要原因之一。此外，国产酱油在酱醪发酵阶段酵母菌和乳酸菌数量少、存活时间短，而日本酱油在酱醪发酵时所添加的酵母菌和乳酸菌数量多、存活时间长，而这两类微生物是乙醇(醇香)、4-乙基愈创木酚(烟熏香)、4-乙烯基愈创木酚(烟熏香)的关键生产菌株。因此，提高米曲霉产蛋白酶酶活(特别是酸性蛋白酶酶活)及酱醪中的酵母菌和乳酸菌数量和存活时间是提高国产酱油原油品质的关键。
对米曲霉进行诱变和定向筛选是提高米曲霉酶活的有效手段，取得了显著成效，但诱变得到的微生物在发酵食品中的应用受到严格限制。酱醪发酵在高盐环境(8-22％，w/v)下进行，因此，乳酸菌和酵母菌在酱醪中的生长、繁殖和代谢受到高盐环境的抑制。高盐稀态酱醪中乳酸菌和酵母菌在发酵的前期数量达到峰值，随后其数量迅速下降，至30天左右时剩余的乳酸菌和酵母菌仅为其峰值的1/10左右。因此，提高米曲霉产蛋白酶酶活及乳酸菌和酵母菌在酱醪中的数量和存活时间，是改善国产酱油滋味和香气的关键技术。
硒在维持人体健康中扮演着重要角色，克山病、大骨节病、心血管疾病等的发生均与硒缺乏高度相关，我国超过三分之一的人处于缺硒或严重缺硒状态。因此，提高我国居民硒摄入量具有重要意义。
发明内容
本发明的目的是为了开发一种通过添加硒、阿拉伯胶、VC和VB等物质改善酱油滋 味和香气物质，同时提高酱油中硒含量的方法。具体地，本发明提供了一种以大豆或豆粕、面粉或粉碎的炒小麦为主要原料、以米曲霉(Aspergillus oryzae)、酱油曲霉(Aspergillus sojae)、乳酸菌和酵母菌为发酵菌种，利用添加硒、阿拉伯胶、VC和VB等物质改善酱油滋味和香气，同时提高酱油硒含量的方法。
具体步骤如下：
(1)硒-阿拉伯胶复合物的制备：以水为溶剂，配制阿拉伯胶与亚硒酸钠或者硒酸钠的混合溶液(即为硒-阿拉伯胶溶液)；所述混合溶液中阿拉伯胶的质量浓度为0.1％-30％；所述混合溶液中亚硒酸钠或硒酸钠的浓度为1-1000mM/L；然后在混合溶液中加入VC(维生素C)、VB1(维生素B1)、蛋氨酸，搅拌均匀，即得到硒-阿拉伯胶复合物；所述亚硒酸钠或硒酸钠与VC、VB1、蛋氨酸的摩尔比为1:(2-10):(0-5):(0-5)；
(2)硒-阿拉伯胶复合物水溶液配制：以硒-阿拉伯胶复合物为原料，配制硒-阿拉伯胶复合物水溶液；
(3)原料浸泡和蒸煮：以大豆或豆粕、面粉或粉碎的炒小麦为原料；以大豆为原料时，使用前先用水清洗干净；然后用步骤(2)制备的硒-阿拉伯胶复合物水溶液浸泡大豆或豆粕，浸泡后淋干水分，将大豆或豆粕在100-140℃进行蒸煮5-180min，蒸煮后降至常温，备用；
(4)制曲：将米曲霉(Aspergillus oryzae)或酱油曲霉孢子(Aspergillus sojae)与面粉或粉碎的炒小麦预混合，得到预混料，然后与步骤(3)中蒸煮后降至常温的大豆或豆粕混合，在20-35℃培养至混合物表面呈现浅黄绿色，即得到大曲；
(5)配制含有甘油、VB(维生素B)、胆碱、肌醇、盐水的盐水混合物：首先配制一定质量浓度的盐水，然后往盐水中添加甘油、VB5(维生素B5)、VB7(维生素B7)、胆碱和肌醇，混合均匀，得到盐水混合物；盐水混合物中盐水、甘油、VB5、VB7、胆碱和肌醇的质量比为1000000:(1000-15000):(0-100):(0-1):(50-5000):(10-1000)；
