阅读说明：本技术 一种具有抗氧化活性的氨基酸类鲜味调味液及其制备方法 (Amino acid delicate flavor seasoning liquid with antioxidant activity and preparation method thereof ) 是由 李迎秋 张明君 孙桂金 赵祥忠 梁艳 王晨莹 于 2021-08-24 设计创作，主要内容包括：本发明涉及一种具有抗氧化活性的氨基酸类鲜味调味液及其制备方法,属于调味品领域。本发明产品以牡丹籽粕、黑豆粕、豆粕、麸皮、玉米粉、果糖为原料,经制曲、发酵原料处理、发酵原料酶解、发酵、美拉德反应处理、过滤提油制得。本发明发酵所得的氨基酸类鲜味调味液酱色浓郁,风味鲜美,含有丰富的氨基酸、维生素、微量元素、多糖、有机酸、酚类、酯类等物质,且本发明制得的氨基酸类鲜味调味液拥有良好的抗氧化特性,具有较强的清除自由基和还原力,在抗衰老、抗疲劳、预防高血压、降血脂、抗癌等方面具有良好的辅助作用。本发明利用高温无盐工艺发酵,方法简单,发酵速率高,发酵周期短,有较大的发展潜力。(The invention relates to an amino acid delicate flavor seasoning liquid with antioxidant activity and a preparation method thereof, belonging to the field of seasonings. The product is prepared by taking peony seed meal, black bean meal, bran, corn flour and fructose as raw materials and performing starter propagation, fermentation raw material treatment, fermentation raw material enzymolysis, fermentation, Maillard reaction treatment and oil extraction by filtration. The amino acid delicious seasoning liquid obtained by fermentation is rich in dark brown color and delicious in flavor, contains rich amino acids, vitamins, trace elements, polysaccharides, organic acids, phenols, esters and other substances, has good oxidation resistance, has strong free radical scavenging and reducing power, and has good auxiliary effects on the aspects of aging resistance, fatigue resistance, hypertension prevention, blood fat reduction, cancer resistance and the like. The invention utilizes high-temperature salt-free process for fermentation, and has the advantages of simple method, high fermentation rate, short fermentation period and great development potential.)

1. An amino acid delicate flavor seasoning liquid with antioxidant activity is characterized in that the seasoning liquid is obtained by fermenting a fermentation raw material which is subjected to enzymolysis in advance at high temperature without salt;

wherein the yeast material comprises the following raw materials in parts by weight: 5-15 parts of soybean meal, 10-20 parts of peony seed meal, 2 parts of corn flour, 10-15 parts of bran, 15-25 parts of water and 0.2-0.5 part of aspergillus oryzae spore powder;

the fermentation raw materials comprise the following raw materials in parts by weight: 5-15 parts of black soybean meal, 5-15 parts of soybean meal, 15-25 parts of peony seed meal, 10-20 parts of bran, 300 parts of water 200, 5-15 parts of salt and 0.3 part of fructose.

2. The amino acid umami seasoning liquid with antioxidant activity according to claim 1, wherein the soybean meal, the black soybean meal or the peony seed meal is obtained by crushing after deoiling.

3. The amino acid-based umami seasoning liquid having antioxidant activity according to claim 1, wherein the corn flour is obtained by crushing after dehydration and drying.

4. A preparation method of amino acid delicate flavor seasoning liquid with antioxidant activity is characterized by comprising the following steps:

mixing soybean meal, peony seed meal, bran and corn flour, adding water to obtain starter material, sterilizing at high temperature, cooling, adding Aspergillus oryzae spore powder, mixing, and culturing to obtain starter;

mixing black bean pulp powder, peony seed pulp powder and bran, adding water, and uniformly mixing to obtain a fermentation raw material;

carrying out enzymolysis on the fermentation raw material, and inactivating to obtain an enzymolysis liquid;

sterilizing and cooling the enzymolysis liquid, adding the finished koji, and fermenting to obtain a fermentation product;

adding fructose into the fermentation product, performing Maillard reaction, centrifuging, filtering, adding salt, sterilizing at high temperature, and storing.

5. The method for producing an amino acid-based umami seasoning liquid having an antioxidant activity according to claim 4, wherein the enzymatic hydrolysis comprises: carrying out primary enzymolysis and secondary enzymolysis; preferably, cellulase is adopted in the primary enzymolysis;

or, the secondary enzymolysis adopts alkaline protease.

6. The method for preparing the amino acid delicate flavor seasoning liquid with antioxidant activity as claimed in claim 5, wherein the condition of the primary enzymolysis is that the pH is 6.0-6.5, the enzymolysis time is 2-3h, and the enzymolysis temperature is 40-45 ℃;

or, the conditions of secondary enzymolysis comprise that the pH is 8.0-8.5, the enzymolysis time is 3-4h, and the enzymolysis temperature is 40-45 ℃.

