Method for efficiently extracting anthocyanin through auxiliary enzymolysis
By employing a multi-step enzymatic hydrolysis and extraction process for fruit and vegetable raw materials, the problems of low anthocyanin extraction rate and high cost in existing technologies have been solved, achieving efficient, green, and safe anthocyanin extraction with an extraction rate of over 87%.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- FUTURE FOOD (BAI MA) RESEARCH INSTITUTE
- Filing Date
- 2023-05-17
- Publication Date
- 2026-04-10
AI Technical Summary
Existing technologies for extracting anthocyanins suffer from high costs, toxicity, large equipment investment, and low extraction rates, making it difficult to meet the needs of industrial production.
After mixing fruit and vegetable raw materials with water and juicing them by cell wall breaking, a high-purity anthocyanin solution is obtained through a multi-step enzymatic hydrolysis process using pectinase, acidic water, and auxiliary enzymatic hydrolysis regulators, including primary, intermediate, and deep enzymatic hydrolysis, combined with repeated extraction and concentration steps.
It achieves efficient, green, and safe anthocyanin extraction with an extraction rate of over 87%, reducing production costs and avoiding the use of organic solvents.
Smart Images
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Abstract
Description
Technical Field
[0001] This invention belongs to the field of bioextraction technology, specifically relating to a method for efficiently extracting anthocyanins through assisted enzymatic hydrolysis. Background Technology
[0002] Current technologies mostly extract anthocyanins by using acidic ethanol or acidic methanol to penetrate cell walls, but this requires large quantities of alcohol, resulting in extremely high costs and significantly limiting its use in food. Furthermore, alcohols are somewhat toxic, and due to their low boiling points, they require high explosion-proof properties. In addition, alcohol recovery and treatment are necessary after extraction, further increasing costs.
[0003] Aqueous solution extraction offers advantages such as being green, safe, and low-cost. However, aqueous solutions cannot disrupt cell walls and cell membrane structures, resulting in extremely low anthocyanin dissolution rates that fail to meet the requirements of industrial production. Enzymatic treatment of the cell wall can improve the anthocyanin extraction rate, but requires large amounts of enzyme and the extraction effect remains unsatisfactory. Some studies have reported that using ultrasound, ultra-high pressure, and pulsed enzymatic hydrolysis techniques has moderately improved the hydrolysis effect, but the equipment investment is substantial and large-scale industrial production is not feasible. Summary of the Invention
[0004] The purpose of this section is to outline some aspects of embodiments of the present invention and to briefly describe some preferred embodiments. Simplifications or omissions may be made in this section, as well as in the abstract and title of this application, to avoid obscuring the purpose of these documents; however, such simplifications or omissions should not be construed as limiting the scope of the invention.
[0005] In view of the problems existing in the above and / or prior art, the present invention is proposed.
[0006] Therefore, the purpose of this invention is to overcome the shortcomings of the prior art and provide a method for efficiently extracting anthocyanins through assisted enzymatic hydrolysis.
[0007] To solve the above-mentioned technical problems, the present invention provides the following technical solution: including,
[0008] Fruit and vegetable raw materials are mixed with water, juiced by cell wall breaking, and filtered to obtain anthocyanin solution I and fruit and vegetable residue I;
[0009] Add pectinase and acidic water to fruit and vegetable residue I for primary enzymatic hydrolysis. After enzymatic hydrolysis at 10-40℃ for 0.5-4 hours, add an auxiliary enzymatic hydrolysis regulator and continue moderate enzymatic hydrolysis at 10-40℃ for 0.5-4 hours. Then add acidic water and perform deep enzymatic hydrolysis at 10-40℃ for 0.5-4 hours.
