Preparation method of cattle bone, salmon bone peptide-oat oligosaccharide Maillard intermediate based on reduced pressure heating

Abstract

The present invention discloses a method for preparing a Maillard intermediate product of cattle bone and salmon bone peptides and oat oligosaccharides using reduced pressure heating, relating to the field of food processing technology. The method comprises the following steps: cutting and dispersing cattle bones and salmon bones, after removing the meat and fascia tissue, followed by ultrasonic cleaning, enzymatic hydrolysis, inactivation, collection of cattle bone peptides, and subsequent Maillard reaction of the peptides with sugars and amino acids to produce a salty and umami intermediate product. The Maillard intermediate product of cattle bone and salmon bone peptides and oat oligosaccharides obtained in the present invention has a low molecular weight and a salty and umami flavor, and can be used as a base ingredient powder in seasonings.

Description

Technical Field

[0001] The invention relates to the technical field of food processing, and in particular to a method for preparing a Maillard intermediate product of cattle bone and salmon bone peptide-oat oligosaccharide based on reduced pressure heating. Background Art

[0002] Maillard products play an important role in enhancing food flavor and have been increasingly used in flavors and seasonings in recent years. However, the market currently lacks flavors with complex tastes and innovative flavors. Industrially produced, compound products with the characteristics of multiple seasonings offer advantages such as nutritional benefits, flavor diversity, and convenience.

[0003] Beef and salmon bones primarily contain water, protein, lipids, calcium, and phosphorus. Over 80% of their protein is collagen, and the enzymatic hydrolysis products contain the eight essential amino acids required by the human body, with high biological value. Furthermore, beef and salmon bones are primarily waste or ground up for use as feed, significantly reducing their utility. However, beef and salmon bones contain a wealth of flavor compounds, which can be further developed into flavorings.

[0004] Current Maillard reaction substrates typically utilize peptides or amino acids to provide amino groups, which react with reducing monosaccharides or disaccharides to provide carbonyl groups. For example, arginine or lysine is added to the reaction system along with xylose, glucose, or lactose. While this method can yield relatively rich flavor compounds, the resulting products are characterized by a predominance of furans and pyrazines, while other flavor compounds such as esters, aldehydes, and ketones are relatively low in content. To enhance the flavor and mouthfeel diversity of Maillard products and increase the content of low-abundance flavor compounds to enhance the richness of the characteristic flavor, optimizing the flavor substrate and the reaction system are key approaches to product improvement. Furthermore, since the Maillard reaction requires prolonged heating, which is energy-intensive for smaller-scale production, optimizing the reaction system should also consider shortening the reaction time. Summary of the Invention

[0005] In order to develop cattle bone and fish bone resources, the purpose of the present invention is to provide a method for preparing a Maillard product of cattle bone and salmon bone peptide-oat oligosaccharide based on reduced pressure heating. The raw materials are cut and dispersed by removing the meat and fascia tissue from the cattle bones, and then ultrasonically cleaned, enzymatically hydrolyzed, and inactivated. The cattle bones and salmon bone peptides are collected, and then the peptides are reacted with oat oligosaccharides and amino acids to obtain a Maillard intermediate product with a salty and fresh taste.

[0006] In order to achieve the technical purpose of the present invention, the present invention provides the following technical solutions:

[0007] The present invention first provides a method for preparing a Maillard intermediate product of bovine bone peptide and salmon bone peptide-oat oligosaccharide by reduced pressure heating, which is characterized by comprising the following steps:

[0008] (1) Remove the meat and fascia tissue from beef and salmon bones, hammer and crush them in equal proportions, add 2.5 times the weight of the raw materials with water and 1.5% of the weight of the raw materials with 52° liquor, and finally heat and steam at 100°C for 30-60 minutes and remove the raw materials;

[0009] (2) The raw material obtained in step (1) is crushed with a grinder, and then mixed and dispersed with water in a mass ratio of 1:20 to 1:25, the pH of the dispersion is adjusted to 8.0 to 9.0, and alkaline protease and bromelain are added for dual enzyme hydrolysis, and the enzymatic hydrolysis is carried out for 4 to 8 hours while stirring throughout the process;

