Composite enzymatic hydrolysis method for improving bioavailability of wolfberry whole fruit puree and application of composite enzymatic hydrolysis method
Through compound enzymatic lysis technology and antioxidant treatment, the problems of insufficient release of nutrients and poor stability in traditional wolfberry processing were solved, and functional drinks of wolfberry with high bioavailability and stability were prepared, which enhanced the nutritional value and market competitiveness of the product.
Patent Information
- Application Number
- CN202510842308.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-06-23
- Publication Date
- 2025-07-29
AI Technical Summary
Traditional wolfberry processing methods are difficult to fully release nutrients, insufficient enzymatic decomposition, single treatment of different nutrients, poor product stability, affecting flavor and shelf life.
Compound enzyme preparations (pectinase, cellulase, lipase, protease) are used to work in a synergistic manner, combining antioxidants and homogenization treatment, and the enzymatic conditions are controlled, including pH, temperature and stirring speed, enzymatic reactions are carried out, and enzyme decomposition and subsequent homogenization are carried out.
It significantly improves the bioavailability of the whole fruit pulp of wolfberry, enhances product stability and taste, expands the application range of wolfberry products, and meets the needs of high-quality functional beverages.
Smart Images
Figure CN120381099A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the technical field of food processing, and specifically provides a composite enzymatic hydrolysis method and application for improving the bioavailability of whole wolfberry fruit pulp. Background Art
[0002] In the field of food processing, wolfberry is a nutritious food ingredient rich in various nutrients such as wolfberry polysaccharides, proteins, and fats, with extremely high nutritional value and health care functions. However, traditional wolfberry processing methods are difficult to fully release the nutrients in wolfberry. For example, in the common production process of wolfberry pulp, simple crushing, pressing, etc. are mostly used, resulting in a large amount of nutrients that cannot be effectively absorbed by the human body, causing waste of resources.
[0003] With the improvement of people's health awareness, higher requirements are put forward for the quality and nutrient absorption efficiency of wolfberry products. Although some existing wolfberry products on the market adopt enzymatic hydrolysis technology, most of them have problems such as insufficient enzymatic hydrolysis and single treatment of different nutrients. For example, some products only focus on the decomposition of pectin to improve the taste, but ignore the optimization of proteins and fats, resulting in great room for improvement in the nutritional balance and bioavailability of the products.
[0004] In addition, in terms of the stability of wolfberry products, traditional processes also face challenges. Since the fat in wolfberry pulp is prone to oxidation and proteins may denature during storage, this not only affects the flavor and quality of the products, but also shortens the shelf life of the products, restricting the development of the wolfberry industry. Therefore, it is urgent to develop an innovative technology that can comprehensively improve the bioavailability of whole wolfberry fruit pulp and enhance the product stability.
[0005] In view of this, the present application is specifically proposed. Summary of the Invention
[0006] The purpose of the present invention is to provide a composite enzymatic hydrolysis method and application for improving the bioavailability of whole wolfberry fruit pulp, so as to solve the problems mentioned in the above background art.
[0007] To solve the above technical problems, a composite enzymatic hydrolysis method for improving the bioavailability of wolfberry whole fruit pulp provided by the present invention includes the following steps: S1: Preparation of wolfberry whole fruit pulp: Select high-quality wolfberries, remove impurities, wash them, and then add deionized water to make a delicate and uniform wolfberry whole fruit pulp; S2: Preparation of composite enzyme preparation: Weigh each enzyme preparation according to a certain mass ratio of pectinase, cellulase, lipase, and protease and mix them evenly; S3: Enzymatic hydrolysis process: Transfer the wolfberry whole fruit pulp to a reaction kettle, adjust the pH to 4.5 - 5.0 with a buffer solution, control the temperature at 35 - 40 °C, add 1.5 - 2.5 g of the composite enzyme preparation per liter of the original pulp, stir and hydrolyze at 100 - 140 r / min for 1.5 - 2.5 hours; Add a natural antioxidant with a mass fraction of 0.05% - 0.15% when the enzymatic hydrolysis proceeds to 0.5 - 1.5 hours; After the enzymatic hydrolysis is completed, quickly raise the temperature to 80 - 90 °C and maintain it for 8 - 12 minutes to inactivate the enzyme; S4: Detection of enzymatic hydrolysis products: Detect the pectin content, dietary fiber content and bioavailability, fat content and stability, protein and small molecule peptide content of the enzymatic hydrolysis products; S5: Subsequent treatment: After filtering the enzymatic hydrolysis solution, add natural sweeteners, acidulants and a stabilizer with a mass fraction of 0.03% - 0.07%, homogenize it 1 - 3 times under a pressure of 20 - 30 MPa, and then perform high-temperature short-time sterilization and aseptic filling to make a functional beverage.
