Preparation method of high-freshness flavor soup base based on synergistic enzymolysis of multi-source byproducts

Abstract

The invention discloses a preparation method of a high-freshness flavor soup base based on synergistic enzymolysis of multi-source byproducts, and belongs to the technical field of food processing. The method comprises the following steps: firstly, screening soybean meal, lentinus edodes stems, chicken skeletons, fish bones and other byproducts, pretreating, cleaning, drying, crushing, performing high-pressure treatment, performing ultrasonic-assisted enzymolysis, mixing with blended marinating soup, and performing enzyme deactivation, filtration treatment, filling and sterilization to obtain the high-freshness flavor soup base. The prepared flavored soup base is bright in color, rich in taste level, strong and mellow in marinating soup flavor, fresh and sweet in taste, strong and diversified in delicate flavor level and free of flavoring agents and preservatives, the delicate flavor level of the flavored soup base is improved, the preparation process of traditional marinated food can be shortened, the preparation cost can be reduced, and high value of resources can be achieved.

Description

Technical Field

[0001] The present invention belongs to the technical field of food processing, and particularly relates to a method for preparing a high-freshness flavored soup base based on the synergistic enzymatic hydrolysis of multi-source by-products. Background Art

[0002] Brine is made by "making brine" with more than a dozen natural spices such as brown sugar, water, salt, light soy sauce, white soy sauce, shallots, galangal, tangerine peel, star anise, lemongrass, etc., and then immersing geese, ducks, or pig's feet, pig's head skin, etc. into the brine pot and simmering over fire. The final marinated flavor is soft yet firm, rich but not salty, fresh but not fishy. Brine is the soul of marinated flavors. Brine has unique regional flavor characteristics, and various marinated flavor processes and products emerge in an endless stream. Many enterprises have embarked on the road of product innovation, and the market share shows a trend of blooming and prosperous development. However, at present, most brines are made by families and small merchants, with unstable quality, complex production processes, cumbersome procedures, and difficult storage. At the same time, soybean meal, mushroom stalks, chicken skeletons, and fish bones, as by-products of plants, fungi, animals, etc., have the characteristics of high nutrition, high freshness, and difficult processing, and can be used as one of the raw materials for flavored soup bases. And using enzymatic hydrolysis of multi-source by-products is a method for making high-freshness flavored soup bases. The preparation of flavored soup bases mostly relies on traditional enzymatic hydrolysis processes, but the cell structure of the raw materials is not fully destroyed, resulting in low enzymatic hydrolysis efficiency and insufficient release of umami substances. In addition, the traditional process is difficult to achieve efficient utilization of raw materials, especially the treatment effect of hard raw materials such as fish bones and chicken skeletons is poor. Therefore, there is an urgent need for an innovative process that can effectively destroy the cell structure of raw materials and improve the enzymatic hydrolysis efficiency.

[0003] Zhang Ding et al. (Patent Application No.: CN201711063716.6) disclosed a method for preparing brine materials. The preparation method includes: (1) putting animal oil, refined salt, monosodium glutamate, light soy sauce, cooking wine, and a brine material packet into water and bringing to a boil over high heat; (2) removing the foam on the surface of the brine soup, adding spicy oil, and simmering over low heat for 1 - 3 hours; (3) adding 5 - 10 parts of five-spice powder, 10 - 13 parts of star anise, 15 - 30 parts of Chinese prickly ash, 3 - 8 parts of ginger, 1 - 2 parts of fennel, 3 - 5 parts of tangerine peel, and 1 - 5 parts of cinnamon leaves by weight to the brine soup and boiling for 2 hours. The present invention only involves the selection of the formula of the brine soup base and does not solve the problem of high-value utilization of by-products.

[0004] Wang Gaoping (Patent Application No.: CN202111402407.3) disclosed a formula for donkey meat hot pot base material and its preparation method. The preparation method includes: Step 1: 1. Take 20 to 25 catties of donkey big bones, put them into the pot with cold water, add cooking wine, bring to a boil over high heat, then skim off the foam, and then soak them in cold water; 2. Add 200 catties of pure water to the brine bucket, put in the soaked big bones, turn to low heat and boil for 8 hours to make the original soup; Step 2: 1. First make the base soup ingredients; 2. Put the base soup ingredients into the pot and add the original soup boiled in Step 1. With donkey big bones as the main ingredient and various auxiliary ingredients, the hot pot base material can have a fresh, fragrant and light taste. However, donkey meat has a relatively higher cost and lower output, making it difficult to meet the needs of consumers.

[0005] Su Kui (Patent Application No.: CN201911388377.8) disclosed a method for making concentrated brine. This method uses minced meat and plant protein meal as protein raw materials, soaks them, then performs steam explosion treatment, adds starch and water, and conducts extrusion processing in a twin-screw extruder. Mix the extruded material and water, add alkaline protease, flavor protease and α-amylase, and enzymolyze for 3 - 5 hours, then inactivate at high temperature to obtain the inactivated enzymolysis solution. After heating, it is spray-dried to make Maillard flavor peptides. Mix the Maillard flavor peptides with spices, add various bones, and then perform ultrasonic-assisted boiling, press filtration, vacuum decompression concentration, and vacuum packaging and high-pressure sterilization to make a concentrated brine product. The concentrated brine made by this method has stable quality, is convenient to use, and has good quality without the need to add additional additives. However, it has the disadvantages of cumbersome steps, a large number of designed equipment, long processing time, and difficulty in achieving automation and quality control.

[0006] Ren Kang (Patent Application No.: CN201810565764.3) disclosed a disposable brine, its preparation method, and a method for preparing marinated dishes. The preparation method first mixes sodium salt, glucose, monosodium glutamate, xanthan gum and water evenly in a pot, heats it with water, then adds chicken meat enzyme degradation products, L-cysteine, alanine, leucine and glutamic acid in the pot and stirs to dissolve evenly. Finally, add five-spice juice in the jacketed pot and stir evenly to obtain the finished product. Through the synergistic effect of five-spice juice, meat enzyme degradation products, amino acid mixtures, flavoring substances, thickeners and sugars, this method reduces the harmful substances generated by lipid peroxidation, and uses the Maillard reaction to generate aroma and color and produce umami components. However, this product has a complex composition, and the Maillard reaction products are complex and prone to produce bitter substances, and thickeners, flavor enhancers and other additives are added, which cannot meet the needs of consumers for safe and additive-free brine.

