Method for preparing bovine bone Maillard reaction product by superfine grinding technology

Abstract

The invention relates to the technical field of bioengineering, in particular to a method for preparing a bovine bone Maillard reaction product by a superfine grinding technology, which comprises the following steps: cleaning bovine bones; baking and drying the cleaned ox bones; crushing by using a coarse crusher, and sieving to obtain cattle bone coarse powder; selecting bovine bone coarse powder, and crushing the bovine bone coarse powder by using an ultrafine crusher to obtain bovine bone ultrafine powder; dissolving the bovine bone ultrafine powder in water, carrying out heat treatment to obtain proteinase, adding protease, mixing, adjusting the pH value, and carrying out an enzymatic hydrolysis reaction; according to the invention, a superfine grinding technology is added to treat the bovine bone, so that the bone structure is destroyed, the structure of protein is changed, the contact area of protein and enzyme is increased, and the enzymolysis effect is promoted; small peptides and amino acids are prepared by adopting multiple proteolysis technologies, then a Maillard reaction product is prepared through a thermal reaction, and meanwhile, the Maillard reaction temperature and pH value are regulated and controlled, so that the overall flavor of the final product is improved, and the unpleasant smell is weakened.

Description

Technical Field

[0001] The invention relates to the technical field of bioengineering, and in particular to a method for preparing a Maillard reaction product of a bovine bone using an ultrafine grinding technique. Background Art

[0002] Beef bones are an important by-product of beef processing and have high nutritional value. Beef bone protein is a complete soluble protein containing 17 amino acids, including 8 essential amino acids for the human body. The small molecule peptides and amino acids generated by enzymatic hydrolysis of beef bones are more conducive to human digestion and absorption, and also have functional properties such as antioxidant, anti-aging, and prevention and treatment of osteoporosis. At present, the food industry has begun to develop various technologies to utilize this by-product. In addition to the development and application of traditional bone products, modern enzyme engineering technology and scientific production processes are also used to carry out deep processing of beef bones.

[0003] However, due to the high toughness and rigidity of cattle bones, it is difficult to extract protein and amino acids from cattle bones. Even if the cattle bones are crushed into particles, the protein extraction efficiency is often insufficient due to the large particle size of cattle bone powder. Ultrafine grinding, as a tool for making ultrafine powder, is used in the cattle bone crushing process, which can reduce the particle size of cattle bone powder, increase the solubility of cattle bone protein, and help enzymatically hydrolyze cattle bone protein and prepare cattle bone Maillard reaction products with characteristic meat flavor.

[0004] The Maillard reaction, also known as the carbonyl-amine reaction, refers to a series of complex reactions between carbonyl compounds (reducing sugars, aldehydes, ketones) and amino compounds (proteins, peptides, free amino acids, amines, etc.); the Maillard reaction plays an important role in food storage and processing, producing a variety of volatile compounds, non-enzymatic intermediates and high molecular weight melanin compounds, which play a vital role in the aroma, color, flavor and antioxidant properties of food.

[0005] At present, beef processing has driven the production of beef bones, but due to low-end processing equipment or technology, the quality of beef bone products is poor, the processing level is low, and the beef bone resources are seriously wasted; at the same time, beef bones have large skeletons and hard bone structures, and ordinary crushing technology cannot achieve the ideal particle size of beef bone powder, so it is difficult to obtain more nutrients. There is an urgent need for a crushing method to effectively crush beef bones and reduce the particle size of beef bone powder so that its soluble protein can be fully dissolved in water; in addition, with the development of the food industry, the demand for meat flavors and spices is increasing, but the current production methods are relatively traditional, resulting in a single category and the characteristic flavor of meat is not ideal. Therefore, new processing technology means are needed to regulate the mechanism of meat flavor. Summary of the invention

[0006] In view of the above-mentioned shortcomings of the prior art, the present invention provides a method for preparing Maillard reaction products of cattle bones by ultrafine grinding technology, which can effectively solve the problems mentioned in the prior art.

