Composite enzymatic hydrolysis aroma enhancement preparation method of seasoning sauce

Abstract

The invention relates to the technical field of seasoning sauce preparation, in particular to a composite enzymatic hydrolysis aroma enhancement preparation method of seasoning sauce, which comprises the following steps: soaking shiitake mushroom particles in a composite buffer solution to obtain a solid A and a flavor liquid B, mixing the solid A with deionized water, grinding into a homogenate, performing three times of enzymatic hydrolysis to obtain an enzymatic hydrolysate C, and adding the sterilized supernate D, the solid A and the flavor liquid B into other stir-fried raw materials to obtain the seasoning sauce with obvious shiitake mushroom flavor and sufficient fragrance. Cellulase in the composite buffer solution can quickly decompose a cellulose skeleton of a cell wall, so that polyphenol oxidase can enter cells conveniently, an enzymatic browning reaction is catalyzed, a Maillard reaction is facilitated, and various flavor substances are generated; and after the three-step enzymolysis reaction, protein in the shiitake mushrooms is hydrolyzed into small molecular amino acids, so that the fresh and sweet flavor is effectively increased, the shiitake mushroom flavor is more obvious, and the method has a wide application prospect.

Description

Technical Field

[0001] This invention relates to the field of seasoning sauce preparation technology, and in particular to a compound enzymatic hydrolysis method for preparing seasoning sauces with enhanced flavor. Background Technology

[0002] Sauces are condiments used to harmonize the flavors of various foods to satisfy the diners' preferences. Sauces can be eaten with food; for example, when eating rice, the sauce can be poured into the rice and stirred before eating. They can also be used as a dip for bread, steak, and other foods. Sauces can also be added to food ingredients to create sauce-infused foods, such as sandwich cookies, sandwich bread, and flavored beverages, to improve their taste. Most commonly used sauces in daily life are made from vegetables or fruits, such as mushroom sauce, chili sauce, salad dressing, tomato sauce, kiwi sauce, and blueberry sauce. Because vegetables and fruits are rich in vitamins, minerals, aromatic substances, pigments, and organic acids, fruit and vegetable sauces are widely popular.

[0003] Shiitake mushrooms, also known as fragrant mushrooms, are fungi belonging to the phylum Basidiomycetes, class Agaricales, order Tricholomatales, family Tricholomataceae, and genus Lentinus. With their delicious flavor and unique taste, shiitake mushrooms are beloved by consumers. They not only contain nutrients such as protein, fat, carbohydrates, minerals, trace elements, amino acids, and vitamins, but also possess medicinal value, including anti-tumor properties, prevention of cardiovascular and cerebrovascular diseases, and liver protection and detoxification.

[0004] Fresh shiitake mushrooms are difficult to preserve due to their high water content. Therefore, deep processing of shiitake mushrooms is an important way to maintain their flavor and nutritional value. In recent years, edible fungi and their products have become increasingly popular, especially edible fungi sauces, which are attracting more and more attention. my country has conducted considerable research on edible fungi sauces, mainly focusing on the process of combining edible fungi with other ingredients and seasonings for stir-frying.

[0005] With the development of shiitake mushroom products in recent years, shiitake mushroom sauce has become increasingly popular among consumers. The main raw materials for making shiitake mushroom sauce include shiitake mushrooms, soybean paste, and soybean oil, while auxiliary materials include salt, white sugar, chili peppers, and spices. The selection and proportion of these raw materials are crucial to the final flavor and quality of the shiitake mushroom sauce.

[0006] The existing technology for preparing shiitake mushroom sauce usually involves soaking raw shiitake mushrooms in water and then stir-frying them directly with other raw materials to obtain the final shiitake mushroom sauce. This process cannot effectively release the flavor compounds in shiitake mushrooms, resulting in the flavors of shiitake mushrooms and other raw materials not blending perfectly in the sauce.

[0007] Therefore, based on the relevant technologies mentioned above, there is an urgent need to develop a compound enzymatic hydrolysis method for preparing flavoring sauces. Summary of the Invention

[0008] In view of this, the purpose of this invention is to propose a compound enzymatic hydrolysis method for preparing flavoring sauces, so as to solve the problem that the flavor of shiitake mushrooms is not obvious in the existing shiitake mushroom seasoning sauce.

[0009] To achieve the above objectives, the present invention provides a method for preparing a flavoring sauce by compound enzymatic hydrolysis.