[0010] (3) The final enzymatic hydrolysate obtained in step (2) was heated at 100° C. for 10 to 15 minutes to inactivate the enzyme, then cooled to room temperature, and the pH was adjusted to 4.0 to 5.0. The supernatant was collected after standing;

[0011] (4) The supernatant obtained in step (3) was filtered using a nanofiltration membrane to retain molecules with a molecular weight cutoff of 3000 Da, and the filtrate was collected and freeze-dried to obtain polypeptide powder;

[0012] (5) Oat polysaccharide was added to water at a material-liquid mass ratio of 1:200, and α-amylase was added for enzymatic hydrolysis. The mixture was heated at 100°C for 10-15 minutes to inactivate the enzyme, and then freeze-dried to obtain oat oligosaccharide powder;

[0013] (6) 10 parts by mass of the polypeptide powder obtained in step (4) and 2 to 3 parts by mass of the oat oligosaccharide obtained in step (5) are dissolved in 50 parts by mass of an edible ethanol-water mixture, and reacted at 70° C. to 90° C., 0.03 to 0.06 MPa under reduced pressure and heating conditions for 0.5 to 1 h; then 1 part by mass of xylose, 1 part by mass of arginine, 0.5 parts by mass of methionine, and 0.5 parts by mass of lysine are added, and reacted at 75 to 95° C., 0.03 to 0.06 MPa under reduced pressure and heating conditions for 0.5 to 1 h to obtain a Maillard intermediate.

[0014] As a preferred embodiment of the present invention, in step (1), the size of the raw material after being hammered and crushed is 3-5 cubic centimeters.

[0015] As a preferred embodiment of the present invention, the medium-frequency ultrasonic method in step (1) is as follows: the ultrasonic frequency is 100-200 kHz, the ultrasonic power is 500-800 W, and the ultrasonic time is 10-30 min.

[0016] As a preferred embodiment of the present invention, in step (2), the raw material is crushed by a crusher to a particle size of less than 1 mm.

[0017] As a preferred embodiment of the present invention, the enzymolysis temperature in step (2) is 50° C. to 60° C., the stirring speed during the enzymolysis process is 100 to 200 rpm, the amount of alkaline protease added is 0.25 to 0.35% of the weight of the crushed raw material, the amount of bromelain added is 0.15 to 0.25% of the weight of the crushed raw material, the enzymatic activity of alkaline protease is 40,000 to 70,000 U / g, and the enzymatic activity of bromelain is 20,000 to 40,000 U / g.

[0018] As a preferred embodiment of the present invention, the purity of oat polysaccharide in step (5) is 80%.

[0019] As a preferred embodiment of the present invention, the addition ratio of α-amylase in step (5) is 0.2% to 0.5% of the mass of oat polysaccharide, the α-amylase activity is 3000U / g to 5000U / g, the enzymatic hydrolysis temperature is 45°C to 60°C, and the enzymatic hydrolysis time is 60min to 80min.

[0020] As a preferred embodiment of the present invention, in step (6), the purity of edible ethanol is 95%; and in the edible ethanol-water mixture, the volume percentage of edible ethanol is 20% to 40%.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] (1) The present invention utilizes controlled enzymatic hydrolysis technology to hydrolyze bone protein, and then ultrafiltration is used to obtain small-molecule beef bone and salmon bone peptides. This is then subjected to a Maillard reaction with oat oligosaccharides, amino acids, and the like to obtain an intermediate product with a rich flavor and a fresh, fragrant, and mellow taste.

[0023] (2) The present invention reduces the fishy odor substances in the beef bones by adding white wine and using ultrasound, adds substrate composite polypeptide and malto oligosaccharide to increase the flavor source, and optimizes the secondary Maillard reaction.

[0024] (3) The method of the present invention is safe and green, and can further improve the functional properties and utilization of raw materials, expand its application field, and the obtained Maillard intermediate product can be applied to the base powder of seasonings. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 It is a production process flow chart of the present invention. DETAILED DESCRIPTION

[0026] The specific embodiments of the present invention are further described below.

[0027] Unless otherwise specified, the oat polysaccharide used in the present invention is purchased from Lanzhou Waterless Biotechnology Co., Ltd., and other raw materials can be purchased from the market or are commonly used in the art. Unless otherwise specified, the methods in the following embodiments are all conventional methods in the art.

