Polygonatum sibiricum extract as well as preparation method and application thereof

Abstract

The invention discloses a rhizoma polygonati extract as well as a preparation method and application thereof, and belongs to the technical field of rhizoma polygonati extraction. According to the preparation method disclosed by the invention, the crushed rhizoma polygonati is soaked and ground to obtain the rhizoma polygonati slurry, and then the rhizoma polygonati slurry is subjected to enzymolysis-fermentation synchronous extraction, so that the extraction rate of rhizoma polygonati polysaccharide and rhizoma polygonati saponin in the rhizoma polygonati extract can be effectively increased, and a remarkable effect is achieved; specifically, the extraction rate of polygonatum sibiricum polysaccharide in the obtained polygonatum sibiricum extract is 70.28% or above, and the extraction rate of polygonatum sibiricum saponin is 22.68% or above. In addition, the preparation method of the rhizoma polygonati extract provided by the invention is simple to operate and beneficial to actual large-scale production.

Description

Technical Field

[0001] The invention belongs to the technical field of polygonatum extraction, and in particular relates to a polygonatum extract and a preparation method and application thereof. Background Art

[0002] Polygonatum sibiricum is a perennial herbaceous plant of the genus Polygonatum in the Liliaceae family. Its rhizome is used as medicine, and it has the effects of replenishing qi and nourishing yin, strengthening the spleen, moistening the lungs, and benefiting the kidneys. Modern medical pharmacology research believes that Polygonatum sibiricum polysaccharides and saponin compounds are the main chemical components of Polygonatum sibiricum's effective ingredients. In addition, flavonoids, alkaloids, vitamins, and a variety of essential amino acids and trace elements are also important components of Polygonatum sibiricum's chemical composition. Together, they determine the wide range of pharmacological activities of Polygonatum sibiricum, including anti-fatigue, anti-oxidation, immune regulation, blood sugar and blood lipid regulation, and anti-tumor.

[0003] In 2002, Polygonatum sibiricum was included in the list of traditional Chinese medicines with medicinal and edible properties by the National Health Commission. In recent years, the continuous improvement of national health awareness has led to the rapid development of the big health industry, and the development and utilization of Polygonatum sibiricum will also be more widely used. Because fresh Polygonatum sibiricum contains calcium oxalate needle crystal bundles, eating it raw has a numb tongue and throat sensation, so Polygonatum sibiricum will be processed nine times before eating, but the nine-steaming and nine-processing process takes a long time, the process is cumbersome, and the content of Polygonatum sibiricum polysaccharides will be reduced during the process. At present, most of the Polygonatum sibiricum slices on the market are directly crushed and added or brewed, which greatly reduces the effective utilization rate of Polygonatum sibiricum. Therefore, it is necessary to efficiently extract Polygonatum sibiricum polysaccharides and other effective ingredients in Polygonatum sibiricum to improve the effective utilization rate of Polygonatum sibiricum. At present, the extraction of active ingredients in Polygonatum sibiricum is mostly hot water extraction or ultrasonic extraction, which has the limitations of low extraction rate, high energy consumption, high equipment requirements, and inability to scale production. In recent years, enzyme extraction and fermentation extraction have been widely studied and applied, but enzymatic hydrolysis or fermentation alone cannot fully release the functional ingredients in Polygonatum sibiricum. Summary of the invention

[0004] The purpose of the present invention is to overcome the shortcomings of the above-mentioned prior art and provide a polygonatum extract with high extraction rate of polygonatum polysaccharide and polygonatum saponin and capable of large-scale production, as well as a preparation method and application thereof.

[0005] To achieve the above object, in a first aspect of the present invention, the present invention provides a method for preparing a polygonatum sibiricum extract, the preparation method comprising the following steps:

[0006] (1) taking the crushed polygonatum, adding it into water and soaking it, and grinding it after the soaking to obtain polygonatum slurry;

[0007] (2) adding enzymes and microorganisms to the polygonatum slurry to perform enzymolysis-fermentation simultaneous extraction;

[0008] (3) After the simultaneous extraction by enzymolysis and fermentation, the mixture is sterilized and filtered, the filtrate is collected and centrifuged, and the supernatant is concentrated to obtain the polygonatum sibiricum extract.

[0009] The invention soaks and grinds the crushed polygonatum to obtain polygonatum slurry, and then performs enzymolysis-fermentation simultaneous extraction on the polygonatum slurry, which can effectively improve the extraction rates of polygonatum polysaccharides and polygonatum saponins in the polygonatum extract and achieve remarkable results.

