Process for extracting arginine fructose from onions
Through gradient enzymatic lysis and multi-layer column elution technology, arginine fructose is efficiently extracted from onions, solving the problems of low concentration, low efficiency and poor stability, and achieving a high purity and environmentally friendly extraction process, reducing the odor impact.
Patent Information
- Application Number
- CN202411278352.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-09-12
- Publication Date
- 2024-12-13
AI Technical Summary
In the prior art, the extraction of arginine fructose from onions has low concentration, low efficiency, poor stability, difficulty in separation and odor problems, and the aqueous ammonia solution used in traditional methods is harmful to the human body and the environment.
After pulverization by wet ball milling, cellulase, α-amylase and complex enzyme A are used for enzymatic dissection through gradient enzymatic dissection process, pH is adjusted and further enzymatic dissection is used for further enzymatic dissection using complex enzyme B, combining anhydrous ethanol precipitation and multilayer columns. Eluted and finally obtained arginine fructose by spray drying, improving extraction efficiency, purity and stability, and reducing odor.
It improves the extraction rate and purity of arginine fructose in onions, improves stability, reduces odor problems, and avoids the use of harmful aqueous ammonia solutions, achieving a more environmentally friendly and efficient extraction process.
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of extracting compounds from processed products, and particularly relates to a process for extracting arginine fructose from onions. Background Art
[0002] Arginine fructose exists in many plants in nature. It is also a Maillard reaction product produced in the process of processing fresh ginseng into red ginseng and raw sun-dried ginseng. It has obvious pharmacological activity and is one of the characteristic and iconic components of red ginseng. However, the price of red ginseng is relatively high. Studies have found that relatively cheap onions also produce functional arginine-fructose during the steaming process. Arginine-fructose derived from onions has the same blocking activity and anti-diabetic activity as isolated α-glucosidase (α-glucosidase).
[0003] When extracting arginine fructose from onions, the following technical problems are faced: (1) Low concentration: The concentration of arginine fructose in food is usually very low, which means that a large amount of raw materials needs to be processed to obtain sufficient extract. This not only increases costs but may also put pressure on the environment. (2) Extraction efficiency: The extraction of arginine fructose requires specific conditions, including appropriate solvents and extraction techniques, to ensure high yields. (3) Stability: arginine fructose may be sensitive to light, heat and pH and easily degrade. During extraction, processing and storage, measures need to be taken to protect the stability of these compounds. (4) Separation and purification: It may be difficult to separate arginine fructose from other components in the food matrix. The strong odor unique to arginine fructose in low-purity onions is also undesirable in practical applications. (5) Biological activity: The extraction process may affect the biological activity of arginine fructose.
[0004] At present, the main extraction method for extracting arginine fructose from onions is to use high-concentration ethanol extraction, remove the supernatant, precipitate water extraction and alcohol precipitation, and then separate it through a cationic resin column. However, this method has many impurities, and the subsequent elution through the cationic resin requires ammonia solution, which has a pungent taste and is harmful to the human body and the environment.
[0005] Therefore, there is an urgent need for a process for extracting arginine fructose from onions. Summary of the invention
[0006] The purpose of the invention is to provide a process for extracting arginine fructose from onions.