[0028] Sensory evaluation

[0029]

[0030]

[0031] Sensory score weighting = color sensory score 30% + smell sensory score 30% + taste sensory score 40%

[0032] UV determination of Maillard products

[0033] The obtained Maillard intermediate product liquid was mixed and diluted with water, and the volume was fixed after dilution by 10 times to 100 times respectively, and the UV-visible absorption value was measured at 290 nm and 420 nm.

[0034] GC-MS flavor compound determination

[0035] Extraction conditions: Dissolve about 2g of Maillard intermediate in 25mL of saturated saline, add 20μL of 2-methyl-3-heptanone (0.8mg / mL), mix well, and seal. Use a syringe to adsorb at 60℃ for 30min before injection; heating program: maintain at 35℃ for 3min, and increase at 3℃·min -1 Heat to 40℃ and hold for 1min, then at 5℃·min -1 Heat to 230°C and maintain for 12 minutes.

[0036] GC-MS conditions: Volatile compounds were separated on a TG-5SICMS column (30 m × 0.25 mm, 0.25 μm); carrier gas was He at a flow rate of 1 mL / min, with a split ratio of 20:1 (V:V); electron impact (EI) ionization source temperature was 285°C, and transfer temperature was 255°C; scanning mass range was 35–450 amu, with a scan rate of 0.1 amu / s. Volatile components were semiquantitated using an internal standard method to determine the mass concentration of each volatile component.

[0037] Example 1

[0038] The process of the embodiment of the present invention refers to Figure 1 , specifically:

[0039] (1) Remove the meat and fascia tissue from beef and salmon bones, hammer and crush them into 3 cubic centimeter blocks in equal proportions, add 2.5 times the weight of water, add 1.5% of the weight of the raw materials in 52° liquor, and remove the fishy odor by ultrasonication at a frequency of 100 kHz, a power of 600 W, and a duration of 20 minutes. Finally, heat at 100°C for 40 minutes and remove the raw materials.

[0040] (2) The raw material obtained in step (1) is pulverized with a pulverizer until the particle size of the raw material after pulverization is less than 1 mm. The pulverized raw material is mixed and dispersed with water at a mass ratio of 1:20, and alkaline protease and flavor protein are added for double enzyme hydrolysis. The pH value of the homogenate is maintained at 8.3, the enzymatic hydrolysis time is 5 hours, the enzymatic hydrolysis temperature is 57° C., the stirring speed is 145 rpm, the amount of alkaline protease added is 0.3% of the mass of the pulverized raw material, the amount of bromelain added is 0.2% of the mass of the pulverized raw material, the alkaline protease activity is 50,000 U / g, and the bromelain activity is 35,000 U / g;

[0041] (3) The final enzymatic hydrolysate of step (2) was heated at 100°C for 12 min to inactivate the enzyme, and the pH was adjusted to 4.1 at room temperature. The supernatant was collected after standing for 3 h;

[0042] (4) The supernatant obtained in step (3) was filtered using a nanofiltration membrane to retain molecules with a molecular weight cutoff of 3000 Da, and the filtrate was collected and freeze-dried to obtain polypeptide powder;

[0043] (5) Oat polysaccharide (purchased from Lanzhou Waterless Biotechnology Co., Ltd., 80% purity) was added with water at a material-liquid mass ratio of 1:200, and α-amylase was added for enzymatic hydrolysis. The addition ratio was 0.35% of the mass of the oat polysaccharide. The enzymatic hydrolysis temperature was 58°C, the α-amylase activity was 3800 U / g, the enzymatic hydrolysis time was 80 min, and the oat oligosaccharide powder was obtained by heating at 100°C for 10-15 min to inactivate the enzyme.