[0007] Yan Shoulei (Patent Application No.: CN201810855724.2) disclosed a method for preparing brine. The preparation method includes (1) the preparation of prefabricated soup, the preparation of caramel solution, the preparation of spice powder and compound powder. The prepared prefabricated soup, caramel solution, spice powder and compound powder are mixed in corresponding proportions by a suitable process and boiled to obtain brine. This method uses traditional caramel frying, directly heating white sugar with water to fry out a bright red caramel solution. This method requires strong control of the heat, cannot guarantee the stability of the product, and is prone to bitterness. The present invention uses an ultrasonic physical field and an enzymatic hydrolysis process, which not only has a short processing time, but also can reduce costs and increase the production stability of products.

[0008] Shu Shuangping (Patent Application No.: CN201510534329.0) disclosed a method for preparing brine for making tea dried bean curd, including: (1) adding lard to a pot, heating until melted, then adding black sesame oil, then adding mushrooms and boiling until dehydrated, fishing out and adding onions, Chinese prickly ash and anise, continuing to boil for 10 - 15 minutes, filtering to obtain sesame oil; (2) first adding the sesame oil to the pot, then adding fermented soybeans and pomelo peel, stir-frying for 5 - 10 minutes, adding water, bringing to a boil, adding cooking wine, coix seed and ginkgo, boiling for 30 - 45 minutes, adding polygonatum odoratum, sophora japonica flower, pogostemon cablin, rattanleaf cudweed herb and Chinese kale, continuing to boil for 15 - 20 minutes, and finally adding coconut milk, konjac gum and fish sauce, mixing evenly and filtering to obtain brine. This production method adds a variety of unique spices, has a strong fragrance and rich nutritional value, has a unique flavor with both the characteristics of tea dried bean curd and a unique aroma, giving consumers a unique taste experience, but the production process is complex, the quality control requirements are high, and the product quality control cost is high. Summary of the Invention

[0009] In order to overcome the problems in the prior art such as complex preparation process of flavor soup base, single flavor, poor storage stability, high cost, etc., the present invention provides a method for preparing a high - freshness flavor soup base based on the synergistic enzymatic hydrolysis of multi - source by - products.

[0010] The technical solution of the present invention is as follows:

[0011] A method for preparing a high - freshness flavor soup base based on the synergistic enzymatic hydrolysis of multi - source by - products mainly includes the following steps:

[0012] (1) Raw material pretreatment: Chicken skeleton: Wash, remove inedible parts, boil in boiling water for 20 minutes to degrease, and cut into pieces smaller than 1 cm 3Bone pieces were freeze-dried and then ground to 80 mesh to obtain chicken skeleton powder; fish bones: soaked in 5% brine for 30 minutes to remove fishy smell, blanched in boiling water for 5 minutes, dried at 60°C until the moisture content was less than 8%, and ground to 80 mesh to obtain fish bone powder; soybean meal: defatted with food-grade 75% ethanol for 2 hours, treated with steam for 10 minutes, ground to 80 mesh, and soaked in NaHCO3 solution with pH 8.5 for 30 minutes for activation to obtain soybean meal powder; mushroom stalks: after washing and removing impurities, soaked in 3% citric acid solution for 15 minutes to remove earthy smell, then dried in hot air at 60°C until the moisture content was less than 10%, and ground to 80 mesh to obtain mushroom stalk powder;

[0013] (2) Mixing and high-pressure treatment of raw materials: Mix the above-mentioned chicken skeleton powder, fish bone powder, soybean meal powder, and mushroom stalk powder and put them into a high-pressure-resistant flexible packaging bag, and perform high-pressure treatment using an ultra-high pressure cell crusher;

[0014] (3) Homogenization of the mixed raw materials: After sieving the mixed raw materials treated above, add pure water, with a material-liquid ratio of 1:12, adjust the pH to 6.5 - 7.0, and perform high-speed shear homogenization for 5 minutes;

[0015] (4) Ultrasonic combined with enzymatic hydrolysis: Add 0.28% of compound protease, 0.24% of cellulase, 0.2% of collagenase, and 0.08% of β-glucanase based on the mass of the mixed raw materials to the homogenized mixed raw materials for enzymatic hydrolysis treatment, and at the same time assist with ultrasonic treatment, and perform constant-temperature enzymatic hydrolysis at 55°C for 2 hours to obtain a flavor soup base enzymatic hydrolysate;

[0016] (5) High-temperature and high-pressure enzyme inactivation and filtration: Use a high-temperature steam sterilization kettle, set the parameters to 121°C, perform high-temperature and high-pressure treatment on the flavor soup base enzymatic hydrolysate for 15 minutes to inactivate the enzyme, and then filter with double-layer 100-mesh gauze to remove residues;

[0017] (6) Preparation of the brine: Prepare the brine according to the following formula: 7.5 g of spice powder, 6 g of edible salt, 8 g of sugar, 4.5 g of soy sauce, 3 g of huadiao wine, 3.5 g of fish sauce, 16 g of sun-dried soy sauce, 1.5 g of caramel color, 5 g of dried galangal, 0.8 g of disodium 5'-ribonucleotide, 6.5 g of monosodium glutamate, 0.1 g of inosinic acid, 0.2 g of scallop essence, and 1000 g of pure water;

[0018] (7) Preparation of the flavor soup base: Mix the enzyme-inactivated and filtered flavor soup base enzymatic hydrolysate obtained in step (5) with the brine obtained in step (6) at a ratio of 1:1, and add 30 g of mixed oil for oil sealing to obtain the flavor soup base;

[0019] (8) Filling and sterilization: Perform high-temperature short-time sterilization on the flavor soup base in step (7), with the treatment conditions of temperature: 121°C, time: 15 s. After cooling to room temperature, use an aluminum foil bag packaging machine to package in a sterile room to make a high-freshness flavor soup base finished product.

[0020] Further, in step (2), the high-pressure parameters of the ultra-high pressure cell crusher are a pressure of 400 MPa, a temperature of 25 °C, applying high pressure and maintaining it for 10 minutes, and then quickly releasing the pressure.