[0010] The product after deep enzymatic hydrolysis was filtered to separate the juice and residue, yielding anthocyanin solution II and fruit and vegetable residue II;
[0011] The fruit and vegetable residues II are added with acidic water for repeated extraction, and the anthocyanin solution III and fruit and vegetable residues III are obtained by filtration and centrifugation;
[0012] The anthocyanin solution I, the anthocyanin solution II and the anthocyanin solution III are mixed to obtain an anthocyanin mixed solution, and the anthocyanin mixed solution is sequentially concentrated, filtered, purified and dried to obtain the anthocyanin product.
[0013] As a preferred scheme of the method for efficiently extracting anthocyanins by assisted enzymolysis, the fruit and vegetable raw material is anthocyanin-rich fruits and vegetables, including one or more of blueberries, grapes, black corn and black chokeberry, and the solid-liquid ratio of the fruit and vegetable raw material to water is 1:1-5 (M / V).
[0014] As a preferred scheme of the method for efficiently extracting anthocyanins by assisted enzymolysis, in the primary enzymolysis, the addition amount of pectinase is 0.1%-5% of the fruit and vegetable residues I, and the addition amount of acidic water is 2-5 times of the fruit and vegetable residues I.
[0015] As a preferred scheme of the method for efficiently extracting anthocyanins by assisted enzymolysis, the assisted enzymolysis regulator includes one or more of erythritol, maltitol, sorbitol and sodium chloride.
[0016] As a preferred scheme of the method for efficiently extracting anthocyanins by assisted enzymolysis, the addition amount of the assisted enzymolysis regulator is 10%-30% of the mass of the acidic water used in the primary enzymolysis.
[0017] As a preferred scheme of the method for efficiently extracting anthocyanins by assisted enzymolysis, the addition amount of acidic water in the deep enzymolysis is 3-25 times of the fruit and vegetable residues I, and the total addition amount of acidic water in the whole enzymolysis process is 5-30 times of the mass of the fruit and vegetable residues I.
[0018] As a preferred scheme of the method for efficiently extracting anthocyanins by assisted enzymolysis, the addition amount of acidic water in the repeated extraction is 5-30 times of the mass of the fruit and vegetable residues II.
[0019] As a preferred scheme of the method for efficiently extracting anthocyanins by assisted enzymolysis, the repeated extraction is carried out at 10-40°C for 1-4h, and the repeated extraction is repeated 1-2 times.
[0020] As a preferred scheme of the method for efficiently extracting anthocyanins by assisted enzymolysis, the concentration is carried out at 50-60°C for 30-60min.
[0021] As a preferred solution of the method for efficiently extracting anthocyanins by assisted enzymatic hydrolysis, the method can efficiently extract anthocyanins, and the extraction rate is above 87%.
[0022] The present application has the following advantages:
[0023] The present application provides a method for efficiently extracting anthocyanins by assisted enzymatic hydrolysis, which does not use organic solvents such as alcohol in the whole process, is green and safe, only uses one enzyme preparation of pectinase, and the addition amount is low, thereby greatly reducing the production cost. The use of the assisted enzymatic hydrolysis regulator greatly improves the extraction rate of anthocyanins and overcomes the defects of low extraction rate in the prior art. DETAILED DESCRIPTION
[0024] In order to make the above-mentioned objects, features and advantages of the present application more apparent and easy to understand, the specific embodiments of the present application will be described in detail below with reference to the embodiments of the present application.
[0025] In the following description, a large number of specific details are set forth in order to provide a thorough understanding of the present application, but the present application can also be implemented in other ways different from those described herein, and those skilled in the art can make similar generalizations without departing from the connotation of the present application, therefore the present application is not limited by the specific embodiments disclosed below.
[0026] Secondly, the "one embodiment" or "embodiment" referred to herein means that the specific features, structures or characteristics can be included in at least one implementation of the present application. "In one embodiment" appearing in different places in the specification does not mean the same embodiment, nor is it an embodiment that is separate or alternative to other embodiments.
[0027] The determination method of the anthocyanin extraction rate in the present application refers to the high performance liquid chromatography method in the announcement of the National Center for Food Safety Risk Assessment.