[0007] To achieve the above objectives, the present invention is implemented through the following technical solutions:

[0008] The present invention provides a method for preparing a Maillard reaction product of cattle bones by ultrafine grinding technology, comprising the following steps:

[0009] Step 1: Clean the beef bones, remove the non-bone components, and rinse with clean water;

[0010] Step 2, take out the cleaned beef bones and put them in an oven to bake and dry;

[0011] Step 3, crushing with a coarse grinder and sieving to obtain coarse beef bone powder;

[0012] Step 4, select the coarse powder of cattle bones and grind it with an ultrafine grinder to obtain ultrafine powder of cattle bones;

[0013] Step 5, dissolving the ultrafine bovine bone powder in water, performing heat treatment to obtain deformed protein, adding protease to mix and adjusting the pH to perform enzymatic hydrolysis; after the reaction is completed, the enzyme is inactivated, cooled and centrifuged to obtain a small molecule bovine bone protein peptide product;

[0014] Step 6: Add sugar source and exogenous amino acid to the small molecule bovine bone protein peptide product, adjust the reaction temperature, pH and reaction time; wait for the reaction to end, and obtain a bovine bone Maillard reaction product with a certain degree of reaction after cooling.

[0015] Preferably, the baking and drying temperature in step 2 is 40-60° C., and the drying time is 4-8 hours.

[0016] Preferably, in step 3, the raw material is crushed in a coarse crusher for 5 to 20 minutes and then passed through a 30 to 80 mesh sieve.

[0017] Preferably, the ultrafine pulverizer in step 4 is a vibrating ultrafine pulverizer, and the pulverization takes 5-30 minutes.

[0018] Preferably, in step 5, the heat treatment temperature is 80-100° C., and the heat treatment time is 20-120 minutes; the protease used is a mixture of one or more of neutral protease, alkaline protease, papain, flavor enzyme, pepsin, and trypsin, which is a conventional protein endo- or protein exo-enzyme; the enzymolysis temperature is 40-60° C., the enzymolysis pH is 6.0-9.0, the enzyme addition amount is 1000-8000 U / g, the enzymolysis time is 0.5-6 hours, the degree of hydrolysis is controlled at 5.0-12.0, the enzyme inactivation temperature is 80-100° C., and the enzyme inactivation time is 15-35 minutes; the centrifugal equipment is a high-speed refrigerated centrifuge, the centrifugation temperature is 3.0-5.0° C., the rotation speed is 6000-10000 rpm / min, and the centrifugation time is 10-40 minutes.

[0019] Preferably, the sugar source in step 6 is at least one of xylose, ribose, fructose and glucose, and the added amount is 0.5-5% (w / w) of the bovine bone protein peptide content; the exogenous amino acid is cysteine, and the added amount is 1.0-10.0% (w / w) of the bovine bone protein peptide, and the pH value is adjusted to 6.0-10.0; the Maillard reaction temperature is 80-140°C, and the reaction time is 0.5-4 hours.

[0020] Compared with the known prior art, the technical solution provided by the present invention has the following beneficial effects:

[0021] (1) The cattle bones are fully crushed, which is conducive to the processing and utilization of cattle bone protein and avoids the waste of cattle bone resources;

[0022] (2) The present invention uses ultrafine grinding technology to process cattle bones, which reduces the particle size of cattle bone powder, destroys the bone structure, changes the structure of protein, and increases the contact area between protein and enzyme during enzymatic hydrolysis, thereby promoting enzymatic hydrolysis and providing sufficient amino acids and peptides for the subsequent Maillard reaction;

[0023] (3) The present invention adopts a variety of proteolysis technologies to enzymatically hydrolyze bovine bone protein to prepare small peptides and amino acids, and then prepare Maillard reaction products through thermal reaction; the overall flavor of the final product is improved and the unpleasant odor is weakened. Ultrafine grinding pretreatment assists the Maillard reaction and has a certain effect;