[0010] Specifically, in the first aspect, the present invention can reduce the particle size of polygonatum by soaking and grinding, so that the contact area between enzyme and microorganism and polygonatum is increased in the subsequent enzymatic hydrolysis-fermentation simultaneous extraction process, thereby improving the efficiency of the interfacial reaction and being more conducive to the release of the effective ingredients in polygonatum. In the second aspect, the present invention uses enzymatic hydrolysis-fermentation simultaneous extraction, and the enzyme and microorganism can work synergistically to further improve the extraction rate of polygonatum polysaccharides and polygonatum saponins.

[0011] As a preferred embodiment of the preparation method of the present invention, in the step (2), before adding enzymes and microorganisms to the polygonatum slurry for enzymatic hydrolysis-fermentation simultaneous extraction, it also includes the steps of adding esterase to the polygonatum slurry for a first enzymatic hydrolysis and enzyme inactivation.

[0012] The present invention has found that further adding esterase to the polygonatum slurry for the first enzymatic hydrolysis before the enzymatic hydrolysis-fermentation simultaneous extraction can effectively achieve the purpose of defatting the polygonatum raw material, improve the interfacial reaction efficiency of the subsequent enzymatic hydrolysis-fermentation simultaneous extraction, and thereby improve the extraction rate of polygonatum polysaccharides and polygonatum saponins.

[0013] Exemplarily, the esterase includes any one of carboxylic ester hydrolase and glycerol ester hydrolase. The esterase can be purchased from Wuhan Xinhuayang Biological Co., Ltd. Jiulong Branch, etc.

[0014] As a preferred embodiment of the preparation method of the present invention, in the step (2), the enzyme includes at least one of cellulase, papain, and β-glucanase.

[0015] For example, the cellulase, papain and β-glucanase can be purchased from Wuhan Xinhuayang Biological Co., Ltd. Jiulong Branch or Nanning Pangbo Bioengineering Co., Ltd.

[0016] Preferably, the enzyme is papain.

[0017] As a preferred embodiment of the preparation method of the present invention, in the step (2), the microorganism includes at least one of yeast and lactic acid bacteria.

[0018] Preferably, the lactic acid bacteria include at least one of Lactobacillus plantarum, Lactobacillus acidophilus, and Lactobacillus paracasei.

[0019] Illustratively, the yeast can be purchased from Angel Yeast Co., Ltd., etc.; the Lactobacillus plantarum, Lactobacillus acidophilus and Lactobacillus paracasei can be purchased from Weikang Probiotics (Suzhou) Co., Ltd., Shandong Zhongke Jiayi Bioengineering Co., Ltd., etc.

[0020] Preferably, the microorganism is yeast.

[0021] As a preferred embodiment of the preparation method of the present invention, in the step (1), the mass ratio of the crushed polygonatum to water is 1:(5-10).

[0022] Exemplarily, the mass ratio of the crushed polygonatum to water can be any point value between 1:(5-10) or any two point range values, such as 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, etc.

[0023] Preferably, in the step (1), the mass ratio of the crushed polygonatum to water is 1:(5-8).

[0024] As a preferred embodiment of the preparation method of the present invention, in the step (1), the soaking time is 0.5-1.5h.

[0025] Exemplarily, the soaking time may be any point value or any two point range values ​​between 0.5-1.5 h, such as 0.5 h, 1 h, 1.5 h, etc.

[0026] Preferably, in step (1), the soaking time is 1-1.5 hours.

[0027] As a preferred embodiment of the preparation method of the present invention, in the step (1), the grinding rate is 5-20 kg / min.

[0028] Exemplarily, the grinding rate may be any point value or any two point range value between 5-20 kg / min, such as 5 kg / min, 10 kg / min, 15 kg / min, 20 kg / min, etc.

[0029] Preferably, in step (1), the grinding rate is 5-10 kg / min.

[0030] As a preferred embodiment of the preparation method of the present invention, in the step (1), the fineness of the polygonatum slurry is ≤5 mm.

[0031] Preferably, in step (1), the fineness of the polygonatum slurry is ≤2 mm.

[0032] The present invention has found that further selecting the mass ratio of polygonatum to water and the soaking time in the soaking process within the above range, and selecting the grinding rate and the fineness of the polygonatum slurry after grinding within the above range, especially within a further preferred range, can better help refine the particle size of polygonatum, improve the interfacial reaction rate of subsequent enzymatic hydrolysis and microbial fermentation, increase the extraction rate of polygonatum polysaccharides and polygonatum saponins, and improve the sensory score of the polygonatum extract.

[0033] As a preferred embodiment of the preparation method of the present invention, in the step (2), the amount of the first enzymatic esterase added is 0.2-1.5% based on the mass of the crushed polygonatum.

[0034] Exemplarily, based on the mass of the crushed Polygonatum, the added amount of the first enzymatic esterase can be any point value or any two point range values ​​between 0.2-1.5%, such as 0.2%, 0.5%, 0.8%, 1%, 1.2%, 1.5%, etc.