7. The method for preparing amino acid delicate flavour seasoning liquid with antioxidant activity as claimed in claim 4, wherein the mass ratio of the enzymolysis liquid to the koji is 10-12: 1.

8. the method for producing the amino acid-based umami seasoning liquid having antioxidant activity according to claim 4, wherein the fermentation is carried out at 50 to 55 ℃ for 35 to 40 days.

9. The method for preparing amino acid-based umami seasoning liquid having antioxidant activity according to claim 4, wherein Maillard reaction conditions are 115 to 120 ℃, 0.1 to 0.3MPa, and 10 to 15 min.

10. Use of the amino acid-based umami seasoning liquid according to any one of claims 1 to 4 in the field of foods.

Technical Field

The invention belongs to the field of seasonings, and particularly relates to an amino acid delicate flavor seasoning liquid with antioxidant activity and a preparation method thereof.

Background

The information in this background section is only for enhancement of understanding of the general background of the invention and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art that is already known to a person of ordinary skill in the art.

The delicate flavor is one of five human tastes, is an important index for evaluating the flavor of food, and plays an important role in the traditional food culture of China. The amino acid delicate flavour seasoning liquid takes amino acid as a main flavour development substance, and also contains abundant organic acid, ester, polysaccharide, melanoid and other substances, which not only endow the product with good taste, color and flavor, but also can meet the requirements of human bodies on nutrient substances as a nutrient component, and in addition, the contained various vitamins, phenols, ketones and other substances have certain physiological activity and play a good role in maintaining the normal physiological functions of the human bodies. The seasoning is widely used by people and becomes a necessity in life of people.

At present, amino acid delicate flavour seasoning liquid mainly comprises two preparation methods of preparation and biological fermentation. The amino acid delicate flavour seasoning liquid such as soy sauce, chicken juice and the like is prepared by taking protein concentrated extract or protein hydrolysate as main raw materials and adding food additives such as salt, chicken soup, fruit and vegetable particles and the like. However, the product is often not rich enough in flavor and low in nutritive value, and a carcinogenic chloropropanol is generated when the process for preparing the protein hydrolysate by hydrolyzing the plant protein with hydrochloric acid is incomplete, so that the potential safety hazard of food exists.

The biological fermentation amino acid delicate flavour seasoning liquid product takes agricultural products such as soybean, wheat and the like as main raw materials, and is prepared by a specific process mode depending on the biological action of fermentation microorganisms such as aspergillus oryzae, aspergillus niger and the like. The traditional biological fermentation mainly comprises a high-salt dilute state fermentation mode and a low-salt solid state fermentation mode. The product prepared by the high-salt dilute state fermentation mode is rich in flavor, bright in color and luster, rich in nutrient substances, long in fermentation time, and in order to ensure that the fermented product does not go bad in the fermentation process, a large amount of salt needs to be added, and excessive edible salt can cause diseases such as hypertension, high cholesterol, sphagitis, osteoporosis and the like, so that the risk of stomach cancer is increased, and the product is more dangerous to the elderly. Although the low-salt solid fermentation method greatly reduces the fermentation period, the protein decomposition degree is low, the amino acid is not sufficiently utilized, and the obtained product has low nutritive value and single flavor. The peony seed meal is solid waste residue left after oil is extracted by squeezing peony seeds, is usually discarded or used as animal feed and fermented into fertilizer, and has very low additional value. Research shows that the peony seed meal contains 32% of protein, contains a large amount of vitamins, polysaccharides, angelica, flavonoids, minerals and other components, and has good auxiliary effects on maintaining normal functions of organisms, promoting growth and development, enhancing digestion and absorption of nutrient substances, preventing cardiovascular and cerebrovascular diseases, resisting aging and the like. The peony seed protein has amino acid content as high as 89.69%, and the 18 kinds of common amino acids including 8 kinds of essential amino acids and 10 kinds of non-essential amino acids are well-balanced. The alkaline protease has strong capability of enzymolysis of peony seed protein, and the enzymolysis product has good capability of eliminating ABTS, DPPH and OH free radicals and chelating Fe²⁺And (3) an antioxidant capacity. The rich acidic amino acid in the alkaline protease enzymolysis product can be used as Fe²⁺The chelating agent can effectively inhibit the generation of superoxide anions and hydroxyl radicals caused by transition metal ions (particularly transition metal iron) serving as a Fenton reaction catalyst, thereby reducing lipid peroxidation. The high content of hydrophobic amino acids can enhance the solubility of the enzymatic hydrolysate in lipid food systems, thereby promoting the enzymatic hydrolysate to react with oxygenThe damaged radicals interact with each other to eliminate the radicals. Therefore, the peony seed meal is a raw material with rich nutrition and good oxidation resistance.