(6)酱醪制备、发酵及酱油制备：往步骤(5)得到的盐水混合物中添加嗜盐四联球菌(Tetragenococcus halophilus)，使其在盐水混合物中的含量达到1×10 4-1×10 7CFU/ml，混合均匀，然后加入步骤(4)制备的大曲混合制醪，发酵数天后，得到醪液；然后再添加鲁氏酵母(Zygosaccharomyces rouxii)，使其在醪液中的含量达到5×10 4-1×10 7CFU/ml，常温发酵30-360天，发酵结束通过压榨或淋油、过滤、杀菌和灌装即得酱油。
进一步的，步骤(1)中所述混合溶液中阿拉伯胶的质量浓度为0.2％-10％；所述混合溶液中亚硒酸钠或硒酸钠的浓度为10-100mM/L。
进一步的，步骤(1)中所述亚硒酸钠或硒酸钠与VC、VB1、蛋氨酸的摩尔比为1:(3-7):(0.5-3):(0.5-3)。
进一步的，步骤(2)中所述硒-阿拉伯胶复合物水溶液中硒含量为0.1-10mg/L；优选的，硒含量为0.5-8mg/L。
进一步的，步骤(3)中所述大豆或豆粕与面粉或粉碎的炒小麦的质量比为100:(15-100)。
进一步的，步骤(3)中所述大豆或豆粕浸泡的时间为0.5h-24h
进一步的，步骤(3)中所述蒸煮的温度为110-130℃，蒸煮时间为30-60min。
进一步的，步骤(4)中所述米曲霉或酱油曲霉孢子与面粉或粉碎的炒小麦的质量比为1：(100-10000)。
进一步的，步骤(4)中所述大豆或豆粕与预混料中面粉或粉碎的炒小麦的质量比为100:(15-500)；优选的质量比为100:(15-40)。
进一步的，步骤(5)中盐水的质量浓度为5％-25％；所述盐水混合物中盐水、甘油、VB5、VB7、胆碱和肌醇的质量比为1000000:(2000-10000):(5-50):(0.05-0.5):(50-1000):(50-500)。
进一步的，步骤(6)中所述嗜盐四联球菌在盐水混合物中的含量为5×10 4-5×10 6CFU/ml；所述鲁氏酵母在醪液中的含量为1×10 5-5×10 6CFU/ml。
进一步的，步骤(6)中所述大曲与盐水混合物的用量比为1g:(1-4)ml。
进一步的，步骤(6)中所述发酵数天具体是3-15天；所述常温发酵30-180天。
本发明方法生产的酱油与传统方法生产的酱油相比，具有如下优点和有益效果：
(1)本团队研究发现在酱油生产过程中添加适量的硒、VC、蛋氨酸、VB1、VB5和甘油，不但可显著提高大曲蛋白酶酶活，而且可显著提高酱醪中乳酸菌和酵母菌的数量和生存时间，进而显著改善酱油滋味和风味品质。
(2)本发明方法可同时显著提高大曲蛋白酶酶活，增加酱醪中的乳酸菌和酵母菌数量及存活时间；
(3)与传统方法生产的酱油相比，本发明生产得到的酱油滋味、香气物质及硒含量显著提高，鲜味、甜味、醇厚感、醇香及烟熏香显著改善，并兼具保健功能；本发明具有重要的经济价值和社会意义。
附图说明
图1为实施例1和其对照发酵过程中鲁氏酵母和嗜盐四联球菌数量变化图；
图2为实施例1和其对照滋味(A)和香气(B)评价结果图；
图3为实施例2和其对照发酵过程中鲁氏酵母和嗜盐四联球菌数量变化图；
图4为实施例2和其对照滋味(A)和香气(B)评价结果图；
图5为实施例3和其对照发酵过程中鲁氏酵母和嗜盐四联球菌数量变化图；
图6为实施例3和其对照滋味(A)和香气(B)评价结果图；
其中，T.halophilus是指实验组中的嗜盐四联球菌，Z.rouxii是指实验组中的鲁氏酵母，D-T.halophilus是指对照组中的嗜盐四联球菌，D-Z.rouxii是指对照组中的鲁氏酵母。