[0035] Preferably, in the step (2), the amount of the first enzymatic esterase added is 0.5-0.8% based on the mass of the crushed polygonatum.

[0036] As a preferred embodiment of the preparation method of the present invention, the temperature of the first enzymatic hydrolysis is 40-60° C. and the time is 1-3 h.

[0037] Exemplarily, the temperature of the first enzymatic hydrolysis can be any point value or any two points in the range between 40-60°C, such as 40°C, 45°C, 50°C, 55°C, 60°C, etc.; the time can be any point value or any two points in the range between 1-3h, such as 1h, 2h, 3h, etc.

[0038] Preferably, the temperature of the first enzymatic hydrolysis is 50-60°C and the time is 2-3h.

[0039] As a preferred embodiment of the preparation method of the present invention, during the first enzymatic hydrolysis, the solid-liquid ratio of the polygonatum slurry is 1: (10-40), and the pH value is 4.0-7.0.

[0040] Illustratively, during the first enzymatic hydrolysis, the solid-to-liquid ratio of the Polygonatum slurry can be any point value or any range value between 1:(10-40), for example, 1:10, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40, etc.; the pH value can be any point value or any range value between 4.0-7.0, for example, 4, 4.5, 5, 5.5, 6, 6.5, 7, etc.

[0041] Preferably, during the first enzymatic hydrolysis, the solid-liquid ratio of the polygonatum slurry is 1:(18-22), and the pH value is 5.0-6.0.

[0042] The present invention has been found that by selecting the amount of esterase added during the first esterase hydrolysis, the temperature and time of the first enzymatic hydrolysis, and the solid-liquid ratio and pH value of the first enzymatic hydrolysis Polygonatum slurry within the above range, especially within a further preferred range, the lipid components in the Polygonatum sibiricum raw material can be better removed to achieve the purpose of degreasing, avoiding the conventional use of chemical reagents for degreasing, and at the same time more effectively improving the interfacial reaction rate of the subsequent enzymatic hydrolysis-microbial synchronous extraction, thereby achieving the effect of increasing the extraction rate of Polygonatum sibiricum polysaccharides and Polygonatum sibiricum saponins.

[0043] As a preferred embodiment of the preparation method of the present invention, in the step (2), in the enzymatic hydrolysis-fermentation simultaneous extraction, based on the mass of the crushed polygonatum, the mass percentage of the enzyme is 0.1-1.5%, and the mass percentage of the microorganism is 1-10%.

[0044] Exemplarily, based on the mass of the crushed Polygonatum, the mass percentage of the enzyme can be any point value or any range value between 0.1-1.5%, such as 0.1%, 0.5%, 1%, 1.5%, etc.; the mass percentage of the microorganism can be any point value or any range value between 1-10%, such as 1%, 2%, 4%, 6%, 8%, 10%, etc.

[0045] Preferably, in the step (2), in the simultaneous enzymatic hydrolysis-fermentation extraction, based on the mass of the crushed polygonatum, the mass percentage of the enzyme is 0.5-1.5%, and the mass percentage of the microorganism is 2-8%.

[0046] As a preferred embodiment of the preparation method of the present invention, in the step (2), in the simultaneous enzymatic hydrolysis-fermentation extraction, the extraction temperature is 25-40°C and the extraction time is 12-60h.

[0047] Exemplarily, in the simultaneous enzymatic hydrolysis-fermentation extraction, the extraction temperature can be any point value or any range value between 25-40°C, for example, 25°C, 30°C, 35°C, 40°C, etc.; the extraction time can be any point value or any range value between 12-60h, for example, 12h, 15h, 20h, 25h, 30h, 35h, 40h, 45h, 50h, 55h, 60h, etc.

[0048] Preferably, in the step (2), in the simultaneous enzymatic hydrolysis-fermentation extraction, the extraction temperature is 25-35° C., and the extraction time is 12-36 h.

[0049] The present invention has found that in the simultaneous enzymatic hydrolysis-fermentation extraction, the amount of enzyme and microorganism added, as well as the temperature and time of extraction will affect the extraction rate of polygonatum polysaccharides and polygonatum saponins. When the parameters are further selected within the above range, especially within a further preferred range, the extraction rate of polygonatum polysaccharides and polygonatum saponins obtained is higher, and the sensory score of the polygonatum extract is also higher.

[0050] As a preferred embodiment of the preparation method of the present invention, the temperature for inactivating the enzyme is 90-100° C. and the time is 10-20 min.

[0051] As a preferred embodiment of the preparation method of the present invention, the sterilization temperature is 85-115° C. and the time is 10-30 min.

[0052] Preferably, the sterilization temperature is 90-100°C and the time is 10-20 minutes.