The black bean is rich in nutrient substances, the protein content of the black bean is more than 40%, the black bean contains nearly 20 amino acids, the 8 essential amino acids comprise 8 essential amino acids required by a human body, the content of the 8 essential amino acids is higher than that of the soybean, and 7 of the essential amino acids are higher than that of the FAO/WTO equilibrium mode standard and are vegetatively reputed as the plant protein king. The black bean has high content of microelements such as zinc, copper, magnesium, molybdenum, selenium, etc., and can delay human body aging and reduce blood viscosity. In addition, the content of vitamin E in the black beans is more than 7 times of that of meat, and the black beans contain abundant anthocyanin, polyphenol, black bean polysaccharide, black bean pigment and other substances, and can clear away free radicals in vivo, resist fatigue, reduce blood pressure, reduce blood fat, enhance immunity, strengthen brain, increase intelligence and the like.

Disclosure of Invention

In order to overcome the problems, the invention aims to provide an amino acid delicious seasoning liquid product with antioxidant activity, which is prepared by fermenting peony seed meal, black soybean meal and soybean meal which are used as main raw materials, and the product not only has rich nutrient substances and good flavor, but also has good antioxidant function.

In order to achieve the technical purpose, the invention adopts the following technical scheme:

the invention provides an amino acid delicate flavor seasoning liquid with antioxidant activity, which is obtained by performing high-temperature salt-free fermentation on a fermentation raw material subjected to enzymolysis in advance;

wherein the yeast material comprises the following raw materials in parts by weight: 5-15 parts of soybean meal, 10-20 parts of peony seed meal, 2 parts of corn flour, 10-15 parts of bran, 15-25 parts of water and 0.2-0.5 part of aspergillus oryzae spore powder;

the fermentation raw materials comprise the following raw materials in parts by weight: 5-15 parts of black soybean meal, 5-15 parts of soybean meal, 15-25 parts of peony seed meal, 10-20 parts of bran, 300 parts of water 200, 5-15 parts of salt and 0.3 part of fructose.

The peony seed meal, the black bean pulp with the peel and the like are used as main raw materials, and the amino acid delicate flavor seasoning liquid with antioxidant activity is prepared by performing biological fermentation on fermentation raw materials in a mode of enzymolysis and high-temperature salt-free fermentation, so that the cost is low, the preparation process is simple and convenient, and the fermentation period is far shorter than that of the traditional high-salt dilute state fermentation mode. The preparation method has the advantages of high utilization rate of protein and amino acid, rich nutrition, flavor and active substance output, low salt content of the obtained delicious seasoning liquid product, rich caramel color, delicious flavor, rich amino acid and a large amount of vitamins, trace elements, polysaccharide, organic acid, phenols, esters and other substances. The product has nutrition and quality superior to those of traditional low-salt solid fermentation and seasoning liquid preparation, has good antioxidant property, strong free radical scavenging and reducing power, has good auxiliary effects on aspects of aging resistance, fatigue resistance, hypertension prevention, blood fat reduction, cancer resistance and the like, and accords with the pursuit of modern people for healthy diet.

In a second aspect of the present invention, there is provided a method for preparing an amino acid-based umami seasoning liquid having antioxidant activity, comprising:

mixing soybean meal, peony seed meal, bran and corn flour, adding water to obtain starter material, sterilizing at high temperature, cooling, adding Aspergillus oryzae spore powder, mixing, and culturing to obtain starter;

mixing black bean pulp powder, peony seed pulp powder and bran, adding water, and uniformly mixing to obtain a fermentation raw material;

carrying out enzymolysis on the fermentation raw material, and inactivating to obtain an enzymolysis liquid;

sterilizing and cooling the enzymolysis liquid, adding the finished koji, and fermenting to obtain a fermentation product;

adding fructose into the fermentation product, performing Maillard reaction, centrifuging, filtering, adding salt, sterilizing at high temperature, and storing.

In order to overcome the defects of products prepared by high-salt dilute state fermentation and low-salt solid state fermentation, the invention uses protease to carry out enzymolysis on protein raw materials before fermentation, is beneficial to improving the utilization rate of protein and amino acid, is convenient for direct utilization of fermentation microorganisms, and shortens the fermentation period; secondly, the high temperature is utilized for fermentation, so that the salt can be replaced to play a role in inhibiting the growth of mixed bacteria, the influence of salt on the activity of a fermentation enzyme system is reduced, and the prepared product has low salt content and meets the requirements of modern people on healthy diet.