[0010] A method for preparing a flavoring sauce using compound enzymatic hydrolysis includes the following steps: Step S1. Take the washed fresh and dried shiitake mushrooms, separate the stems and caps, and crush them to 0.1-0.5cm. Then soak them in a compound buffer solution at 18-23℃ for 40-60min, and centrifuge to obtain solid A and flavor liquid B. Step S2. Wash solid A with deionized water, then add deionized water to solid A and continue grinding into a homogenate. Then add a compound enzyme for enzymatic hydrolysis to obtain hydrolysate C. Step S3. Heat the enzymatic hydrolysate C to 88-95℃ for sterilization, centrifuge, and filter to obtain the supernatant D; Step S4. Add rapeseed oil to the pot and heat it to 120-130℃. Then add onion, garlic, scallion, and minced ginger and stir-fry for 8-15 minutes. Then add solid A, supernatant D and flavor liquid B and continue to stir-fry for 5-10 minutes. Finally, add Pixian chili bean paste, sweet bean sauce, soybean paste, white sugar, salt, MSG and thirteen spices and stir-fry for 4-7 minutes to obtain the compound enzymatically hydrolyzed flavoring sauce. The composite buffer solution in step S1 is prepared from polyphenol oxidase with an enzyme activity of 20,000 U / g, cellulase with an enzyme activity of 10,000 U / g, citrate-disodium hydrogen phosphate buffer with a concentration of 0.05 mol / L, and disodium hydrogen phosphate solution with a concentration of 0.1 mol / L.

[0011] Preferably, the preparation method of the composite buffer solution is as follows: Mix 0.05 mol / L citrate-disodium hydrogen phosphate buffer and 0.1 mol / L disodium hydrogen phosphate solution and control the pH of the system to 5.2-5.4 to obtain a buffer solution. Then add polyphenol oxidase with an enzyme activity of 20000 U / g and cellulase with an enzyme activity of 10000 U / g to the buffer solution and stir until completely dissolved to obtain a composite buffer solution.

[0012] Preferably, the mass ratio of the polyphenol oxidase, cellulase and buffer solution is 0.04-0.06:0.01-0.03:100-200.

[0013] Preferably, the enzymatic hydrolysis in step S2 includes a first enzymatic hydrolysis, a second enzymatic hydrolysis, and a third enzymatic hydrolysis; The first enzymatic hydrolysis of the compound enzyme 1 is prepared by mixing Bacillus subtilis protease with an enzyme activity of 10000 U / g and cellulase with an enzyme activity of 10000 U / g at a mass ratio of 5.2-8:3.7-6. The temperature of the first enzymatic hydrolysis is 50-55℃, the hydrolysis time is 1.5-2.5h, and the pH of the first enzymatic hydrolysis is adjusted to 5.3-5.5.

[0014] Preferably, the mass ratio of compound enzyme 1 to homogenate during the first enzymatic hydrolysis is 0.5-0.8:190-230.

[0015] Preferably, the compound enzyme 2 in the second enzymatic hydrolysis is obtained by mixing leucine aminopeptidase with an enzyme activity of 8000 U / g and flavor protease with an enzyme activity of 50000 U / g at a mass ratio of 1-2:2.5-5.7. The temperature of the second enzymatic hydrolysis is 48-52℃, the hydrolysis time is 1-2h, and the pH of the second enzymatic hydrolysis is adjusted to 6.5-6.8.

[0016] Preferably, the mass ratio of compound enzyme 2 to homogenate during the second enzymatic hydrolysis is 0.7-1:190-230.

[0017] Preferably, the compound enzyme 3 used in the third enzymatic hydrolysis is a mixture of 5′ phosphodiesterase with an enzyme activity of 20000 U / g and glutaminase with an enzyme activity of 10000 U / g at a mass ratio of 2-5:1-2. The temperature of the third enzymatic hydrolysis is 43-47℃, the hydrolysis time is 1-1.5h, and the pH of the third enzymatic hydrolysis is adjusted to 5.2-5.5.

[0018] Preferably, the mass ratio of compound enzyme 3 to homogenate during the third enzymatic hydrolysis is 1-1.2:190-230.

[0019] Preferably, the mass ratio of rapeseed oil, onion, garlic, scallion, minced ginger, solid A, supernatant D, flavor liquid B, Pixian chili bean paste, sweet bean sauce, soybean paste, white sugar, salt, monosodium glutamate and thirteen-spice powder in step S4 is 50-70:20-30:5-7:8-13:1-3:5-8:13-17:5-8:6-8:2-3:8-10:8-10:3-5:3-5:2-4.

[0020] The beneficial effects of this invention are: This invention provides a method for preparing a flavor-enhancing sauce using compound enzymatic hydrolysis. The method involves soaking shiitake mushroom granules in a compound buffer solution to obtain solid A and flavor liquid B. Solid A is then mixed with deionized water and ground into a homogenate. This homogenate is subjected to three enzymatic hydrolysates to obtain hydrolysate C. The supernatant D obtained after sterilization is then added to other raw materials during stir-frying, along with solid A and flavor liquid B, to obtain a flavor-enhancing sauce with a distinct shiitake mushroom flavor and rich aroma. The compound buffer solution is prepared by mixing polyphenol oxidase, cellulase, and the buffer solution. Cellulase rapidly decomposes the cellulose skeleton of the cell wall, which is beneficial for polyphenol oxidase. Entering the cell interior, it catalyzes an enzymatic browning reaction, generating quinone compounds that spontaneously undergo Maillard reactions with amino acids in shiitake mushrooms, producing a variety of flavor compounds. In the three-step enzymatic hydrolysis reaction, the first enzymatic hydrolysis effectively breaks down the proteins in shiitake mushrooms into small molecule peptides and disrupts the cell wall skeleton structure. The second enzymatic hydrolysis further hydrolyzes the small molecule peptides into amino acids, removing bitterness while increasing umami flavor. The third enzymatic hydrolysis increases the 5'-nucleotide content in the seasoning sauce, further enhancing aroma and umami. Compared with existing technologies, this method has broad application prospects. Detailed Implementation

[0021] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to specific embodiments.