[0008] Through enzymatic hydrolysis with a composite enzyme preparation under certain conditions, combined with antioxidant addition, enzyme inactivation treatment, and subsequent detection and processing techniques, the overall technical solution can flexibly adapt to different raw material characteristics and production requirements, efficiently degrade pectin and fiber, stabilize fat components, optimize protein structure, significantly improve the bioavailability of wolfberry whole fruit pulp, and make high-quality functional beverages, solving many problems existing in traditional wolfberry pulp.
[0009] Further, in the above S2, the mass ratio of pectinase, cellulase, lipase, and protease in the composite enzyme preparation is 3:3:2:2. This composite enzyme preparation with a specific mass ratio can make the enzymes synergistically act to achieve the best effect, more effectively process pectin, fiber, fat, and protein in wolfberry whole fruit pulp, and improve the bioavailability.
[0010] Further, in the above S3, the pH is adjusted to 4.8 with a citric acid - sodium citrate buffer solution. pH 4.8 is a relatively optimal pH environment for the synergistic action of pectinase, cellulase, lipase, and protease, which can make each enzyme exhibit nearly the best activity, promote the efficient progress of the enzymatic hydrolysis reaction, and better realize the treatment of each nutrient component.
[0011] Further, in step S3, the enzymatic hydrolysis temperature is controlled at 37°C. 37°C is close to the human body temperature and is the suitable working temperature for most enzymes. At this temperature, the enzyme activity is relatively high, which can accelerate the enzymatic hydrolysis reaction rate, improve the enzymatic hydrolysis efficiency, and ensure that all nutritional components are fully processed.
[0012] Further, in step S3, 2 g of compound enzyme preparation is added per liter of the original pulp. This addition amount can ensure that the compound enzyme fully reacts with the substrates in the whole-fruit wolfberry original pulp. While effectively degrading pectin, promoting the decomposition of dietary fiber, stabilizing fats, and optimizing the protein structure, it can avoid the increase in cost and unnecessary side reactions caused by excessive enzyme amount.
[0013] Further, in step S3, the stirring speed during enzymatic hydrolysis is 120 r / min, the enzymatic hydrolysis time is 2 hours, and rosemary extract with a mass fraction of 0.1% is added when the enzymatic hydrolysis proceeds for 1 hour. The stirring speed of 120 r / min can enable the enzyme to fully mix and contact with the original pulp, and the enzymatic hydrolysis time of 2 hours can ensure that the enzymatic hydrolysis reaction proceeds fully; adding 0.1% rosemary extract when the enzymatic hydrolysis proceeds for 1 hour can timely inhibit fat oxidation, ensure the stability of fat during enzymatic hydrolysis and subsequent storage of the product, and maintain the quality and flavor consistency of the product.
[0014] Further, in step S3, after the enzymatic hydrolysis, the temperature is rapidly raised to 85°C and maintained for 10 minutes to inactivate the enzyme. This enzyme inactivation condition can effectively terminate the enzyme activity, prevent the enzyme from continuing to act in the subsequent processing, resulting in excessive decomposition of nutritional components or adverse changes, and at the same time, retain the effective nutritional components in the product to the greatest extent.
