Preparation method of flavor enhancing base powder of bovine bone peptide Maillard reaction product

Abstract

The present invention relates to the technical field of food processing, and discloses a preparation method of a flavor-enhancing base powder of a bovine bone peptide Maillard reaction product, comprising the following steps: cutting and crushing beef bones with meat, subjecting them to ultrasonic cleaning, enzymatic hydrolysis, inactivation, freeze-drying to collect bovine bone peptides, then reacting the peptides with oligosaccharides and amino acids through Maillard reaction to obtain a umami intermediate product, and then adding an excipient and spray-drying to obtain the base powder. The present invention obtains a preparation method of a Maillard intermediate product using bovine bone peptides and wheat bran oligosaccharides as the main reaction raw materials. This method is safe and efficient, and the prepared product has a strong mellow flavor characteristic, and can be used as a basic ingredient powder in seasonings.

Description

Technical Field

[0001] The present invention relates to the technical field of food processing, and relates to a preparation method of a flavor base powder of a Maillard reaction product of bovine bone peptides. Background Art

[0002] Maillard products play an important role in enhancing food flavors and have been gradually applied to the production of flavors or seasonings in recent years. However, there is a lack of flavor products with compounded taste and innovative flavors on the market. Industrialized composite products with the characteristics of various seasonings have the advantages of nutrition, flavor diversity, and convenience.

[0003] Bovine bones mainly contain substances such as water, protein, lipids, calcium, and phosphorus. More than 80% of its protein is collagen, and the enzymatic hydrolysis products contain 8 essential amino acids required by the human body, with high biological potency. CN201911276918.8 discloses a preparation method of "Bovine Bone Peptides and Their Preparation Process". However, the polypeptides prepared by this method have a plain flavor and cannot be applied to food seasonings. Currently, the substrates for the Maillard reaction mostly use peptides or amino acids to provide amino groups and react with reducing monosaccharides or disaccharides to provide carbonyl groups in a single reaction. For example, arginine, lysine, xylose, glucose, fructose, etc. are added to the reaction system. Although this method can obtain relatively strong flavor substances, the resulting products are mainly rich in furan and pyrazine flavor substances, while the contents of other flavor substances such as esters, aldehydes, and ketones are relatively low. Therefore, broadening the source of reaction substrates and optimizing the reaction system to increase the contents of products such as lipids and ketones and improve the balance of reaction products are the keys to obtaining products with strong flavors and rich aromas.

[0004] Regarding the preparation of seasonings using bovine bones as raw materials, CN108968004A, "A High-Temperature-Resistant Microencapsulated Beef Flavor and Its Preparation Method", discloses a preparation method of a Maillard flavor with a ratio of 20 - 40 parts of beef enzymatic hydrolysis solution, 0.2 - 2 parts of bovine bone oil, 6 - 20 parts of gum arabic, 0.5 - 2 parts of xylose, 0.5 - 2 parts of hydrolyzed vegetable protein seasoning powder, 2 - 7 parts of table salt, 2 - 5 parts of monosodium glutamate, 1 - 3 parts of white sugar, 1 - 6 parts of β-cyclodextrin, 0.1 - 0.4 parts of thiamine hydrochloride, 0.1 - 0.2 parts of cinnamon powder, 10 - 30 parts of maltodextrin, 0.5 - 5 parts of beef flavor base, and 50 - 60 parts of deionized water. However, this method uses beef shanks, and the required production cost is relatively high. Summary of the Invention

[0005] To develop bovine bone resources, the objective of the present invention is to provide a preparation method for a flavor-enhancing base powder of a Maillard reaction product of bovine bone peptides. After cutting and dispersing bovine bones with meat and fascia tissues removed, they are ultrasonically cleaned, enzymatically hydrolyzed, inactivated, and bovine bone oligopeptides are collected. Then, the oligopeptides are subjected to a Maillard reaction with wheat bran oligosaccharides and amino acids to obtain a Maillard intermediate product with more flavor substances such as esters, aldehydes, and ketones. Finally, an excipient is added and spray-dried to obtain the base powder.

[0006] To achieve the technical objectives of the present invention, the following technical solutions are provided:

[0007] A preparation method for a flavor-enhancing base powder of a Maillard reaction product of bovine bone peptides, comprising the following steps:

[0008] (1) After hammering and crushing the beef-containing bovine bones, add 2.5 times the weight of the raw materials in water and 1.5% by weight of the raw materials of 52° liquor, use ultrasonic waves to remove the fishy substances, and finally heat and cook at 100°C for 30 - 60 min and remove the raw materials.

[0009] (2) Crush the raw materials obtained in step (1) using a pulverizer, then mix and disperse them with water at a mass ratio of 1:20 - 1:25, adjust the pH of the dispersion to 8.0 - 9.0, add alkaline protease and flavor protease for double-enzyme hydrolysis, and continuously stir and hydrolyze for 4 - 8 h throughout the process.

[0010] (3) Heat the final enzyme hydrolysis solution obtained in step (2) at 100°C for 10 - 15 min for enzyme inactivation treatment, then cool to room temperature and adjust the pH to 4.0 - 5.0, and take the supernatant after standing.

[0011] (4) The supernatant obtained in step (3) is filtered through a nanofiltration membrane to retain molecules with a molecular weight of 1000 Da, the filtrate is collected, and freeze-dried to obtain a polypeptide powder.

[0012] (5) Add wheat bran polysaccharide to water at a mass ratio of 1:200 of the feed liquid, add α-amylase for enzymatic hydrolysis, and after heat treatment at 100°C for 10 - 15 min for enzyme inactivation, a wheat bran oligosaccharide solution is obtained.