[0053] As a preferred embodiment of the preparation method of the present invention, the filtration is carried out using a 200-500 mesh filter.

[0054] Exemplarily, the filter can be any point value or any two point range value between 200-500 mesh, for example, 200 mesh, 300 mesh, 400 mesh, 500 mesh, etc.

[0055] As a preferred embodiment of the preparation method of the present invention, the centrifugal speed is 3000-5000 rpm and the time is 10-20 min.

[0056] Exemplarily, the centrifugal speed can be any point value or any two point range values ​​between 3000-5000rpm, such as 3000rpm, 3500rpm, 4000rpm, 4500rpm, 5000rpm, etc.; the time can be any point value or any two point range values ​​between 10-20min, such as 10min, 15min, 20min, etc.

[0057] As a preferred embodiment of the preparation method of the present invention, the concentration temperature is ≤80°C, and the concentration vacuum degree is ≤-0.07MPa.

[0058] In a second aspect of the present invention, the present invention provides a polygonatum sibiricum extract, which is prepared by the preparation method of the present invention.

[0059] In the third aspect of the present invention, the present invention provides the use of the polygonatum sibiricum extract in the preparation of immunity enhancing products, blood sugar and blood lipid regulating products, anti-tumor products, and antioxidant products.

[0060] As a preferred embodiment of the application of the present invention, the products include food and health products.

[0061] Compared with the prior art, the present invention has the following beneficial effects:

[0062] 1. The present invention adopts a colloid mill to prepare the raw material of polygonatum into polygonatum slurry. After the colloid mill treatment, the particle size of the raw material of polygonatum becomes smaller, so that the specific surface area for the action of enzymes and microorganisms becomes larger, which is more conducive to the release of functional components in polygonatum;

[0063] 2. The present invention utilizes esterase enzymolysis to achieve the purpose of degreasing the raw material of polygonatum, avoiding the cumbersome process of degreasing with chemical reagents, improving the interface reaction efficiency of simultaneous synergistic extraction of enzymes and microorganisms, and being more conducive to the extraction of functional components in polygonatum;

[0064] 3. The present invention optimizes the enzyme system and microorganism suitable for extracting polygonatum polysaccharide and polygonatum saponin from polygonatum, and uses exogenous enzymes and microbial fermentation to perform enzymolysis-fermentation synchronous synergistic extraction, thereby improving the extraction rate of polygonatum polysaccharide and polygonatum saponin. The extraction rate of polygonatum polysaccharide is more than 70.28%, and the extraction rate of polygonatum saponin is more than 22.68%, thereby improving the effective utilization rate of polygonatum;

[0065] 4. The preparation method of the polygonatum sibiricum extract provided by the present invention is simple to operate and is conducive to actual large-scale production. BRIEF DESCRIPTION OF THE DRAWINGS

[0066] Figure 1 This is a sensory evaluation result diagram of the polygonatum sibiricum extracts prepared in Example 7 and Comparative Example 12;

[0067] Figure 2 The present invention is a schematic diagram of the preparation process of the polygonatum sibiricum extract. DETAILED DESCRIPTION

[0068] In order to better illustrate the purpose, technical solutions and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments.

[0069] Unless otherwise specified, the reagents, methods and equipment used in the present invention are all conventional reagents, methods and equipment in the art; and unless otherwise specified, the raw materials used in parallel experiments are from the same batch of raw materials.

[0070] Example 1

[0071] The present invention provides a polygonatum extract, and the schematic diagram of the preparation process of the polygonatum extract is as follows: Figure 2 As shown, the following steps are included:

[0072] (1) removing moldy, worm-infested parts and impurities from polygonatum sibiricum and crushing them with a pulverizer to obtain polygonatum sibiricum, then adding 1.0 kg of the crushed polygonatum sibiricum to 5 times the mass of purified water and soaking for 1 hour, grinding with a colloid mill at a grinding rate of 5 kg / min until the fineness is ≤2 mm, and obtaining polygonatum sibiricum slurry;

[0073] (2) adding water to adjust the solid-liquid ratio of Polygonatum sibiricum slurry to 1:20 and the pH value to 5.0, then adding 0.6 wt % glycerol ester hydrolase for the first enzymatic hydrolysis at 50° C. for 2 h. After the first enzymatic hydrolysis is completed, the enzyme is inactivated at 95° C. for 10 min and cooled to 25° C.;

[0074] (3) Then, 1 wt% papain and 2 wt% yeast were added to the system cooled to 25°C and subjected to simultaneous enzymatic hydrolysis and fermentation extraction at 30°C for 20 h. After the simultaneous enzymatic hydrolysis and fermentation extraction, the temperature was raised to 95°C for sterilization for 10 min.