[0021] Further, in step (2), the ratio of the mixed raw materials is chicken bone powder: fish bone powder: soybean meal powder: mushroom stalk powder = 5:3:2:1.

[0022] Further, in step (4), the compound protease in the enzymatic hydrolysis reaction is composed of flavor protease, neutral protease, alkaline protease, papain, and trypsin, accounting for 35%, 25%, 20%, 15%, and 5% of the mass of the compound protease respectively.

[0023] Further, in step (4), the ultrasonic power of the ultrasonic treatment is 500 - 700 W, the ultrasonic frequency is 20 kHz, and the total ultrasonic time is 120 min.

[0024] Further, in step (6), the proportion of the spice powder includes: star anise 0.75 g, sand ginger 0.75 g, tsaoko 0.75 g, white cardamom 0.75 g, dried lemongrass 0.81 g, monk fruit 0.38 g, geranium leaves 0.38 g, fennel 0.38 g, gardenia yellow 0.38 g, white pepper 0.38 g, broken cinnamon 0.6 g, Chinese prickly ash 0.6 g, dried tangerine peel 0.15 g, broken licorice 0.22 g, cloves 0.22 g, and the total amount of spices is 7.5 g.

[0025] Further, in step (7), the mixed oil is 17.37 g of soybean oil, 11.05 g of lard, and 1.58 g of sesame oil.

[0026] The beneficial effects of the present invention compared with the prior art are as follows:

[0027] (1) High-value utilization of raw materials and environmental protection; the present invention uses multi-source by-products such as chicken bones, fish bones, soybean meal, and mushroom stalks as raw materials, which are economically accessible and environmentally friendly, effectively enhancing the added value of livestock and poultry by-products, aquatic processing waste, and plant residues. Through the compound enzymatic hydrolysis technology, rich umami substances (such as free amino acids, flavor nucleotides, and polysaccharides) are released, realizing the efficient utilization of resources, which is in line with the concept of circular economy and sustainable development.

[0028] (2) Crushing high pressure improves the utilization rate of raw materials: the present invention effectively destroys the cell structure of the raw materials and releases more umami precursor substances by introducing the crushing high-pressure pretreatment process, significantly improving the enzymatic hydrolysis efficiency. The crushing high-pressure treatment can not only improve the utilization rate of raw materials, but also improve the flavor, texture, and nutritional value of the soup base.

[0029] (3) Synergistic enhancement of ultrasound-enzymolysis; The invention adopts low-frequency high-field ultrasound technology. Through the cavitation effect and mechanical action of ultrasonic waves, the enzymolysis process is effectively enhanced. Under the action of ultrasound, the hydrogen bonds of proteins are effectively broken, the structure changes, promoting their degradation into polypeptides and amino acids, releasing rich umami components and soluble polypeptides, effectively improving the taste of the soup base, making it more delicious and mellow. At the same time, high-intensity ultrasound induces chemical reactions and biological effects, accelerating the depolymerization and degradation of polymers, effectively improving the enzymolysis efficiency. Compared with traditional processes, ultrasound effectively improves the activity of enzymes and enhances the enzymolysis efficiency of proteins.

[0030] (4) Efficient release of umami substances by compound protease; The invention adopts the synergistic action of compound protease (flavor protease, neutral protease, alkaline protease, papain and trypsin) to release umami substances according to the characteristics of different raw materials: chicken skeletons and fish bones release flavor nucleotides such as IMP and AMP; soybean meal releases glutamic acid and aspartic acid; mushroom stalks release GMP and polysaccharides. Compared with traditional single enzymolysis, compound enzymolysis effectively improves the umami level and nutritional value of the flavor soup base, and effectively increases the content of free amino acids and soluble proteins.

[0031] (5) Flavor regulation and antioxidant capacity: The invention adds a variety of plant spices (such as star anise, sand ginger, lemongrass, etc.) to endow the flavor soup base with unique flavors and aromas, while reducing the fishy smell, bitterness and strange smell of animal and plant raw materials. The active ingredients in spices (such as polyphenols, flavonoids) have effective antioxidant capacity, which can inhibit the growth of total colony count and delay the oxidation process of proteins and fats, so that the color, flavor and nutritional quality of the brine can be better maintained and the shelf life can be extended.

[0032] In summary, the invention uses a variety of cheap by-products such as chicken skeletons, fish bones, soybean meal and mushroom stalks, introduces a pre-treatment process of high-pressure crushing and adopts ultrasonic-assisted enzymolysis technology to improve the utilization rate of raw materials, effectively increase the content of umami substances in the flavor soup base, enhance the Maillard reaction, reduce the generation of bitter substances, reduce the addition of caramel color and endow the flavor soup base with a fresh, fragrant, mellow and rich taste. At the same time, the shelf life is extended to meet the needs of consumers for high-quality flavor soup bases. Brief Description of the Drawings

[0033] Figure 1 Shows the effects of different treatment methods on the contents of soluble proteins and soluble peptides in the flavor soup base.

[0034] Figure 2 Shows the effects of different sample groups on the flavor of the flavor soup base.

[0035] Figure 3 Shows the effects of different sample groups on the calcium content of the flavor soup base. Detailed Description of the Invention

[0036] In order to make the objectives, technical solutions and advantages of the present invention clearer and more understandable, the present invention will be further described in detail below in combination with comparative examples and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0037] The chicken skeletons, fish heads and fish bones, mushrooms, and spices used in the following embodiments were freshly purchased.