[0013] (6) Dissolve 10 parts by mass of the polypeptide powder obtained in step (4) in 20 - 30 parts by mass of the wheat bran oligosaccharide solution obtained in step (5), add edible ethanol, adjust the pH to 8.0 - 9.0, and heat and react at 110°C - 130°C for 1 - 2 h; then add 1 part by mass of xylose, 1 part by mass of arginine, and 0.5 part by mass of lysine, and heat and react at 130 - 155°C for 1 - 2 h to obtain a Maillard intermediate product solution of bovine bone oligopeptide - wheat bran oligosaccharide.

[0014] (7) Mix 100 parts by mass of the Maillard intermediate product solution obtained in step (6) with 0.5 part by mass of alginate, 0.5 part by mass of corn flour, 0.5 part by mass of cyclodextrin, 0.5 part by mass of arginine, 0.2 part by mass of lysine, 0.3 part by mass of sodium chloride, 0.2 part by mass of potassium chloride, and 0.5 part by mass of shrimp oil, and homogenize at 3000 rpm for 10 - 20 minutes to obtain a homogenized liquid;

[0015] (8) Spray-dry the homogenized liquid obtained in step (7) to obtain a flavor-enhancing base powder of the Maillard reaction product of bovine bone peptides; the inlet temperature of spray-drying is 140 - 165 °C, the outlet temperature is 100 - 110 °C, and the feeding flow rate is 4 - 6 mL / min.

[0016] 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.

[0017] As a preferred embodiment of the present invention, in step (1), the ultrasonic method is: the ultrasonic frequency is 100 - 200 kHz, the ultrasonic power is 500 - 800 W, and the ultrasonic action time is 10 - 30 min.

[0018] As a preferred embodiment of the present invention, in step (2), the particle size of the raw material after being pulverized by a pulverizer is less than 1 mm

[0019] As a preferred embodiment of the present invention, in step (2), the enzymatic hydrolysis temperature is 50 °C - 60 °C, the stirring speed during the enzymatic hydrolysis process is 100 - 200 rpm, the addition amount of alkaline protease is 0.25 - 0.35% of the mass of the pulverized raw material, the addition amount of flavor protease is 0.15 - 0.25% of the mass of the pulverized raw material, the enzymatic activity of alkaline protease is 40000 - 70000 U / g, and the enzymatic activity of flavor protease is 20000 - 40000 U / g.

[0020] The purity of the wheat bran polysaccharide in step (5) is 80%.

[0021] As a preferred embodiment of the present invention, in step (5), the addition ratio of α-amylase is 0.2% - 0.5% of the mass of the wheat bran polysaccharide, the activity of α-amylase is 3000 U / g - 5000 U / g, the enzymatic hydrolysis temperature is 45 °C - 60 °C, and the enzymatic hydrolysis time is 60 min - 80 min.

[0022] As a preferred embodiment of the present invention, in step (6), the purity of the edible ethanol is 95%; in the mixed solution of polypeptide powder, wheat bran oligosaccharide solution and edible ethanol, the volume percentage of edible ethanol is 20 - 40%.

[0023] The present invention also provides a flavor-enhancing base powder of the Maillard reaction product of bovine bone peptides prepared by the above method.

[0024] The present invention also provides the application of the flavor-enhancing base powder of the Maillard reaction product of bovine bone peptides in the preparation of flavor conditioning powder.

[0025] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0026] (1) In the present invention, through the Maillard reaction of bovine bone hydrolyzate and wheat bran oligosaccharide under reduced pressure heating, the preparation time is shortened, the production cost of the product is low, and the obtained Maillard intermediate product has a salty and umami taste.

[0027] (2) After adding white liquor and using ultrasonic treatment in the present invention, the fishy substances in bovine bone are effectively reduced. Adding exogenous amino acids is to enhance the reaction intensity and enrich the flavor; adding shrimp oil is to promote the generation of flavor substances and utilize lipid aroma production.

[0028] (3) The base powder obtained by spray-drying the Maillard reaction product mainly composed of bovine bone peptides and wheat bran oligosaccharide in the present invention, after adding excipients, has the characteristics of rich fragrance, strong and lasting flavor, and can be further applied to flavor conditioning powder.

[0029] (4) In the present invention, adding white liquor and ultrasonic treatment are used to remove the fishy smell of bovine bone raw materials, improving the product flavor from the source.

[0030] (5) The wheat bran oligosaccharide used in the present invention mainly contains more than 30% glucose, and also contains various other reducing sugars that are the sources of Maillard flavor products, which can make the product flavor more mellow. In addition, a variety of compound flavor amino acids are added during the reaction to promote flavor enhancement, and the obtained intermediate product has a strong flavor. This Maillard method is safe, green, and can further improve the functional characteristics and utilization rate of raw materials, expanding its application fields. Description of the Drawings

[0031] Figure 1 It is the production process flow chart of the present invention. Detailed Embodiments

[0032] The following further describes the detailed embodiments of the present invention.

[0033] Unless otherwise specified, the wheat bran polysaccharide used in the present invention is purchased from Lanzhou Wateles Biotech Co., Ltd., and other raw materials can be obtained from the market or are commonly used in the art. Unless otherwise specified, the methods in the following examples are all conventional methods in the art.

[0034] Determination of Degree of Protein Hydrolysis

[0035] Dissolve the reagent of 2,4,6-trinitrobenzenesulfonic acid (TNBS) with a content of 0.1% (w / w) in water. All undigested samples, digested samples, and standard solutions are dissolved in 0.1% (w / w) SDS. In a test tube, add 2.0 mL of sodium phosphate buffer with a concentration of 0.2125 M and pH 8.2, and then add 0.25 mL of the sample or standard solution with the same concentration. Add 2.0 mL of TNBS reagent to each centrifuge tube, then mix with an oscillator and incubate in a dark water bath at 50 °C for 60 min. After covering the incubation, add 4.0 mL of 0.1 M HCl to each tube to terminate the reaction. Then cool at room temperature for 30 min, and finally measure the absorbance value at 340 nm with a spectrophotometer. Calculate the proteolysis rate according to the following formula:

[0036]

[0037] AN1: Nitrogen content of the undigested matrix;

[0038] AN2: Nitrogen content of the matrix in the digested sample;

[0039] Npb: Nitrogen content of peptide bonds in matrix proteins. The unit is mg / g protein.