The invention also aims to provide a preparation method of the amino acid delicious seasoning liquid with antioxidant activity, which effectively shortens the fermentation period by carrying out enzymolysis and high-temperature salt-free fermentation on the fermentation raw materials in advance, and the prepared amino acid delicious seasoning liquid has fresh and fragrant flavor, rich color and luster and rich nutrient substances.

In a third aspect of the invention, the application of any one of the amino acid delicate flavor seasoning liquids in the food field is provided.

The invention has the beneficial effects that:

(1) the main raw materials of the invention are peony seed meal and black bean meal, which contain abundant protein, amino acid and trace elements, and also contain anti-oxidation active ingredients such as polysaccharide, polyphenol, vitamin E, flavone, anthocyanin and isoflavone, and the prepared amino acid delicate flavor seasoning liquid has abundant nutrition, has good anti-oxidation effect, and has auxiliary effects on the aspects of clearing free radicals in vivo, preventing cancer, resisting aging, regulating immunologic function, and the like.

(2) According to the invention, before fermentation, cellulase and protease are used for treating peony seed meal, black bean meal and bean meal, and substances such as protein, polysaccharide and the like in the peony seed meal, black bean meal and bean meal are subjected to enzymolysis, and related fermentation microorganisms such as aspergillus oryzae directly utilize enzymolysis products in the initial fermentation stage, so that the fermentation period is shortened, the later-stage enrichment of flavor substances is facilitated, and the mouthfeel is optimized; in the fermentation process, a high-temperature fermentation mode is adopted, and salt is not added, so that the aim of inhibiting the growth of mixed bacteria is fulfilled while salt influences the activity of fermentation-related enzyme systems such as protease are avoided.

(3) The amino acid delicate flavour seasoning liquid prepared by the invention has low salt content and meets the requirement of the contemporary people on healthy diet.

(4) The peony seed meal is used as a main raw material, so that the added value of the peony seeds can be increased, the raw material cost is reduced, economic benefits are created, and the problem of resource waste of the peony seeds can be solved.

(5) The operation method is simple, low in cost, universal and easy for large-scale production.

Detailed Description

It is to be understood that the following detailed description is exemplary and is intended to provide further explanation of the invention as claimed. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

An amino acid delicate flavor seasoning liquid with antioxidant activity and a preparation method thereof, wherein the yeast material comprises the following components by mass: 5-15g of soybean meal, 10-20g of peony seed meal, 2g of corn flour, 10-15g of bran, 15-25g of water and 0.2-0.5g of aspergillus oryzae spore powder.

An amino acid delicate flavor seasoning liquid with antioxidant activity and a preparation method thereof, wherein the fermentation raw materials comprise the following components by mass: 5-15g of black soybean meal, 5-15g of soybean meal, 15-25g of peony seed meal, 10-20g of bran, 300g of water 200, 5-15g of salt and 0.3g of fructose.

In some embodiments, the soybean meal, the black soybean meal and the peony seed meal are obtained by deoiling and crushing by a crusher.

In some embodiments, the corn meal is obtained by dehydrating, drying, and crushing with a crusher.

An amino acid delicate flavor seasoning liquid with antioxidant activity and a preparation method thereof, the specific steps comprise:

(1) preparing yeast: taking appropriate amount of soybean meal, peony seed meal, bran and corn flour, mixing uniformly according to a certain proportion, adding distilled water to obtain a starter material, then sterilizing at high temperature, cooling, adding aspergillus oryzae spore powder, mixing uniformly, and culturing for a period of time to obtain the finished starter.

(2) Fermentation raw material treatment: sieving semen Sojae Atricolor cake powder, and peony seed cake powder, mixing with testa Tritici, adding a certain amount of water, and stirring.

(3) Enzymolysis of fermentation raw materials: adjusting the pH value of the raw material mixed liquor, and then adding enzyme to carry out constant temperature enzymolysis twice. And (4) inactivating enzyme in a high-temperature water bath after enzymolysis to obtain an enzymolysis liquid.

(4) Fermentation: sterilizing under high pressure, cooling, adding yeast, stirring, fermenting at constant temperature, and stirring twice per day.

(5) And (3) Maillard reaction treatment: adding appropriate amount of fructose, and treating at high temperature and high pressure.

(6) Centrifugal filtration: centrifuging at high speed, filtering, adding salt, sterilizing at high temperature, and storing.

In some embodiments, in the step (1), the soybean meal is added in an amount of 5-15g, the peony seed meal is added in an amount of 10-20g, the corn flour is added in an amount of 2g, the bran is added in an amount of 10-15g, and 15-25g of water is added to mix uniformly.

In some embodiments, the sterilization conditions in step (1) are 121 ℃ and 0.1MPa for 20 min.