[0015] Further, in step S5, xanthan gum with a mass fraction of 0.05% is added as a stabilizer, the homogenization pressure is 25 MPa, homogenization is carried out 2 times, and the high-temperature short-time sterilization condition is 121°C for 3 s. Xanthan gum with a mass fraction of 0.05% can effectively enhance the stability of the product; homogenization 2 times under a pressure of 25 MPa can make all components in the product fully mixed and uniform, refine the particles, and make the product taste more delicate and smooth; high-temperature short-time sterilization at 121°C for 3 s can, while ensuring the killing of microorganisms, minimize the impact on the nutritional components and flavor of the product.
[0016] Application of a functional beverage prepared by a compound enzymatic hydrolysis method for improving the bioavailability of whole-fruit wolfberry original pulp in the food field. This functional beverage has high bioavailability and is rich in easily absorbable nutritional components. Applying it in the food field can provide consumers with healthy beverages with rich nutrition and good taste, expand the application scope of wolfberry products, and meet the market demand for high-quality functional foods.
[0017] Compared with the prior art, the beneficial effects of the present invention are: 1. Improve the bioavailability of nutrients: By precisely controlling the proportions of pectinase, cellulase, lipase, and protease in the complex enzyme preparation and conducting enzymatic hydrolysis reactions under suitable temperature and pH conditions, the pectin and fiber in the wolfberry puree were effectively degraded, making dietary fibers such as wolfberry polysaccharides more easily absorbed by the human body. At the same time, the structures of proteins and fats were optimized. After detection, the bioavailability of wolfberry polysaccharides was increased to 70%, and the bioavailability of proteins was increased to 80%, greatly improving the nutritional value of the whole wolfberry fruit puree and meeting the human body's high-efficiency nutrient intake requirements.
[0018] 2. Stabilize the product quality and flavor: Add natural antioxidant rosemary extract during the enzymatic hydrolysis process, combined with the precise enzymatic hydrolysis of lipase on fats, effectively inhibiting fat oxidation and avoiding the layering phenomenon of the product during storage. After a 7-day accelerated oxidation experiment at 40°C, the peroxide value and acid value of the product still remained at a low level, ensuring the quality stability and flavor consistency of the product, extending the product's shelf life, and enhancing the consumer's drinking experience.
[0019] 3. Optimize the product taste and texture: The enzymatic hydrolysis treatment reduced the pectin content of the wolfberry puree. Subsequently, adding stabilizers and conducting homogenization treatment made the product taste more delicate and smooth. Blending natural sweeteners and acidulants further improved the product flavor, meeting the consumer's taste requirements while enhancing the overall quality of the product and making it more competitive in the market.
[0020] 4. Expand the application scope of wolfberry products: The high-bioavailability wolfberry whole fruit puree functional beverage prepared by this invention enriches the variety of wolfberry products. Its characteristics of rich nutrition and easy absorption make it not only suitable as a daily drink but also applicable to fields such as health foods and foods for special medical purposes, providing technical support for the diversified development of the wolfberry industry and creating greater economic value. Description of the Drawings
[0021] Figure 1 It is a flowchart of a complex enzymatic hydrolysis method for improving the bioavailability of wolfberry whole fruit puree. Detailed Embodiments
[0022] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.
[0023] Please refer to Figure 1 , the present invention provides a technical solution: A complex enzymatic hydrolysis method and application for improving the bioavailability of wolfberry whole fruit puree, including: Example 1: (I) Raw material preparation: Select 500 g of super-grade Chinese wolfberries from Zhongning, Ningxia, carefully pick out the impurities and bad fruits, and rinse them 3 times with deionized water. Mix the washed Chinese wolfberries with 500 g of deionized water, pour them into a high-speed pulping machine, and pulp at a speed of 8000 r / min for 5 min to obtain a delicate and uniform whole-fruit Chinese wolfberry pulp.
[0024] (II) Preparation of compound enzyme preparation: Accurately weigh 3 g of pectinase, 3 g of cellulase, 2 g of lipase, and 2 g of protease, mix them thoroughly, and prepare 10 g of compound enzyme preparation.