[0024] (4) The production process of the present invention is simple, efficient and low-cost. Compared with the Maillard reaction product of coarsely ground beef bones, the new product has a richer and mellower overall taste. At the same time, by regulating the Maillard reaction temperature and pH, the product can obtain a rich characteristic aroma, which can be used to increase the meaty aroma, umami and salty taste in seasonings, and can meet consumers' pursuit of flavors, spices and condiments. The market prospects are broad. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the prior art descriptions are briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention, and for ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0026] Figure 1 This is a comparison chart of particle sizes after coarse grinding and ultrafine grinding for 5, 10, 15, 20, and 25 minutes;

[0027] Figure 2 The effect of ultrafine grinding time on the hydrolysis degree of bovine bone protein;

[0028] Figure 3 To study the effect of flavor protease addition on hydrolysis degree and sensory quality;

[0029] Figure 4 The effect of the amount of neutral protease added on the hydrolysis degree and sensory quality;

[0030] Figure 5 The effect of enzymatic hydrolysis time on hydrolysis degree and sensory quality;

[0031] Figure 6 The effect of enzymatic hydrolysis temperature on the degree of hydrolysis and sensory quality;

[0032] Figure 7 The effect of enzymatic hydrolysis pH on the degree of hydrolysis and sensory quality;

[0033] Figure 8 The effect of substrate concentration on the degree of hydrolysis and sensory quality;

[0034] in, Figure 3-8 This is an experiment on the protease hydrolysis of bovine bone. DETAILED DESCRIPTION

[0035] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0036] The present invention will be further described below in conjunction with the embodiments.

[0037] Embodiment 1:

[0038] Step 1: Clean the beef bones, remove the non-bone components (residual meat, skin, tendons, cartilage and other non-bone components), and rinse with clean water (remove residue and some fat);

[0039] Step 2, take out the cleaned beef bones and bake them in an oven at 40°C for 8 hours;

[0040] Step 3, using a coarse grinder to grind for 10 minutes and then pass through a 60-mesh sieve to obtain coarse beef bone powder;

[0041] Step 4, select the coarse powder of cattle bones and grind it with a vibrating superfine grinder for 20 minutes to obtain superfine powder of cattle bones;

[0042] Step 5, dissolving 20g of ultrafine bovine bone powder in water at a ratio of 1:5 (w / w), and placing in a water bath at 80°C for 40 minutes to obtain deformed protein, adding neutral protease at an enzyme amount of 5500U / g, mixing, adjusting the pH value to 7.0, and adjusting the temperature to 50°C for enzymatic hydrolysis for 2 hours; after the reaction, inactivating the enzyme in a water bath at 90°C for 25 minutes, cooling at 4°C and centrifuging at 10,000 rpm for 20 minutes, to obtain a small molecule bovine bone protein peptide product;

[0043] Step 6: Add 1% xylose and 2% cysteine ​​by mass to the product obtained in step 5, adjust the reaction temperature to 120° C., pH to 7.0, and the reaction time to 120 minutes; wait for the reaction to end, and obtain a Maillard reaction product of beef bone with a certain degree of reaction after cooling.

[0044] Embodiment 2:

[0045] Step 1: Clean the beef bones, remove the non-bone components (residual meat, skin, tendons, cartilage and other non-bone components), and rinse with clean water (remove residue and some fat);

[0046] Step 2, take out the cleaned beef bones and bake them in an oven at 50°C for 6 hours;

[0047] Step 3, using a coarse grinder to grind for 15 minutes and then pass through an 80-mesh sieve to obtain coarse beef bone powder;

[0048] Step 4, select the coarse powder of cattle bones and grind it with a vibrating superfine grinder for 15 minutes to obtain superfine powder of cattle bones;

[0049] Step 5, dissolving 25g of ultrafine bovine bone powder in water at a ratio of 1:5 (w / w), and heating at 90°C for 35 minutes to obtain deformed protein, adding alkaline protease at an enzyme dosage of 8000U / g, adjusting the pH value to 9.0, and adjusting the temperature to 45°C for enzymatic hydrolysis for 4 hours. After the reaction, inactivate the enzyme in a 100°C water bath for 20 minutes, cool at 3°C, and centrifuge at 8000rpm for 30 minutes to obtain a small molecule bovine bone protein peptide product;

[0050] Step 6: Add 0.5% ribose and 1% cysteine ​​by mass to the product obtained in step 5, adjust the reaction temperature to 110° C., pH to 7.5, and the reaction time to 100 minutes, wait for the reaction to end, and obtain a bovine bone Maillard reaction product with a certain degree of reaction after cooling.