[0075] (4) After sterilization, the mixture was filtered through a 300-mesh filter cloth, and the filtrate was collected. The filtrate was centrifuged at 4000 rpm for 15 min, and the supernatant was collected after centrifugation. The supernatant was then concentrated at a temperature of ≤80°C and a vacuum degree of ≤-0.07 MPa to obtain a polygonatum sibiricum extract.

[0076] Example 2

[0077] The embodiment of the present invention provides a polygonatum extract, and the only difference between the preparation method of the polygonatum extract and that of embodiment 1 is that cellulase is used instead of papain.

[0078] Example 3

[0079] The embodiment of the present invention provides a polygonatum extract. The only difference between the preparation method of the polygonatum extract and that of embodiment 1 is that β-glucanase is used instead of papain.

[0080] Example 4

[0081] The embodiment of the present invention provides a polygonatum extract, and the only difference between the preparation method of the polygonatum extract and that of Example 1 is that a mixture of cellulase, papain and β-glucanase (wherein the mass ratio of cellulase, papain and β-glucanase is 1:8:1) is used to replace papain.

[0082] Example 5

[0083] The embodiment of the present invention provides a polygonatum extract. The only difference between the preparation method of the polygonatum extract and that of embodiment 1 is that plant lactobacillus is used instead of yeast.

[0084] Example 6

[0085] The embodiment of the present invention provides a polygonatum extract, and the only difference between the preparation method of the polygonatum extract and that of embodiment 1 is that the first enzymatic hydrolysis in step (2) is omitted.

[0086] Example 7

[0087] The embodiment of the present invention provides a polygonatum extract. The only difference between the preparation method of the polygonatum extract and that of Example 1 is that the mass of the crushed polygonatum is 50.0 kg for scaled-up production.

[0088] Comparative Example 1

[0089] The comparative example of the present invention provides a polygonatum extract, and the only difference between the preparation method of the polygonatum extract and that of Example 1 is that no colloid mill grinding is performed after soaking.

[0090] Comparative Example 2

[0091] The comparative example of the present invention provides a polygonatum extract, and the preparation method of the polygonatum extract is different from that of Example 1 in that papain enzymolysis is first performed, and then fermentation extraction is performed, specifically:

[0092] (1) removing moldy, worm-infested parts and impurities from polygonatum sibiricum and crushing them with a pulverizer to obtain polygonatum sibiricum, then adding 1.0 kg of the crushed polygonatum sibiricum to 5 times the mass of purified water and soaking for 1 hour, grinding with a colloid mill at a grinding rate of 5 kg / min until the fineness is ≤2 mm, and obtaining polygonatum sibiricum slurry;

[0093] (2) adding water to adjust the solid-liquid ratio of Polygonatum sibiricum slurry to 1:20 and the pH value to 5.0, then adding 0.6 wt % glycerol ester hydrolase for the first enzymatic hydrolysis at 50° C. for 2 h;

[0094] (3) After the first enzymatic hydrolysis, 1 wt% papain was added and enzymatic hydrolysis was carried out at 50°C for 2 h. After the enzymatic hydrolysis, the enzyme was inactivated at 95°C for 10 min and cooled to 25°C;

[0095] (4) then adding 2 wt% yeast to the system cooled to 25°C and fermenting and extracting at 30°C for 20 hours, and then heating to 95°C for sterilization for 10 minutes;

[0096] (5) After sterilization, the mixture was filtered through a 300-mesh filter cloth, and the filtrate was collected. The filtrate was centrifuged at 4000 rpm for 15 min, and the supernatant was collected after centrifugation. The supernatant was then concentrated at a temperature of ≤80°C and a vacuum degree of ≤-0.07 MPa to obtain a polygonatum sibiricum extract.

[0097] Comparative Example 3

[0098] The comparative example of the present invention provides a polygonatum extract, and the preparation method of the polygonatum extract is different from that of Example 1 in that fermentation extraction is performed first and then enzymolysis is performed, specifically:

[0099] (1) removing moldy, worm-infested parts and impurities from polygonatum sibiricum and crushing them with a pulverizer to obtain polygonatum sibiricum, then adding 1.0 kg of the crushed polygonatum sibiricum to 5 times the mass of purified water and soaking for 1 hour, grinding with a colloid mill at a grinding rate of 5 kg / min until the fineness is ≤2 mm, and obtaining polygonatum sibiricum slurry;

[0100] (2) adding water to adjust the solid-liquid ratio of Polygonatum sibiricum slurry to 1:20 and the pH value to 5.0, then adding 0.6 wt % glycerol ester hydrolase for the first enzymatic hydrolysis at 50° C. for 2 h. After the first enzymatic hydrolysis is completed, the enzyme is inactivated at 95° C. for 10 min and cooled to 25° C.;

[0101] (3) Then, 2 wt% yeast was added to the system cooled to 25°C and fermented and extracted at 30°C for 20 h. After fermentation and extraction, the temperature was raised to 95°C for sterilization for 10 min. The temperature was then lowered to 50°C and 1 wt% papain was enzymolyzed at 50°C for 2 h. After the enzymolysis was completed, the enzyme was inactivated at 95°C for 10 min.