[0040] Sensory evaluation

[0041]

[0042]

[0043] Weighted sensory score = 30% of color sensory score + 30% of odor sensory score + 40% of taste sensory score

[0044] UV determination of Maillard products

[0045] Mix and dilute the obtained Maillard intermediate product liquid with water, dilute it 10 to 100 times respectively and make up the volume, and measure the ultraviolet-visible absorption value at 290 nm and 420 nm.

[0046] GC-MS determination of flavor substances

[0047] Extraction conditions: Dissolve about 2 g of the Maillard intermediate product in 25 mL of saturated brine, add 20 μL of 2-methyl-3-heptanone (0.8 mg / mL), mix well and seal. Use a syringe to adsorb for 30 min at 60 °C and then inject the sample; Temperature programming: Keep at 35 °C for 3 min, increase the temperature at 3 °C·min -1 Increase the temperature to 40 °C and keep for 1 min, then increase the temperature at 5 °C·min -1 Increase the temperature to 230 °C and keep for 12 min.

[0048] GC-MS conditions: Volatile compounds were separated on a TG-5SICMS column (30m×0.25mm, 0.25μm); carrier gas He, flow rate 1mL / min, split ratio 20:1 (V:V); electron impact (EI) ionization source, ion source temperature: 285℃, transmission temperature: 255℃; scanning mass range: 35~450amu, scanning rate 0.1amu / s. The internal standard method was used to semi-quantitatively analyze the volatile components and determine the mass concentration of each volatile component.

[0049] Example 1

[0050] (1) Hammer and crush beef bones into 3 cubic centimeter blocks, add 2.5 times the weight of the raw material water and 1.5% of the weight of the raw material 52° liquor, use ultrasound to remove the fishy odor substances, the ultrasound frequency is 100 kHz, the ultrasound power is 600 W, the ultrasound method is ultrasound for 20 minutes, and finally heat at 100° C. for 40 minutes to remove the raw material;

[0051] (2) The raw material obtained in step (1) is crushed by a crusher, and the particle size of the raw material after crushing is less than 1 mm. The crushed raw material is dispersed in water at a solid-liquid ratio of 1:20 (w / w), and alkaline protease and flavor protease are added for double enzymatic hydrolysis. The pH value of the homogenate is maintained at 8.3, the enzymatic hydrolysis time is 5 h, the enzymatic hydrolysis temperature is 57° C., the stirring speed is 130 rpm, the amount of alkaline protease added is 0.3% of the mass of the crushed raw material, the amount of flavor protease added is 0.2% of the mass of the crushed raw material, the alkaline protease activity is 50,000 U / g, and the flavor protease activity is 35,000 U / g;

[0052] (3) The final enzymatic hydrolyzate 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;

[0053] (4) The supernatant obtained in step (3) is filtered through a nanofiltration membrane to retain molecules with a molecular weight cutoff of 1000 Da, the filtrate is collected, and lyophilized to obtain polypeptide powder;

[0054] (5) adding water to wheat bran polysaccharide (80% purity, purchased from Lanzhou Waters Biotechnology Co., Ltd.) at a material-liquid mass ratio of 1:200, adding α-amylase for enzymolysis, the addition ratio being 0.35% of the mass of the wheat bran polysaccharide, the enzymolysis temperature being 58° C., the α-amylase activity being 3800 U / g, the enzymolysis time being 80 min, and heating at 100° C. for 10 to 15 min to inactivate the enzyme to obtain wheat bran oligosaccharides;

[0055] (6) Dissolve 10 parts by mass of the polypeptide powder obtained in step (4) in 30 parts by mass of the wheat bran oligosaccharide obtained in step (5), and add edible ethanol; in the mixed solution of the polypeptide powder, the wheat bran oligosaccharide solution and the edible ethanol, the volume percentage of the edible ethanol is 25%; in this example, the purity of the edible ethanol is 95%, and heat and react at 115 °C for 1.5 h; then add 1 part by mass of xylose, 1 part by mass of arginine, and 0.5 part by mass of lysine, and heat and react at 150 °C for 1 h to obtain a Maillard product solution.

[0056] (7) For the Maillard product solution obtained in step (6), according to 100 parts by mass of the Maillard product solution, 0.5 part by mass of alginate, 0.5 part by mass of corn flour, 0.5 part by mass of cyclodextrin, 0.5 part by mass of arginine, 0.2 part by mass of lysine, 0.3 part by mass of sodium chloride, 0.2 part by mass of potassium chloride, and 0.5 part by mass of shrimp oil, homogenize at 3000 rpm for 10 - 20 minutes;

[0057] (8) Spray-dry the homogenate obtained in step (7), with the inlet temperature of spray drying being 145 °C, the outlet temperature being 105 °C, and the feeding flow rate being 4.5 mL / min.