[0038] Example 1

[0039] A method for preparing a high-freshness flavored soup base based on the synergistic enzymatic hydrolysis of multi-source by-products, the main steps are as follows:

[0040] (1) Raw material pretreatment: Chicken skeletons: Wash, remove inedible parts, boil in boiling water for 20 minutes to degrease, cut into bone pieces smaller than 1 cm, freeze-dry and then crush to 80 mesh to obtain chicken skeleton powder; Fish bones: Soak in 5% brine for 30 minutes to remove fishy smell, blanch in boiling water for 5 minutes, dry at 60°C until the moisture content is less than 8%, and crush to 80 mesh to obtain fish bone powder; Soybean meal: Use food-grade 75% ethanol to degrease for 2 hours, steam-treat for 10 minutes, grind to 80 mesh, and soak in a NaHCO3 solution with a pH of 8.5 for 30 minutes for activation to obtain soybean meal powder; Mushroom stalks: Wash, remove impurities, then dry in hot air at 60°C until the moisture content is less than 10%, and crush to 80 mesh to obtain mushroom stalk powder; 3 Bone pieces, freeze-dry and then crush to 80 mesh to obtain chicken skeleton powder; Fish bones: Soak in 5% brine for 30 minutes to remove fishy smell, blanch in boiling water for 5 minutes, dry at 60°C until the moisture content is less than 8%, and crush to 80 mesh to obtain fish bone powder; Soybean meal: Use food-grade 75% ethanol to degrease for 2 hours, steam-treat for 10 minutes, grind to 80 mesh, and soak in a NaHCO3 solution with a pH of 8.5 for 30 minutes for activation to obtain soybean meal powder; Mushroom stalks: Wash, remove impurities, then dry in hot air at 60°C until the moisture content is less than 10%, and crush to 80 mesh to obtain mushroom stalk powder;

[0041] (2) Pretreatment by crushing and high pressure: Mix the above-mentioned chicken skeleton powder: fish bone powder: soybean meal powder: mushroom stalk powder in a ratio of 5:3:2:1, crush the raw materials into small particles to increase the surface area, and put the crushed raw materials into a flexible packaging bag resistant to high pressure; Use an ultra-high pressure cell crusher, set the high pressure parameter to 400 MPa, the treatment time to 10 minutes, the temperature to 25°C, apply high pressure and maintain it for a certain time, and then quickly release the pressure;

[0042] (3) Homogenization of the mixed raw materials: After the mixed raw materials after high-pressure treatment are sieved, add pure water (the ratio of material to liquid is 1:12), adjust the pH to 6.5 - 7.0, and perform high-speed shear homogenization for 5 minutes;

[0043] (4) Ultrasonic combined enzymatic hydrolysis: In the raw materials after the above homogenization, add 0.28% of compound protease, 0.24% of cellulase, 0.2% of collagenase, and 0.08% of β-glucanase based on the mass of the mixed raw materials. Using a temperature control device and an enzyme reactor, while the enzymatic hydrolysis treatment is carried out, assist with 700W ultrasonic treatment, and perform constant-temperature enzymatic hydrolysis at 55°C for 2 hours to obtain the enzymatic hydrolysis solution of the flavored soup base;

[0044] (5) High temperature and high pressure inactivation and filtration: The flavor soup base enzymatic hydrolysate is treated with a high temperature and high pressure device at 121° C. for 15 minutes to inactivate the enzyme and further sterilize it, and then filtered through a double layer of 100 mesh gauze to remove the residue;

[0045] (6) Preparation of marinade: The marinade is prepared according to the following formula: 6g edible salt, 8g sugar, 4.5g soy sauce, 3g Huadiao wine, 3.5g fish sauce, 16g sun-dried soy sauce, 1.5g caramel color, 5g dried galangal, 7.5g spice powder, 0.8g disodium nucleotide, 6.5g monosodium glutamate, 0.1g inosinic acid, 0.2g scallop extract, 1000g purified water; spice powder ratio: star anise 0.75g, galangal 0.75g, grass fruit 0.75g, white cardamom 0.75g, dried lemongrass 0.81g, monk fruit 0.38g, bay leaf 0.38g, fennel 0.38g, yellow gardenia 0.38g, white pepper 0.38g, cinnamon bark 0.6g, pepper 0.6g, tangerine peel 0.15g, licorice 0.22g, cloves 0.22g, total spices 7.5g;

[0046] (7) Preparation of flavor soup base: The enzyme-killed and filtered flavor soup base enzymatic hydrolyzate obtained in step 5 is mixed with the marinade obtained in step 6 at a ratio of 1:1, and 30 g of mixed oil (17.37 g soybean oil, 11.05 g lard, and 1.58 g sesame oil) is added for oil sealing;

[0047] (8) Filling and sterilization: The flavor soup base is fully mixed and homogenized, and the flavor soup base is sterilized at high temperature for a short time (121°C, 15S), cooled to room temperature, and filled in sterile aluminum foil bags.

[0048] Example 2

[0049] A method for preparing a high-fresh flavor soup base based on synergistic enzymatic hydrolysis of multi-source by-products, the main steps of which are as follows:

[0050] (1) Raw material pretreatment: Chicken skeleton: clean and remove unusable parts, boil in boiling water for 20 minutes to remove fat, and cut into pieces less than 1 cm 3 Bone blocks: freeze-dried and crushed to 80 mesh to obtain chicken skeleton powder; fish bones: soaked in 5% salt water for 30 minutes to remove the fishy smell, blanched in boiling water for 5 minutes, dried at 60℃ until the moisture is less than 8%, crushed to 80 mesh to obtain fish bone powder; soybean meal: defatted with food-grade 75% ethanol for 2 hours, steamed for 10 minutes, ground to 80 mesh, and soaked in NaHCO3 solution with pH 8.5 for 30 minutes to activate, to obtain soybean meal powder; mushroom stems: washed and impurities removed, dried with hot air at 60℃ until the moisture is less than 10%, crushed to 80 mesh to obtain mushroom stem powder;

[0051] (2) Crushing and high-pressure pretreatment: the above-mentioned chicken skeleton powder: fish bone powder: soybean meal powder: mushroom stem powder are mixed in a ratio of 5:3:2:1, and the raw materials are crushed into small particles to increase the surface area. The crushed raw materials are packed into a high-pressure resistant flexible packaging bag; an ultra-high pressure cell crusher is used, the high pressure parameters are set to 400 MPa, the processing time is 10 minutes, the temperature is 25°C, high pressure is applied and maintained for a certain period of time, and then the pressure is quickly released;

[0052] (3) Homogenizing the mixed raw materials: After the high-pressure treated mixed raw materials are sieved, purified water is added (material-liquid ratio 1:12), the pH is adjusted to 6.5-7.0, and high-speed shear homogenization is performed for 5 minutes;