[0102] (5) After the enzyme is inactivated, the mixture is filtered through a 300-mesh filter cloth, the filtrate is collected, and the filtrate is centrifuged at 4000 rpm for 15 min. After centrifugation, the supernatant is collected and then concentrated at a temperature of ≤80°C and a vacuum degree of ≤-0.07 MPa to obtain a polygonatum sibiricum extract.

[0103] Comparative Example 4

[0104] The comparative example of the present invention provides a polygonatum extract, and the only difference between the preparation method of the polygonatum extract and that of Example 1 is that yeast is not added in step (3), and the enzymatic hydrolysis temperature of papain is maintained at 30° C. for 20 hours.

[0105] Comparative Example 5

[0106] The comparative example of the present invention provides a polygonatum extract, and the only difference between the preparation method of the polygonatum extract and that of Example 1 is that yeast is not added in step (3), and the enzymatic hydrolysis temperature of papain is adjusted to 50° C. for 2 h.

[0107] Comparative Example 6

[0108] The comparative example of the present invention provides a polygonatum extract, and the only difference between the preparation method of the polygonatum extract and that of Example 1 is that yeast is not added in step (3), and cellulase is used for enzymolysis at a temperature of 50° C. for 2 h.

[0109] Comparative Example 7

[0110] The comparative example of the present invention provides a polygonatum extract, and the only difference between the preparation method of the polygonatum extract and that of Example 1 is that yeast is not added in step (3), and β-glucanase is used for enzymolysis at a temperature of 50° C. for 2 h.

[0111] Comparative Example 8

[0112] The comparative example of the present invention provides a polygonatum extract, and the only difference between the preparation method of the polygonatum extract and that of Example 1 is that papain is not added in step (3).

[0113] Comparative Example 9

[0114] The comparative example of the present invention provides a polygonatum extract, the preparation method of the polygonatum extract is different from that of Example 1 only in that papain is not added in step (3), 4wt% plant lactobacillus is used for fermentation, the fermentation temperature is 37°C, and the fermentation time is 24h.

[0115] Comparative Example 10

[0116] The comparative example of the present invention provides a polygonatum extract, and the preparation method of the polygonatum extract adopts a hot water extraction method, comprising the following steps:

[0117] (1) removing moldy, worm-infested parts and impurities from polygonatum sibiricum and crushing them with a pulverizer to obtain polygonatum sibiricum, then adding 1.0 kg of the crushed polygonatum sibiricum to 5 times the mass of purified water and soaking for 1 hour, grinding with a colloid mill at a grinding rate of 5 kg / min until the fineness is ≤2 mm, and obtaining polygonatum sibiricum slurry;

[0118] (2) adding water to adjust the solid-liquid ratio of the polygonatum slurry to 1:20, extracting at 100°C for 2 hours, then filtering with a 300-mesh filter cloth, adding water 8 times the mass of the crushed polygonatum slurry, extracting at 100°C for 1 hour, and combining the two extraction filtrates;

[0119] (3) The filtrate was centrifuged at 4000 rpm for 15 min, the supernatant was collected after centrifugation, and then concentrated at a temperature of ≤80°C and a vacuum degree of ≤-0.07 MPa, and then sterilized at 110°C for 20 min to obtain a polygonatum sibiricum extract.

[0120] Comparative Example 11

[0121] The comparative example of the present invention provides a polygonatum sibiricum extract, and the only difference between the preparation method of the polygonatum sibiricum extract and the comparative example 10 is that no grinding process is performed in step (1).

[0122] Comparative Example 12

[0123] The comparative example of the present invention provides a polygonatum sibiricum extract, and the only difference between the preparation method of the polygonatum sibiricum extract and the comparative example 10 is that no crushing and grinding treatment is performed in step (1).