[0044] (6) 10 parts by mass of the polypeptide powder obtained in step (4) and 3 parts by mass of the oat oligosaccharide obtained in step (5) were dissolved in 50 parts by mass of a 35% edible ethanol-water solution (the purity of the edible ethanol used in this embodiment is 95%, and the edible ethanol is mixed with water to obtain the edible ethanol-water solution. The subsequent embodiments all adopt this method to obtain the edible ethanol-water solution, which will not be repeated here). The mixture was reacted at 70°C, 0.05 MPa under reduced pressure and heating conditions for 1 hour. 1 part by mass of xylose, 1 part by mass of arginine, 0.5 parts by mass of methionine, and 0.5 parts by mass of lysine were then added, and the mixture was reacted at 80°C, 0.05 MPa under reduced pressure and heating conditions for 0.5 hour to obtain a Maillard product with a salty and fresh taste.

[0045] Comparative Example 1

[0046] (1) The meat and fascia tissue of beef and salmon bones were crushed with a grinder to a particle size of 1 mm. The crushed raw materials were mixed and dispersed with water at a mass ratio of 1:20, and alkaline protease and flavor protein were added for double enzyme hydrolysis. The pH value of the homogenate was maintained at 8.3, the hydrolysis time was 5 h, the hydrolysis temperature was 57° C., the stirring speed was 145 rpm, the amount of alkaline protease added was 3% of the protein content, the amount of bromelain added was 2% of the protein content, the alkaline protease activity was 50,000 U / g, and the bromelain activity was 35,000 U / g;

[0047] (2) The final enzymatic hydrolysate of step (1) was heated at 100°C for 12 minutes to inactivate the enzyme, and the pH was adjusted to 4.1 at room temperature. The supernatant was collected after standing for 3 hours;

[0048] (3) The supernatant obtained in step (2) was filtered using a nanofiltration membrane to retain molecules with a molecular weight cutoff of 3000 Da, the filtrate was collected, and the polypeptide powder was obtained after lyophilization;

[0049] (4) Oat polysaccharide (purchased from Lanzhou Waterless Biotechnology Co., Ltd., 80% purity) was added with water at a material-liquid mass ratio of 1:200, and α-amylase was added for enzymatic hydrolysis. The addition ratio was 0.35% of the mass of the oat polysaccharide. The enzymatic hydrolysis temperature was 58°C, the α-amylase activity was 3800 U / g, the enzymatic hydrolysis time was 80 min, and the oat oligosaccharide powder was obtained by heating at 100°C for 10-15 min to inactivate the enzyme.

[0050] (5) 10 parts by mass of the polypeptide powder obtained in step (3) and the oat oligosaccharide obtained in step (4) are dissolved in 50 parts by mass of a 35% volume fraction edible ethanol aqueous solution (the purity of the edible ethanol is 95%), and reacted at 70° C. to 90° C., 0.03 to 0.06 MPa under reduced pressure and heating conditions for 0.5 to 1 h; then, 1 part by mass of xylose, 1 part by mass of arginine, 0.5 parts by mass of methionine, and 0.5 parts by mass of lysine are added, and reacted at 75° C. to 95° C., 0.03 to 0.06 MPa under reduced pressure and heating conditions for 0.5 to 1 h to obtain a Maillard intermediate.

[0051] Table 1 Sensory scores

[0052]

[0053] Table 1 shows the sensory scores of Example 1 and Comparative Example 1. Compared with Comparative Example 1, Example 1 adopts ultrasonic cooking to remove the fishy smell, while Comparative Example 1 does not take this step. The results show that the sensory evaluation of the product prepared in Comparative Example 2 is poor.

[0054] Comparative Example 2

[0055] (1) Remove the meat and fascia tissue from beef and salmon bones, hammer and crush them into 3 cubic centimeter blocks in equal proportions, add 2.5 times the weight of water, add 1.5% of the weight of the raw materials in 52° liquor, and remove the fishy odor by ultrasonication at a frequency of 100 kHz, a power of 600 W, and a duration of 20 minutes. Finally, heat at 100°C for 40 minutes and remove the raw materials.