[0007] In order to achieve the above object, the present invention provides the following technical solutions: A process for extracting arginine fructose from onions, the extraction process comprising the following steps: (1) mixing onion and water, performing wet ball milling and sieving to obtain onion pulverizer; (2) heating the crushed onion at 80-90°C for 2-3 hours, cooling to room temperature, and obtaining an onion water extract; (3) Adding cellulase to the onion water extract, enzymolyzing at 35-37°C for 3-5 hours, and then inactivating the enzyme; continuing to add α-amylase, enzymolyzing at 35-37°C for 1-2 hours, and then inactivating the enzyme; continuing to add complex enzyme A, enzymolyzing at 35-37°C for 2-3 hours, and then inactivating the enzyme to obtain the first enzymolysis product of onion; (4) filtering the first enzymatic hydrolysis product of onion to obtain onion juice; (5) adjusting the pH of the onion juice to 8-8.2 to obtain a mixture, treating the mixture with a composite enzyme B, and then stirring and enzymolyzing the mixture at a stirring speed of 120-140 rpm, an enzymolysis temperature of 45-50°C, and a time of 2-3 hours; inactivating the enzyme to obtain a second enzymolysis product of the onion; the composite enzyme B is a mixture of alkaline protease, neutral protease, and flavor enzyme in a mass ratio of (1.5-1.7):1:(0.3-0.5), and the mass of the composite enzyme B accounts for 2-3% of the mass of the onion; (6) The second enzymatic hydrolysate of onion is precipitated with anhydrous ethanol, the supernatant is removed, the precipitate is passed through a 200-300 mesh silica gel column, and eluted with a 70-75% ethanol aqueous solution, the first eluate is collected, and after the first eluate is concentrated, it is continued to pass through a polyacrylamide gel column, eluted with a 0.15-0.17% acetic acid aqueous solution, and the second eluate is collected; (7) The second eluate is spray-dried to obtain arginine fructose in onion.
[0008] Furthermore, the onions can be freshly picked onions.
[0009] Furthermore, in the step (3), the mass of the cellulase accounts for 0.8-1.2% of the mass of the onion.
[0010] Furthermore, in step (3), the mass of the α-amylase accounts for 0.8-1.2% of the mass of the onion.
[0011] Furthermore, in step (3), the mass of the complex enzyme A accounts for 0.8-1.2% of the mass of the onion.
[0012] At present, the onion on the market uses water extraction to improve the extraction rate of fructose, and then uses a single enzyme for enzymolysis, thereby improving the yield of onion arginine fructose. In order to further improve the yield, the present invention selects different types of enzymes to perform gradient enzymolysis in sequence after water extraction of onions, which can improve the yield and purity of arginine fructose in onions. The use of enzymes in stages in the present invention can more efficiently and thoroughly decompose the complex structure of onions, thereby improving extraction efficiency and product purity. However, the arginine fructose in the onions prepared under this condition has a relatively strong smell, which affects the research and development and use of later products.
[0013] Furthermore, in step (1), the product is sieved through a 200-300 mesh sieve.
[0014] Furthermore, in step (1), the mass ratio of onion to water is 1:(7-10).
[0015] Furthermore, in the step (1), the wet ball milling time is 2-4 h, the ball milling speed is 400-500 rpm, and the temperature is controlled at 40-45°C during the milling process.
[0016] Furthermore, the complex enzyme A is a mixture of hemicellulase, pectinase and catalase.
[0017] Furthermore, the complex enzyme A is a mixture of hemicellulase, pectinase and catalase in a mass ratio of 1: (1.3-1.5): (0.4-0.6).
[0018] After the cellulase and α-amylase are enzymatically hydrolyzed, the composite enzyme A mixed with hemicellulase, pectinase and catalase is used for enzymatic hydrolysis in the third stage, and the odor problem of arginine fructose in onions can be improved. The composite enzyme has a synergistic effect and can decompose the components causing the odor problem in the extract after the enzymatic hydrolysis of cellulase and α-amylase, while maintaining a high extraction rate and purity.
[0019] The stability of arginine fructose in the onion obtained by the above steps is poor, and the smell and color will change after long-term storage. Analysis shows that some components produced by the above enzymatic hydrolysis will affect the stability of AF. The present invention further uses compound enzyme B for enzymatic hydrolysis to improve the stability of arginine fructose in onions. After enzymatic hydrolysis by compound enzyme B, the original unstable substances are eliminated in the product, and the effective substances produced at the same time can protect arginine fructose.
[0020] Furthermore, the enzyme inactivation condition is heating at 100-121°C for 10-15 minutes.
[0021] Compared with the prior art, the advantages and beneficial effects of the present invention are: 1. The present invention provides a process for extracting arginine fructose from onions. After water extraction of onions, different types of enzymes are selected to perform gradient enzymolysis in sequence, thereby improving the yield and purity of arginine fructose in onions.