[0053] (4) Ultrasonic combined enzymatic hydrolysis: Add 0.28% of the mass of the mixed raw material composite protease, 0.24% of the mass of the mixed raw material cellulase, 0.2% of the mass of the mixed raw material collagenase and 0.08% of the mass of the mixed raw material β-glucanase to the above-mentioned homogenized raw materials, use a temperature control device and an enzyme reactor, and simultaneously perform 600W ultrasonic treatment at 55°C for 2 hours to obtain a flavor soup base enzymatic hydrolyzate;

[0054] (5) High temperature and high pressure inactivation and filtration: The flavor soup base enzymatic hydrolysate is treated with high temperature and high pressure equipment (such as an autoclave) at 121° C. for 15 minutes to inactivate the enzyme and further sterilize it, and then filtered through a double layer of 100 mesh gauze to remove the residue;

[0055] (6) Preparation of marinade: The marinade is prepared according to the following formula: 6g edible salt, 8g sugar, 4.5g soy sauce, 3g Huadiao wine, 3.5g fish sauce, 16g sun-dried soy sauce, 1.5g caramel color, 5g dried galangal, 7.5g spice powder, 0.8g disodium nucleotide, 6.5g monosodium glutamate, 0.1g inosinic acid, 0.2g scallop extract, 1000g purified water; spice powder ratio: star anise 0.75g, galangal 0.75g, grass fruit 0.75g, white cardamom 0.75g, dried lemongrass 0.81g, monk fruit 0.38g, bay leaf 0.38g, fennel 0.38g, yellow gardenia 0.38g, white pepper 0.38g, cinnamon bark 0.6g, pepper 0.6g, tangerine peel 0.15g, licorice 0.22g, cloves 0.22g, total spices 7.5g;

[0056] (7) Preparation of flavor soup base: The enzyme-killed and filtered flavor soup base enzymatic hydrolyzate obtained in step 5 is mixed with the marinade obtained in step 6 at a ratio of 1:1, and 30 g of mixed oil (17.37 g soybean oil, 11.05 g lard, and 1.58 g sesame oil) is added for oil sealing;

[0057] (8) Filling and sterilization: Thoroughly mix and homogenize the flavored soup base, subject the flavored soup base to high-temperature short-time sterilization (121 °C, 15 s), cool it to room temperature, and fill it into sterile aluminum foil bags.

[0058] Use the flavored soup base that has not been ultrasonically treated and enzymatically hydrolyzed as blank group 1 (do not perform the ultrasonic treatment and enzymatic hydrolysis processes in step 4 of Example 2, only keep it at a constant temperature of 55 °C for 2 h; the other steps are the same).

[0059] Use the flavored soup base that has not been ultrasonically treated and enzymatically hydrolyzed and only contains chicken bone powder without fish bone powder, soybean meal powder, and shiitake mushroom stalk powder as blank group 2 (only add chicken bone powder in step 2 of Example 2, do not add other types of raw materials; do not perform step 4 of Example 2; the other steps are the same).

[0060] Use the flavored soup base that has not been subjected to high-pressure crushing treatment as blank group 3 (only perform raw material mixing in step 2 of Example 2, do not perform crushing; the other steps are the same).

[0061] Use the flavored soup base that has not been enzymatically hydrolyzed and only been ultrasonically treated as the ultrasonic group (the same as step 4 of Example 2, but do not add any enzyme preparations and do not perform the enzymatic hydrolysis process; the other steps are the same).

[0062] Use the flavored soup base that has not been ultrasonically treated and only been enzymatically hydrolyzed as the enzymatic hydrolysis group (the same as step 4 of Example 2, but do not undergo ultrasonic treatment; the other steps are the same).

[0063] Use Example 2 as the ultrasonic combined enzymatic hydrolysis group.

[0064] Example 3

[0065] A method for preparing a high-freshness flavored soup base based on the synergistic enzymatic hydrolysis of multi-source by-products, the main steps are as follows:

[0066] (1) Raw material pretreatment: Chicken bones: Wash, remove the unusable parts, boil in boiling water for 20 minutes to degrease, cut into bone pieces smaller than 1 cm 3 Bone pieces, freeze-dry and then pulverize to 80 mesh to obtain chicken bone powder; Fish bones: Soak in 5% brine for 30 minutes to remove fishy smell, blanch in boiling water for 5 minutes, dry at 60 °C until the moisture content is less than 8%, and pulverize to 80 mesh to obtain fish bone powder; Soybean meal: Degrease with 75% food-grade ethanol for 2 hours, steam-treat for 10 minutes, grind to 80 mesh, and soak in a NaHCO3 solution with a pH of 8.5 for 30 minutes for activation to obtain soybean meal powder; Shiitake mushroom stalks: Wash, remove impurities, then dry at 60 °C in hot air until the moisture content is less than 10%, and pulverize to 80 mesh to obtain shiitake mushroom stalk powder;

[0067] (2) Crushing and high-pressure pretreatment: the above-mentioned chicken skeleton powder: fish bone powder: soybean meal powder: mushroom stem powder are mixed in a ratio of 4:3:1:2, and the raw materials are crushed into small particles to increase the surface area. The crushed raw materials are packed into a high-pressure resistant flexible packaging bag; an ultra-high pressure cell crusher is used, the high pressure parameters are set to 400 MPa, the processing time is 10 minutes, the temperature is 25°C, high pressure is applied and maintained for a certain period of time, and then the pressure is quickly released;

[0068] (3) Homogenizing the mixed raw materials: After the high-pressure treated mixed raw materials are sieved, purified water is added (material-liquid ratio 1:12), the pH is adjusted to 6.5-7.0, and high-speed shear homogenization is performed for 5 minutes;

[0069] (4) Ultrasonic combined enzymatic hydrolysis: Add 0.28% of the mass of the mixed raw material composite protease, 0.24% of the mass of the mixed raw material cellulase, 0.2% of the mass of the mixed raw material collagenase and 0.08% of the mass of the mixed raw material β-glucanase to the above-mentioned homogenized raw materials, use a temperature control device and an enzyme reactor, and simultaneously perform 500W ultrasonic treatment, and perform enzymatic hydrolysis at a constant temperature of 55°C for 2h to obtain a flavor soup base enzymatic hydrolyzate;