In some embodiments, the Aspergillus oryzae spore powder is inoculated in the step (1) in an amount of 0.2-0.5g, and the inoculation environment is required to be sterile. The starter propagation temperature is 37 ℃, and the starter propagation time is 48 h.

In some embodiments, in the step (2), the black soybean meal is added in an amount of 5-15g, the peony seed meal is added in an amount of 15-25g, the bran is added in an amount of 10-20g, and the water is added in an amount of 200-300g, and the mixture is stirred uniformly.

The research finds that: the problems of too long fermentation time and low utilization rate of protein and amino acid exist in pure high-temperature salt-free fermentation of peony seed meal, black bean meal and bean meal. For this reason, the present application finds through systematic research and experimental investigation: the peony seed meal, the black bean meal and the bean meal are subjected to two-step enzymolysis treatment and then fermented, so that the subsequent fermentation time can be obviously shortened, and the content of amino acid nitrogen and the antioxidant activity in a fermentation product are effectively improved. Therefore, in some embodiments, in the step (3), the first enzymolysis pH is 6.0, the enzyme used is cellulase, the addition amount is 2g, the enzymolysis time is 2-3h, and the enzymolysis temperature is 40 ℃; the pH of the second enzymolysis is 8.0, the used enzyme is alkaline protease, the addition amount is 2g, the enzymolysis time is 3-4h, and the enzymolysis temperature is 40 ℃. After the two times of enzymolysis, the enzymolysis liquid is moved to a water bath with the temperature of 95 ℃ for 5min to inactivate the enzyme. In the enzymolysis process, cellulose in the peony seed meal, the black bean meal and the bean meal is subjected to enzymolysis through cellulase to release more protein, and then the protein is subjected to enzymolysis through alkaline protease to form amino acid, so that the amino acid and carbohydrate in the raw materials are effectively released, and each small molecular substance forms a good fermentation substrate which can be directly utilized by fermentation microorganisms.

In some embodiments, the sterilization conditions in step (4) are 121 ℃, 0.1MPa, and the sterilization time is 20 min.

In some embodiments, the step (4) comprises the following steps: finished koji is 10: 1 (by mass) adding yeast, and fermenting at 50 deg.C for 35 days.

In some embodiments, the fructose is added in an amount of 0.3g in step (5), and the treatment conditions are 115 ℃ and 0.1MPa for 10 min.

In some embodiments, the salt is added in an amount of 5% by mass of the filtrate in step (6).

In some embodiments, in the step (6), the rotation speed of the centrifuge is 8000r/min, the centrifugation time is 20min, and the amino acid delicate flavor seasoning liquid with antioxidant activity is obtained after filtering and sterilizing in a water bath kettle at 95 ℃ for 5 min.

The present invention is described in further detail below with reference to specific examples, which are intended to be illustrative of the invention and not limiting.

Example 1

A preparation method of amino acid delicate flavor seasoning liquid with antioxidant activity comprises the following steps:

(1) preparing yeast: taking 7g of soybean meal, 13g of peony seed meal, 10g of bran and 2g of corn flour, adding 15g of distilled water, and uniformly mixing to obtain the yeast material. Sterilizing the koji material at 121 deg.C and 0.1Mpa for 20min, cooling to room temperature, inoculating 0.25g Aspergillus oryzae spore powder under aseptic condition, stirring, culturing at 37 deg.C under aseptic condition for 48 hr, shaking the fermentation bottle every 12 hr to disperse the caked koji material, and making into koji when yellow mycelium is covered with the koji material.

(2) Fermentation raw material treatment: sieving black soybean meal, soybean meal and peony seed meal with a 60-mesh sieve, taking 10g of black soybean meal, 10g of soybean meal, 20g of peony seed meal, 20g of bran and 240g of water, and uniformly mixing for later use.

(3) Enzymolysis of fermentation raw materials: adjusting pH of the fermentation raw material to 6.0, adding 2g cellulase, performing enzymolysis at 40 deg.C for 2 hr, and stirring once every 20 min. After the enzymolysis of the cellulase is finished, the pH value is adjusted to 8.0, 2g of alkaline protease is added, and the enzymolysis is carried out for 3 hours at the constant temperature of 40 ℃, and the stirring is carried out once every 20 min. After the enzymolysis is finished, the enzymolysis liquid is put in a water bath at 95 ℃ for 5min to inactivate the enzyme.

(4) Fermentation: autoclaving the enzymolysis solution at 121 deg.C and 0.1Mpa for 20 min. And (3) after the enzymolysis liquid is cooled, adding 30g of finished yeast into the enzymolysis liquid, uniformly stirring, culturing for 35d at 50 ℃, and uniformly shaking twice every day.

(5) And (3) Maillard reaction treatment: adding 0.3g fructose into the fermentation broth, mixing well, and keeping under 0.1Mpa at 115 deg.C for 10 min.