[0028] Embodiment 1
[0029] 1) Juice residue separation: 100 parts of blueberry fruit were weighed, water was added according to a material liquid ratio of 1:2 (M / V), and the wall was broken and the juice was squeezed, and after filtration, anthocyanin solution I and fruit and vegetable residue I were obtained;
[0030] 2) Primary enzymatic hydrolysis: blueberry fruit residue I was weighed, 0.5% of pectinase based on the mass of blueberry fruit residue I was added, 2 times the weight of blueberry fruit residue I of acidic water was added, and enzymatic hydrolysis was carried out at 25 DEG C for 1h, wherein the acidic water was a citric acid aqueous solution with a pH of 2.8;
[0031] 3) Moderate enzymatic hydrolysis: erythritol was added, the addition amount was 20% of the acidic water in the primary enzymatic hydrolysis, and the enzymatic hydrolysis was continued at 25 DEG C for 1h;
[0032] 4) Deep enzymolysis: continue to add 28 times the weight of acidic aqueous solution of blueberry pomace I, continue to enzymolysis at 30°C for 1h;
[0033] 5) The product after deep enzymolysis is filtered to separate juice and residue, to obtain anthocyanin solution II and fruit and vegetable residue II;
[0034] 6) Soaking extraction: add 30 times the weight of acidic water of fruit and vegetable residue II, extract at 25°C for 1h, filter to obtain anthocyanin solution III and fruit and vegetable residue III;
[0035] 7) Concentration: mix anthocyanin solution II and anthocyanin solution III to obtain anthocyanin mixed solution, purify and remove impurities from the anthocyanin mixed solution in turn, vacuum concentrate at 50°C for 30min, mix the obtained solution with anthocyanin solution I to obtain anthocyanin concentrate;
[0036] 8) Drying: freeze-dry the anthocyanin concentrate to obtain powder to obtain anthocyanin product.
[0037] Example 2
[0038] The difference between this example and Example 1 is that the number of times of leaching in step 6) is 4, which is specifically:
[0039] 6) Leaching: add 30 times the weight of acidic water of fruit and vegetable residue II, extract at 25°C for 1h, filter to obtain anthocyanin solution, repeat the above steps for 3 times, and perform leaching for 4 times in total;
[0040] The remaining process steps are the same as those of Example 1.
[0041] Example 3
[0042] The difference between this example and Example 1 is that multiple enzymes are used for primary enzymolysis in step 2), which is specifically:
[0043] 2) Primary enzymolysis: take blueberry pomace I, add 0.5% pectinase, cellulase and protease by weight of blueberry pomace I, add 2 times the weight of acidic water of blueberry pomace I, and enzymolysis at 25°C for 1h, wherein the acidic water is a citric acid aqueous solution with pH 2.8;
[0044] The remaining process steps are the same as those of Example 1.
[0045] Comparative Example 1
[0046] This comparative example is a conventional organic solvent extraction method, which is different from Example 1 in that acidic water and auxiliary enzymolysis agent are not used for adjustment, but acidic ethanol is used for extraction, and the remaining process steps are the same as those of Example 1.
[0047] Comparative Example 2
[0048] The difference between the present comparative example and example 1 is that the primary enzymolysis in step 2) is performed with multiple enzymes, and no auxiliary enzymolysis is performed by adding an auxiliary enzymolysis agent, specifically:
[0049] 2) Primary enzymolysis: blueberry pomace I is weighed, 0.5% pectinase, cellulase and protease are added respectively based on the mass of the blueberry pomace I, 2 times the weight of the blueberry pomace I of acidic water is added, and enzymolysis is performed at 25°C for 1h, wherein the acidic water is a citric acid aqueous solution with a pH of 2.8;
[0050] 3) Moderate enzymolysis: no auxiliary enzymolysis regulator is added, and enzymolysis is continued at 25°C for 1h;
[0051] The remaining process steps are the same as those of example 1.