[0058] Comparative Example 1

[0059] (1) Hammer the beef bones with meat attached, then crush them with a pulverizer. After crushing, the particle size of the raw material is less than 1 mm. Disperse the crushed raw material in water at a material-liquid ratio (w / w) of 1:20, add alkaline protease and flavor protease for double enzyme hydrolysis, keep the pH value of the homogenate at 8.3, the enzyme hydrolysis time is 5 h, the enzyme hydrolysis temperature is 57 °C, the stirring speed is 130 rpm, the addition amount of alkaline protease is 0.3% of the mass of the crushed raw material, the addition amount of flavor protease is 0.2% of the mass of the crushed raw material, the enzyme activity of alkaline protease is 50000 U / g, and the enzyme activity of flavor protease is 35000 U / g;

[0060] (2) Heat the final enzyme hydrolysis solution obtained in step (1) at 100 °C for 12 min for enzyme inactivation treatment, adjust the pH to 4.1 at room temperature, and take the supernatant after standing for 3 h;

[0061] (3) Filter the supernatant obtained in step (2) through a nanofiltration membrane to retain molecules with a molecular weight cut-off of 1000 Da, collect the filtrate, and obtain the polypeptide powder after freeze-drying;

[0062] (4) Add 1 part by mass of xylose, 1 part by mass of arginine, and 0.5 part by mass of lysine to 10 parts by mass of the polypeptide powder obtained in step (3), and heat and react at 150 °C for 1 h to obtain a Maillard product solution.

[0063] (5) Homogenize the Maillard product solution obtained in step (4) at 3000 rpm for 10 - 20 minutes according to 100 parts by mass of the Maillard product solution, 0.5 parts by mass of alginate, 0.5 parts by mass of corn starch, 0.5 parts by mass of cyclodextrin, 0.5 parts by mass of arginine, 0.2 parts by mass of lysine, 0.3 parts by mass of sodium chloride, 0.2 parts by mass of potassium chloride, and 0.5 parts by mass of shrimp oil.

[0064] (6) Spray - dry the homogenate obtained in step (5). The inlet temperature of spray - drying is 145 °C, the outlet temperature is 105 °C, and the feeding flow rate is 4.5 mL / min.

[0065] Table 1 Sensory Score

[0066] Group Color Sensory Score Odor Sensory Score Taste Sensory Score Weighted Score Example 1 90 90 90 90 Comparative Example 1 80 70 70 73

[0067] The results show that compared with Comparative Example 1, in Example 1, after chopping the beef bones, 52° liquor with a content of 1.5% was added for ultrasonic treatment to remove fishy substances. The ultrasonic frequency was 100 kHz, the ultrasonic power was 600 W, and the ultrasonic treatment time was 20 min. And wheat bran oligosaccharide was added in advance during the Maillard reaction, and then xylose, arginine, and lysine were added for the secondary Maillard reaction. While in Comparative Example 1, the above two steps were missing. The finally obtained Maillard product had a fishy smell and a little bitter substance. The scores of the odor sensory evaluation and the taste sensory evaluation were both 70 points, and the sensory score was significantly lower than that of Example 1. Therefore, adding 1.5% of 52° liquor, using 100 kHz, 600 W, ultrasonic treatment for 20 min, and adding wheat bran oligosaccharide for pre - Maillard reaction are beneficial to improving the odor and taste of the product.

[0068] Comparative Example 2

[0069] (1) Hammer and break the beef bones with meat into pieces of 3 cubic centimeters in size, add 2.5 times the mass of the raw materials of water and 1.5% of the mass of the raw materials of 52° liquor, use ultrasonic to remove fishy substances. The ultrasonic frequency is 100 kHz, the ultrasonic power is 600 W, and the ultrasonic treatment method is ultrasonic treatment for 20 min. Finally, heat at 100 °C for 40 min and then fish out the raw materials; (2) Crush the raw materials obtained in step (1) with a pulverizer. The particle size of the crushed raw materials is less than 1 mm. Disperse the crushed raw materials in water at a solid - liquid ratio (w / w) of 1:20, add alkaline protease for enzymatic hydrolysis. Keep the pH value of the homogenate at 8.3, the enzymatic hydrolysis time is 5 h, the enzymatic hydrolysis temperature is 57 °C, the stirring speed is 130 rpm, the addition amount of alkaline protease is 3% of the mass of the crushed raw materials, and the enzymatic activity of alkaline protease is 50000 U / g. (3) Heat the final enzymatic hydrolysate obtained in step (2) at 100 °C for 12 min to inactivate the enzyme, adjust the pH to 4.1 at room temperature, let it stand for 3 h, and then take the supernatant.

[0070] (4) The supernatant obtained in step (3) is filtered through a nanofiltration membrane to retain molecules with a molecular weight cut-off of 1000 Da. The filtrate is collected and freeze-dried to obtain a polypeptide powder;

[0071] (5) Wheat bran polysaccharide (80% purity, purchased from Lanzhou Woteles Biotechnology Co., Ltd.) is added to water at a solid-liquid mass ratio of 1:200, and α-amylase is added for enzymatic hydrolysis. The addition ratio is 0.35% of the mass of wheat bran polysaccharide. The enzymatic hydrolysis temperature is 58 °C, the activity of α-amylase is 3800 U / g, and the enzymatic hydrolysis time is 80 min. After heat inactivation at 100 °C for 10 - 15 min, wheat bran oligosaccharide is obtained;

[0072] (6) 10 parts by mass of the polypeptide powder obtained in step (4) is dissolved in 30 parts by mass of the wheat bran oligosaccharide obtained in step (5), and edible ethanol is added. In the mixed solution of the polypeptide powder, the wheat bran oligosaccharide solution, and edible ethanol, the volume percentage of edible ethanol is 25%; the purity of edible ethanol in this comparative example is 95%. The mixture is heated and reacted at 115 °C for 1.5 h; then 1 part by mass of xylose, 1 part by mass of arginine, and 0.5 part by mass of lysine are added, and the mixture is heated and reacted at 150 °C for 1 h to obtain a Maillard reaction product solution.