[0124] Effect example

[0125] Effect Example of the Invention The extraction rates of polygonatum polysaccharides and polygonatum saponins in the polygonatum extracts prepared in Examples 1-7 and Comparative Examples 1-12 were investigated, and the test method included the following contents:

[0126] 1. The determination of the content of Polygonatum sibiricum polysaccharide refers to the determination method of the 2020 edition of the Chinese Pharmacopoeia, specifically:

[0127] (1) Preparation of reference solution: Take about 38 mg of anhydrous glucose reference substance dried to constant weight at 105°C, accurately weigh it, place it in a 100 mL volumetric flask, add water to dissolve and dilute to the mark, shake well, and obtain the solution;

[0128] (2) Preparation of standard curve: Accurately measure 0.1mL, 0.2mL, 0.3mL, 0.4mL, and 0.5mL of the reference solution, respectively, and place them in 10mL stoppered graduated test tubes, add water to 2.0mL each, shake well, slowly add 0.05% anthrone-sulfuric acid solution to the scale in an ice water bath, mix well, cool and then place in a water bath for 10 minutes, take out, immediately place in an ice water bath to cool for 10 minutes, take out, and use the corresponding reagent as a blank. Measure the absorbance at a wavelength of 582nm, and draw a standard curve with absorbance as the ordinate and concentration as the abscissa;

[0129] (3) Determination method: Accurately measure 5 mL of sample, place in a round-bottom flask, add 25 mL of water, mix well, slowly add 120 mL of anhydrous ethanol, mix well, place in a 90°C water bath and heat to reflux for 0.5 hour, filter while hot, wash the residue twice with 80% hot ethanol, 10 mL each time, place the residue and filter paper in a flask, add 150 mL of water, place in a boiling water bath and heat to reflux for 1 hour, filter while hot, wash the residue and flask with hot water 4 times, 10 mL each time, combine the filtrate and washing liquid, cool, transfer to a 250 mL volumetric flask, add water to the scale, shake well, accurately measure 2 mL, place in a 10 mL stoppered graduated test tube, and measure the absorbance according to the method under the preparation of the standard curve, starting from "slowly add 0.05% anthrone-sulfuric acid solution to the scale in an ice water bath", read the amount of anhydrous glucose in the test solution from the standard curve, and calculate.

[0130] 2. Determination of Polygonatum sibiricum saponin content

[0131] (1) Preparation of standard curve: Pipette 0.0 mL, 0.4 mL, 0.6 mL, 0.8 mL, 1.0 mL, and 1.2 mL of ginsenoside Re standard solution into a 10 mL stoppered colorimetric tube, place in a water bath to evaporate the solvent, add 0.2 mL of vanillin solution, then add 0.8 mL of perchloric acid, mix well to dissolve all the residue, place in a 60°C water bath and heat for 10 min, take out, cool in an ice bath, add 5.0 mL of glacial acetic acid, shake well, use the corresponding reagent as blank, and immediately measure the absorbance at a wavelength of 560 nm;

[0132] (2) Determination method: Add 1.0 mL of sample to a 3 cm activated and eluted macroporous resin in a glass chromatography column with an inner diameter of 1.5 cm, elute with 25 mL of water, discard the eluent, and then elute ginsenosides with 25 mL of 70% ethanol at a rate not exceeding 3 mL / min until the eluent is colorless. Collect the eluent in an evaporating dish and evaporate it in a 60°C water bath. Dissolve the residue in a small amount of methanol and transfer it to a 10 mL stoppered colorimetric tube. According to the method under the preparation of the standard curve, from "evaporating the solvent in a water bath", determine the absorbance in the same way as the standard solution, and calculate the total saponin content based on the standard curve.

[0133] 3. Sensory evaluation of Polygonatum sibiricum extract

[0134] The sensory evaluation was conducted by 12 people who met the sensory evaluation requirements. The full score was 100 points. The evaluation indicators included taste (30 points), smell (30 points), color (20 points), and tissue morphology (20 points). The specific scoring criteria are shown in Table 1.

[0135] Table 1 Sensory evaluation standards

[0136]

[0137] Through the analysis of the recorded data, the results of the extraction rates of polygonatum polysaccharides and polygonatum saponins of various embodiments and comparative examples are shown in Table 2;

[0138] Table 2 Extraction rate of polygonatum polysaccharide and polygonatum saponin

[0139]

[0140]

[0141] It can be seen from Table 2 that when the preparation method provided by the present invention is adopted, the extraction rates of polygonatum polysaccharide and polygonatum saponin in the obtained polygonatum extract are high; specifically, the extraction rate of polygonatum polysaccharide is above 70.28%, and the extraction rate of polygonatum saponin is above 22.68%;

[0142] It can be seen from Example 1 and Comparative Example 1 that when the colloid mill grinding and pulping treatment is not carried out, the extraction rates of polygonatum polysaccharides and polygonatum saponins in the obtained polygonatum extract show a significant downward trend; it can be seen from Example 1 and Comparative Examples 2-3 that when the enzymatic hydrolysis-fermentation simultaneous extraction is not carried out, the extraction rates of polygonatum polysaccharides and polygonatum saponins in the obtained polygonatum extract also show a significant downward trend; it can be seen from Example 1 and Comparative Examples 4-9 that when only enzymatic hydrolysis or fermentation is selected in the enzymatic hydrolysis-fermentation simultaneous extraction, the effect of the present invention cannot be achieved; it can be seen from Example 1 and Comparative Examples 10-12 that when traditional hot water extraction is used, the extraction rates of polygonatum polysaccharides and polygonatum saponins in the polygonatum extract show a significant downward trend.