[0022] The sources and properties of some of the raw materials used in this invention are as follows: Flavor protease was purchased from Henan Wanbang Industrial Co., Ltd.; cellulase was purchased from Henan Wanbang Industrial Co., Ltd.

[0023] Example 1: A method for preparing a flavoring sauce using compound enzymatic hydrolysis, comprising the following steps: S1. Mix 0.05 mol / L citrate-disodium hydrogen phosphate buffer and 0.1 mol / L disodium hydrogen phosphate solution and control the pH of the system to 5.2 to obtain a buffer solution. Then add 0.04 g of polyphenol oxidase with an enzyme activity of 20000 U / g and 0.01 g of cellulase with an enzyme activity of 10000 U / g to 100 g of buffer solution and stir until completely dissolved to obtain a composite buffer solution. S2. Take the washed fresh and dried shiitake mushrooms, separate the stems and caps, and crush them to 0.1cm. Then soak them in a compound buffer solution at 18℃ for 40min, centrifuge, and obtain solid A and flavor liquid B. S3. Wash solid A with deionized water, then add more deionized water to solid A and continue grinding to form a homogenate. Next, add 0.5g of compound enzyme 1 to 190g of homogenate and hydrolyze at 50℃ for 1.5h. Adjust the pH to 5.3 during the first hydrolysis. Compound enzyme 1 is obtained by mixing 5.2g of Bacillus subtilis protease with an activity of 10000U / g and 3.7g of cellulase with an activity of 10000U / g. After the first hydrolysis, add 0.7g of compound enzyme 2 to the homogenate and hydrolyze at 48℃ for 1h. Adjust the pH to 5.3 during the second hydrolysis. The pH during enzymatic hydrolysis was 6.5. Complex enzyme 2 was obtained by mixing 1g of leucine aminopeptidase with an activity of 8000U / g and 2.5g of flavor protease with an activity of 50000U / g. After the second enzymatic hydrolysis, 1g of complex enzyme 3 was added to the homogenate and enzymatic hydrolysis was carried out at 43℃ for 1h. The pH during the third enzymatic hydrolysis was adjusted to 5.2. Complex enzyme 3 was obtained by mixing 2g of 5′ phosphodiesterase with an activity of 20000U / g and 1g of glutaminase with an activity of 10000U / g. After the third enzymatic hydrolysis, enzymatic hydrolysate C was obtained. S4. Heat the enzymatic hydrolysate C to 88°C for sterilization, centrifuge, and filter to obtain the supernatant D; S5. Add 50g of rapeseed oil to the pot and heat it to 120℃. Then add 20g of onion, 5g of garlic, 8g of scallion, and 1g of minced ginger and stir-fry for 8 minutes. Then add 5g of solid A, 13g of supernatant D and 5g of flavor liquid B and continue to stir-fry for 5 minutes. Finally, add 6g of Pixian chili bean paste, 2g of sweet bean sauce, 8g of soybean paste, 8g of white sugar, 3g of salt, 3g of MSG and 2g of thirteen-spice powder and stir-fry for 4 minutes to obtain the compound enzymatically hydrolyzed flavoring sauce.

[0024] Example 2: A method for preparing a flavoring sauce using compound enzymatic hydrolysis, comprising the following steps: S1. Mix 0.05 mol / L citrate-disodium hydrogen phosphate buffer and 0.1 mol / L disodium hydrogen phosphate solution and control the pH of the system to 5.3 to obtain a buffer solution. Then add 0.045 g of polyphenol oxidase with an enzyme activity of 20000 U / g and 0.015 g of cellulase with an enzyme activity of 10000 U / g to 125 g of buffer solution and stir until completely dissolved to obtain a composite buffer solution. S2. Take the washed fresh and dried shiitake mushrooms, separate the stems and caps, and crush them to 0.2cm. Then soak them in a compound buffer solution at 20℃ for 45min, centrifuge, and obtain solid A and flavor liquid B. S3. Wash solid A with deionized water, then add more deionized water to solid A and continue grinding to form a homogenate. Next, add 0.6g of compound enzyme 1 to 200g of the homogenate and hydrolyze at 52℃ for 2 hours, adjusting the pH to 5.3 during the first hydrolysis. Compound enzyme 1 is obtained by mixing 5.8g of Bacillus subtilis protease with an enzyme activity of 10000U / g and 4.3g of cellulase with an enzyme activity of 10000U / g. After the first hydrolysis, add 0.8g of compound enzyme 2 to the homogenate and hydrolyze at 50℃ for 1 hour, adjusting the pH to 5.3 during the second hydrolysis. The pH of the solution was 6.6. Complex enzyme 2 was obtained by mixing 1.4g of leucine aminopeptidase with an enzyme activity of 8000U / g and 3g of flavor protease with an enzyme activity of 50000U / g. After the second enzymatic hydrolysis, 1.1g of complex enzyme 3 was added to the homogenate and enzymatically hydrolyzed at 45℃ for 1h. The pH of the third enzymatic hydrolysis was adjusted to 5.3. Complex enzyme 3 was obtained by mixing 3g of 5′ phosphodiesterase with an enzyme activity of 20000U / g and 1.4g of glutaminase with an enzyme activity of 10000U / g. After the third enzymatic hydrolysis, the enzymatic hydrolysate C was obtained. S4. Sterilize the enzymatic hydrolysate C by heating it to 90°C, centrifuging, and filtering to obtain the supernatant D. S5. Add 55g rapeseed oil to the pot and heat it to 125℃. Then add 24g onion, 5.5g garlic, 9g scallion, and 1.5g minced ginger and stir-fry for 10 minutes. Then add 6g solid A, 14g supernatant D and 6g flavor liquid B and continue to stir-fry for 6 minutes. Finally, add 6.5g Pixian chili bean paste, 2.5g sweet bean sauce, 8.5g soybean paste, 8.5g white sugar, 3.5g salt, 3.5g MSG and 3g thirteen-spice powder and stir-fry for 5 minutes to obtain the compound enzymatically hydrolyzed flavoring sauce.