[0073] (7) For the Maillard reaction product solution obtained in step (6), according to 100 parts by mass of the Maillard reaction product solution, 0.5 part by mass of alginate, 0.5 part by mass of corn starch, 0.5 part by mass of cyclodextrin, 0.5 part by mass of arginine, 0.2 part by mass of lysine, 0.3 part by mass of sodium chloride, 0.2 part by mass of potassium chloride, and 0.5 part by mass of shrimp oil, homogenize at 3000 rpm for 10 - 20 minutes;

[0074] (8) The homogenate obtained in step (7) is spray-dried. The inlet temperature of the spray dryer is 145 °C, the outlet temperature is 105 °C, and the feed flow rate is 4.5 mL / min.

[0075] Comparative Example 3

[0076] (1) The beef bones with meat are broken and crushed into blocks of 3 cubic centimeters in size, 2.5 times the weight of the raw material of water and 1.5% of the weight of the raw material of 52° liquor are added, and the fishy smell substances are removed by ultrasonic wave. The ultrasonic frequency is 100 kHz, the ultrasonic power is 600 W, the ultrasonic mode is ultrasonic for 20 min, and finally the raw materials are fished out after heating at 100 °C for 40 min; (2) The beef bones obtained in step (1) are pulverized by a pulverizer. After pulverization, the particle size of the raw materials is less than 1 mm. The pulverized raw materials are dispersed in water at a solid-liquid ratio of 1:20 (w / w), and flavor protease is added for enzymatic hydrolysis. The pH value of the homogenate is maintained at 8.3, the enzymatic hydrolysis time is 5 h, the enzymatic hydrolysis temperature is 57 °C, the stirring speed is 130 rpm, the addition amount of flavor protease is 2% of the mass of the pulverized raw materials, and the enzyme activity of flavor protease is 35000 U / g;

[0077] (3) The final enzymatic hydrolyzate 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;

[0078] (4) The supernatant obtained in step (3) is filtered through a nanofiltration membrane to retain molecules with a molecular weight cutoff of 1000 Da, the filtrate is collected, and lyophilized to obtain polypeptide powder;

[0079] (5) adding water to wheat bran polysaccharide (80% purity, purchased from Lanzhou Waters Biotechnology Co., Ltd.) at a material-liquid mass ratio of 1:200, adding α-amylase for enzymolysis, the addition ratio being 0.35% of the mass of the wheat bran polysaccharide, the enzymolysis temperature being 58° C., the α-amylase activity being 3800 U / g, the enzymolysis time being 80 min, and heating at 100° C. for 10 to 15 min to inactivate the enzyme to obtain wheat bran oligosaccharides;

[0080] (6) 10 parts by weight of the polypeptide powder obtained in step (4) are dissolved in 30 parts by weight of the wheat bran oligosaccharides obtained in step (5), and edible ethanol is added; in the mixed solution of the polypeptide powder, the wheat bran oligosaccharide solution and the edible ethanol, the volume percentage of the edible ethanol is 25%; the purity of the edible ethanol in this comparative example is 95%, and the mixture is heated at 115° C. for 1.5 h; then 1 part by weight of xylose, 1 part by weight of arginine and 0.5 part by weight of lysine are added, and the mixture is heated at 150° C. for 1 h to obtain a Maillard product solution.

[0081] (7) homogenizing the Maillard product solution obtained in step (6) at 3000 rpm for 10 to 20 minutes according to 100 parts by mass of the Maillard product solution, 0.5 parts by mass of alginate, 0.5 parts by mass of corn flour, 0.5 parts by mass of cyclodextrin, 0.5 parts by mass of arginine, 0.2 parts by mass of lysine, 0.3 parts by mass of sodium chloride, 0.2 parts by mass of potassium chloride, and 0.5 parts by mass of shrimp oil;

[0082] (8) The homogenized liquid obtained in step (7) was spray dried, with an inlet temperature of 145° C., an outlet temperature of 105° C., and a feed flow rate of 4.5 mL / min.

[0083] Table 2 Hydrolysis degree and sensory score

[0084]

[0085]

[0086] The results are shown in Table 2. Compared with Comparative Examples 2 and 3, Example 1 uses alkaline protease and flavor protease for dual enzymatic hydrolysis, while Comparative Examples 2 and 3 use alkaline protease and flavor protease for single enzymatic hydrolysis, respectively. The hydrolysis degrees of Comparative Examples 2 and 3 are lower than those of Example 1, and the Maillard product of Comparative Example 2 tastes slightly bitter. The odor sensory scores and taste sensory scores of Comparative Examples 2 and 3 are lower than those of Example 1. Therefore, dual enzymatic hydrolysis with alkaline protease and flavor protease not only improves the protein hydrolysis degree, but also enhances the sensory flavor of the Maillard product.

[0087] Comparative Example 4

[0088] (1) Hammer and crush beef bones into 3 cubic centimeter blocks, add 2.5 times the weight of the raw material water and 1.5% of the weight of the raw material 52° liquor, use ultrasound to remove the fishy odor substances, the ultrasound frequency is 100 kHz, the ultrasound power is 600 W, the ultrasound method is ultrasound for 20 minutes, and finally heat at 100° C. for 40 minutes to remove the raw material;

[0089] (2) The raw material obtained in step (1) is crushed by a crusher, and the particle size of the raw material after crushing is less than 1 mm. The crushed raw material is dispersed in water at a solid-liquid ratio of 1:20 (w / w), and alkaline protease and flavor protease are added for double enzymatic hydrolysis. The pH value of the homogenate is maintained at 8.3, the enzymatic hydrolysis time is 5 h, the enzymatic hydrolysis temperature is 57° C., the stirring speed is 130 rpm, the amount of alkaline protease added is 0.3% of the mass of the crushed raw material, the amount of flavor protease added is 0.2% of the mass of the crushed raw material, the alkaline protease activity is 50,000 U / g, and the flavor protease activity is 35,000 U / g;