(6) Filtering and extracting oil: after the fermentation liquid is cooled to room temperature, centrifuging for 20min at the rotation speed of 8000r/min, filtering, adding salt accounting for 5% of the mass of the filtrate into the filtrate, fully dissolving, and sterilizing at 95 ℃ for 5min in a water bath kettle to obtain the amino acid delicate flavor seasoning liquid with antioxidant activity.

Example 2

A preparation method of amino acid delicate flavor seasoning liquid with antioxidant activity comprises the following steps:

(1) preparing yeast: taking 7g of soybean meal, 13g of peony seed meal, 15g of bran and 2g of corn flour, adding 20g of distilled water, and uniformly mixing to obtain the yeast material. Sterilizing the koji material at 121 deg.C and 0.1Mpa for 20min, cooling to room temperature, inoculating 0.3g Aspergillus oryzae spore powder under aseptic condition, stirring, culturing at 37 deg.C under aseptic condition for 48 hr, shaking the fermentation bottle every 12 hr to disperse the caked koji material, and making into koji when yellow mycelium is covered with the koji material.

(2) Fermentation raw material treatment: sieving black soybean meal, soybean meal and peony seed meal with a 60-mesh sieve, taking 5g of black soybean meal, 5g of soybean meal, 30g of peony seed meal, 20g of bran and 240g of water, and uniformly mixing for later use.

(3) Enzymolysis of fermentation raw materials: adjusting pH of the fermentation raw material to 6.0, adding 2g cellulase, performing enzymolysis at 40 deg.C for 2 hr, and stirring once every 20 min. After the enzymolysis of the cellulase is finished, the pH value is adjusted to 8.0, 2g of alkaline protease is added, and the enzymolysis is carried out for 3 hours at the constant temperature of 40 ℃, and the stirring is carried out once every 20 min. After the enzymolysis is finished, the enzymolysis liquid is put in a water bath at 95 ℃ for 5min to inactivate the enzyme.

(4) Fermentation: autoclaving the enzymolysis solution at 121 deg.C under 0.1Mpa for 20 min. And (3) after the enzymolysis liquid is cooled, adding 30g of finished yeast into the enzymolysis liquid, uniformly stirring, culturing for 35d at 50 ℃, and uniformly shaking twice every day.

(5) And (3) Maillard reaction treatment: adding 0.3g fructose into the fermentation broth, mixing well, and keeping under 0.1Mpa at 115 deg.C for 10 min.

(6) Filtering and extracting oil: after the fermentation liquid is cooled to room temperature, centrifuging for 20min at the rotation speed of 8000r/min, filtering, adding salt accounting for 5% of the mass of the filtrate into the filtrate, fully dissolving, and sterilizing for 5min at 95 ℃ in a water bath kettle to obtain the amino acid delicate flavor seasoning liquid with antioxidant activity.

Example 3

A preparation method of amino acid delicate flavor seasoning liquid with antioxidant activity comprises the following steps:

(1) preparing yeast: taking 13g of soybean meal, 7g of peony seed meal, 10g of bran and 2g of corn flour, adding 15g of distilled water, and uniformly mixing to obtain the yeast material. Sterilizing the koji material at 121 deg.C and 0.1Mpa for 20min, cooling to room temperature, inoculating 0.25g Aspergillus oryzae spore powder under aseptic condition, stirring, culturing at 37 deg.C under aseptic condition for 48 hr, shaking the fermentation bottle every 12 hr to disperse the caked koji material, and making into koji when yellow mycelium is covered with the koji material.

(2) Fermentation raw material treatment: sieving black soybean meal, soybean meal and peony seed meal with a 60-mesh sieve, taking 5g of black soybean meal, 15g of soybean meal, 20g of peony seed meal, 15g of bran and 200g of water, and uniformly mixing for later use.

(3) Enzymolysis of fermentation raw materials: adjusting pH of the fermentation raw material to 6.0, adding 2g cellulase, performing enzymolysis at 40 deg.C for 2 hr, and stirring once every 20 min. After the enzymolysis of the cellulase is finished, the pH value is adjusted to 8.0, 2g of alkaline protease is added, and the enzymolysis is carried out for 3 hours at the constant temperature of 40 ℃, and the stirring is carried out once every 20 min. After the enzymolysis is finished, the enzymolysis liquid is put in a water bath at 95 ℃ for 5min to inactivate the enzyme.

(4) Fermentation: autoclaving the enzymolysis solution at 121 deg.C under 0.1Mpa for 20 min. And (3) after the enzymolysis liquid is cooled, adding 30g of finished yeast into the enzymolysis liquid, uniformly stirring, culturing for 35d at 50 ℃, and uniformly shaking twice every day.