具体实施方式
下面结合实例对本发明的具体实施方式作进一步说明，但本发明的实施和保护范围不限于此。
本发明所涉及的蛋白酶酶活、氨基酸态氮含量、酱油中风味物质含量及乳酸菌和酵母菌活菌数参照文献SB/T 10317-1999和Gao Xianli等方法(Accelerating aroma formation of raw soy sauce using low intensity sonication[J].Food Chemistry,2020,329,127118)。总硒、有机硒和无机硒含量的测定方法参照GB 5009.93-2017和Jing Dawei等方法(Selenium enrichment,fruit quality and yield of winter jujube as affected by addition of sodium selenite[J].Scientia Horticulturae,2017,225,1-5)。
实施例1：以大豆为蛋白质原料采用本发明方法生产酱油
配制阿拉伯水溶液，向阿拉伯胶水溶液中加入亚硒酸钠得到混合溶液；混合溶液中阿拉伯胶的质量浓度为0.1％，亚硒酸钠溶液浓度1mM/L；向上述混合溶液中缓慢加入VC，调整亚硒酸钠与VC的摩尔比为1:2，搅拌均匀即为硒-阿拉伯胶复合物；以硒-阿拉伯胶复合物为原料，配制硒-阿拉伯胶复合物水溶液，使硒含量达到0.1mg/L。
以大豆、面粉为原料，大豆与面粉的比例为100:15(w/w)，大豆先用水清洗干净，然后用硒-阿拉伯胶复合物水溶液浸泡大豆，至大豆充分吸水膨胀，淋干的大豆在100℃下蒸煮180min。将米曲霉(Aspergillus oryzae)与面粉预混合，然后与降至常温的大豆混合，大豆与面粉的比例为100:15(w/w)，在20℃培养至混合物表面呈现浅黄绿色，即得到大曲。
配制浓度为5％(w/w)盐水，往盐水中添加甘油、VB5、VB7、胆碱和肌醇，混合均匀，使盐水:甘油:胆碱:肌醇＝1000000:1000:50:10(w/w)。往含有甘油、VB、胆碱和肌醇的盐水中添加嗜盐四联球菌(Tetragenococcus halophilus)，使其在盐水中的含量达到1×10 4CFU/ml，混合均匀，以大曲与盐水比为1:1(w/v)混合制醪，发酵至第5天时，添加鲁氏酵母(Zygosaccharomyces rouxii)，使其在醪液中的含量达到5×10 4CFU/ml，常温发酵30天，发酵结束通过压榨、过滤、杀菌和灌装即得酱油。
对照组1：对照组1为未使用硒-阿拉伯胶复合物水溶液浸泡大豆，同时盐水中未添加甘油、胆碱和肌醇，原料配比和生产工艺与实施例1相同。
由表1和表2可知，采用本发明方法生产的大曲其中性和碱性蛋白酶酶活显著高于对照样品大曲酶活，采用本发明方法生产的酱油其氨基酸态氮、总糖和还原糖含量显著高于对照酱油相应值。感官评价结果显示(图2A)，与对照酱油相比，采用本发明方法生产的酱油其鲜味、甜味和醇厚感均显著改善。由图1可知，采用本发明专利方法可显著提高发酵过程中酱醪中乳酸菌和酵母菌数量，并显著提高终产品中关键风味化合物的含量(见表3)。感官评价结果显示(图2B)，采用本发明专利方法显著改善了酱油的醇香和烟熏香气。综上，采用本发明方法显著改善了酱油的滋味和香气。采用本发明方法生产的酱油总硒和有机硒含量分别为0.049mg/L和0.034mg/L，其对照总硒和有机硒含量分别为0.021mg/L和0.016mg/L，说明采用本发明方法可显著提高酱油总硒和有机硒含量。