[0052] Comparative example 3
[0053] The difference between the present comparative example and example 1 is that the repeated extraction in step 6) is omitted, and the remaining process steps are the same as those of example 1.
[0054] The extraction rate of anthocyanins in the above examples and comparative examples is determined, and the results are shown in Table 1.
[0055] Table 1 Anthocyanin extraction rate of different extraction processes
[0056]
[0057]
[0058] As can be seen from Table 1, the method of the present application can greatly improve the extraction rate of anthocyanins by adding an auxiliary enzymolysis regulator during enzymolysis, and the extraction effect is comparable to that of conventional organic solvent alcohol extraction. On this basis, the extraction rate of anthocyanins can be further improved by optimizing the extraction times and increasing the types of enzymes, but considering the overall cost, the scheme of example 1 is the optimal scheme.
[0059] Example 4
[0060] This example is used to explore the effect of pectinase addition amount on the extraction effect in the primary enzymolysis process. Compared with example 1, the pectinase addition amount in step 2) is adjusted to 0.05%, 0.5% and 6% respectively, and the remaining process steps are the same as those of example 1.
[0061] The anthocyanin extraction rate under different pectinase addition amounts is determined, and the results are shown in Table 2.
[0062] Table 2 Anthocyanin extraction rate of different pectinase addition amounts
[0063]
[0064] As can be seen from Table 2, in the present application scheme, pectinase within the scope of the present claim (0.5%) can achieve excellent extraction effect, too low addition (0.05%) of enzyme hydrolysis of cell wall pectin is insufficient, resulting in too low extraction rate of anthocyanin extraction, too high addition (6%) has little contribution to anthocyanin extraction rate, but leads to high production cost.
[0065] Example 5
[0066] This example is used to explore the influence of the addition amount of auxiliary enzyme hydrolysis agent on the extraction effect. Compared with Example 1, the addition amount of erythritol in the secondary enzymatic hydrolysis process of step 3) is adjusted to 3%, 10% and 40% of the weight of the primary enzymatic hydrolysis acid aqueous solution of step 2) respectively, and the remaining process steps are the same as those of Example 1. The anthocyanin extraction rate under different erythritol addition amounts is determined, and the results are shown in Table 3.
[0067] Table 3 Anthocyanin extraction rate under different erythritol addition amounts
[0068]
[0069] As can be seen from Table 3, within the scope of the present technical requirement (10% to 30%), the addition of erythritol can greatly improve the anthocyanin extraction rate, but too low addition (3%) of erythritol cannot significantly improve the anthocyanin extraction rate. On the basis of the present application scheme, further increasing the addition amount of erythritol cannot improve the anthocyanin extraction rate. Therefore, the addition amount of erythritol is preferably 10% to 30%.
[0070] Example 6
[0071] This example is used to explore the influence of the type of auxiliary enzyme hydrolysis agent on the extraction effect. Compared with Example 1, the type of auxiliary enzyme hydrolysis agent in the secondary enzymatic hydrolysis process of step 3) is adjusted to maltitol, sorbitol + sodium chloride, and the remaining process steps are the same as those of Example 1.
[0072] The anthocyanin extraction rate of different auxiliary enzyme hydrolysis agents is determined, and the results are shown in Table 4.
[0073] Table 3 Anthocyanin extraction rate under different erythritol addition amounts
[0074]
[0075] As can be seen from Table 4, in the present application scheme, the addition of one or more of erythritol, maltitol, sorbitol and sodium chloride can greatly improve the anthocyanin extraction rate, and the technical effect achieved by selecting erythritol is the best.
[0076] In conclusion, the present application provides a method for efficiently extracting anthocyanins through auxiliary enzymolysis, which does not use organic solvents such as alcohol in the whole process, is green and safe, only uses one enzyme preparation of pectinase, and has a low addition amount, greatly reduces the production cost, greatly improves the extraction yield of anthocyanins through the use of auxiliary enzymolysis regulator, and overcomes the defects of low extraction rate in the prior art.