[0022] 2. After the cellulase and α-amylase of the present invention are enzymatically hydrolyzed, the composite enzyme A mixed with hemicellulase, pectinase and catalase is used for enzymatic hydrolysis in the third stage. The enzymatic hydrolysis can improve the odor problem of arginine fructose in onions.
[0023] 3. The present invention further uses complex enzyme B for enzymolysis to improve the stability of arginine fructose in onions. DETAILED DESCRIPTION
[0024] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. 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.
[0025] The enzyme used in the present invention is a commercially available product, and the specific information is as follows: Hemicellulase, enzyme activity 200u / mg, Shanghai Yuanye Biotechnology Co., Ltd.
[0026] Catalase, enzyme activity 100u / mg, Shanghai Yuanye Biotechnology Co., Ltd., S25070.
[0027] α-Amylase, enzyme activity 50u / mg, Shanghai Yuanye Biotechnology Co., Ltd.
[0028] Cellulase, enzyme activity 50u / mg, Shanghai Yuanye Biotechnology Co., Ltd.
[0029] Neutral protease, enzyme activity 100u / mg, Shanghai Yuanye Biotechnology Co., Ltd.
[0030] Pectinase, enzyme activity 50u / mg, Shanghai Yuanye Biotechnology Co., Ltd.
[0031] Bromelain, enzyme activity 300u / mg, Shanghai Yuanye Biotechnology Co., Ltd.
[0032] Fig proteinase, enzyme activity 300u / mg, Shanghai Yuanye Biotechnology Co., Ltd.
[0033] Alkaline protease, enzyme activity 200u / mg, Shanghai Yuanye Biotechnology Co., Ltd.
[0034] Flavor enzyme, enzyme activity 20u / mg, Shanghai Yuanye Biotechnology Co., Ltd.
[0035] Example 1 This embodiment provides a process for extracting arginine fructose from onions, and the extraction process comprises the following steps: (1) Mix fresh onion and water in a mass ratio of 1:8, and perform wet ball milling. The wet ball milling time is 3 h, the ball milling speed is 450 rpm, and the temperature is controlled at 42°C during the milling process. The onion powder is passed through a 200-mesh sieve to obtain onion powder; (2) heating the crushed onion at 85°C for 2.5 hours and cooling to room temperature to obtain an onion water extract product; (3) Add cellulase to the onion water extract, hydrolyze at 36°C for 4 hours, and then inactivate the enzyme; continue to add α-amylase, hydrolyze at 36°C for 1.5 hours, and then inactivate the enzyme; continue to add compound enzyme A, hydrolyze at 36°C for 2.5 hours, and then inactivate the enzyme to obtain the first onion hydrolysis product; the mass of cellulase accounts for 1% of the mass of fresh onion; the mass of α-amylase accounts for 1% of the mass of fresh onion; the mass of compound enzyme A accounts for 1% of the mass of fresh onion; the compound enzyme A is a mixture of hemicellulase, pectinase and catalase in a mass ratio of 1:1.4:0.5; (4) filtering the first enzymatic hydrolysis product of onion to obtain onion juice; (5) The pH of the onion juice is adjusted to 8 to obtain a mixture, and the mixture is treated with complex enzyme B, followed by stirring and enzymolysis at a stirring speed of 130 rpm, an enzymolysis temperature of 46°C, and a time of 2.5 h; the enzyme is inactivated to obtain a second enzymolysis product of onion; the complex enzyme B is a mixture of alkaline protease, neutral protease, and flavor enzyme in a mass ratio of 1.6:1:0.4, and the mass of the complex enzyme B accounts for 2.4% of the mass of fresh onion; (6) The second enzymatic hydrolysate of onion was precipitated with anhydrous ethanol, the supernatant was removed, the precipitate was passed through a 200-300 mesh silica gel column, and eluted with a 75% ethanol aqueous solution. The first eluate was collected. After the first eluate was concentrated, it was continued to pass through a polyacrylamide gel column, eluted with a 0.15% acetic acid aqueous solution, and the second eluate was collected; (7) The second eluate is spray-dried to obtain arginine fructose in onion.