[0025] (III) Enzymolysis process: Transfer the whole-fruit Chinese wolfberry pulp to a 5-L reaction kettle, and adjust the pH of the pulp to 4.8 with citric acid-sodium citrate buffer solution. Turn on the temperature control system of the reaction kettle, stabilize the temperature at 37 °C, and stir the pulp at a speed of 120 r / min. Add 10 g of compound enzyme preparation and start timing for enzymolysis for 2 h. When the enzymolysis proceeds to 1 h, add 0.5 g (mass fraction of 0.1%) of natural antioxidant rosemary extract. After the enzymolysis is completed, quickly raise the temperature in the reaction kettle to 85 °C and maintain it for 10 min for enzyme inactivation treatment.
[0026] (IV) Detection of enzymolysis products: 1. Detection of pectin content: Take 10 mL of enzymolysis product, centrifuge at 3000 r / min for 10 min, and take the supernatant. Using the carbazole colorimetric method, sequentially add 0.5 mL of 0.15% carbazole ethanol solution and 5 mL of concentrated sulfuric acid to 5 mL of the supernatant, shake well, heat in a boiling water bath for 15 min, and cool to room temperature. Measure the absorbance at a wavelength of 530 nm with a spectrophotometer, and calculate through the standard curve. The pectin content is reduced by 60% compared with that before enzymolysis.
[0027] 2. Evaluation of dietary fiber (Chinese wolfberry polysaccharide) content and bioavailability: The content of Chinese wolfberry polysaccharide is determined by the sulfuric acid-phenol method. The content of Chinese wolfberry polysaccharide after enzymolysis is 80 mg / g, which is increased by 20% compared with that before enzymolysis. Through a simulated human digestion experiment, the content of absorbable Chinese wolfberry polysaccharide after digestion is measured, and the bioavailability is calculated to be increased to 70%, while the bioavailability of the non-enzymolyzed pulp is only 30%.
[0028] 3. Detection of fat content and stability: The fat content is determined by the Soxhlet extraction method, and the fat content after enzymolysis is 2 g / 100 g. After a 7-day accelerated oxidation experiment at 40 °C, the peroxide value is measured to be 0.08 meq / kg, and the acid value is 1.2 mg KOH / g, indicating good fat stability and no obvious oxidation and stratification phenomena.
[0029] 4. Detection of protein and small molecule peptide content: The total protein content measured by the Kjeldahl method was 12 g / 100 g, and the small molecule peptide content measured by the Folin-phenol method was 8 g / 100 g. After calculation, the protein decomposition rate reached 66.7%, and the biological utilization rate increased to 80%, which was much higher than 50% of the non-enzymatically hydrolyzed original pulp.
[0030] (V) Subsequent treatment: The enzymolysis solution after inactivating the enzyme was filtered through a 100-mesh filter screen to remove the residues that were not completely enzymolyzed. 2 g of xylitol was added to the filtrate as a sweetener, 0.5 g of citric acid as an acidulant, and 2.5 g (mass fraction of 0.05%) of xanthan gum as a stabilizer. The prepared solution was transferred to a homogenizer and homogenized twice at a pressure of 25 MPa. Finally, high-temperature short-time sterilization (121 °C, 3 s) was carried out, and it was filled into 200-mL glass bottles under aseptic conditions to obtain a functional beverage with high-efficiency absorption of wolfberry whole fruit original pulp compound enzymolysis.
[0031] Example 2: Single enzymolysis (only using pectinase): 1. Raw materials and preparation: Prepare 500 g of wolfberry whole fruit original pulp as in the optimal example.
[0032] 2. Enzymolysis process: Adjust the pH of the original pulp to 4.8, control the temperature at 37 °C, add 3 g of pectinase, stir and enzymolyze at 120 r / min for 2 h, add 0.5 g of rosemary extract at 1 h of enzymolysis, and inactivate the enzyme at 85 °C for 10 min after the enzymolysis ends.