[0025] Example 3: A method for preparing a flavoring sauce using compound enzymatic hydrolysis, comprising the following steps: S1. Mix 0.05 mol / L citrate-disodium hydrogen phosphate buffer and 0.1 mol / L disodium hydrogen phosphate solution and control the pH of the system to 5.3 to obtain a buffer solution. Then add 0.05 g of polyphenol oxidase with an enzyme activity of 20000 U / g and 0.02 g of cellulase with an enzyme activity of 10000 U / g to 150 g of the buffer solution and stir until completely dissolved to obtain a composite buffer solution. S2. Take the washed fresh shiitake mushrooms and dried shiitake mushrooms, separate the stems and caps, and crush them to 0.3cm. Then soak them in a compound buffer solution at 20℃ for 50min, and centrifuge to obtain solid A and flavor liquid B. S3. Wash solid A with deionized water, then add more deionized water to solid A and continue grinding to form a homogenate. Next, add 0.7g of compound enzyme 1 to 210g of the homogenate and hydrolyze at 53℃ for 2 hours, adjusting the pH to 5.4 during the first hydrolysis. Compound enzyme 1 is obtained by mixing 6.5g of Bacillus subtilis protease with an activity of 10000U / g and 5g of cellulase with an activity of 10000U / g. After the first hydrolysis, add 0.9g of compound enzyme 2 to the homogenate and hydrolyze at 50℃ for 1.5 hours, adjusting the pH to 5.4 during the second hydrolysis. The pH of the solution was 6.7. Complex enzyme 2 was obtained by mixing 1.7g of leucine aminopeptidase with an enzyme activity of 8000U / g and 4g of flavor protease with an enzyme activity of 50000U / g. After the second enzymatic hydrolysis, 1.15g of complex enzyme 3 was added to the homogenate and enzymatically hydrolyzed at 45℃ for 1h. The pH of the third enzymatic hydrolysis was adjusted to 5.4. Complex enzyme 3 was obtained by mixing 4g of 5′ phosphodiesterase with an enzyme activity of 20000U / g and 1.7g of glutaminase with an enzyme activity of 10000U / g. After the third enzymatic hydrolysis, the enzymatic hydrolysate C was obtained. S4. Sterilize the enzymatic hydrolysate C by heating it to 90°C, centrifuging, and filtering to obtain the supernatant D. S5. Add 60g of rapeseed oil to the pot and heat it to 125℃. Then add 27g of onion, 6g of garlic, 11g of scallion, and 2g of minced ginger and stir-fry for 12 minutes. Then add 7g of solid A, 15g of supernatant D and 7g of flavor liquid B and continue to stir-fry for 7 minutes. Finally, add 7g of Pixian chili bean paste, 2.7g of sweet bean sauce, 9g of soybean paste, 9g of white sugar, 4g of salt, 4g of MSG and 3g of thirteen-spice powder and stir-fry for 5 minutes to obtain the compound enzymatically hydrolyzed flavoring sauce.