[0070] (5) High temperature and high pressure inactivation and filtration: The flavor soup base enzymatic hydrolysate is treated with high temperature and high pressure equipment (such as an autoclave) at 121° C. for 15 minutes to inactivate the enzyme and further sterilize it, and then filtered through a double layer of 100 mesh gauze to remove the residue;

[0071] (6) Preparation of marinade: The marinade is prepared according to the following formula: 6g edible salt, 8g sugar, 4.5g soy sauce, 3g Huadiao wine, 3.5g fish sauce, 16g sun-dried soy sauce, 1.5g caramel color, 5g dried galangal, 7.5g spice powder, 0.8g disodium nucleotide, 6.5g monosodium glutamate, 0.1g inosinic acid, 0.2g scallop extract, 1000g purified water; spice powder ratio: star anise 0.75g, galangal 0.75g, grass fruit 0.75g, white cardamom 0.75g, dried lemongrass 0.81g, monk fruit 0.38g, bay leaf 0.38g, fennel 0.38g, yellow gardenia 0.38g, white pepper 0.38g, cinnamon bark 0.6g, pepper 0.6g, tangerine peel 0.15g, licorice 0.22g, cloves 0.22g, total spices 7.5g;

[0072] (7) Preparation of flavor soup base: The enzyme-killed and filtered flavor soup base enzymatic hydrolyzate obtained in step 5 is mixed with the marinade obtained in step 6 at a ratio of 1:1, and 30 g of mixed oil (17.37 g soybean oil, 11.05 g lard, and 1.58 g sesame oil) is added for oil sealing;

[0073] (8) Filling and sterilization: Thoroughly mix and homogenize the flavored soup base, perform high-temperature short-time sterilization on the flavored soup base (121 °C, 15 s), cool it to room temperature, and fill it into sterile aluminum foil bags.

[0074] Experimental methods:

[0075] 1. Soluble protein content

[0076] Measure and evaluate the quality of the above-prepared marinated soup products. Experimental method: Centrifuge 5 mL of the sample at 10,000 rpm for 15 min at 4 °C, and collect the supernatant after centrifugation. Use the biuret method to measure the absorbance of the supernatant of different samples at a wavelength of 540 nm. Take 1 mL of the supernatant, add 4 mL of biuret reagent, and use bovine serum albumin as an external standard method to quantitatively analyze the water-soluble protein content in the sample.

[0077] 2. Soluble peptide content

[0078] Measure and evaluate the quality of the above-prepared marinated soup products. Experimental method: Mix 5 mL of the sample with 5 mL of trichloroacetic acid (15%) and let it stand for 30 min, centrifuge at 10,000 rpm for 15 min at 4 °C, and collect the supernatant after centrifugation. Use the biuret method to measure the absorbance of the supernatant of different samples at a wavelength of 540 nm. Take 1 mL of the supernatant, add 4 mL of biuret reagent, and use bovine serum albumin as an external standard method to quantitatively analyze the water-soluble peptide content in the sample.

[0079] 3. Electronic nose

[0080] Place the sample in a glass bottle, take 5 mL of the filtered supernatant and put it into the electronic nose bottle, and keep it in a water bath at 45 °C for 30 min to allow the odor to completely volatilize and fill the entire electronic nose bottle. Before measurement, first clean the electronic nose for 3000 s, clean it for 120 s before each measurement, after the end of cleaning for 100 s, the measurement time for each time is 120 s, and the gas flow rate is 1.0 L / min.

[0081] 4. Determination of 5'-nucleotides content

[0082] Take 10 mL of the sample, after standing at 4 °C for 12 h, filter off the upper floating foam using double-layer filter cloth, centrifuge (10,000 r / min, 20 min, 4 °C), dilute the obtained supernatant by 5 times, filter it through a 0.22 μm syringe filter membrane, and use the filtrate for

[0083] Analysis of 5'-flavor nucleotides. The 5'-flavor nucleotides were analyzed using an Ultimate 3000 HPLC system, a Sunfire C18 (4.6 mm x 250 mm, 5 μm) chromatographic column, and a DAD detector. The mobile phase (A) was KH2PO4 (0.05 mol / L, pH 4.5): the mobile phase (B) was methanol = 98%: 2%, the flow rate was fixed at 1.0 mL / min, and the detection wavelength was 254 nm. The content of 5'-flavor nucleotides was quantitatively analyzed using an external standard curve.

[0084] 5. Sensory evaluation

[0085] The sensory evaluation was determined by the observation method. An evaluation panel consisting of 10 professionals was trained and then used to conduct an overall sensory score on the qualified samples, and the average value of the comprehensive evaluation results was taken from the three aspects of vision, smell, and taste.

[0086] 6. Determination of calcium content

[0087] According to GB 5009.268-2016 "National Food Safety Standard - Determination of Multiple Elements in Foods", the second method, inductively coupled plasma optical emission spectrometry, was used to detect the calcium content in the flavor soup base. An appropriate amount of the flavor soup base sample was taken, stirred evenly, digested with concentrated nitric acid in a closed container, and then the calcium content was determined using an inductively coupled plasma optical emission spectrometer. The operating conditions of the ICP-OES were a radio frequency power of 1150 W, a plasma flow rate of 0.5 L / min, an auxiliary flow rate of 0.5 L / min, a nebulizer flow rate of 12.5 L / min, and an integration time of 30 seconds. The calcium characteristic spectral line was 315.8 nm.

[0088] 7. Determination of total colony count

[0089] According to the determination of total colony count in GB 4789.2-2022, the plate pour method was used to determine the total colony count of the flavor soup base at 0 d, 10 d, 30 d, and 90 d.

[0090] The scores are shown in the following table:

[0091] Table 1 Sensory scoring rules

[0092]

[0093] Water-soluble proteins and water-soluble peptides are important for the nutrition, appearance, and sensory experience of the flavor soup base. They are continuously released from multi-source by-product powder during the ultrasonic combined enzymatic hydrolysis process and enter the aqueous solution. They can endow the flavor soup base with nutritional components, increase the viscosity of the soup base to make its color more attractive, and improve the flavor stability and storage stability of the soup base.