[0051] Embodiment 3:

[0052] Step 1: Clean the beef bones, remove the non-bone components (residual meat, skin, tendons, cartilage and other non-bone components), and rinse with clean water (remove residue and some fat);

[0053] Step 2: Take out the cleaned beef bones and bake them in an oven at 60°C for 4.5 hours;

[0054] Step 3, using a coarse grinder to grind for 8 minutes and then pass through a 50-mesh sieve to obtain coarse beef bone powder;

[0055] Step 4, select the coarse powder of cattle bones and grind it with a vibrating superfine grinder for 25 minutes to obtain superfine powder of cattle bones;

[0056] Step 5, dissolving 100g of ultrafine bovine bone powder in water at a ratio of 1:5 (w / w), and placing in a water bath at 80°C for 40 minutes to obtain deformed protein, and adding flavor protease and neutral protease at an enzyme amount of 6000U / g, mixing and adjusting the pH value to 7.0, adjusting the temperature to 50°C for enzymatic hydrolysis for 6 hours, and after the reaction, inactivating the enzyme in a water bath at 100°C for 20 minutes, cooling at 4°C and centrifuging at 9000 rpm for 25 minutes to obtain a small molecule bovine bone protein peptide product;

[0057] Step 6: Add 2% fructose and 4% cysteine ​​by mass to the product obtained in step 5, adjust the reaction temperature to 130° C., pH to 7.2, and the reaction time to 140 minutes, wait for the reaction to end, and obtain a bovine bone Maillard reaction product with a certain degree of reaction after cooling.

[0058] In combination with Examples 1-3, Figure 1 It can be seen that ultrafine grinding can effectively reduce the particle size of cattle bone powder; Figure 2 It can be seen that the most suitable time for ultrafine grinding is 20 minutes. After 20 minutes, the increase in hydrolysis degree is no longer obvious. Figure 3 —8 It can be seen that the more suitable hydrolysis conditions for protease are mixed protease hydrolysis time of 4h, flavor protease addition of 1400U / g, neutral protease addition of 5000U / g, substrate concentration of 10%, enzymolysis temperature of 50℃, and pH of about 7.0.

[0059] As shown in Table 1, the effects of different proteases on the hydrolysis of bovine bone protein were investigated using the degree of hydrolysis and sensory quality of the hydrolyzate as indicators:

[0060] Table 1 Screening of protease hydrolysis of bovine bone powder

[0061]

[0062]

[0063] As shown in Table 1, neutral protease and flavor protease have higher hydrolysis degree and better sensory quality, so the hydrolysis effect is better.

[0064] Table 2 shows the sensory evaluation of Maillard reaction products obtained by ultrafine grinding of bovine bone protein hydrolysate for 20 minutes at different temperatures:

[0065] Table 2 Sensory evaluation of Maillard reaction products of cattle bones at different Maillard reaction temperatures

[0066]

[0067] As shown in Table 2, after the Maillard reaction, the oily smell of the enzymatic hydrolyzate decreases, while the meaty and roasted nutty smells increase; at the same time, the higher the temperature, the stronger the caramel smell, and the more unpleasant odors are produced; when the temperature is controlled at 120°C, the meaty smell is the strongest.

