Method for preparing narrow-distribution small molecules by targeting enzymolysis of a medicinal and edible raw material

Abstract

This invention provides a method for preparing narrow-distribution small molecules from food-derived raw materials through targeted enzymatic hydrolysis, relating to the fields of food biotechnology, preparation of functional components from food-derived raw materials, and plant extraction and purification. The method is characterized by the following steps: raw material pretreatment; gentle extraction; targeted enzymatic hydrolysis; enzyme inactivation and stabilization; and narrow-distribution small molecule enrichment. The advantages of this invention are: through the synergistic technology of targeted enzymatic hydrolysis and membrane separation, the method achieves the directional release and narrow-distribution enrichment (molecular weight ≥70% of 300-800 Da, PDI <1.5) of non-protein active small molecules such as polyphenols and flavonoids from food-derived raw materials. The process is gentle and efficient, significantly improving product uniformity, batch stability, and bioavailability, and is adaptable to various end-product dosage forms, facilitating large-scale production.

Description

Technical Field

[0001] This invention relates to the fields of food biotechnology, preparation of medicinal and edible functional components and plant extraction and purification technology, and particularly to a method for preparing narrowly distributed small molecules by targeted controlled enzymatic hydrolysis of medicinal and edible raw materials. Background Technology

[0002] Food-medicine homology active ingredients refer to natural compounds that can be consumed as everyday food and also possess specific pharmacological effects. These are commonly found in substances listed in the national catalogue of "substances that are both food and traditional Chinese medicine," such as goji berries, hawthorn, and jujubes. These components mainly include polysaccharides, flavonoids, polyphenols, and alkaloids, exhibiting various physiological activities such as antioxidant, anti-inflammatory, immunomodulatory, and hypoglycemic effects. They originate from the traditional Chinese medicine concept of "food and medicine sharing the same origin," emphasizing the commonality between food and medicine in terms of source and efficacy, and have also been scientifically validated in modern nutritional and pharmacological research.

[0003] Currently, the extraction and preparation technologies for active ingredients from food and medicine homology still have shortcomings. First, existing extraction methods typically involve boiling, alcohol extraction, and coarse grinding, resulting in insufficient dissolution of active ingredients, a high proportion of macromolecular substances, and a high proportion of macromolecular impurities in the resulting extracts, significantly restricting the bioavailability and functional stability of the products. Second, conventional enzymatic hydrolysis is mostly a non-directional and extensive process, resulting in products with a wide molecular weight distribution and insufficient uniformity, making it difficult to consistently obtain small molecule components with good consistency. In addition, current research on the preparation of small molecule active substances focuses mainly on protein sources and peptide components, while targeted release, structural modification, and narrow molecular weight distribution regulation technologies for non-protein active small molecules such as polyphenols, flavonoids, and oligosaccharides widely present in food and medicine homology materials remain weak. Furthermore, some purification and enrichment processes have high equipment requirements and complex procedures, which are not conducive to large-scale, low-cost application on conventional food production lines.

[0004] Therefore, there is an urgent need to develop an efficient, targeted, scalable extraction and preparation technology applicable to various types of active ingredients in order to break through existing bottlenecks and promote the high-value utilization of medicinal and edible resources. Summary of the Invention

[0005] The purpose of this invention is to provide a method for preparing narrowly distributed small molecules by targeted enzymatic hydrolysis of medicinal and edible raw materials, which solves the technical problems in the prior art of insufficient dissolution of active ingredients in the extraction of medicinal and edible raw materials, and the wide distribution, poor uniformity and difficulty in large-scale enrichment of small molecule products caused by non-directional enzymatic hydrolysis.

[0006] To achieve the above-mentioned objectives, the technical solution adopted by this invention is as follows:

[0007] The technical solution adopted in this invention is as follows: The method for preparing narrowly distributed small molecules by targeted enzymatic hydrolysis of medicinal and edible raw materials in this invention includes the following steps: (1) Raw material pretreatment: Cleaning and crushing the medicinal and edible plant raw materials and passing them through a 20-40 mesh sieve;

[0008] (2) Mild extraction: Add water at a material-to-liquid ratio of 1:8-12, extract at 75-90℃ for 1-1.5 hours, and filter to obtain the extract;

[0009] (3) Targeted enzymatic hydrolysis: Adjust the pH of the extract to 4.5-6.5, add the compound enzyme system, and enzymatically hydrolyze at 45-55℃ for 2-4 hours;

[0010] (4) Enzyme inactivation and stabilization: Heat to 85-95℃ and keep warm for 10-15 min to terminate enzymatic hydrolysis;

[0011] (5) Enrichment of narrow-distribution small molecules: Membrane separation and fractionation are used to retain macromolecular impurities and collect target small molecule components.