表1实施例1和对照组1制备的大曲酶活对比
Figure PCTCN2021081108-appb-000001
注：(1)同一列数值上标同一个字母表示两组数值之间不存在显著差异(p>0.05)，同一列数值上标不同字母表示两组数值之间存在显著差异(p<0.05)。
表2实施例1和其对照组1制备酱油的滋味物质含量
Figure PCTCN2021081108-appb-000002
注：(1)同一列数值上标同一个字母表示两组数值之间不存在显著差异(p>0.05)，同一列数值 上标不同字母表示两组数值之间存在显著差异(p<0.05)。
表3实施例1和对照组1制备酱油中的关键香气化合物
Figure PCTCN2021081108-appb-000003
Figure PCTCN2021081108-appb-000004
a,b,c化合物鉴定方法：(a)通过与相同条件下标准化合物的保留指数(RI)和GC-MS色谱图比对；(b)通过与NIST05数据库图谱比对；(c)通过与GC-O所嗅闻到的标准化合物的香气特征进行比对。
d在DB-wax和TR-5MS色谱柱上分别计算RI。
ND：未检测到，上述数据为3次平行实验平均值。
实施例2：以豆粕为蛋白质原料采用本发明方法生产酱油
先配制硒酸钠溶液，再向该硒酸钠溶液添加阿拉伯胶得到混合溶液；混合溶液中阿拉伯胶的质量浓度为30％，硒酸钠的浓度1000mM/L；向上述混合溶液中缓慢加入VC、VB1、蛋氨酸，调整硒酸钠与VC、VB1、蛋氨酸的摩尔比为1:10:5:5，搅拌均匀即为硒-阿拉伯胶复合物。以硒-阿拉伯胶复合物为原料，配制硒-阿拉伯胶复合物水溶液，使硒含量达到10mg/L。以豆粕、粉碎的炒小麦为原料，豆粕与粉碎的炒小麦的比例为100:100(w/w)，豆粕用硒-阿拉伯胶复合物水溶液充分浸润，润水后的豆粕在140℃下蒸煮5min。将酱油曲霉孢子(Aspergillus sojae)与粉碎的炒小麦预混合，然后与降至常温的豆粕混合，豆粕与粉碎的炒小麦的比例为100:100(w/w)，在35℃培养至混合物表面呈现浅黄绿色，即得到大曲。配制浓度为25％(w/w)盐水，往盐水中添加甘油、VB5、VB7、胆碱和肌醇，混合均匀，使盐水:甘油:VB5:VB7:胆碱:肌醇＝1000000:15000:100:1:5000:1000(w/w)。往含有甘油、VB、胆碱和肌醇的盐水中添加嗜盐四联球菌(Tetragenococcus halophilus)，使其在盐水中的含量达到1×10 7CFU/ml，混合均匀，以大曲与盐水比为1:4(w/v)混合制醪，发酵至第5天时，添加鲁氏酵母(Zygosaccharomyces rouxii)，使其在醪液中的含量达到1×10 7CFU/ml，常温发酵360天，发酵结束通过淋油、过滤、杀菌和灌装即得酱油。
对照组2：对照组2为未使用硒-阿拉伯胶复合物水溶液浸泡大豆，同时盐水中未添加甘油、胆碱和肌醇，原料配比和生产工艺与实施例2相同。