[0090] (3) The final enzymatic hydrolyzate 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;

[0091] (4) The supernatant obtained in step (3) is filtered through a nanofiltration membrane to retain molecules with a molecular weight cutoff of 1000 Da, the filtrate is collected, and lyophilized to obtain polypeptide powder;

[0092] (5) adding water to wheat bran polysaccharide (80% purity, purchased from Lanzhou Waters Biotechnology Co., Ltd.) at a material-liquid mass ratio of 1:200, and heating at 100° C. for 10 to 15 minutes to obtain a wheat bran polysaccharide solution;

[0093] (6) Dissolve 10 parts by mass of the polypeptide powder obtained in step (4) in 30 parts by mass of the wheat bran polysaccharide solution obtained in step (5). In the mixed solution of the polypeptide powder, wheat bran polysaccharide and edible ethanol, the volume percentage of edible ethanol is 25%; the purity of edible ethanol in this comparative example is 95%. Heat and react at 115 °C for 1.5 h; then add 1 part by mass of xylose, 1 part by mass of arginine, and 0.5 part by mass of lysine, and heat and react at 150 °C for 1 h to obtain a Maillard product solution.

[0094] (7) For the Maillard product solution obtained in step (6), according to 100 parts by mass of the Maillard product solution, 0.5 part by mass of alginate, 0.5 part by mass of corn starch, 0.5 part by mass of cyclodextrin, 0.5 part by mass of arginine, 0.2 part by mass of lysine, 0.3 part by mass of sodium chloride, 0.2 part by mass of potassium chloride, and 0.5 part by mass of shrimp oil, homogenize at 3000 rpm for 10 - 20 minutes;

[0095] (8) Spray - dry the homogenate obtained in step (7), with an inlet temperature of 145 °C, an outlet temperature of 105 °C, and a feed flow rate of 4.5 mL / min.

[0096] Table 3 Sensory Evaluation

[0097] Group Color Sensory Score Odor Sensory Score Taste Sensory Score Weighted Score Example 1 90 90 90 90 Comparative Example 4 70 80 80 76

[0098] When comparing Example 1 with Comparative Example 4, in Example 1, the wheat bran polysaccharide was hydrolyzed by α - amylase to obtain reducing sugars and then Maillard reaction was carried out, while in Comparative Example 4, Maillard reaction was directly carried out using wheat bran polysaccharide. As shown in Table 3, the product obtained by directly carrying out Maillard reaction using wheat bran polysaccharide has a lighter color, and the sensory evaluation of both smell and taste is lower than that of the Maillard product of wheat bran oligosaccharide in Example 1.

[0099] Example 2

[0100] (1) Hammer and break the beef bones with meat into pieces of 3 cubic centimeters in size, add 2.5 times the weight of the raw materials of water and 1.5% of the weight of the raw materials of 52° liquor, use ultrasonic waves to remove the fishy smell substances, the ultrasonic frequency is 100 kHz, the ultrasonic power is 600 W, the ultrasonic mode is ultrasonic for 20 min, and finally heat at 100 °C for 40 min and then fish out the raw materials;

[0101] (2) The raw material obtained in step (1) is crushed by a crusher, and the particle size of the raw material after crushing is less than 1 mm. The crushed raw material is dispersed in water at a solid-liquid ratio of 1:20 (w / w), and alkaline protease and flavor protease are added for enzymolysis. The pH value of the homogenate is maintained at 8.3, the enzymolysis time is 3 hours, the enzymolysis temperature is 57° C., the stirring speed is 130 rpm, the amount of alkaline protease added is 0.3% of the mass of the crushed raw material, the amount of flavor protease added is 0.2% of the mass of the crushed raw material, the alkaline protease activity is 50,000 U / g, and the flavor protease activity is 35,000 U / g;

[0102] (3) The final enzymatic hydrolyzate 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;

[0103] (4) The supernatant obtained in step (3) is filtered through a nanofiltration membrane to retain molecules with a molecular weight cutoff of 1000 Da, the filtrate is collected, and lyophilized to obtain polypeptide powder;

[0104] (5) adding water to wheat bran polysaccharide (80% purity, purchased from Lanzhou Waters Biotechnology Co., Ltd.) at a material-liquid mass ratio of 1:200, adding α-amylase for enzymolysis, the addition ratio being 0.35% of the mass of the wheat bran polysaccharide, the enzymolysis temperature being 58° C., the α-amylase activity being 3800 U / g, the enzymolysis time being 80 min, and heating at 100° C. for 10 to 15 min to inactivate the enzyme to obtain wheat bran oligosaccharides;

[0105] (6) Dissolve 10 parts by weight of the polypeptide powder obtained in step (4) in 30 parts by weight of the wheat bran oligosaccharides obtained in step (5), and add edible ethanol; in the mixed solution of the polypeptide powder, the wheat bran oligosaccharide solution and the edible ethanol, the volume percentage of the edible ethanol is 25%; the purity of the edible ethanol in this embodiment is 95%, and heat the reaction at 115° C. for 1.5 h; then add 1 part by weight of xylose, 1 part by weight of arginine, and 0.5 part by weight of lysine, and heat the reaction at 150° C. for 1 h to obtain a Maillard product solution.