[0077] It should be noted that the above examples are only used to illustrate the technical solutions of the present application and are not limiting. Although the present application has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solutions of the present application can be modified or replaced by equivalents without departing from the spirit and scope of the technical solutions of the present application, which should be covered in the scope of the claims of the present application.
Claims
1. A method for efficient extraction of anthocyanins by assisted enzymatic hydrolysis, characterized by: The application relates to a method for extracting anthocyanins from fruit and vegetable raw materials. The fruit and vegetable raw materials are mixed with water, broken wall juice is extracted, and filtration is performed to obtain an anthocyanin solution I and fruit and vegetable residues I; Pectinase and acidic water are added to the fruit and vegetable residues I to perform primary enzymolysis, auxiliary enzymolysis regulators are added after enzymolysis at 10-40 DEG C for 0.5-4h to continue moderate enzymolysis at 10-40 DEG C for 0.5-4h, and then acidic water is added to perform deep enzymolysis at 10-40 DEG C for 0.5-4h; The product after deep enzymolysis is filtered to separate juice and residues, and an anthocyanin solution II and fruit and vegetable residues II are obtained; Acidic water is added to the fruit and vegetable residues II to perform leaching, and filtration and centrifugation are performed to obtain an anthocyanin solution III and fruit and vegetable residues III; The anthocyanin solution I, the anthocyanin solution II and the anthocyanin solution III are mixed to obtain an anthocyanin mixed solution, and the anthocyanin mixed solution is sequentially concentrated, filtered, purified and dried to obtain an anthocyanin product.
2. The method for improving the extraction of anthocyanins by assisted enzymatic hydrolysis according to claim 1, characterized in that: The fruit and vegetable raw materials are fruit and vegetable materials rich in anthocyanins, and the fruit and vegetable raw materials are one or more of blueberries, grapes, black corn and black fruit and vegetable materials.
3. The method for improving the extraction of anthocyanins by assisted enzymatic hydrolysis according to claim 1, characterized in that: In the primary enzymolysis, the addition amount of pectinase is 0.1%-5% of the fruit and vegetable residues I, and the addition amount of acidic water is 2-5 times of the fruit and vegetable residues I.
4. The method for improving the extraction of anthocyanins by assisted enzymatic hydrolysis according to claim 1, characterized in that: The auxiliary enzymolysis regulator comprises one or more of erythritol, maltitol, sorbitol and sodium chloride.
5. The method for improving the extraction of anthocyanins by assisted enzymatic hydrolysis according to claim 1 or 4, characterized in that: The addition amount of the auxiliary enzymolysis regulator is 10%-30% of the mass of the acidic water used in the primary enzymolysis.
6. The method for improving the extraction of anthocyanins by assisted enzymatic hydrolysis according to claim 1, characterized in that: In the deep enzymolysis, the addition amount of acidic water is 3-25 times of the fruit and vegetable residues I, and the total addition amount of acidic water in the whole enzymolysis process is 5-30 times of the mass of the fruit and vegetable residues I.
7. The method for improving the extraction of anthocyanins by assisted enzymatic hydrolysis according to claim 1, characterized in that: In the leaching process, the addition amount of acidic water is 5-30 times of the mass of the fruit and vegetable residues II.
8. The method for improving the extraction of anthocyanins by assisted enzymatic hydrolysis according to claim 7, characterized in that: The leaching is performed at 10-40 DEG C for 1-4h, and the leaching is repeated for 1-2 times.
9. The method for improving the extraction of anthocyanins by assisted enzymatic hydrolysis according to claim 1, characterized in that: The concentration is performed at 50-60 DEG C for 30-60 min.
10. The method for improving the extraction of anthocyanins by assisted enzymatic hydrolysis according to any one of claims 1 to 9, characterized in that: The method can efficiently extract anthocyanins, and the extraction rate is above 87%.