[0094] From Figure 1It can be seen that for the samples without ultrasonic and enzymatic hydrolysis treatment, the contents of soluble proteins and peptides are relatively low, and the flavor and nutrition are relatively poor. However, the ultrasonic enzymatic hydrolysis process increases the protein dissolution rate and the content of small molecular peptides, making the soluble protein in the example group about twice that of blank group 1, and the soluble peptide content about 2.5 times. The soluble protein content and soluble peptide content of blank group 3 are lower than those of the example group and the enzymatic hydrolysis group, indicating that subjecting the raw material powder to high-pressure crushing before enzymatic hydrolysis can improve the enzymatic hydrolysis efficiency, evenly crush the substrate of the enzymatic hydrolysate, and release more umami and nutrients. During the process of braising meat, the soluble proteins and polypeptides in the flavor soup base can interact with other substances to delay the flavor release and continuously degrade, forming oligopeptides and free amino acids, which endow the braised products with unique flavors, enhance the color and luster, and strengthen the sensory experience.

[0095] Table 2 Effects of different sample groups on the content of 5'-flavor nucleotides in the flavor soup base

[0096]

[0097] 5'-flavor nucleotides include inosine monophosphate (IMP), guanosine monophosphate (GMP), cytidine monophosphate (CMP), and adenosine monophosphate (AMP). There is a synergistic effect among them. Using them in combination has a stronger umami taste than consuming them alone and can endow food with rich flavors. At the same time, they can act synergistically with monosodium glutamate to form a mellow taste and a stronger umami. Enzymatic treatment will damage the cell membranes in the four by-products and the cell walls in plant cells, resulting in more nucleotides being dissolved. The umami substances dissolved from different by-products have a synergistic effect, making the umami more mellow and the taste more rich. As can be seen from the table, ultrasonic combined with enzymatic hydrolysis significantly increases the content of the four flavor nucleotides, especially IMP and GMP, reaching 20.26 mg / 100 g and 8.13 mg / 100 g respectively, which are 2-3 times that of the control group. The cavitation effect of ultrasound enhances the effect of enzymatic hydrolysis and thermal degradation on the decomposition of nucleic acids into nucleotides, promotes the enrichment of a large amount of umami nucleotides in the brine, and the thermal decomposition of ATP during boiling can form a large amount of AMP, giving the flavor soup base a unique sweet taste. Compared with the ultrasonic-enzymatic hydrolysis group, blank group 3 without high-pressure crushing shows a lower content of 5'-flavor nucleotides, indicating that the high-pressure crushing process makes the raw material mixing degree higher, improves the efficiency in subsequent enzymatic hydrolysis, and releases more umami substances.

[0098] From Figure 2It can be seen that the differences between different groups are obvious. However, the overall readings are arranged in ascending order as follows: Blank Group 1 < Blank Group 2 < Ultrasonic Group < Enzymolysis Group < Blank Group 3 < Ultrasonic Enzymolysis Group. The reason why the reading of the Ultrasonic Group 2 is greater than that of the Ultrasonic Group 1 is that the by-products of the Ultrasonic Group 2 come from not only chicken skeletons but also fish bones, soybean meal, and mushroom stalks, making the smell richer and more harmonious. The ultrasonic-assisted enzymolysis process increases the readings of each sensor. Enzymolysis makes the smell more intense, and ultrasonic cavitation makes the macromolecules more completely thermally degraded. During the enzymolysis process, macromolecular proteins are enzymolyzed into small-molecular peptides, and fats are gradually oxidized and decomposed to produce a unique smell. The comparison between the ultrasonic combined enzymolysis group and Blank Group 3 shows that the high-pressure crushing process before enzymolysis enhances the flavor of the subsequent enzymolysis solution, making the smell produced by enzymolysis more intense. Among them, the response values of S4 (sulfur-containing compounds), S5 (organic amines), S9 (aromatic compounds), and S16 (sulfur-containing compounds) are relatively high. This indicates that sulfur-containing and nitrogen-containing compounds, as well as short-chain alkanes, may make significant contributions to the flavor of marinated flavors, indicating that there are relatively more protein-based and fat-containing aroma substances.

[0099] In terms of the sensory evaluation scores, there is no significant difference among the three groups of examples, and the total scores are all significantly higher than those of the control group, indicating that the flavor soup base of ultrasonic combined enzymolysis of multi-source by-products is higher in sensory experience than the flavor soup base without ultrasonic enzymolysis. Slightly changing the proportion of by-products and the proportion of compound protease has a weak impact on the sensory evaluation. As shown in Table 3, subjecting the enzymolysis substrate to high-pressure crushing before enzymolysis can improve the sensory properties. Similarly, ultrasonic has a weak impact on the sensory evaluation, while the combined effect of high-pressure crushing and enzymolysis on improving the sensory properties of the flavor soup base is obvious.

[0100] Table 3 Sensory evaluation scores of different sample groups

[0101]

[0102] It can be Figure 3 seen that the calcium contents of the three examples are similar, indicating that the source of by-products has little impact on the calcium content of the flavor soup base. The calcium content of Example 1 reaches 541.56 mg / L, while that of Blank Group 1 without ultrasonic and enzymolysis processes is only 115.26 mg / L. Among them, the enzymolysis process significantly increases the concentration of calcium ions. The improvement effect of high-pressure crushing is second, and the improvement effect of ultrasonic on calcium content is the weakest. The combination of high-pressure crushing, ultrasonic, and enzymolysis enables the calcium in the multi-source by-products to be released and enriched into the flavor soup base. High-pressure treatment can destroy the cell structure of raw materials and promote the dissolution and release of calcium salts. During the enzymolysis process, protease can decompose the calcium bound to proteins and release calcium ions, while cellulase and β-glucanase can help the calcium ions in soybean meal and mushrooms dissociate into the soup base and be enriched in the flavor soup base as free calcium ions and soluble calcium salts. Thermal processes and ultrasonic cavitation accelerate the dissolution of calcium.

[0103] Table 4 Total number of colonies in different sample groups

[0104]

[0105] The determination of the total number of colonies can indicate the shelf life of the flavor soup base and the microbial reproduction situation. As can be seen from Table 4, with the extension of the storage time, the total number of colonies in all groups increased significantly. The total number of colonies in the enzymatic hydrolysis group and the ultrasonic group were both less than that in the blank group, while the ultrasonic combined with enzymatic hydrolysis group showed the lowest total colony count value. Both ultrasonic and enzymatic hydrolysis treatments can reduce the growth of colonies. Due to the hurdle effect, the ultrasonic combined with enzymatic hydrolysis treatment can extend the shelf life and microbial safety of the flavor soup base.