[0106] (7) homogenizing the Maillard product solution obtained in step (6) at 3000 rpm for 10 to 20 minutes according to 100 parts by mass of the Maillard product solution, 0.5 parts by mass of alginate, 0.5 parts by mass of corn flour, 0.5 parts by mass of cyclodextrin, 0.5 parts by mass of arginine, 0.2 parts by mass of lysine, 0.3 parts by mass of sodium chloride, 0.2 parts by mass of potassium chloride, and 0.5 parts by mass of shrimp oil;

[0107] (8) The homogenized liquid obtained in step (7) was spray dried, with an inlet temperature of 145° C., an outlet temperature of 105° C., and a feed flow rate of 4.5 mL / min.

[0108] Example 3

[0109] (1) Hammer and crush beef bones into 3 cubic centimeter blocks, add 2.5 times the weight of the raw material water and 1.5% of the weight of the raw material 52° liquor, use ultrasound to remove the fishy odor substances, the ultrasound frequency is 100 kHz, the ultrasound power is 400 W, the ultrasound method is ultrasound for 20 minutes, and finally heat at 100° C. for 40 minutes to remove the raw material;

[0110] (2) The raw material obtained in step (1) is crushed by a crusher, and the particle size of the raw material after crushing is less than 1 mm. The crushed raw material is dispersed in water at a solid-liquid ratio of 1:30 (w / w), and alkaline protease and flavor protease are added for double enzymatic hydrolysis. The pH value of the homogenate is maintained at 8.3, the enzymatic hydrolysis time is 5 h, the enzymatic hydrolysis temperature is 57° C., the stirring speed is 130 rpm, the amount of alkaline protease added is 0.3% of the mass of the crushed raw material, the amount of flavor protease added is 0.2% of the mass of the crushed raw material, the alkaline protease activity is 50,000 U / g, and the flavor protease activity is 35,000 U / g;

[0111] (3) The final enzymatic hydrolyzate 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;

[0112] (4) The supernatant obtained in step (3) is filtered through a nanofiltration membrane to retain molecules with a molecular weight cutoff of 1000 Da, the filtrate is collected, and lyophilized to obtain polypeptide powder;

[0113] (5) adding water to wheat bran polysaccharide (80% purity, purchased from Lanzhou Waters Biotechnology Co., Ltd.) at a material-liquid mass ratio of 1:200, adding α-amylase for enzymolysis, the addition ratio being 0.35% of the mass of the wheat bran polysaccharide, the enzymolysis temperature being 58° C., the α-amylase activity being 3800 U / g, the enzymolysis time being 80 min, and heating at 100° C. for 10 to 15 min to inactivate the enzyme to obtain wheat bran oligosaccharides;

[0114] (6) Dissolve 10 parts by weight of the polypeptide powder obtained in step (4) in 20 parts by weight of the wheat bran oligosaccharides obtained in step (5), and add edible ethanol; in the mixed solution of the polypeptide powder, the wheat bran oligosaccharide solution and the edible ethanol, the volume percentage of the edible ethanol is 25%; the purity of the edible ethanol in this embodiment is 95%, and heat the reaction at 115° C. for 1.5 h; then add 1 part by weight of xylose, 1 part by weight of arginine, and 0.5 part by weight of lysine, and heat the reaction at 150° C. for 1 h to obtain a Maillard product solution.

[0115] (7) Homogenize the Maillard product solution obtained in step (6) at 3000 rpm for 10 - 20 minutes according to 100 parts by mass of the Maillard product solution, 0.5 parts by mass of alginate, 0.5 parts by mass of corn starch, 0.5 parts by mass of cyclodextrin, 0.5 parts by mass of arginine, 0.2 parts by mass of lysine, 0.3 parts by mass of sodium chloride, 0.2 parts by mass of potassium chloride, and 0.5 parts by mass of shrimp oil.

[0116] (8) Spray - dry the homogenate obtained in step (7) with an inlet temperature of 145 °C, an outlet temperature of 105 °C, and a feed flow rate of 4.5 mL / min.

[0117] Table 4 Sensory Score

[0118] Group Degree of Hydrolysis Color Sensory Score Odor Sensory Score Taste Sensory Score Weighted Score Example 1 27.8 90 90 90 90 Example 2 25.3 90 90 80 87 Example 3 27.2 90 90 80 87

[0119] Compared with Example 1, the enzymatic hydrolysis time of the protein and wheat bran polysaccharide in Example 2 is slightly shorter than that in Example 1. As shown in Table 4, the degree of hydrolysis and the sensory score of the taste in Example 2 are slightly lower than those in Example 1. Compared with Example 1, the ultrasonic power for removing fishy smell from beef bones in Example 3 is slightly lower than that in Example 1, the liquid - to - solid ratio of the beef bone cooking liquid is slightly lower than that in Example 1, and the addition ratio of wheat bran oligosaccharide in the Maillard reaction is slightly lower than that in Example 1. The results show that the sensory score of the taste in Example 3 is slightly lower than that in Example 1. Therefore, by comparison, Example 1 is the optimal preparation process scheme.

[0120] Table 5 UV Absorption of Maillard Products

[0121] Group 290nm 420nm Example 1 0.861 0.992 Example 2 0.786 0.901 Example 3 0.825 0.925

[0122] As shown in Table 5, compared with Example 2 and Example 3, Example 1 has the highest absorbance at 290 nm and 420 nm. Among them, 290 nm is the absorption peak of hydroxymethylfurfural products, and 420 nm is the maximum absorption peak of melanoidins generated by the carbonyl - amine reaction, indicating that the content of Maillard intermediate products in Example 1 is high.

[0123] Experimental Results:

[0124] The main flavor substances of the Maillard product of beef bone peptide and wheat bran oligosaccharide are shown in Table 6. Table 6 shows the flavor substances of the Maillard product in Example 1 determined by GC - MS. Comparative Example 1 is a conventional method, a single Maillard reaction without the addition of wheat bran oligosaccharide.