[0026] Example 4: A method for preparing a flavoring sauce using compound enzymatic hydrolysis, comprising the following steps: S1. Mix 0.05 mol / L citrate-disodium hydrogen phosphate buffer and 0.1 mol / L disodium hydrogen phosphate solution and control the pH of the system to 5.4 to obtain a buffer solution. Then add 0.055 g of polyphenol oxidase with an enzyme activity of 20000 U / g and 0.025 g of cellulase with an enzyme activity of 10000 U / g to 175 g of the buffer solution and stir until completely dissolved to obtain a composite buffer solution. S2. Take the washed fresh shiitake mushrooms and dried shiitake mushrooms, separate the stems and caps, and crush them to 0.4 cm. Then soak them in a compound buffer solution at 22℃ for 55 min, and centrifuge to obtain solid A and flavor liquid B. S3. Wash solid A with deionized water, then add more deionized water to solid A and continue grinding to form a homogenate. Next, add 0.7g of compound enzyme 1 to 220g of the homogenate and hydrolyze at 53℃ for 2.5h. Adjust the pH to 5.5 during the first hydrolysis. Compound enzyme 1 is obtained by mixing 7g of Bacillus subtilis protease with an enzyme activity of 10000U / g and 5.5g of cellulase with an enzyme activity of 10000U / g. After the first hydrolysis, add 0.9g of compound enzyme 2 to the homogenate and hydrolyze at 50℃ for 1.5h. Adjust the pH to 5.5 during the second hydrolysis. The pH of the solution was 6.8. Complex enzyme 2 was obtained by mixing 1.8g of leucine aminopeptidase with an enzyme activity of 8000U / g and 5g of flavor protease with an enzyme activity of 50000U / g. After the second enzymatic hydrolysis, 1.2g of complex enzyme 3 was added to the homogenate and enzymatically hydrolyzed at 47℃ for 1.5h. The pH of the third enzymatic hydrolysis was adjusted to 5.5. Complex enzyme 3 was obtained by mixing 4g of 5′ phosphodiesterase with an enzyme activity of 20000U / g and 1.8g of glutaminase with an enzyme activity of 10000U / g. After the third enzymatic hydrolysis, the enzymatic hydrolysate C was obtained. S4. Heat the enzymatic hydrolysate C to 93°C for sterilization, centrifuge, and filter to obtain the supernatant D; S5. Add 65g of rapeseed oil to the pot and heat it to 128℃. Then add 27g of onion, 6.5g of garlic, 12g of scallion, and 2.5g of minced ginger and stir-fry for 13 minutes. Then add 7.5g of solid A, 16g of supernatant D and 7.5g of flavor liquid B and continue to stir-fry for 8 minutes. Finally, add 7.5g of Pixian chili bean paste, 2.8g of sweet bean sauce, 9.5g of soybean paste, 9.5g of white sugar, 4.5g of salt, 4.5g of MSG and 3.5g of thirteen-spice powder and stir-fry for 6 minutes to obtain the compound enzymatically hydrolyzed flavoring sauce.

[0027] Example 5: A method for preparing a flavoring sauce using compound enzymatic hydrolysis, comprising the following steps: S1. Mix 0.05 mol / L citrate-disodium hydrogen phosphate buffer and 0.1 mol / L disodium hydrogen phosphate solution and control the pH of the system to 5.4 to obtain a buffer solution. Then add 0.06 g of polyphenol oxidase with an enzyme activity of 20000 U / g and 0.03 g of cellulase with an enzyme activity of 10000 U / g to 200 g of buffer solution and stir until completely dissolved to obtain a composite buffer solution. S2. Take the washed fresh shiitake mushrooms and dried shiitake mushrooms, separate the stems and caps, and crush them to 0.5cm. Then soak them in a compound buffer solution at 23℃ for 60min, centrifuge, and obtain solid A and flavor liquid B. S3. Wash solid A with deionized water, then add more deionized water to solid A and continue grinding to form a homogenate. Next, add 0.8g of compound enzyme 1 to 230g of the homogenate and hydrolyze at 55℃ for 2.5h. Adjust the pH to 5.5 during the first hydrolysis. Compound enzyme 1 is obtained by mixing 8g of Bacillus subtilis protease with an activity of 10000U / g and 6g of cellulase with an activity of 10000U / g. After the first hydrolysis, add 1g of compound enzyme 2 to the homogenate and hydrolyze at 52℃ for 2h. Adjust the pH to 5.5 during the second hydrolysis. The pH was 6.8. Complex enzyme 2 was obtained by mixing 2g of leucine aminopeptidase with an activity of 8000U / g and 5.7g of flavor protease with an activity of 50000U / g. After the second enzymatic hydrolysis, 1.2g of complex enzyme 3 was added to the homogenate and enzymatically hydrolyzed at 47℃ for 1.5h. The pH was adjusted to 5.5 for the third enzymatic hydrolysis. Complex enzyme 3 was obtained by mixing 5g of 5′ phosphodiesterase with an activity of 20000U / g and 2g of glutaminase with an activity of 10000U / g. After the third enzymatic hydrolysis, enzymatic hydrolysate C was obtained. S4. Heat the enzymatic hydrolysate C to 95°C for sterilization, centrifuge, and filter to obtain the supernatant D; S5. Add 70g of rapeseed oil to the pot and heat it to 130℃. Then add 30g of onion, 7g of garlic, 3g of scallion, and 3g of minced ginger and stir-fry for 15 minutes. Then add 8g of solid A, 17g of supernatant D and 8g of flavor liquid B and continue to stir-fry for 10 minutes. Finally, add 8g of Pixian chili bean paste, 3g of sweet bean sauce, 10g of soybean paste, 10g of white sugar, 5g of salt, 5g of MSG and 4g of thirteen-spice powder and stir-fry for 7 minutes to obtain the compound enzymatically hydrolyzed flavoring sauce.