[0143] It can be seen from Example 1 and Examples 2-4 that in the enzymatic hydrolysis-fermentation simultaneous extraction, the selection of enzyme will also affect the comprehensive performance of the extract. When papain is further selected as the enzyme, the extraction rates of polygonatum polysaccharide and polygonatum saponin in the obtained polygonatum extract are higher; it can be seen from Example 1 and Example 5 that in the enzymatic hydrolysis-fermentation simultaneous extraction, the selection of microorganism will also affect the comprehensive performance of the extract. When yeast is further selected as the microorganism, the extraction rates of polygonatum polysaccharide and polygonatum saponin in the obtained polygonatum extract are higher; it can be seen from Example 1 and Example 6 that when the first enzymatic hydrolysis by esterase is not introduced, although the extraction rates of polygonatum polysaccharide and polygonatum saponin in the polygonatum extract are better than those in the comparative example, there is a certain degree of decrease compared with Example 1, indicating that the first enzymatic hydrolysis by esterase can effectively help improve the extraction rate of the effective ingredients in polygonatum.

[0144] In addition, the sensory evaluation of the polygonatum extracts prepared in Example 7 and Comparative Example 12 was performed. The sensory evaluation results are as follows: Figure 1 As shown, from Figure 1 It can be clearly seen that the polygonatum extract prepared by the preparation method provided by the present invention has a higher sensory score.

[0145] Finally, it should be noted that the above embodiments are intended to illustrate the technical solution of the present invention rather than to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention may be modified or replaced by equivalents without departing from the essence and scope of the technical solution of the present invention.

Claims

1. A method for preparing a polygonatum extract, characterized in that: The preparation method comprises the following steps: (1) taking the crushed polygonatum, adding it into water and soaking it, and grinding it after the soaking to obtain polygonatum slurry; (2) adding enzymes and microorganisms to the polygonatum slurry to perform enzymolysis-fermentation simultaneous extraction; (3) After the simultaneous extraction by enzymolysis and fermentation, the mixture is sterilized and filtered, the filtrate is collected and centrifuged, and the supernatant is concentrated to obtain the polygonatum sibiricum extract.

2. The preparation method according to claim 1, characterized in that: In the step (2), before adding enzymes and microorganisms to the polygonatum slurry for enzymatic hydrolysis-fermentation simultaneous extraction, the step also includes adding esterase to the polygonatum slurry for a first enzymatic hydrolysis and enzyme inactivation.

3. The preparation method according to claim 1, characterized in that: In the step (2), the enzyme includes at least one of cellulase, papain, and β-glucanase; And / or, in step (2), the microorganism includes at least one of yeast and lactic acid bacteria.

4. The preparation method according to claim 1, characterized in that: In the step (1), the mass ratio of the crushed polygonatum to water is 1:(5-10); and / or, the soaking time is 0.5-1.5h; and / or, the grinding rate is 5-20 kg / min; And / or, the fineness of the polygonatum slurry is ≤5mm.

5. The preparation method according to claim 2, characterized in that: In the step (2), the amount of the first enzymatic esterase added is 0.2-1.5% based on the mass of the crushed polygonatum; and / or, the temperature of the first enzymatic hydrolysis is 40-60° C. and the time is 1-3 h; And / or, during the first enzymatic hydrolysis, the solid-liquid ratio of the polygonatum slurry is 1:(10-40), and the pH value is 4.0-7.

0.

6. The preparation method according to claim 1, characterized in that: In the step (2), in the simultaneous enzymatic hydrolysis-fermentation extraction, the mass percentage of the enzyme is 0.1-1.5%, and the mass percentage of the microorganism is 1-10% based on the mass of the crushed polygonatum; And / or, in the step (2), in the simultaneous enzymatic hydrolysis-fermentation extraction, the extraction temperature is 25-40° C., and the extraction time is 12-60 h.

7. The preparation method according to claim 2, characterized in that: The temperature for inactivating the enzyme is 90-100°C and the time is 10-20 minutes; And / or, the sterilization temperature is 85-115°C and the time is 10-30 minutes.

8. A polygonatum extract, characterized in that The polygonatum sibiricum extract is prepared by the preparation method according to any one of claims 1 to 7.

9. Use of the polygonatum sibiricum extract as claimed in claim 8 in the preparation of products for enhancing immunity, regulating blood sugar and blood lipids, anti-tumor products, and antioxidant products.

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