CN114158725A - Preparation method of ginseng zymolyte containing polypeptide polysaccharide - Google Patents

Abstract

The invention provides a preparation method of ginseng zymolyte containing polypeptide polysaccharide, belonging to the field of deep processing of traditional Chinese medicinal materials. The method of the invention comprises the following steps: pulverizing 100 weight parts of dried Ginseng radix, adding 500-2000 weight parts of water, adding 1-8 weight parts of carbohydrase at 30-65 deg.C, and reacting for 0.5-12 hr; 2) simultaneously adding 0.1-8 parts by weight of alkaline protease and 0.1-8 parts by weight of neutral protease to react for 0.5-12 hours at the same temperature, or adding 0.1-8 parts by weight of alkaline protease to react for 0.5-12 hours, and then adding 0.1-8 parts by weight of neutral protease to react for 0.5-12 hours; step 2) maintaining the pH value to be 6-9; 3) terminating the enzyme reaction of step 2). The method of the invention can improve the yield of effective components in the ginseng zymolyte, and the zymolyte has good health care and medicinal values.

Description

Preparation method of ginseng zymolyte containing polypeptide polysaccharide

Technical Field

The invention belongs to the field of deep processing of traditional Chinese medicinal materials, and particularly relates to an enzymolysis process of ginseng.

Background

Ginseng (Panax ginseng) is the dried root of Panax ginseng C.A.Meyer of Araliaceae family, commonly known as herba Caesalpiniae, herba Bidentis Bipinnatae, herba Gei Piloselloidis, and radix Ginseng Indici.

The medicinal history of ginseng is long, and ginseng is listed as the top grade in Shen nong Ben Cao Jing, and is well known as Bai Cao Yao Wang. The research on the chemical composition of ginseng has been conducted for hundreds of years. Ginseng contains many components such as ginsenoside, polysaccharide, polypeptide, fatty acid, amino acid, volatile oil, flavone, vitamin, trace elements, etc., and ginsenoside is considered as one of its main active components. At present, ginseng is reported to contain more than 40 ginsenosides, and ginsenosides can be classified into three types according to the difference of aglycone structures: panaxadiol type, panaxatriol type and oleanolic acid type M. The composition of ginsenoside varies to different extent during processing, such as ginsenoside Rh2 and ginsenoside Rg3 which are not contained in Ginseng radix during processing into Ginseng radix Rubri. The determination result of amino acids in ginseng by using Weiyongdi and the like shows that the ginseng contains more than 17 amino acids, and the highest content of arginine. Monosaccharides found in ginseng include fructose, glucose, arabinose, etc., disaccharides include sucrose, maltose, etc., trisaccharides include raffinose, ginseng trisaccharide A, B, C, D, etc., and ginseng polysaccharides.

The application of the modern ginseng is mainly to accept western medicine theory and extract the saponin in the ginseng by extracting and purifying the ginsenoside by selecting an organic solvent or under the action of acid. For example, in the 'method for extracting ginsenoside and its application' disclosed in the patent application No. 201811610107.2, ginseng root is dried in an oven, crushed, extracted under microwave conditions with organic alcohol as a solvent, then adsorbed by macroporous resin, eluted with deionized water and ethanol, concentrated under reduced pressure, and dried to obtain total ginsenoside.

In addition, there is a method for extracting ginseng juice by treating ginseng using an enzyme engineering technique, such as "an enzymatic preparation method of full-ingredient ginseng juice" disclosed in patent application No. 201310355915.X, which is a method for extracting ginseng juice by hydrolyzing ginseng using complex enzyme. The operation is that the ginseng and water are pulped; finely grinding the ginseng pulp by using a colloid mill and adjusting the pH value to 2.5-4.5. And (3) steaming the ginseng pulp by using high-pressure steaming equipment at the steaming temperature of 102-115 ℃, putting the steamed ginseng into an enzymolysis tank, and carrying out enzymolysis twice under an acidic condition to obtain ginseng whole-component hydrolysate.

In addition, "enzymatic hydrolysis of ginseng and its activity study, the study of the Neugua, Jilin university of agriculture, the master academic paper of 2013" discusses the use of cellulase and amylase for enzymatic hydrolysis of ginseng. The cellulase is used for cracking cells and improving the dissolution of ginsenoside; the amylase is used for generating reducing sugar, provides raw materials for Maillard reaction, and increases the content of arginine monoglycoside and arginine diglycoside in the enzymatic hydrolysate.

However, the products obtained by the prior art lose a lot of active ingredients and cannot fully exert the medicinal function of the ginseng.

Disclosure of Invention

The invention aims to provide a preparation method of ginseng zymolyte containing polypeptide polysaccharide.