[0028] Comparative Example 1: S1. Take the washed fresh shiitake mushrooms and dried shiitake mushrooms, separate the stems and caps, and crush them to 0.1cm. Then soak them in deionized water at 50℃ for 40min, centrifuge, and obtain solid A and flavor liquid B. S2. Wash solid A with deionized water, then add more deionized water to solid A and continue grinding to form a homogenate. Next, add 0.5g of compound enzyme 1 to 190g of homogenate and hydrolyze at 50℃ for 1.5h. Adjust the pH to 5.3 during the first hydrolysis. Compound enzyme 1 is obtained by mixing 5.2g of Bacillus subtilis protease with an enzyme activity of 10000U / g and 3.7g of cellulase with an enzyme activity of 10000U / g. After the first hydrolysis, add 0.7g of compound enzyme 2 to the homogenate and hydrolyze at 48℃ for 1h. Adjust the pH to 5.3 during the second hydrolysis. The pH during enzymatic hydrolysis was 6.5. Complex enzyme 2 was obtained by mixing 1g of leucine aminopeptidase with an activity of 8000U / g and 2.5g of flavor protease with an activity of 50000U / g. After the second enzymatic hydrolysis, 1g of complex enzyme 3 was added to the homogenate and enzymatic hydrolysis was carried out at 43℃ for 1h. The pH during the third enzymatic hydrolysis was adjusted to 5.2. Complex enzyme 3 was obtained by mixing 2g of 5′ phosphodiesterase with an activity of 20000U / g and 1g of glutaminase with an activity of 10000U / g. After the third enzymatic hydrolysis, enzymatic hydrolysate C was obtained. S3. Heat the enzymatic hydrolysate C to 88°C for sterilization, centrifuge, and filter to obtain the supernatant D; S4. Add 50g of rapeseed oil to the pot and heat it to 120℃. Then add 20g of onion, 5g of garlic, 8g of scallion, and 1g of minced ginger and stir-fry for 8 minutes. Then add 5g of solid A, 13g of supernatant D and 5g of flavor liquid B and continue to stir-fry for 5 minutes. Finally, add 6g of Pixian chili bean paste, 2g of sweet bean sauce, 8g of soybean paste, 8g of white sugar, 3g of salt, 3g of MSG and 2g of thirteen-spice powder and stir-fry for 4 minutes to obtain the seasoning sauce.

[0029] Comparative Example 2: Compared with Example 1, this comparative example did not add polyphenol oxidase during the preparation of the complex buffer solution. All other steps and parameters were the same, and will not be repeated here. The final product was a seasoning sauce.

[0030] Comparative Example 3: Compared with Example 1, this comparative example did not add cellulase during the preparation of the complex buffer solution. All other steps and parameters were the same, and will not be repeated here. The final product was a seasoning sauce.

[0031] Comparative Example 4: Compared with Example 1, this comparative example did not add solid A in the S5 preparation process, and the remaining steps and parameters were the same. The details of this comparative example will not be repeated here. The final product is a seasoning sauce.

[0032] Comparative Example 5: Compared with Example 1, this comparative example did not add flavor liquid B in the preparation process of S5. All other steps and parameters were the same, and will not be repeated here. The final product was a seasoning sauce.

[0033] Comparative Example 6: S1. Mix 0.05 mol / L citrate-disodium hydrogen phosphate buffer and 0.1 mol / L disodium hydrogen phosphate solution and control the pH of the system to 5.2 to obtain a buffer solution. Then add 0.04 g of polyphenol oxidase with an enzyme activity of 20000 U / g and 0.01 g of cellulase with an enzyme activity of 10000 U / g to 100 g of buffer solution and stir until completely dissolved to obtain a composite buffer solution. S2. Take the washed fresh and dried shiitake mushrooms, separate the stems and caps, and crush them to 0.1cm. Then soak them in a compound buffer solution at 18℃ for 40min, centrifuge, and obtain solid A and flavor liquid B. S3. Wash solid A with deionized water, then add deionized water to solid A and continue grinding into a homogenate. Add 50g of rapeseed oil to a pot and heat to 120℃. Then add 20g of onion, 5g of garlic, 8g of scallion, and 1g of minced ginger and stir-fry for 8 minutes. Then add 10g of homogenate and 10g of flavor liquid B and continue stir-frying for 5 minutes. Finally, add 6g of Pixian chili bean paste, 2g of sweet bean sauce, 8g of soybean paste, 8g of white sugar, 3g of salt, 3g of MSG, and 2g of thirteen-spice powder and stir-fry for 4 minutes to obtain the seasoning sauce.