[0012] Furthermore, the medicinal and edible plant materials are those listed in the catalog of both food and traditional Chinese medicine published by the National Health Commission, preferably one or more of the following: kudzu root, hawthorn, wolfberry, polygonatum, poria cocos, mulberry leaf, and chrysanthemum.

[0013] Furthermore, the complex enzyme system described in step (3) is at least one of complex carbohydrate enzyme, pectinase, and cellulase, used for the targeted degradation of plant cell walls and macromolecular carbohydrates.

[0014] Furthermore, the targeted enzymatic hydrolysis achieves the directed release and transformation of non-protein active small molecules such as polyphenols, flavonoids, and oligosaccharides by controlling the type of enzyme, pH, and temperature.

[0015] Furthermore, the membrane separation in step (5) is ultrafiltration or nanofiltration, the membrane molecular weight cutoff is 500 to 3000 Da, and the molecular weight distribution of the obtained small molecule components is concentrated in 300 to 800 Da, accounting for no less than 70%.

[0016] Furthermore, the molecular weight distribution coefficient (PDI) of the narrow-distribution small molecule component is less than 1.5, and the component uniformity and batch stability are significantly better than those of conventional enzymatic hydrolysis processes.

[0017] Furthermore, after enrichment of narrowly distributed small molecules, subsequent molding steps include concentration, blending, filling, and sterilization to obtain the final product.

[0018] Furthermore, the dosage form of the end product is an oral liquid, a solid beverage, a health drink, a granule, or a paste.

[0019] Furthermore, the gentle extraction temperature does not exceed 90°C, and the targeted enzymatic hydrolysis adopts closed-loop control of pH and temperature, ensuring a gentle process throughout and protecting the heat-sensitive active ingredients.

[0020] Furthermore, this component has good solubility, high absorption efficiency, and high bioavailability, making it suitable for functional foods, health drinks, and tonic foods.

[0021] The beneficial effects of this invention are as follows:

[0022] (i) By targeting enzymatic hydrolysis to destroy the plant cell wall structure, the dissolution and conversion efficiency of active small molecules such as polyphenols, flavonoids and oligosaccharides can be significantly improved, thereby increasing the utilization rate of raw materials and the bioavailability of products.

[0023] (ii) By using the synergistic regulation of enzymatic hydrolysis and membrane separation, narrow distribution control of small molecule products is achieved, resulting in higher component uniformity and stronger batch stability.

[0024] (III) With non-protein plant bioactive small molecules as the main enrichment target, the technology route is unique and the application scenarios are more diverse, which can expand the space for high-value utilization of medicinal and edible raw materials.

[0025] (iv) The overall process is mild and simple, and can be implemented using conventional extraction, enzymatic hydrolysis and membrane separation equipment. The process is highly controllable and suitable for large-scale production.

[0026] (v) The prepared small molecule components have good solubility and high absorption efficiency, and can be adapted to various dosage forms such as oral liquids, solid beverages, and drinks, with a wide range of applications. Detailed Implementation

[0027] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0028] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Every smaller range between any stated value or intermediate value within a stated range, and any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0029] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0030] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be obvious to those skilled in the art. This application specification and embodiments are merely exemplary.

[0031] The present invention provides a method for preparing narrowly distributed small molecules by targeted enzymatic hydrolysis of medicinal and edible raw materials, comprising the following steps: (1) Raw material pretreatment: cleaning and pulverizing the medicinal and edible plant raw materials and passing them through a 20-40 mesh sieve;

[0032] (2) Mild extraction: Add water at a material-to-liquid ratio of 1:8-12, extract at 75-90℃ for 1-1.5 hours, and filter to obtain the extract;

[0033] (3) Targeted enzymatic hydrolysis: Adjust the pH of the extract to 4.5-6.5, add the compound enzyme system, and enzymatically hydrolyze at 45-55℃ for 2-4 hours;

[0034] (4) Enzyme inactivation and stabilization: Heat to 85-95℃ and keep warm for 10-15 min to terminate enzymatic hydrolysis;

[0035] (5) Enrichment of narrow-distribution small molecules: Membrane separation and fractionation are used to retain macromolecular impurities and collect target small molecule components.

[0036] The medicinal and edible plant raw materials are those listed in the National Health Commission's catalogue of plants that are both food and traditional Chinese medicine, preferably one or more of the following: kudzu root, hawthorn, wolfberry, polygonatum, poria cocos, mulberry leaf, and chrysanthemum. The complex enzyme system in step (3) consists of at least one of a complex carbohydrate enzyme, pectinase, and cellulase, used for the targeted degradation of plant cell walls and macromolecular carbohydrates. The targeted enzymatic hydrolysis, by controlling the enzyme type, pH, and temperature, achieves the targeted release and transformation of non-protein active small molecules such as polyphenols, flavonoids, and oligosaccharides.