[0068] Table 3 shows the determination of volatile compounds in Maillard reaction products obtained from the enzyme hydrolyzate of bovine bone by ultrafine grinding for 20 minutes at different temperatures:

[0069] Table 3 Effect of reaction temperature on the volatile compound content of Maillard reaction products of cattle bones

[0070]

[0071]

[0072]

[0073]

[0074]

[0075]

[0076] From Table 3, it can be found that after the Maillard reaction, the content of volatile compounds increases, with the highest volatile substances at 140°C, and decreasing as the temperature decreases; it is further found that when the Maillard reaction temperature is controlled at 120°C, the content of characteristic aroma substances (2-methyl-3-furanthiol, 2-furfurylthiol, 2-methylfuran, 3-methylfuran, 3,3'-dithiobis[2-methyl]furan, pyrrole, etc.) is the highest, which is the main reason why the Maillard reaction product has a characteristic meaty flavor. In summary, the Maillard reaction product of beef bones is effective in increasing the characteristic meaty flavor, and it is more appropriate to control the reaction temperature at 120°C.

[0077] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements will not cause the essence of the corresponding technical solutions to deviate from the protection scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for preparing Maillard reaction products of cattle bones by ultrafine grinding technology, characterized in that: The following steps are involved: Step 1: Clean the beef bones, remove the non-bone components, and rinse with clean water; Step 2, take out the cleaned beef bones and put them in an oven to bake and dry; Step 3, crushing with a coarse grinder and sieving to obtain coarse beef bone powder; Step 4, select the coarse powder of cattle bones and grind it with an ultrafine grinder to obtain ultrafine powder of cattle bones; Step 5, dissolving the ultrafine bovine bone powder in water, performing heat treatment to obtain deformed protein, adding protease to mix and adjusting the pH to perform enzymatic hydrolysis; after the reaction is completed, the enzyme is inactivated, cooled and centrifuged to obtain a small molecule bovine bone protein peptide product; Step 6: adding sugar source and exogenous amino acid to the small molecule bovine bone protein peptide product in step 5, adjusting the reaction temperature, pH and reaction time; waiting for the reaction to end, and obtaining a bovine bone Maillard reaction product with a certain degree of reaction after cooling.

2. The method for preparing Maillard reaction products of cattle bones by ultrafine grinding technology according to claim 1, characterized in that: The baking and drying temperature in step 2 is 40-60° C., and the drying time is 4-8 hours.

3. The method for preparing Maillard reaction products of cattle bones by ultrafine grinding technology according to claim 1, characterized in that: In the step 3, the coarse grinder is used to grind for 5-20 minutes, and then the mixture is passed through a 30-80 mesh sieve.

4. The method for preparing Maillard reaction products of cattle bones by ultrafine grinding technology according to claim 1, characterized in that: The ultrafine pulverizer in step 4 is a vibrating ultrafine pulverizer, and the pulverization takes 5-30 minutes.

5. The method for preparing Maillard reaction products of cattle bones by ultrafine grinding technology according to claim 1, characterized in that: In the step 5, the heat treatment temperature is 80-100° C., and the heat treatment time is 20-120 minutes; the protease used is a mixture of one or more of neutral protease, alkaline protease, papain, flavor enzyme, pepsin, and trypsin, and is a conventional protein endo- or protein exo-enzyme; the enzymolysis temperature is 40-60° C., the enzymolysis pH value is 6.0-9.0, the enzyme addition amount is 1000-8000 U / g, the enzymolysis time is 0.5-6 hours, the hydrolysis degree is controlled at 5.0-12.0, the enzyme inactivation temperature is 80-100° C., and the enzyme inactivation time is 15-35 minutes; the centrifugal equipment is a high-speed refrigerated centrifuge, the centrifugation temperature is 3.0-5.0° C., the rotation speed is 6000-10000 rpm / min, and the centrifugation time is 10-40 minutes.

6. The method for preparing Maillard reaction products of cattle bones by ultrafine grinding technology according to claim 1, characterized in that: The sugar source in step 6 is at least one of xylose, ribose, fructose and glucose, and the added amount is 0.5-5% (w / w) of the bovine bone protein peptide content; the exogenous amino acid is cysteine, and the added amount is 1.0-10.0% (w / w) of the bovine bone protein peptide, and the pH value is adjusted to 6.0-10.0; the Maillard reaction temperature is 80-140°C, and the reaction time is 0.5-4 hours.

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