[0056] (2) The raw material obtained in step (1) was pulverized with a pulverizer to a particle size of 1 mm. The pulverized raw material was mixed and dispersed with water at a mass ratio of 1:20, and alkaline protease and flavor protein were added for double enzyme hydrolysis. The pH value of the homogenate was maintained at 8.3, the hydrolysis time was 5 h, the hydrolysis temperature was 57° C., the stirring speed was 145 rpm, the amount of alkaline protease added was 3% of the protein content, the amount of bromelain added was 2% of the protein content, the alkaline protease activity was 50,000 U / g, and the bromelain activity was 35,000 U / g;

[0057] (3) The final enzymatic hydrolysate of step (2) was heated at 100°C for 12 min to inactivate the enzyme, and the pH was adjusted to 4.1 at room temperature. The supernatant was collected after standing for 3 h;

[0058] (4) The supernatant obtained in step (3) was filtered using a nanofiltration membrane to retain molecules with a molecular weight cutoff of 3000 Da, and the filtrate was collected and freeze-dried to obtain polypeptide powder;

[0059] (5) Oat polysaccharide (purchased from Lanzhou Waterless Biotechnology Co., Ltd., 80% purity) was added with water at a material-liquid mass ratio of 1:200, and α-amylase was added for enzymatic hydrolysis. The addition ratio was 0.35% of the mass of the oat polysaccharide. The enzymatic hydrolysis temperature was 58°C, the α-amylase activity was 3800 U / g, the enzymatic hydrolysis time was 80 min, and the oat oligosaccharide powder was obtained by heating at 100°C for 10-15 min to inactivate the enzyme.

[0060] (6) 10 parts by mass of the polypeptide powder obtained in step (4) and 3 parts by mass of the oat oligosaccharides obtained in step (5) are dissolved in 50 parts by mass of a 35% edible ethanol-water solution, and heated at 115° C. for 1.5 hours; then 1 part by mass of xylose, 1 part by mass of arginine, 0.5 parts by mass of methionine, and 0.5 parts by mass of lysine are added, and heated at 150° C. for 1 hour to obtain a Maillard product with a salty and fresh taste.

[0061] Table 2 Sensory scores

[0062]

[0063] Table 1 shows the sensory scores of Example 1 and Comparative Example 2. Compared with Comparative Example 2, Example 1 adopts the Maillard reaction under reduced pressure heating at 70°C, 0.05 MPa and 80°C, 0.05 MPa. The results show that the sensory scores of the two are consistent, but the reaction time of reduced pressure heating in Example 1 is shortened.

[0064] Comparative Example 3

[0065] (1) Remove the meat and fascia tissue from beef and salmon bones, hammer and crush them into 3 cubic centimeter blocks in equal proportions, add 2.5 times the weight of water, add 1.5% of the weight of the raw materials in 52° liquor, and remove the fishy odor by ultrasonication at a frequency of 100 kHz, a power of 600 W, and a duration of 20 minutes. Finally, heat at 100°C for 40 minutes and remove the raw materials.

[0066] (2) The raw material obtained in step (1) was crushed with a crusher to a particle size of less than 1 mm. The crushed raw material was mixed and dispersed with water at a mass ratio of 1:20, and alkaline protease was added for enzymatic hydrolysis. The pH value of the homogenate was maintained at 8.3, the enzymatic hydrolysis time was 5 h, the enzymatic hydrolysis temperature was 57° C., the stirring speed was 145 rpm, the amount of alkaline protease added was 3% of the protein content, and the alkaline protease activity was 50,000 U / g.

[0067] (3) The final enzymatic hydrolysate of step (2) was heated at 100°C for 12 min to inactivate the enzyme, and the pH was adjusted to 4.1 at room temperature. The supernatant was collected after standing for 3 h;

[0068] (4) The supernatant obtained in step (3) was filtered using a nanofiltration membrane to retain molecules with a molecular weight cutoff of 3000 Da, and the filtrate was collected and freeze-dried to obtain polypeptide powder;

[0069] (5) Oat polysaccharide (purchased from Lanzhou Waterless Biotechnology Co., Ltd., 80% purity) was added with water at a material-liquid mass ratio of 1:200, and α-amylase was added for enzymatic hydrolysis. The addition ratio was 0.35% of the mass of the oat polysaccharide. The enzymatic hydrolysis temperature was 58°C, the α-amylase activity was 3800 U / g, the enzymatic hydrolysis time was 80 min, and the oat oligosaccharide powder was obtained by heating at 100°C for 10-15 min to inactivate the enzyme.