[0037] The membrane separation in step (5) is ultrafiltration or nanofiltration, with a membrane molecular weight cutoff of 500–3000 Da. The resulting small molecule components have a molecular weight distribution concentrated in the range of 300–800 Da, accounting for no less than 70%. The molecular weight distribution coefficient (PDI) of the narrowly distributed small molecule components is <1.5, and the component uniformity and batch stability are significantly better than conventional enzymatic hydrolysis processes. After the narrowly distributed small molecules are enriched, subsequent shaping steps include concentration, blending, filling, and sterilization to obtain the final product. The dosage form of the final product is oral liquid, solid beverage, health drink, granules, or ointment. The mild extraction temperature does not exceed 90°C, and the targeted enzymatic hydrolysis adopts closed-loop control of pH and temperature. The entire process is mild, protecting the heat-sensitive active ingredients. This component has good solubility, high absorption efficiency, and high bioavailability, making it suitable for functional foods, health drinks, and tonic foods.

[0038] Example 1: Preparation of narrow-distribution small molecules from kudzu root

[0039] Raw material pretreatment: Take dried kudzu root, clean it to remove impurities, pulverize it and pass it through a 30-mesh sieve to obtain kudzu root powder.

[0040] Mild extraction: Add water at a material-to-liquid ratio of 1:10, extract at 85℃ for 1.2 hours, and filter to obtain kudzu root extract.

[0041] Targeted enzymatic hydrolysis: Adjust the pH of the kudzu root extract to 5.5, add a compound enzyme system (containing pectinase and cellulase in a mass ratio of 1:1), and enzymatically hydrolyze at 50°C for 3 hours.

[0042] Enzyme inactivation and stabilization: Heat to 90℃ and hold for 12 minutes to terminate the enzymatic hydrolysis reaction.

[0043] Enrichment of narrow-distribution small molecules: Separation is performed using an ultrafiltration membrane with a molecular weight cutoff of 1000 Da, and the permeate is collected as the target small molecule component.

[0044] Product Formulation: Concentrate the target small molecule component to a solid content of 20%, add appropriate amounts of sweetener and preservative, mix evenly, fill, and sterilize at 121℃ for 15 minutes to obtain kudzu root oral liquid.

[0045] The results showed that the molecular weight distribution of the obtained kudzu root small molecule components was concentrated in the range of 400-700 Da, accounting for 78.5%, and the molecular weight distribution coefficient (PDI) was 1.32.

[0046] Example 2: Preparation of narrowly distributed small molecules from hawthorn-goji berry complex

[0047] Raw material pretreatment: Take dried hawthorn and wolfberry (mass ratio 1:1), clean them separately, mix them together, and pulverize them through a 25-mesh sieve.

[0048] Mild extraction: Add water at a material-to-liquid ratio of 1:9, extract at 80℃ for 1.5 hours, and filter to obtain the extract.

[0049] Targeted enzymatic hydrolysis: Adjust the pH of the extract to 5.0, add a complex carbohydrate enzyme, and hydrolyze at 48°C for 3.5 hours.

[0050] Enzyme inactivation and stabilization: Heat to 88℃ and hold for 15 minutes to terminate enzymatic hydrolysis.

[0051] Narrowly distributed small molecule enrichment: Separation was performed using a nanofiltration membrane with a molecular weight cutoff of 800 Da, and the permeate was collected.

[0052] Product Formation: The permeate is spray-dried to obtain hawthorn-goji berry composite small molecule powder, which can be used in solid beverages or granules.

[0053] The obtained composite small molecule components were found to have a molecular weight distribution concentrated in the range of 350–750 Da, accounting for 72.3%, with a molecular weight distribution coefficient (PDI) of 1.41.

[0054] Example 3: Preparation of narrowly distributed small molecules from a Polygonatum sibiricum-Poria cocos-mulberry leaf complex

[0055] Raw material pretreatment: Take dried Polygonatum sibiricum, Poria cocos and mulberry leaves (mass ratio 2:1:1), clean and mix them, then pulverize them through a 35-mesh sieve.

[0056] Mild extraction: Add water at a material-to-liquid ratio of 1:11, extract at 78℃ for 1.3 hours, and filter to obtain the extract.

[0057] Targeted enzymatic hydrolysis: Adjust the pH of the extract to 6.0, add a complex enzyme system (containing pectinase, cellulase and complex carbohydrate enzyme in a mass ratio of 1:1:2), and enzymatically hydrolyze at 52℃ for 2.5 h.