[0125] Table 6 Contents of Main Flavor Substances of Maillard Reaction between Beef Bone Peptide and Wheat Bran Oligosaccharide

[0126]

[0127]

[0128] As shown in the GC-MS measurement results in Table 6, the main flavor substances of the Maillard product of the bovine bone peptide and wheat bran oligosaccharide of the present invention are alcohols, aldehydes, ketones, esters, phenols, nitrogen-containing compounds, acids, furans, alkanes and other main products. Among them, the alcohol with the highest content is 1-undecanol, the aldehyde with the highest content is phenylacetaldehyde, the phenol is p-ethylphenol, and the nitrogen-containing compound with the highest content is tetramethylpyrazine. The total content of flavor substances is 135.23 μg / g, which is higher than the flavor substance content of Example 2 and Example 3. The preparation method of Example 1 significantly increases the content of aldehyde, ketone and ester flavor substances compared with the conventional method. Therefore, through the method of the present invention, a product with rich flavor, fresh and mellow taste can be obtained safely and greenly, and the functional characteristics and utilization rate of the raw materials can be improved.

[0129] The above embodiments merely represent several implementation manners of the present invention. The description thereof is relatively specific and detailed, but it should not be construed as a limitation to the scope of the patent of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present invention, several modifications and improvements can still be made, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the appended claims.

Claims

1. A preparation method of a flavor-enhancing base powder of a bovine bone peptide Maillard reaction product, characterized in that, It includes the following steps: (1) After hammer-breaking and crushing the beef bones with meat, add water 2.5 times the weight of the raw materials and 52° liquor 1.5% of the weight of the raw materials, use ultrasonic waves to remove the fishy substances, and finally heat and steam at 100 °C for 30 - 60 min and fish out the raw materials; (2) Crush the raw materials obtained in step (1) with a pulverizer, then mix and disperse them with water according to a mass ratio of 1:20 - 1:25, adjust the pH of the dispersion liquid to 8.0 - 9.0, add alkaline protease and flavor protease for double-enzyme hydrolysis, and hydrolyze while stirring throughout the process for 4 - 8 h; the hydrolysis temperature is 50 °C - 60 °C, the stirring speed during the hydrolysis process is 100 - 200 rpm, the addition amount of alkaline protease is 0.25 - 0.35% of the mass of the pulverized raw materials, the addition amount of flavor protease is 0.15 - 0.25% of the mass of the pulverized raw materials, the enzyme activity of alkaline protease is 40000 - 70000 U / g, and the enzyme activity of flavor protease is 20000 - 40000 U / g (3) Heat the final hydrolysis liquid obtained in step (2) at 100 °C for 10 - 15 min for enzyme inactivation treatment, then cool it to room temperature and adjust the pH to 4.0 - 5.0, and take the supernatant after standing; (4) The supernatant obtained in step (3) is filtered through a nanofiltration membrane to retain molecules with a molecular weight of 1000 Da, the filtrate is collected, and a polypeptide powder is obtained after freeze-drying; (5) Take wheat bran polysaccharide and add it to water according to a mass ratio of 1:200 of the feed liquid, add α-amylase for hydrolysis, and obtain a wheat bran oligosaccharide solution after enzyme inactivation treatment by heating at 100 °C for 10 - 15 min; the addition ratio of α-amylase is 0.2% - 0.5% of the mass of wheat bran polysaccharide, the activity of α-amylase is 3000 U / g - 5000 U / g, the hydrolysis temperature is 45 °C - 60 °C, and the hydrolysis time is 60 min - 80 min; (6) Dissolve 10 parts by mass of the polypeptide powder obtained in step (4) in 20 - 30 parts by mass of the wheat bran oligosaccharide solution obtained in step (5), add edible ethanol, adjust the pH to 8.0 - 9.0, and heat and react at 110 °C - 130 °C for 1 - 2 h; then add 1 part by mass of xylose, 1 part by mass of arginine, and 0.5 part by mass of lysine, and heat and react at 130 - 155 °C for 1 - 2 h to obtain a Maillard intermediate product solution of bovine bone oligopeptide - wheat bran oligosaccharide; (7) Mix 100 parts by mass of the Maillard intermediate product solution obtained in step (6) with 0.5 part by mass of alginate, 0.5 part by mass of corn starch, 0.5 part by mass of cyclodextrin, 0.5 part by mass of arginine, 0.2 part by mass of lysine, 0.3 part by mass of sodium chloride, 0.2 part by mass of potassium chloride, and 0.5 part by mass of shrimp oil, and homogenize at 3000 rpm for 10 - 20 minutes to obtain a homogenate; (8) Spray-dry the homogenate obtained in step (7) to obtain a flavor-enhancing base powder of Maillard reaction product of bovine bone peptide; the inlet temperature of spray-drying is 140 - 165 °C, the outlet temperature is 100 - 110 °C, and the feed flow rate is 4 - 6 mL / min.

2. The preparation method according to claim 1, wherein In step (1), the size of the raw materials after hammer-breaking and crushing is 3 - 5 cubic centimeters.

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

4. The preparation method according to claim 1, wherein In step (2), after the raw materials are crushed by a crusher, the particle size is less than 1 mm.

5. The preparation method according to claim 1, wherein In step (5), the purity of the wheat bran polysaccharide is 80%.

6. The preparation method according to claim 1, wherein, In step (6), the purity of the edible ethanol is 95%; in the mixed solution of the polypeptide powder, the wheat bran oligosaccharide solution and the edible ethanol, the volume percentage of the edible ethanol is 20 - 40%.

7. A flavor enhancer powder of a bovine bone peptide Maillard reaction product prepared by the method according to any one of claims 1 - 6.

8. Use of the flavor enhancer powder of the bovine bone peptide Maillard reaction product according to claim 7 in the preparation of a flavor conditioning powder.

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