[0070] (6) 10 parts by mass of the polypeptide powder obtained in step (4) and 3 parts by mass of the oat oligosaccharide obtained in step (5) are dissolved in 50 parts by mass of a 35% edible ethanol-water solution, and reacted at 70°C, 0.05 MPa, reduced pressure heating conditions for 0.5 h; then, 1 part by mass of xylose, 1 part by mass of arginine, 0.5 parts by mass of methionine, and 0.5 parts by mass of lysine are added, and the mixture is reacted at 80°C, 0.05 MPa, reduced pressure heating conditions for 0.5 h to obtain a Maillard product with a salty and fresh taste.

[0071] Comparative Example 4

[0072] (1) Remove the meat and fascia tissue from beef and salmon bones, hammer and crush them into 3 cubic centimeter blocks in equal proportions, add 2.5 times the weight of water, add 1.5% of the weight of the raw materials in 52° liquor, and remove the fishy odor by ultrasonication at a frequency of 100 kHz, a power of 600 W, and a duration of 20 minutes. Finally, heat at 100°C for 40 minutes and remove the raw materials.

[0073] (2) The raw material obtained in step (1) is crushed with a crusher to a particle size of less than 1 mm after crushing. The crushed raw material is mixed and dispersed with water at a mass ratio of 1:20, and flavor protein is added for double enzyme hydrolysis. The pH value of the homogenate is maintained at 8.3, the hydrolysis time is 5 h, the hydrolysis temperature is 57° C., the stirring speed is 145 rpm, the amount of bromelain added is 2% of the protein content, and the bromelain activity is 35,000 U / g;

[0074] (3) The final enzymatic hydrolysate of step (2) was heated at 100°C for 12 min to inactivate the enzyme, and the pH was adjusted to 4.1 at room temperature. The supernatant was collected after standing for 3 h;

[0075] (4) The supernatant obtained in step (3) was filtered using a nanofiltration membrane to retain molecules with a molecular weight cutoff of 3000 Da, and the filtrate was collected and freeze-dried to obtain polypeptide powder;

[0076] (5) 10 parts by mass of the polypeptide powder obtained in step (4) and 3 parts by mass of oat polysaccharide are dissolved in 50 parts by mass of a 35% edible ethanol-water solution, and reacted at 70°C, 0.05 MPa under reduced pressure and heating conditions for 1 hour; then 1 part by mass of xylose, 1 part by mass of arginine, 0.5 parts by mass of methionine, and 0.5 parts by mass of lysine are added, and the mixture is reacted at 80°C, 0.05 MPa under reduced pressure and heating conditions for 0.5 hour to obtain a Maillard product with a salty and fresh taste.

[0077] Table 3 Hydrolysis degree and sensory score

[0078] Group Degree of hydrolysis Color sensory score Odor sensory score Taste sensory score Weighted Scoring Example 1 29.1 90 90 90 90 Comparative Example 3 19.2 80 70 70 73 Comparative Example 4 29.3 70 70 80 74

[0079] The results showed that compared with Example 1 and Comparative Example 3, Example 1 used alkaline protease and bromelain dual enzymatic hydrolysis, while Comparative Example 3 used alkaline protease, and its hydrolysis degree was lower than that of Example 1, and the Maillard taste of Comparative Example 3 was slightly bitter, and the odor sensory score and taste sensory score of Comparative Example 3 were both lower than those of Example 1. Therefore, alkaline protease and bromelain dual enzymatic hydrolysis can increase the degree of protein hydrolysis and also enhance the sensory flavor of the Maillard product; compared with Example 1, Example 1 hydrolyzed oat polysaccharides with α-amylase to obtain reducing sugars and then subjected to the Maillard reaction, while Comparative Example 4 directly used oat polysaccharides for the Maillard reaction. The results showed that the product of the Maillard reaction in Comparative Example 4 was lighter in color, and the sensory odor and taste were lower than those of the Maillard product of oat oligosaccharides in Example 1.

[0080] Example 2

[0081] (1) Remove the meat and fascia tissue from beef and salmon bones, hammer and crush them into 3 cubic centimeter blocks in equal proportions, add 2.5 times the weight of water, add 1.5% of the weight of the raw materials in 52° liquor, and remove the fishy odor by ultrasonication at a frequency of 100 kHz, a power of 600 W, and a duration of 20 minutes. Finally, heat at 100°C for 40 minutes and remove the raw materials.