[0058] Enzyme inactivation and stabilization: Heat to 92℃ and hold for 10 minutes to terminate enzymatic hydrolysis.

[0059] Enrichment of narrow-distribution small molecules: Ultrafiltration membrane with a molecular weight cutoff of 1500 Da is used for separation, and the permeate is collected.

[0060] Product formation: The permeate is concentrated to an appropriate concentration, honey is added for flavoring, and after bottling, it is pasteurized to obtain a healthy beverage.

[0061] The results showed that the molecular weight distribution of the obtained composite small molecule components was concentrated in the range of 500 to 800 Da, accounting for 75.8%, and the molecular weight distribution coefficient (PDI) was 1.38.

[0062] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A method for preparing narrowly distributed small molecules by targeted and controlled enzymatic hydrolysis of medicinal and edible raw materials, characterized in that, Includes the following steps: (1) Raw material pretreatment: Clean and crush the medicinal and edible plant raw materials and pass them through a 20-40 mesh sieve; (2) Mild extraction: Add water at a material-to-liquid ratio of 1:8-12, extract at 75-90℃ for 1-1.5 hours, and filter to obtain the extract; (3) Targeted enzymatic hydrolysis: Adjust the pH of the extract to 4.5-6.5, add the compound enzyme system, and enzymatically hydrolyze at 45-55℃ for 2-4 hours; (4) Enzyme inactivation and stabilization: Heat to 85-95℃ and keep warm for 10-15 min to terminate enzymatic hydrolysis; (5) Enrichment of narrow-distribution small molecules: Membrane separation and fractionation are used to retain macromolecular impurities and collect target small molecule components.

2. The method for preparing narrowly distributed small molecules by targeted controlled enzymatic hydrolysis of medicinal and edible raw materials according to claim 1, characterized in that, The medicinal and edible plant materials are those listed in the catalog of food and traditional Chinese medicine published by the National Health Commission, with a preference for one or more of the following: kudzu root, hawthorn, wolfberry, polygonatum, poria cocos, mulberry leaf, and chrysanthemum.

3. The method for preparing narrowly distributed small molecules by targeted controlled enzymatic hydrolysis of medicinal and edible raw materials according to claim 2, characterized in that, The complex enzyme system described in step (3) is at least one of complex carbohydrate enzyme, pectinase, and cellulase, used for the targeted degradation of plant cell walls and macromolecular carbohydrates.

4. The method for preparing narrowly distributed small molecules by targeted controlled enzymatic hydrolysis of medicinal and edible raw materials according to claim 3, characterized in that, The targeted enzymatic hydrolysis achieves the directed release and transformation of non-protein active small molecules such as polyphenols, flavonoids, and oligosaccharides by controlling the type of enzyme, pH, and temperature.

5. The method for preparing narrowly distributed small molecules by targeted controlled enzymatic hydrolysis of medicinal and edible raw materials according to claim 4, characterized in that, The membrane separation in step (5) is ultrafiltration or nanofiltration, with a membrane molecular weight cutoff of 500 to 3000 Da. The molecular weight distribution of the obtained small molecule components is concentrated in the range of 300 to 800 Da, accounting for no less than 70%.

6. The method for preparing narrowly distributed small molecules by targeted controlled enzymatic hydrolysis of medicinal and edible raw materials according to claim 5, characterized in that, The molecular weight distribution coefficient (PDI) of the narrowly distributed small molecule components is less than 1.5, and the component uniformity and batch stability are significantly better than those of conventional enzymatic hydrolysis processes.

7. The method for preparing narrowly distributed small molecules by targeted controlled enzymatic hydrolysis of medicinal and edible raw materials according to claim 6, characterized in that, After enrichment of narrowly distributed small molecules, subsequent molding steps include concentration, blending, filling, and sterilization to obtain the final product.

8. The method for preparing narrowly distributed small molecules by targeted controlled enzymatic hydrolysis of medicinal and edible raw materials according to claim 7, characterized in that, The dosage form of the end product is oral liquid, solid beverage, health drink, granules or ointment.

9. The method for preparing narrowly distributed small molecules by targeted controlled enzymatic hydrolysis of medicinal and edible raw materials according to claim 8, characterized in that, The extraction temperature does not exceed 90℃, and the targeted enzymatic hydrolysis adopts closed-loop control of pH and temperature. The entire process is gentle and protects the heat-sensitive active ingredients.

10. The method for preparing narrowly distributed small molecules by targeted controlled enzymatic hydrolysis of medicinal and edible raw materials according to claim 9, characterized in that, This component has good solubility, high absorption efficiency, and high bioavailability, making it suitable for functional foods, health drinks, and tonic foods.