[0036] The above enzyme inactivation conditions are heating at 121°C for 15 min.
[0037] Example 2 This embodiment provides a process for extracting arginine fructose from onions, and the extraction process comprises the following steps: (1) Mix fresh onion and water in a mass ratio of 1:10, and perform wet ball milling. The wet ball milling time is 2 h, the ball milling speed is 500 rpm, and the temperature is controlled at 40°C during the milling process. Pass the mixture through a 300-mesh sieve to obtain onion powder; (2) heating the crushed onion at 80°C for 3 hours, cooling to room temperature, and obtaining an onion water extract product; (3) Adding cellulase to the onion water extract, enzymolysis at 35°C for 3 hours, and then inactivating the enzyme; continuing to add α-amylase, enzymolysis at 35°C for 2 hours, and then inactivating the enzyme; continuing to add complex enzyme A, enzymolysis at 35°C for 2 hours, and then inactivating the enzyme to obtain the first enzymolysis product of onion; (4) filtering the first enzymatic hydrolysis product of onion to obtain onion juice; (5) The pH of the onion juice is adjusted to 8.2 to obtain a mixture, and the mixture is treated with complex enzyme B, followed by stirring and enzymolysis at a stirring speed of 120 rpm, an enzymolysis temperature of 50°C, and a time of 2 h; the enzyme is inactivated to obtain a second enzymolysis product of onion; the complex enzyme B is a mixture of alkaline protease, neutral protease and flavor enzyme in a mass ratio of 1.5:1:0.5, and the mass of the complex enzyme B accounts for 2% of the mass of fresh onion; the mass of cellulase accounts for 1.2% of the mass of fresh onion; the mass of α-amylase accounts for 0.8% of the mass of fresh onion; the mass of complex enzyme A accounts for 1.2% of the mass of fresh onion; the complex enzyme A is a mixture of hemicellulase, pectinase and catalase in a mass ratio of 1:1.3:0.4.
[0038] (6) The second enzymatic hydrolysate of onion was precipitated with anhydrous ethanol, the supernatant was removed, the precipitate was passed through a 200-300 mesh silica gel column, and eluted with a 70% ethanol aqueous solution. The first eluate was collected. After the first eluate was concentrated, it was continued to pass through a polyacrylamide gel column, eluted with a 0.17% acetic acid aqueous solution, and the second eluate was collected; (7) The second eluate is spray-dried to obtain arginine fructose in onion.
[0039] The above enzyme inactivation conditions are heating at 121°C for 15 min.
[0040] Comparative Example 1 The difference between this comparative example and Example 1 is that step (3) is different.
[0041] Specifically: (3) Adding mixed enzymes to the onion water extract, hydrolyzing at 36℃ for 8h, inactivating the enzymes, and obtaining the first onion hydrolysis product; the mass of the mixed enzymes accounts for 3% of the mass of the fresh onion. The mixed enzymes are a mixture of cellulase, α-amylase, and complex enzyme A in a mass ratio of 15:25:36, and the complex enzyme A is a mixture of hemicellulase, pectinase, and catalase in a mass ratio of 1:1.4:0.5.
[0042] Comparative Example 2 The difference between this comparative example and Example 1 is that the order of adding the enzymes in step (3) is different.
[0043] Specifically: (3) Add α-amylase to the onion water extract, hydrolyze at 36°C for 4 hours, and inactivate the enzyme; continue to add complex enzyme A, hydrolyze at 36°C for 1.5 hours, and inactivate the enzyme; continue to add cellulase, hydrolyze at 36°C for 2.5 hours, and inactivate the enzyme to obtain the first onion hydrolysis product; the mass of cellulase accounts for 1% of the mass of fresh onion; the mass of α-amylase accounts for 1% of the mass of fresh onion; the mass of complex enzyme A accounts for 1% of the mass of fresh onion; the complex enzyme A is a mixture of hemicellulase, pectinase and catalase in a mass ratio of 1:1.4:0.5.