(5) And (3) Maillard reaction treatment: adding 0.3g fructose into the fermentation broth, mixing well, and keeping under 0.1Mpa at 115 deg.C for 10 min.

(6) Filtering and extracting oil: after the fermentation liquid is cooled to room temperature, centrifuging for 20min at the rotation speed of 8000r/min, filtering, adding salt accounting for 5% of the mass of the filtrate into the filtrate, fully dissolving, and sterilizing for 5min at 95 ℃ in a water bath kettle to obtain the amino acid delicate flavor seasoning liquid with antioxidant activity.

Example 4

A preparation method of amino acid delicate flavor seasoning liquid with antioxidant activity comprises the following steps:

(1) preparing yeast: taking 10g of bean flour, 10g of peony seed meal, 15g of bran and 2g of corn flour, adding 20g of distilled water, and uniformly mixing to obtain the yeast material. Sterilizing the koji material at 121 deg.C and 0.1Mpa for 20min, cooling to room temperature, inoculating 0.3g Aspergillus oryzae spore powder under aseptic condition, stirring, culturing at 37 deg.C under aseptic condition for 48 hr, shaking the fermentation bottle every 12 hr to disperse the caked koji material, and making into koji when yellow mycelium is covered with the koji material.

(2) Fermentation raw material treatment: sieving black soybean meal, soybean meal and peony seed meal with a 60-mesh sieve, taking 15g of black soybean meal, 5g of soybean meal, 20g of peony seed meal, 20g of bran and 200g of water, and uniformly mixing for later use.

(3) Enzymolysis of fermentation raw materials: adjusting pH of the fermentation raw material to 6.0, adding 2g cellulase, performing enzymolysis at 40 deg.C for 2 hr, and stirring once every 20 min. After the enzymolysis of the cellulase is finished, the pH value is adjusted to 8.0, 2g of alkaline protease is added, and the enzymolysis is carried out for 3 hours at the constant temperature of 40 ℃, and the stirring is carried out once every 20 min. After the enzymolysis is finished, the enzymolysis liquid is put in a water bath at 95 ℃ for 5min to inactivate the enzyme.

(4) Fermentation: autoclaving the enzymolysis solution at 121 deg.C under 0.1Mpa for 20 min. And (3) after the enzymolysis liquid is cooled, adding 30g of finished yeast into the enzymolysis liquid, uniformly stirring, culturing for 35d at 50 ℃, and uniformly shaking twice every day.

(5) And (3) Maillard reaction treatment: adding 0.3g fructose into the fermentation broth, mixing well, and keeping under 0.1Mpa at 115 deg.C for 10 min.

(6) Filtering and extracting oil: after the fermentation liquid is cooled to room temperature, centrifuging for 20min at the rotation speed of 8000r/min, filtering, adding salt accounting for 5% of the mass of the filtrate into the filtrate, fully dissolving, and sterilizing for 5min at 95 ℃ in a water bath kettle to obtain the amino acid delicate flavor seasoning liquid with antioxidant activity.

Experimental example 1 evaluation of nutritional characteristics of amino acid-based savory seasoning liquid having antioxidant activity prepared in example

(1) The content determination of amino acid nitrogen in the amino acid fresh-taste seasoning liquid with antioxidant activity prepared in the examples

Weighing 5.0mL of sample in a 50mL beaker, washing the sample in a 100mL volumetric flask with water several times, adding water to the scale, mixing, sucking 20.0mL of the sample, placing the sample in a 200mL beaker, adding 60mL of water, starting a magnetic stirrer, titrating the sample with a sodium hydroxide standard solution [ c (NaOH) ═ 0.050mol/L ] until the pH value is 8.2 according to an acidimeter indication, adding 10.0mL of formaldehyde solution, and mixing. Titration was continued with sodium hydroxide standard titration solution to pH 9.2 and the number of milliliters of sodium hydroxide standard titration solution consumed was recorded. Meanwhile, 80mL of water is taken as a reagent blank test. The calculation formula of the content of the amino acid nitrogen is as follows:

in the formula:

V₁to determine the volume in milliliters (mL) of standard titration solution of sodium hydroxide consumed after formaldehyde was added to the sample diluent;

V₂the volume of the sodium hydroxide standard titration solution in milliliters (mL) was consumed after adding formaldehyde for the reagent blank experiment;

c is the concentration of the sodium hydroxide standard titration solution in units of moles per liter (mol/L);

0.014 is the mass of nitrogen in grams (g) equivalent to 1.00mL of sodium hydroxide standard titration solution [ c (naoh) ═ 1.000mol/L ];

m is the weight of the weighed sample, and the unit is gram (g);

v is the volume of the aspirated sample in milliliters (mL);

V₃the sample dilution was taken in milliliters (mL);

V₄the volumetric volume of the sample diluent is in milliliters (mL);

100 is a unit scaling factor.