由表4和表5可知，采用本发明方法生产的大曲其酸性、中性和碱性蛋白酶酶活显著高于对照样品大曲酶活，采用本发明方法生产的酱油其总氮氨基酸态氮、总糖和还原糖含 量显著高于对照酱油相应值。感官评价结果显示(图4A)，与对照酱油相比，采用本发明方法生产的酱油其鲜味、甜味和醇厚感均显著改善。由图3可知，采用本发明专利方法可显著提高发酵过程中酱醪中乳酸菌和酵母菌数量，并显著提高终产品中关键风味化合物的含量(见表6)。感官评价结果显示(图4B)，采用本发明专利方法显著改善了酱油的醇香和烟熏香气。综上，采用本发明方法显著改善了酱油的滋味和香气。采用本发明方法生产的酱油总硒和有机硒含量分别为0.95mg/L和0.60mg/L，其对照总硒和有机硒含量分别为0.011mg/L和0.07mg/L，说明采用本发明方法可显著提高酱油总硒和有机硒含量。
表4实施例2和对照组2制备的大曲酶活
Figure PCTCN2021081108-appb-000005
注：(1)同一列数值上标同一个字母表示两组数值之间不存在显著差异(p>0.05)，同一列数值上标不同字母表示两组数值之间存在显著差异(p<0.05)。
表5实施例2和对照组2制备酱油的滋味物质含量
Figure PCTCN2021081108-appb-000006
注：(1)同一列数值上标同一个字母表示两组数值之间不存在显著差异(p>0.05)，同一列数值上标不同字母表示两组数值之间存在显著差异(p<0.05)。
表6实施例2和对照组2制备酱油中的关键香气化合物
Figure PCTCN2021081108-appb-000007
Figure PCTCN2021081108-appb-000008
a,b,c化合物鉴定方法：(a)通过与相同条件下标准化合物的保留指数(RI)和GC-MS色谱图比对；(b)通过与NIST05数据库图谱比对；(c)通过与GC-O所嗅闻到的标准化合物的香气特征进行比对。
d在DB-wax和TR-5MS色谱柱上分别计算RI。
ND：未检测到，上述数据为3次平行实验平均值。
实施例3：以大豆为蛋白质原料采用本发明方法生产酱油
配制阿拉伯水溶液，向阿拉伯胶水溶液中加入亚硒酸钠得到混合溶液；混合溶液中阿拉伯胶的质量浓度为1％，亚硒酸钠的浓度25mM/L；向上述混合溶液中缓慢加入VC、VB1、蛋氨酸，调整亚硒酸钠与VC、VB1、蛋氨酸的摩尔比为1:5:2.5:2.5，搅拌均匀即为硒-阿拉伯胶复合物。以硒-阿拉伯胶复合物为原料，配制硒-阿拉伯胶复合物水溶液，使硒含量达到5mg/L。以大豆、粉碎的炒小麦为原料，大豆与粉碎的炒小麦的比例为100:500(w/w)，大豆先用水清洗干净，然后用硒-阿拉伯胶复合物水溶液浸泡大豆，至大豆充分吸水膨胀，豆粕用硒-阿拉伯胶复合物水溶液充分浸润，淋干的大豆在130℃下蒸煮20min。将米曲霉(Aspergillus oryzae)与粉碎的炒小麦预混合，然后与降至常温的大豆混合，大豆与粉碎的炒小麦的比例为100:500(w/w)，在30℃培养至混合物表面呈现浅黄绿色，即得到大曲。配制浓度为18％(w/w)盐水，往盐水中添加甘油、VB5、VB7、胆碱和肌醇，混合均匀，使盐水:甘油:VB5:VB7:胆碱:肌醇＝1000000:5000:50:0.5:2000:100(w/w)。往含有甘油、VB、胆碱和肌醇的盐水中添加嗜盐四联球菌(Tetragenococcus halophilus)，使其在盐水中的含量达到5×10 5CFU/ml，混合均匀，以大曲与盐水比为1:2.5(w/v)混合制醪，发酵至第5天时，添加鲁氏酵母(Zygosaccharomyces rouxii)，使其在醪液中的含量达到1×10 6CFU/ml，常温发酵180天，发酵结束通过淋油、过滤、杀菌和灌装即得酱油。
对照组3：对照组3为未使用硒-阿拉伯胶复合物水溶液浸泡大豆，盐水中未添加甘油、胆碱和肌醇，原料配比和生产工艺与实施例3相同的酱油。