[0033] 3. Detection results: The pectin content decreased by 40%, the biological utilization rate of wolfberry polysaccharide increased to 40%, the fat peroxide value was 0.12 meq / kg, the acid value was 1.5 mg KOH / g, there was a slight oxidation phenomenon, the protein decomposition rate was 20%, the small molecule peptide content was 3 g / 100 g, and the biological utilization rate was 55%.
[0034] 4. Result analysis: Single pectinase can only partially degrade pectin, and the treatment effect on other components is not good, resulting in limited improvement in the comprehensive quality of the product.
[0035] Example 3: Improper enzyme ratio: 1. Raw materials and preparation: Prepare 500 g of wolfberry whole fruit original pulp as in the optimal example.
[0036] 2. Enzymolysis process: Prepare a compound enzyme preparation, including 5 g of pectinase, 1 g of cellulase, 1 g of lipase, and 3 g of protease. Adjust the pH of the original pulp to 4.8, stir and enzymolyze at 37 °C and 120 r / min for 2 h, add 0.5 g of rosemary extract at 1 h of enzymolysis, and inactivate the enzyme at 85 °C for 10 min.
[0037] 3. Detection results: The pectin content decreased by 50%, the bioavailability of wolfberry polysaccharide increased to 50%, the peroxide value of fat was 0.1 meq / kg, the acid value was 1.3 mg KOH / g, the protein decomposition rate was 40%, the content of small molecule peptides was 5 g / 100 g, and the bioavailability was 65%.
[0038] 4. Result analysis: The improper ratio of enzymes affected the synergistic effect of each enzyme, resulting in a less effective treatment of nutritional components compared to the best example, and the improvement of product quality was not obvious.
[0039] Example 4: Different enzymatic hydrolysis temperatures: 1. Raw materials and preparation: Prepare 500 g of original wolfberry fruit pulp in the same way as the best example.
[0040] 2. Enzymatic hydrolysis process: Adjust the pH of the original pulp to 4.8, control the temperature at 30 °C, add 10 g of compound enzyme preparation, stir and hydrolyze at 120 r / min for 2 h, add 0.5 g of rosemary extract when hydrolyzing for 1 h, and inactivate the enzyme at 85 °C for 10 min.
[0041] 3. Detection results: The pectin content decreased by 50%, the bioavailability of wolfberry polysaccharide increased to 55%, the peroxide value of fat was 0.09 meq / kg, the acid value was 1.25 mg KOH / g, the protein decomposition rate was 50%, the content of small molecule peptides was 6 g / 100 g, and the bioavailability was 70%.
[0042] 4. Result analysis: The lower temperature limited the enzyme activity, and the enzymatic hydrolysis reaction rate became slower, resulting in a less effective treatment of each nutritional component compared to the best example, and the improvement of bioavailability was insufficient.
[0043] Example 5: No antioxidant added: 1. Raw materials and preparation: Prepare 500 g of original wolfberry fruit pulp in the same way as the best example.
[0044] 2. Enzymatic hydrolysis process: Adjust the pH of the original pulp to 4.8, 37 °C, add 10 g of compound enzyme preparation, stir and hydrolyze at 120 r / min for 2 h, do not add antioxidants during the whole enzymatic hydrolysis process, and inactivate the enzyme at 85 °C for 10 min.
[0045] 3. Detection results: The pectin content decreased by 60%, the bioavailability of wolfberry polysaccharide increased to 70%, the peroxide value of fat was 0.15 meq / kg, the acid value was 1.8 mg KOH / g, obvious oxidation occurred, the protein decomposition rate was 66.7%, the content of small molecule peptides was 8 g / 100 g, and the bioavailability was 80%.
[0046] 4. Result analysis: The lack of antioxidant addition led to an increase in fat oxidation, affecting the stability and flavor of the product, indicating that antioxidants play an important role in stabilizing fat components.