The results are shown in Table 1. The amino acid nitrogen content of example 4 was higher than that of the other examples, and all of the examples reached the national secondary soy sauce standard.

(2) Measurement of Total Nitrogen content of amino acid-based savory seasoning liquid having antioxidant Activity

Weighing 2g of fully and uniformly mixed amino acid delicate flavor seasoning liquid sample. The amount of the copper sulfate is accurate to 0.001g, and then 0.3g of copper sulfate, 3g of potassium sulfate and 10mL of sulfuric acid are added into a digestion tube for digestion. And (3) after the temperature of the digestion furnace reaches 420 ℃, continuing to digest for 2 hours, taking out the liquid in the digestion tube, cooling, and then automatically adding liquid, distilling, titrating and recording titration data on an automatic Kjeldahl azotometer. The results are shown in Table 1.

Experimental example 2 research on antioxidant activity of an amino acid-based savory seasoning liquid having antioxidant activity, prepared in example.

(1) Example A taste and flavor amino acid based seasoning having antioxidant activity was tested for DPPH radical scavenging

2.0mL of the amino acid seasoning liquid with antioxidant activity obtained in example was added with 2.0mL of LDPPH free radical solution (0.2mmol/L), mixed well, and treated at room temperature (25 ℃ C.) for 30min in the dark. Adjusting to zero with absolute ethyl alcohol, uniformly mixing 2.0mL of sample extracting solution and 2.0mL of ethanol solution to obtain a blank group, uniformly mixing 2.0mL of DPPH solution and ethanol solution to obtain a control group, and measuring the absorbance value at the wavelength of 517 nm. Each set of measurements was repeated 3 times and the results averaged. The DPPH free radical clearance is calculated by the formula:

in the formula:

A₀blank absorbance;

A₁absorbance for the sample set;

A₂absorbance was taken as the control.

(2) ABTS removal of amino acid umami seasoning liquid with antioxidant activity prepared in example⁺Free radical test.

ABTS⁺Preparing a free radical solution: 200mL of ABTS with a concentration of 7mmol/L were added to the beaker⁺The solution and 3.2mL of potassium persulfate with the concentration of 2.45mmol/L are fully mixed by ultrasonic treatment for 2min, the mixture is placed in a dark place to be protected from light for reaction for 12 to 16h, and then the mixture is diluted by phosphate buffer solution with the concentration of 0.05mol/L and the pH of 7.4 to ensure that the absorbance value of the mixture under the condition of 734nm wavelength is 0.70 +/-0.02, namely ABTS, the ABTS is the product⁺Standing the free radical solution at room temperature for 30 min. 2mL of the amino acid-based umami seasoning liquid having antioxidant activity obtained in example was diluted 50-fold in a 100mL volumetric flask. 0.8mL of the diluted solution was taken, and 3.2mL of ABTS was added⁺The free radical solution is mixed evenly, reacted for 10min at room temperature, and the absorbance value of the sample is measured at 734 nm. And replacing the sample extracting solution with absolute ethyl alcohol to be used as a blank group, and measuring the light absorption value of the blank group. Each set of measurements was repeated 3 times and the results averaged. ABTS⁺The formula for the free radical clearance is:

in the formula:

A₀blank absorbance;

A₁absorbance for the sample set;

(3) example A reducing force test of amino acid-based umami seasoning liquid having antioxidant activity

The antioxidant activity of the obtained product2mL of the amino acid-based seasoning liquid (2) was diluted 50-fold in a 100mL volumetric flask. 1mL of the diluted solution was added with 2.5mL of 0.2mol/L phosphate buffer (pH6.6) and 2.5mL of 1% K₃Fe(CN)₆Reacting in 50 deg.C water bath for 30min, cooling to room temperature (25 deg.C), adding 2.5mL of 10% TCA solution into the mixture, centrifuging at 3000rpm for 10min, collecting supernatant 2.5mL, adding 2.5mL of distilled water and 0.5mL of 0.1% FeCl₃The solution is mixed evenly, stored in dark at room temperature (25 ℃) for 10min, the absorbance value is measured at 700nm and adjusted to zero by distilled water, the absorbance (700nm) of the sample represents reducibility, the measurement of each sample is repeated for 3 times, and the measurement result is averaged.

The results of measuring the antioxidant activity of the amino acid-based savory seasoning liquid having antioxidant activity obtained in the examples are shown in Table 2.

TABLE 1

TABLE 2

It should be noted that the above-mentioned embodiments are only preferred embodiments of the present invention, and the present invention is not limited thereto, and although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications and equivalents can be made in the technical solutions described in the foregoing embodiments, or equivalents thereof. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.