由表7和表8可知，采用本发明方法生产的大曲其酸性、中性和碱性蛋白酶酶活显著 高于对照样品大曲酶活，采用本发明方法生产的酱油其总氮氨基酸态氮、总糖和还原糖含量显著高于对照酱油相应值。感官评价结果显示(图6A)，与对照酱油相比，采用本发明方法生产的酱油其鲜味、甜味和醇厚感均显著改善。由图5可知，采用本发明专利方法可显著提高发酵过程中酱醪中乳酸菌和酵母菌数量，并显著提高终产品中关键风味化合物的含量(见表9)。感官评价结果显示(图6B)，采用本发明专利方法显著改善了酱油的醇香和烟熏香气。综上，采用本发明方法显著改善了酱油的滋味和香气。采用本发明方法生产的酱油总硒和有机硒含量分别为0.81mg/L和0.52mg/L，其对照总硒和有机硒含量分别为0.017mg/L和0.012mg/L，说明采用本发明方法可显著提高酱油总硒和有机硒含量。
表7实施例3和对照组3制备的大曲酶活
Figure PCTCN2021081108-appb-000009
注：(1)同一列数值上标同一个字母表示两组数值之间不存在显著差异(p>0.05)，同一列数值上标不同字母表示两组数值之间存在显著差异(p<0.05)。
表8实施例3和对照组3制备酱油中的滋味物质含量
Figure PCTCN2021081108-appb-000010
注：(1)同一列数值上标同一个字母表示两组数值之间不存在显著差异(p>0.05)，同一列数值上标不同字母表示两组数值之间存在显著差异(p<0.05)。
表9实施例3和对照组3制备酱油中的关键香气化合物
Figure PCTCN2021081108-appb-000011
Figure PCTCN2021081108-appb-000012
a,b,c化合物鉴定方法：(a)通过与相同条件下标准化合物的保留指数(RI)和GC-MS色谱图比对；(b)通过与NIST05数据库图谱比对；(c)通过与GC-O所嗅闻到的标准化合物的香气特征进行比对。
d在DB-wax和TR-5MS色谱柱上分别计算RI。
ND：未检测到，上述数据为3次平行实验平均值。
本发明是利用硒与维生素、胆碱、肌醇和甘油，显著提高了酱油大曲酶活、酱醪中的乳酸菌和酵母菌数量，提高了滋味和香气物质含量，同时也提高了酱油中的总硒和有机硒含量。该方法生产的酱油，不但改善了酱油风味，而且赋予了酱油保健功能，进而提高了酱油的附加值，具有良好的市场前景。因此本发明具有重要的经济价值和社会意义。
虽然本发明已以较佳实施例公开如上，但其并非用以限定本发明，任何熟悉此技术的人，在不脱离本发明的精神和范围内，都可做出各种改动与修饰，因此本发明的保护范围应该以权利要求书所界定范围为准。

Claims (10)

  1. 一种同时改善酱油风味和提高酱油硒含量的方法，其特征在于，步骤如下：
    (1)以水为溶剂，配制阿拉伯胶与亚硒酸钠或者硒酸钠的混合溶液；所述混合溶液中阿拉伯胶的质量浓度为0.1％-30％；所述混合溶液中亚硒酸钠或硒酸钠的浓度为1-1000mM/L；然后在混合溶液中加入VC、VB1、蛋氨酸，搅拌均匀，即得到硒-阿拉伯胶复合物；所述亚硒酸钠或硒酸钠与VC、VB1、蛋氨酸的摩尔比为1:(2-10):(0-5):(0-5)；
    (2)以硒-阿拉伯胶复合物为原料，配制硒-阿拉伯胶复合物水溶液；