[0047] In summary, by using a composite enzyme preparation in a specific ratio, precisely controlling conditions such as temperature and pH value during the enzymatic hydrolysis process, and combining means such as antioxidant addition and homogenization treatment, the present invention realizes the functions of efficiently degrading pectin and fiber in wolfberry puree, stabilizing fat components, and optimizing protein structure. A functional beverage of wolfberry whole fruit puree with high bioavailability, stable quality, and excellent taste is successfully prepared, effectively solving problems such as insufficient utilization of nutrients and poor product stability existing in traditional wolfberry processing, and promoting the innovative development of wolfberry processing technology.
Claims
1. A composite enzymatic hydrolysis method for improving the bioavailability of whole wolfberry fruit pulp, characterized in that: It includes the following steps: S1: Prepare wolfberry whole fruit puree: Select high-quality wolfberries, remove impurities, wash them, and then add deionized water to make a delicate and uniform wolfberry whole fruit puree; S2: Prepare a complex enzyme preparation: Weigh each enzyme preparation according to a certain mass ratio of pectinase, cellulase, lipase, and protease and mix them evenly; S3: Enzymolysis process: Transfer the wolfberry whole fruit puree to a reaction kettle, adjust the pH to 4.5 - 5.0 with a buffer solution, control the temperature at 35 - 40 °C, add 1.5 - 2.5 g of the complex enzyme preparation per liter of puree, stir and enzymolyze at 100 - 140 r / min for 1.5 - 2.5 hours; Add a natural antioxidant with a mass fraction of 0.05% - 0.15% when the enzymolysis proceeds to 0.5 - 1.5 hours; After the enzymolysis ends, quickly raise the temperature to 80 - 90 °C and maintain it for 8 - 12 minutes to inactivate the enzyme; S4: Detection of enzymolysis products: Detect the pectin content, dietary fiber content and bioavailability, fat content and stability, protein and small molecule peptide content of the enzymolysis products; S5: Subsequent treatment: After filtering the enzymolysis solution, add a natural sweetener, acidulant and a stabilizer with a mass fraction of 0.03% - 0.07%, homogenize 1 - 3 times under a pressure of 20 - 30 MPa, and then perform high-temperature short-time sterilization and aseptic filling to make a functional beverage.
2. The composite enzymatic hydrolysis method for improving the bioavailability of wolfberry whole fruit pulp according to claim 1, characterized in that: In S2, the mass ratio of pectinase, cellulase, lipase, and protease in the complex enzyme preparation is 3:3:2:
2.
3. The composite enzymatic hydrolysis method for improving the bioavailability of whole wolfberry fruit puree according to claim 1, characterized in that: In S3, use a citric acid - sodium citrate buffer solution to adjust the pH to 4.
8.
4. The composite enzymatic hydrolysis method for improving the bioavailability of whole wolfberry fruit puree according to claim 1, characterized in that: In S3, control the enzymolysis temperature at 37 °C.
5. A composite enzymatic hydrolysis method for improving the bioavailability of whole wolfberry pulp as described in claim 1, characterized in that: In S3, add 2 g of the complex enzyme preparation per liter of puree.
6. The composite enzymatic hydrolysis method for improving the bioavailability of whole wolfberry fruit pulp according to claim 1, characterized in that: In S3, the stirring speed during enzymolysis is 120 r / min, the enzymolysis time is 2 hours, and add rosemary extract, a natural antioxidant with a mass fraction of 0.1%, when the enzymolysis proceeds to 1 hour.
7. A composite enzymatic hydrolysis method for improving the bioavailability of whole wolfberry fruit pulp as claimed in claim 1, characterized in that: In S3, quickly raise the temperature to 85 °C and maintain it for 10 minutes to inactivate the enzyme after the enzymolysis ends.
8. A composite enzymatic hydrolysis method for improving the bioavailability of wolfberry whole fruit pulp, characterized in that: In S5, add xanthan gum with a mass fraction of 0.05% as a stabilizer, the homogenization pressure is 25 MPa, homogenize 2 times, and the high-temperature short-time sterilization conditions are 121 °C, 3 s.
9. Application of a functional beverage prepared by a complex enzymolysis method for improving the bioavailability of wolfberry whole fruit puree as described in any one of claims 1 - 8 in the food field.