[0044] Comparative Example 3 The difference between this comparative example and Example 1 is that the complex enzyme A is a mixture of hemicellulase, pectinase and catalase in a mass ratio of 1:1:1.
[0045] Comparative Example 4 The difference between this comparative example and Example 1 is that step (5) is not included.
[0046] The onion juice in step (4) is precipitated with anhydrous ethanol, the supernatant is removed, the precipitate is passed through a 200-300 mesh silica gel column, and the precipitate is eluted with a 75% ethanol aqueous solution, and the first eluate is collected. After the first eluate is concentrated, it is continued to pass through a polyacrylamide gel column, and the elution is performed with a 0.15% acetic acid aqueous solution, and the second eluate is collected.
[0047] Comparative Example 5 The difference between this comparative example and Example 1 is that the compound enzyme B is a mixture of alkaline protease, neutral protease and flavor enzyme in a ratio of 1:1:1 by mass.
[0048] Comparative Example 6 The difference between this comparative example and Example 1 is that the compound enzyme B is a mixture of bromelain, ficin and flavor enzyme in a mass ratio of 1.6:1:0.4.
[0049] Comparative Example 7 The difference between this comparative example and Example 1 is as follows: (6) The second enzymatic hydrolysate of onion was precipitated with anhydrous ethanol, the supernatant was removed, the precipitate was passed through a 200-300 mesh silica gel column, eluted with a 70% by mass ethanol aqueous solution, the first eluate was collected, the first eluate was concentrated, and then passed through a polyacrylamide gel column, eluted with a 0.2% by mass acetic acid aqueous solution, and the second eluate was collected.
[0050] Performance Testing 1. The yield and purity of arginine-fructose in the products of Examples 1-2 and Comparative Examples 1-7 were measured. The yield of AF was: the mass of arginine-fructose in onion / the mass of fresh onion*100%, and the purity of AF was: the mass of AF / the mass of arginine-fructose in onion*100%.
[0051] 2. 30 volunteers were selected to score the smell of arginine fructose in the onions of Examples 1-2 and Comparative Examples 1-7, with a score of 1-10, and the average value was calculated. The higher the score, the lower the pungent smell of arginine fructose in the onions, and the better the product quality.
[0052] 3. The arginine fructose in onions of Examples 1-2 and Comparative Examples 1-7 were stored in dark conditions at 25° C. and 50% relative humidity for six months, and the arginine fructose in onions was observed to see whether the odor and color of the arginine fructose in onions changed. If the arginine fructose in onions did not change in odor and color, it was recorded as passed, otherwise it was recorded as failed.
[0053] Table 1 Performance test results AF yield% AF purity% stability Odor Rating Example 1 1.25 98.9 pass 9.9 Example 2 1.24 98.7 pass 9.8 Comparative Example 1 1.09 97.6 Failed 9.4 Comparative Example 2 1.13 98.0 pass 8.9 Comparative Example 3 1.16 98.3 pass 8.4 Comparative Example 4 1.11 97.8 Failed 8.8 Comparative Example 5 1.14 98.4 Failed 9.0 Comparative Example 6 1.10 98.1 Failed 9.1 Comparative Example 7 1.20 97.9 pass 9.5 From the above performance test results, it can be seen that the onions of Examples 1-2 have high arginine fructose extraction rates and high purities, especially the comprehensive performance of Example 1 is the most outstanding, which is mainly due to the synergistic effect of each step.