    (3)以大豆或豆粕、面粉或粉碎的炒小麦为原料；以大豆为原料时，使用前先用水清洗干净；然后用步骤(2)制备的硒-阿拉伯胶复合物水溶液浸泡大豆或豆粕，浸泡后淋干水分，将大豆或豆粕在100-140℃进行蒸煮5-180min，蒸煮后降至常温，备用；
    (4)将米曲霉或酱油曲霉孢子与面粉或粉碎的炒小麦预混合，得到预混料，然后与步骤(3)中蒸煮后降至常温的大豆或豆粕混合，在20-35℃培养至混合物表面呈现浅黄绿色，即得到大曲；
    (5)首先配制一定质量浓度的盐水，然后往盐水中添加甘油、VB5、VB7、胆碱和肌醇，混合均匀，得到盐水混合物；盐水混合物中盐水、甘油、VB5、VB7、胆碱和肌醇的质量比为1000000:(1000-15000):(0-100):(0-1):(50-5000):(10-1000)；
    (6)往步骤(5)得到的盐水混合物中添加嗜盐四联球菌，使其在盐水混合物中的含量达到1×10 4-1×10 7CFU/mL，混合均匀，然后加入步骤(4)制备的大曲混合制醪，发酵数天后，得到醪液；然后再添加鲁氏酵母，使其在醪液中的含量达到5×5×10 4-1×10 7CFU/mL，常温发酵30-360天，发酵结束通过压榨或淋油、过滤、杀菌和灌装即得酱油。
  2. 根据权利要求1所述的同时改善酱油风味和提高酱油硒含量的方法，其特征在于，步骤(1)中所述混合溶液中阿拉伯胶的质量浓度为0.2％-10％；所述混合溶液中亚硒酸钠或硒酸钠的浓度为10-100mM/L。
  3. 根据权利要求1所述的同时改善酱油风味和提高酱油硒含量的方法，其特征在于，步骤(1)中所述亚硒酸钠或硒酸钠与VC、VB1、蛋氨酸的摩尔比为1:(3-7):(0.5-3):(0.5-3)。
  4. 根据权利要求1所述的同时改善酱油风味和提高酱油硒含量的方法，其特征在于，步骤(2)中所述硒-阿拉伯胶复合物水溶液中硒含量为0.1-10mg/L。
  5. 根据权利要求1所述的同时改善酱油风味和提高酱油硒含量的方法，其特征在于，步骤(3)中所述大豆或豆粕浸泡的时间为0.5h-24h；所述蒸煮的温度为110-130℃，蒸煮时间为30-60min。
  6. 根据权利要求1所述的同时改善酱油风味和提高酱油硒含量的方法，其特征在于，步骤(4)中所述米曲霉或酱油曲霉孢子与面粉或粉碎的炒小麦的质量比为1：(100-10000)。
  7. 根据权利要求1所述的同时改善酱油风味和提高酱油硒含量的方法，其特征在于，步骤(4)中所述大豆或豆粕与预混料中面粉或粉碎的炒小麦的质量比为100:(15-500)。
  8. 根据权利要求1所述的同时改善酱油风味和提高酱油硒含量的方法，其特征在于，步骤(5)中盐水的质量浓度为5％-25％；所述盐水混合物中盐水、甘油、VB5、VB7、胆碱和肌醇的质量比为1000000:(2000-10000):(5-50):(0.05-0.5):(50-1000):(50-500)。
  9. 根据权利要求1所述的同时改善酱油风味和提高酱油硒含量的方法，其特征在于，步骤(6)中所述嗜盐四联球菌在盐水混合物中的含量为5×10 4-5×10 6CFU/mL；所述鲁氏酵母在醪液中的含量为1×10 5-5×10 6CFU/mL。
  10. 根据权利要求1所述的同时改善酱油风味和提高酱油硒含量的方法，其特征在于，步骤(6)中所述大曲与盐水混合物的用量比为1g:(1-4)mL；所述发酵数天具体是3-15天；所述常温发酵30-180天。
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