[0034] Comparative Example 7: S1. Mix 0.05 mol / L citrate-disodium hydrogen phosphate buffer and 0.1 mol / L disodium hydrogen phosphate solution and control the pH of the system to 5.2 to obtain a buffer solution. Then add 0.04 g of polyphenol oxidase with an enzyme activity of 20000 U / g and 0.01 g of cellulase with an enzyme activity of 10000 U / g to 100 g of buffer solution and stir until completely dissolved to obtain a composite buffer solution. S2. Take the washed fresh and dried shiitake mushrooms, separate the stems and caps, and crush them to 0.1cm. Then soak them in a compound buffer solution at 18℃ for 40min, centrifuge, and obtain solid A and flavor liquid B. S3. Wash solid A with deionized water, then add deionized water to solid A and continue grinding to form a homogenate. Then add 0.5g of compound enzyme 1, 0.7g of compound enzyme 2 and 1g of compound enzyme 3 to 190g of homogenate. Compound enzyme 1 is obtained by mixing 5.2g of Bacillus subtilis protease with an enzyme activity of 10000U / g and 3.7g of cellulase with an enzyme activity of 10000U / g. Compound enzyme 2 is obtained by mixing 1g of leucine aminopeptidase with an enzyme activity of 8000U / g and 2.5g of flavor protease with an enzyme activity of 50000U / g. Compound enzyme 3 is obtained by mixing 2g of 5′ phosphodiesterase with an enzyme activity of 20000U / g and 1g of glutaminase with an enzyme activity of 10000U / g. Enzymatic hydrolysis is carried out at 50℃ for 2h, and the pH is adjusted to 5.2. After the enzymatic hydrolysis is completed, the enzymatic hydrolysate C is obtained. S4. Heat the enzymatic hydrolysate C to 88°C for sterilization, centrifuge, and filter to obtain the supernatant D; S5. Add 50g of rapeseed oil to the pot and heat it to 120℃. Then add 20g of onion, 5g of garlic, 8g of scallion, and 1g of minced ginger and stir-fry for 8 minutes. Then add 5g of solid A, 13g of supernatant D and 5g of flavor liquid B and continue to stir-fry for 5 minutes. Finally, add 6g of Pixian chili bean paste, 2g of sweet bean sauce, 8g of soybean paste, 8g of white sugar, 3g of salt, 3g of MSG and 2g of thirteen-spice powder and stir-fry for 4 minutes to obtain the compound enzymatically hydrolyzed flavoring sauce.

[0035] Performance testing: Sensory evaluation was conducted on the seasoning sauces prepared in Examples 1-5 and Comparative Examples 1-7. Thirty volunteers from different regions of my country were selected to taste the sauces. They scored the seasoning sauces prepared in Examples 1-5 and Comparative Examples 1-7 on four dimensions: taste, aroma, color, and appearance. Before the experiment, the participants rinsed their mouths, observed the samples, smelled their aroma at a distance of 1 cm, tasted the sauces, and then scored them. The average score was then taken and recorded for each group.

[0036] Table 1 Scoring Criteria Table 2. Tasting results of the seasoning sauces prepared in Examples 1-5 and Comparative Examples 1-7. Data Analysis: As can be seen from Table 1, the seasoning sauce prepared by this invention has a higher sensory evaluation. This may be because the present invention soaks shiitake mushroom particles in a compound buffer solution to obtain solid A and flavor liquid B. Then, solid A is mixed with deionized water and ground into a homogenate. After three compound enzymatic hydrolysations, enzymatic hydrolysate C is obtained. The supernatant D obtained after sterilization is added to other raw materials for stir-frying along with solid A and flavor liquid B, resulting in a seasoning sauce with a distinct shiitake mushroom flavor and rich aroma. Among them, the compound buffer solution is obtained by mixing polyphenol oxidase, cellulase and buffer solution. Cellulase can rapidly decompose the cellulose skeleton of the cell wall, which is conducive to the entry of polyphenol oxidase into the cell. The enzyme catalyzes the browning reaction, producing quinone compounds that spontaneously react with amino acids in shiitake mushrooms in subsequent Maillard reactions, generating a variety of flavor compounds. In the three-step enzymatic hydrolysis reaction, the first enzymatic hydrolysis effectively breaks down the proteins in shiitake mushrooms into small polypeptide molecules and disrupts the cell wall skeleton structure. The second enzymatic hydrolysis further hydrolyzes the small polypeptide molecules into amino acids, removing bitterness while increasing umami flavor. The third enzymatic hydrolysis increases the 5'-nucleotide content in the seasoning sauce, further enhancing aroma and umami. After the three-step complex enzymatic hydrolysis reaction, the shiitake mushroom homogenate can efficiently release flavor compounds, resulting in a seasoning sauce with a distinct aroma and harmonious flavor.

[0037] Those skilled in the art should understand that the discussion of any of the above embodiments is merely exemplary and is not intended to imply that the scope of the invention is limited to these examples; within the framework of the invention, the technical features of the above embodiments or different embodiments can also be combined, the steps can be implemented in any order, and there are many other variations of the different aspects of the invention as described above, which are not provided in detail for the sake of brevity.

[0038] This invention is intended to cover all such substitutions, modifications, and variations that fall within the broad scope of the appended claims. Therefore, any omissions, modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this invention should be included within the scope of protection of this invention.