Claims

1. A method for preparing a high-freshness flavored soup base based on the coenzymatic hydrolysis of multi-source by-products, characterized in that, It mainly includes the following steps: (1)Raw material pretreatment: Chicken skeletons: Wash, remove unusable parts, boil in boiling water for 20 minutes to degrease, cut into bone pieces smaller than 1 cm 3 , freeze-dry and crush to 80 mesh to obtain chicken skeleton powder; Fish bones: Soak in 5% brine for 30 minutes to remove fishy smell, blanch in boiling water for 5 minutes, dry at 60 °C until the moisture content is less than 8%, and crush to 80 mesh to obtain fish bone powder; Soybean meal: Degrease with 75% food-grade ethanol for 2 hours, steam-treat for 10 minutes, grind to 80 mesh, soak in a NaHCO3 solution with pH 8.5 for 30 minutes for activation to obtain soybean meal powder; Lentinula edodes stalks: After washing and removing impurities, soak in a 3% citric acid solution for 15 minutes to remove the earthy smell, then dry in hot air at 60°C until the moisture content is less than 10%, and crush to 80 mesh to obtain Lentinula edodes stalk powder; (2) Crushing and high-pressure treatment of the mixed raw materials: Mix the above-mentioned chicken bone powder, fish bone powder, soybean meal powder, and Lentinula edodes stalk powder and put them into a high-pressure-resistant flexible packaging bag, and perform high-pressure treatment using a high-pressure cell crusher; (3) Homogenization of the mixed raw materials: After sieving the mixed raw materials after the above high-pressure treatment, add pure water with a material-liquid ratio of 1:12, adjust the pH to 6.5 - 7.0, and perform high-speed shear homogenization for 5 minutes; (4) Ultrasonic combined with enzymatic hydrolysis: Add 0.28% of compound protease, 0.24% of cellulase, 0.2% of collagenase, and 0.08% of β-glucanase based on the mass of the mixed raw materials in the homogenized mixed raw materials for enzymatic hydrolysis treatment, and at the same time assist with ultrasonic treatment, and perform constant-temperature enzymatic hydrolysis at 55°C for 2 hours to obtain the enzymatic hydrolysis solution of the flavor soup base; (5) High-temperature and high-pressure enzyme inactivation and filtration: Use a high-temperature steam sterilization kettle, set the parameters to 121°C, perform high-temperature and high-pressure treatment on the enzymatic hydrolysis solution of the flavor soup base for 15 minutes to inactivate the enzyme, and then filter with double-layer 100-mesh gauze to remove residues; (6) Preparation of the brine: Prepare the brine according to the following formula: 7.5 g of spice powder, 6 g of edible salt, 8 g of sugar, 4.5 g of soy sauce, 3 g of huadiao wine, 3.5 g of fish sauce, 16 g of sun-dried soy sauce, 1.5 g of caramel color, 5 g of dried galangal, 0.8 g of disodium 5'-ribonucleotide, 6.5 g of monosodium glutamate, 0.1 g of inosinic acid, 0.2 g of scallop essence, 1000 g of pure water; (7) Preparation of the flavor soup base: Mix the enzyme-inactivated and filtered enzymatic hydrolysis solution of the flavor soup base obtained in step (5) with the brine obtained in step (6) at a ratio of 1:1, and add 30 g of mixed oil for oil sealing to obtain the flavor soup base; (8) Filling and sterilization: Perform high-temperature short-time sterilization on the flavor soup base in step (7), the treatment conditions are temperature: 121°C, time: 15 s, after cooling to room temperature, use an aluminum foil bag packaging machine to package in a sterile room to make a finished product of high-freshness flavor soup base.

2. The preparation method of a high-freshness flavor soup base based on the coenzymatic hydrolysis of multi-source by-products according to claim 1, characterized in that, In step (2), the high-pressure parameters of the high-pressure cell crusher are a pressure of 400 MPa, a temperature of 25°C, apply high pressure and maintain it for 10 minutes, and then quickly release the pressure.

3. A method for preparing a high-freshness flavor soup base based on the coenzymatic hydrolysis of multi-source by-products according to claim 1, characterized in that, In step (2), the ratio of the mixed raw materials is chicken bone powder: fish bone powder: soybean meal powder: Lentinula edodes stalk powder = 5:3:2:

1.

4. A method for preparing a high-freshness flavor soup base based on the coenzymatic hydrolysis of multi-source by-products according to claim 1, characterized in that, In step (4), the compound protease in the enzymatic hydrolysis reaction is composed of flavor protease, neutral protease, alkaline protease, papain, and trypsin, accounting for 35%, 25%, 20%, 15%, and 5% of the mass of the compound protease respectively.

5. A method for preparing a high-freshness flavor soup base based on the coenzymatic hydrolysis of multi-source by-products according to claim 1, characterized in that: When the ultrasonic treatment in step (4) is carried out synchronously with the enzymatic hydrolysis treatment, the ultrasonic power of the ultrasonic treatment is 500 - 700 W, the ultrasonic frequency is 20 kHz, and the total ultrasonic time is 120 min.

6. The preparation method of a high-freshness flavor soup base based on the synergistic enzymatic hydrolysis of multi-source by-products according to claim 1, characterized in that, The proportion of the spice powder in step (6) includes: 0.75 g of star anise, 0.75 g of sand ginger, 0.75 g of tsaoko, 0.75 g of white cardamom, 0.81 g of dried lemongrass, 0.38 g of monk fruit, 0.38 g of cinnamon leaves, 0.38 g of fennel, 0.38 g of gardenia yellow, 0.38 g of white pepper, 0.6 g of broken cinnamon, 0.6 g of Chinese prickly ash, 0.15 g of tangerine peel, 0.22 g of broken licorice, and 0.22 g of cloves.

7. A method for preparing a high-freshness flavor soup base based on the synergistic enzymatic hydrolysis of multi-source by-products according to claim 1, characterized in that, The mixed oil in step (7) is 17.37 g of soybean oil, 11.05 g of lard, and 1.58 g of sesame oil.

Images