[0082] (2) The raw material obtained in step (1) is pulverized with a pulverizer until the particle size of the raw material is less than 1 mm. The pulverized raw material is mixed and dispersed with water at a mass ratio of 1:20, and alkaline protease and flavor protein are added for double enzyme hydrolysis. The pH value of the homogenate is maintained at 8.3, the hydrolysis time is 3 hours, the hydrolysis temperature is 57° C., the stirring speed is 145 rpm, the amount of alkaline protease added is 3% of the protein content, the amount of bromelain added is 2% of the protein content, the alkaline protease activity is 50,000 U / g, and the bromelain activity is 35,000 U / g;

[0083] (3) The final enzymatic hydrolysate of step (2) was heated at 100°C for 12 min to inactivate the enzyme, and the pH was adjusted to 4.1 at room temperature. The supernatant was collected after standing for 3 h;

[0084] (4) The supernatant obtained in step (3) was filtered using a nanofiltration membrane to retain molecules with a molecular weight cutoff of 3000 Da, and the filtrate was collected and freeze-dried to obtain polypeptide powder;

[0085] (5) Oat polysaccharide (purchased from Lanzhou Waterless Biotechnology Co., Ltd., 80% purity) was added with water at a material-liquid mass ratio of 1:200, and α-amylase was added for enzymatic hydrolysis. The addition ratio was 0.35% of the mass of the oat polysaccharide. The enzymatic hydrolysis temperature was 58°C, the α-amylase activity was 3800 U / g, the enzymatic hydrolysis time was 60 min, and the oat oligosaccharide powder was obtained by heating at 100°C for 10-15 min to inactivate the enzyme.

[0086] (6) 10 parts by mass of the polypeptide powder obtained in step (4) and 3 parts by mass of the oat oligosaccharide obtained in step (5) are dissolved in 50 parts by mass of a 35% edible ethanol-water solution, and the mixture is reacted at 70° C. and 0.05 MPa under reduced pressure and heating conditions for 1 hour; then 1 part by mass of xylose, 1 part by mass of arginine, 0.5 parts by mass of methionine, and 0.5 parts by mass of lysine are added, and the mixture is reacted at 80° C. and 0.05 MPa under reduced pressure and heating conditions for 0.5 hour to obtain a Maillard product with a salty and fresh taste.

[0087] Table 4 Sensory scores

[0088] Group Color sensory score Odor sensory score Taste sensory score Weighted Scoring Example 1 90 90 90 90 Example 2 90 90 80 87

[0089] Compared with Example 1, the enzymatic hydrolysis time of the protein and oat polysaccharide in Example 2 was slightly shorter than that in Example 1. The results showed that the hydrolysis degree and taste sensory score of Example 2 were slightly lower than those of Example 1. Therefore, Example 1 was the best preparation process scheme in comparison.

[0090] Table 5 Maillard product UV absorption

[0091] Group 290nm 420nm Example 1 0.799 1.020 Example 2 0.786 0.934

[0092] Compared with Example 2, Example 1 has the highest absorbance at 290nm and 420nm, wherein 290nm is the absorption peak of the hydroxymethylfurfural product, and 420nm is the maximum absorption peak of the melanoidin generated by the carbonylamine reaction, indicating that Example 1 has a high content of Maillard intermediates.

[0093] Experimental results:

[0094] The main flavor substances of the Maillard intermediate products of beef bone and salmon bone peptide-oat oligosaccharide are shown in Table 1. Table 5 shows the flavor substances of the Maillard product in Example 1 determined by GC-MS.

[0095] Table 6 Types and contents of Maillard products

[0096]

[0097]

[0098] As shown in the GC-MS results in Table 6, the main flavor compounds in the Maillard intermediates of the present invention's ox bone, tripterygium peptide, and oat oligosaccharides include alcohols, aldehydes, ketones, esters, phenols, nitrogen-containing compounds, acids, furans, and alkanes. Among these, 1-undecanol is the highest content among alcohols, phenylacetaldehyde is the highest among aldehydes, p-ethylphenol is the highest among phenols, and tetramethylpyrazine is the highest among nitrogen-containing compounds. The total flavor compound content is 150.34 μg / g, significantly higher than that of products produced by conventional methods. Therefore, the present method can safely and environmentally friendly produce products with a rich flavor and a fresh, fragrant, and mellow taste, improving the functional properties and utilization of the raw materials.