Claims

1. A method for preparing a flavoring sauce using compound enzymatic hydrolysis, characterized in that, Includes the following steps: Step S1. Take the washed fresh and dried shiitake mushrooms, separate the stems and caps, and crush them to 0.1-0.5cm. Then soak them in a compound buffer solution at 18-23℃ for 40-60min, and centrifuge to obtain solid A and flavor liquid B. Step S2. Wash solid A with deionized water, then add deionized water to solid A and continue grinding into a homogenate. Then add a compound enzyme for enzymatic hydrolysis to obtain hydrolysate C. Step S3. Heat the enzymatic hydrolysate C to 88-95℃ for sterilization, centrifuge, and filter to obtain the supernatant D; Step S4. Add rapeseed oil to the pot and heat it to 120-130℃. Then add onion, garlic, scallion, and minced ginger and stir-fry for 8-15 minutes. Then add solid A, supernatant D and flavor liquid B and continue to stir-fry for 5-10 minutes. Finally, add Pixian chili bean paste, sweet bean sauce, soybean paste, white sugar, salt, MSG and thirteen spices and stir-fry for 4-7 minutes to obtain the compound enzymatically hydrolyzed flavoring sauce. The composite buffer solution in step S1 is prepared from polyphenol oxidase with an enzyme activity of 20,000 U / g, cellulase with an enzyme activity of 10,000 U / g, citrate-disodium hydrogen phosphate buffer with a concentration of 0.05 mol / L, and disodium hydrogen phosphate solution with a concentration of 0.1 mol / L.

2. The method for preparing a flavoring sauce using compound enzymatic hydrolysis according to claim 1, characterized in that, The preparation method of the composite buffer solution is as follows: Mix 0.05 mol / L citrate-disodium hydrogen phosphate buffer and 0.1 mol / L disodium hydrogen phosphate solution and control the pH of the system to 5.2-5.4 to obtain a buffer solution. Then add polyphenol oxidase with an enzyme activity of 20000 U / g and cellulase with an enzyme activity of 10000 U / g to the buffer solution and stir until completely dissolved to obtain a composite buffer solution.

3. The method for preparing a flavoring sauce using compound enzymatic hydrolysis according to claim 2, characterized in that, The mass ratio of polyphenol oxidase, cellulase and buffer solution is 0.04-0.06:0.01-0.03:100-200.

4. The method for preparing a flavoring sauce using compound enzymatic hydrolysis according to claim 1, characterized in that, The enzymatic hydrolysis described in step S2 includes a first enzymatic hydrolysis, a second enzymatic hydrolysis, and a third enzymatic hydrolysis; The first enzymatic hydrolysis of the compound enzyme 1 is prepared by mixing Bacillus subtilis protease with an enzyme activity of 10000 U / g and cellulase with an enzyme activity of 10000 U / g at a mass ratio of 5.2-8:3.7-6. The temperature of the first enzymatic hydrolysis is 50-55℃, the hydrolysis time is 1.5-2.5h, and the pH of the first enzymatic hydrolysis is adjusted to 5.3-5.

5.

5. The method for preparing a flavoring sauce using compound enzymatic hydrolysis according to claim 4, characterized in that, During the first enzymatic hydrolysis, the mass ratio of compound enzyme 1 to homogenate was 0.5-0.8:190-230.

6. The method for preparing a flavoring sauce using compound enzymatic hydrolysis according to claim 4, characterized in that, The second enzymatic hydrolysis of the compound enzyme 2 is obtained by mixing leucine aminopeptidase with an enzyme activity of 8000 U / g and flavor protease with an enzyme activity of 50000 U / g at a mass ratio of 1-2:2.5-5.

7. The temperature of the second enzymatic hydrolysis is 48-52℃, the hydrolysis time is 1-2h, and the pH of the second enzymatic hydrolysis is adjusted to 6.5-6.

8.

7. The method for preparing a flavoring sauce using compound enzymatic hydrolysis according to claim 4, characterized in that, During the second enzymatic hydrolysis, the mass ratio of compound enzyme 2 to homogenate was 0.7-1:190-230.

8. The method for preparing a flavoring sauce using compound enzymatic hydrolysis according to claim 4, characterized in that, The third enzymatic hydrolysis involves mixing 5′ phosphodiesterase (20,000 U / g) and glutaminase (10,000 U / g) at a mass ratio of 2-5:1-2. The third enzymatic hydrolysis is performed at a temperature of 43-47°C for 1-1.5 hours, with the pH adjusted to 5.2-5.

5.

9. The method for preparing a flavoring sauce using compound enzymatic hydrolysis according to claim 4, characterized in that, During the third enzymatic hydrolysis, the mass ratio of compound enzyme 3 to homogenate was 1-1.2:190-230.

10. The method for preparing a flavoring sauce using compound enzymatic hydrolysis according to claim 1, characterized in that, The mass ratio of rapeseed oil, onion, garlic, scallion, minced ginger, solid A, supernatant D, flavor liquid B, Pixian chili bean paste, sweet bean sauce, soybean paste, white sugar, salt, monosodium glutamate, and thirteen-spice powder in step S4 is 50-70:20-30:5-7:8-13:1-3:5-8:13-17:5-8:6-8:2-3:8-10:8-10:3-5:3-5:2-4.

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