[0054] However, the comparative examples were significantly inferior to the embodiments in corresponding performance tests because they did not adopt the necessary technical solutions. In comparative example 1, the gradient enzymatic hydrolysis method was changed, and it can be seen that the extraction rate and purity effect decreased, proving that the gradient enzymatic hydrolysis scheme using a specific type of enzyme in the present invention has an important influence on the extraction rate and purity effect. In comparative example 3, the composite enzyme A scheme was not used, resulting in a stronger odor in the product. From the results, it can be seen that the changes in the composite enzyme A scheme failed to well decompose the components in the product that cause the odor to increase. The above experimental conclusions can be seen from comparative examples 4-6 that the stability of the product is improved after enzymatic hydrolysis using a composite enzyme B with a specific composition, further proving the importance of the technical scheme defined in the present invention for its technical effect.
[0055] The above is a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.
Claims
1. A process for extracting arginine fructose from onions, characterized in that: The extraction process comprises the following steps: (1) mixing onion and water, performing wet ball milling and sieving to obtain onion pulverizer; (2) heating the crushed onion at 80-90°C for 2-3 hours, cooling to room temperature, and obtaining an onion water extract; (3) Adding cellulase to the onion water extract, enzymolyzing at 35-37°C for 3-5 hours, and then inactivating the enzyme; continuing to add α-amylase, enzymolyzing at 35-37°C for 1-2 hours, and then inactivating the enzyme; continuing to add complex enzyme A, enzymolyzing at 35-37°C for 2-3 hours, and then inactivating the enzyme to obtain the first enzymolysis product of onion; (4) filtering the first enzymatic hydrolysis product of onion to obtain onion juice; (5) adjusting the pH of the onion juice to 8-8.2 to obtain a mixture, treating the mixture with a composite enzyme B, and then stirring and enzymolyzing the mixture at a stirring speed of 120-140 rpm, an enzymolysis temperature of 45-50°C, and a time of 2-3 hours; inactivating the enzyme to obtain a second enzymolysis product of the onion; the composite enzyme B is a mixture of alkaline protease, neutral protease, and flavor enzyme in a mass ratio of (1.5-1.7):1:(0.3-0.5), and the mass of the composite enzyme B accounts for 2-3% of the mass of the onion; (6) The second enzymatic hydrolysate of onion is precipitated with anhydrous ethanol, the supernatant is removed, the precipitate is passed through a 200-300 mesh silica gel column, and eluted with a 70-75% ethanol aqueous solution, the first eluate is collected, and after the first eluate is concentrated, it is continued to pass through a polyacrylamide gel column, eluted with a 0.15-0.17% acetic acid aqueous solution, and the second eluate is collected; (7) The second eluate is spray-dried to obtain arginine fructose in onion.
2. The process for extracting arginine fructose from onions according to claim 1, characterized in that: The mass of the cellulase in step (3) accounts for 0.8-1.2% of the mass of the onion.
3. The process for extracting arginine fructose from onion according to claim 1, characterized in that: The mass of the α-amylase in step (3) accounts for 0.8-1.2% of the mass of the onion.
4. The process for extracting arginine fructose from onion according to claim 1, characterized in that: The mass of the complex enzyme A in step (3) accounts for 0.8-1.2% of the mass of the onion.
5. The process for extracting arginine fructose from onion according to claim 1, characterized in that: In the step (1), the product is sieved through a 200-300 mesh sieve.
6. The process for extracting arginine fructose from onion according to claim 1, characterized in that: In the step (1), the mass ratio of onion to water is 1:(7-10).
7. The process for extracting arginine fructose from onion according to claim 1, characterized in that: In the step (1), the wet ball milling time is 2-4 h, the ball milling speed is 400-500 rpm, and the temperature is controlled at 40-45°C during the milling process.
8. The process for extracting arginine fructose from onion according to claim 1, characterized in that: Complex enzyme A is a mixture of hemicellulase, pectinase and catalase.
9. The process for extracting arginine fructose from onion according to claim 8, characterized in that: The complex enzyme A is a mixture of hemicellulase, pectinase and catalase in a mass ratio of 1: (1.3-1.5): (0.4-0.6).
10. The process for extracting arginine fructose from onion according to claim 1, characterized in that: The conditions for inactivating the enzyme are heating at 100-121°C for 10-15 minutes.