[0099] The above-described embodiments merely represent several implementation methods of the present invention. While the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that a person skilled in the art would be able to make various modifications and improvements without departing from the spirit of the present invention, all of which fall within the scope of protection of the present invention. Therefore, the scope of protection of the present invention shall be determined by the appended claims.

Claims

1. A method for preparing a Maillard intermediate of cattle bone, salmon bone peptide and oat oligosaccharide based on reduced pressure heating, characterized in that: The following steps are involved: (1) Remove the meat and fascia from beef and salmon bones, hammer and crush them in equal proportions, add 2.5 times the weight of the raw materials with water and 1.5% of the weight of the raw materials with 52° liquor, remove the fishy odor with ultrasonication, and finally heat and steam at 100°C for 30-60 minutes and remove the raw materials; (2) The raw material obtained in step (1) is crushed with a grinder, and then mixed and dispersed with water in a mass ratio of 1:20 to 1:25, the pH of the dispersion is adjusted to 8.0 to 9.0, and alkaline protease and bromelain are added for dual enzyme hydrolysis, and the enzymatic hydrolysis is carried out for 4 to 8 hours while stirring throughout the process; In step (2), the enzymolysis temperature is 50° C. to 60° C., the stirring speed during the enzymolysis process is 100 to 200 rpm, the amount of alkaline protease added is 0.25 to 0.35% of the weight of the crushed raw material, the amount of bromelain added is 0.15 to 0.25% of the weight of the crushed raw material, the alkaline protease activity is 40,000 to 70,000 U / g, and the bromelain activity is 20,000 to 40,000 U / g; (3) The final enzymatic hydrolyzate obtained in step (2) was heated at 100° C. for 10 to 15 minutes to inactivate the enzyme, then cooled to room temperature, and the pH was adjusted to 4.0 to 5.

0. The supernatant was collected after standing; (4) The supernatant obtained in step (3) was filtered using a nanofiltration membrane to retain molecules with a molecular weight cutoff of 3000 Da, the filtrate was collected, and the polypeptide powder was obtained after lyophilization; (5) Oat polysaccharide was added to water at a material-liquid mass ratio of 1:200, and α-amylase was added for enzymatic hydrolysis. The mixture was heated at 100°C for 10-15 minutes to inactivate the enzyme, and then freeze-dried to obtain oat oligosaccharide powder; The purity of oat polysaccharide in step (5) is 80%; In step (5), the addition ratio of α-amylase is 0.2% to 0.5% of the mass of oat polysaccharide, the activity of α-amylase is 3000U / g to 5000U / g, the enzymolysis temperature is 45°C to 60°C, and the enzymolysis time is 60min to 80min; (6) dissolving 10 parts by mass of the polypeptide powder obtained in step (4) and 2 to 3 parts by mass of the oat oligosaccharide obtained in step (5) in 50 parts by mass of an edible ethanol-water mixture, and reacting them at 70° C. to 90° C., 0.03 to 0.06 MPa under reduced pressure and heating conditions for 0.5 to 1 hour; then adding 1 part by mass of xylose, 1 part by mass of arginine, 0.5 parts by mass of methionine, and 0.5 parts by mass of lysine, and reacting them at 75 to 95° C., 0.03 to 0.06 MPa under reduced pressure and heating conditions for 0.5 to 1 hour to obtain a Maillard intermediate; In step (6), the purity of edible ethanol is 95%; in the edible ethanol-water mixture, the volume percentage of edible ethanol is 20% to 40%.

2. The preparation method according to claim 1, characterized in that In step (1), the size of the raw material after being hammered and crushed is 3-5 cubic centimeters.

3. The preparation method according to claim 1, characterized in that: The ultrasonic method in step (1) is: ultrasonic frequency is 100-200kHz, ultrasonic power is 500-800W, and ultrasonic time is 10-30min.

4. The preparation method according to claim 1, characterized in that In step (2), the raw material is crushed by a crusher to a particle size of less than 1 mm.

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