The technical scheme of the invention is as follows:

a ginseng enzymolysis method comprises the following steps:

1) pulverizing 100 weight parts of dry root of Panax plant, adding 500-2000 weight parts of water, adding 1-8 weight parts of carbohydrase at 30-65 deg.C, and reacting for 0.5-12 hr;

2) simultaneously adding 0.1-8 parts by weight of alkaline protease and 0.1-8 parts by weight of neutral protease to react for 0.5-12 hours at the same temperature, or adding 0.1-8 parts by weight of alkaline protease to react for 0.5-12 hours, and then adding 0.1-8 parts by weight of neutral protease to react for 0.5-12 hours;

3) terminating the enzyme reaction of step 2);

step 2) maintaining the pH value of 6-9.

According to the ginseng enzymolysis method, the ginseng plants in the step 1) are: ginseng (Panax ginseng), american ginseng (Panax quinquefolius), pseudoginseng (Panax pseudoginseng), or Panax notoginseng (Panax zingiberensis);

and/or the reaction time of step 1) is 3 hours;

and/or, the carbohydrases of step 1) are amylases and cellulases; preferably, the amylase in step 1) is used in an amount of 1-2 parts by weight, and the cellulase is used in an amount of 0.5-2 parts by weight.

The ginseng enzymolysis method is as described above, and the pH value of the step 2) is 8.

The ginseng enzymolysis method is characterized in that the alkaline protease in the step 2) accounts for 0.5-3 parts by weight; and/or the reaction time is 2-4 h.

According to the ginseng enzymolysis method, the neutral protease in the step 3) is 0.2-0.5 weight part; and/or the reaction time is 2 h.

According to the ginseng enzymolysis method, the step 3) is followed by the following steps:

4) filtering to obtain supernatant, concentrating under reduced pressure, and freeze drying to obtain dry powder.

A ginseng zymolyte is prepared by the method.

The ginseng zymolyte is prepared from the ginseng zymolyte, and the number average molecular weight of the ginseng zymolyte is 346;

and/or a number average molecular weight of 274;

and/or, Z average molecular weight is 515;

and/or, Z +1 has an average molecular weight of 878.

The ginseng zymolyte is used for preparing anti-fatigue or kidney-tonifying and yang-strengthening nutritional food, health-care products or medicines.

The nutritional food, the health product or the medicament also contains fish scale collagen;

preferably, the fish scale collagen is prepared by taking tilapia scales as raw materials, wherein the weight percentage of hydroxylysine is more than 1.3%; the weight average molecular weight is 800-2400; the hydroxyproline accounts for more than 7.3 percent by weight;

preferably, the weight ratio of the ginseng zymolyte to the fish scale collagen is 1: 9.

The invention has the following beneficial effects:

1) the product yield is high. Filtering the zymolyte (taking supernatant), and freeze-drying to obtain zymolyte dry powder with the content of the ginseng raw material being more than 40%.

2) The zymolyte has rich active substances. The polysaccharide, flavone, total nitrogen, protein and total saponin have high proportion in lyophilized powder.

3) Has high health and medicinal value. The zymolyte of the invention has good functions of resisting fatigue, tonifying kidney and strengthening yang.

Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.

The foregoing aspects of the present invention are explained in further detail below with reference to specific embodiments. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.

Drawings

FIG. 1: HPLC peak chart for saponin detection of comparative example 1.

FIG. 2: HPLC peak pattern for saponin detection of example 1.

FIG. 3: HPLC peak pattern (220nm wavelength) of the substrate of example 1.

Detailed Description

Example 1 preparation of ginseng zymolyte of the present invention

Crushing 100 g of ginseng sun-dried ginseng to 60-80 meshes, adding 1000 ml of water, keeping the temperature at 30-65 ℃, adding 2 g of cellulase and 1 g of amylase, and reacting for 3 hours; then, under the same temperature, keeping the pH value constant to 8, adding 0.5 g of alkaline protease to react for 2 hours under constant-temperature stirring, and then adding 0.5 g of neutral protease to react for 2 hours; then heating to 90 deg.C, reacting at constant temperature for half an hour to inactivate enzyme, and filtering. And concentrating the supernatant under reduced pressure at the vacuum degree of 0.06-0.10 Mpa and the temperature of 40-65 ℃, and freeze-drying to obtain solid ginseng zymolyte dry powder. The yield (weight of ginseng zymolyte dry powder/initial weight of ginseng zymolyte feed, the same below) is 51.2%.

Example 2 preparation of ginseng zymolyte of the present invention

Crushing 100 g of ginseng root hair to 60-80 meshes, adding 1000 ml of water, keeping the temperature at 30-65 ℃, adding 0.5 g of cellulase and 1 g of amylase, and reacting for 3 hours; then, under the same temperature, the pH value is kept constant to 8, 0.5 g of alkaline protease is added under constant temperature stirring for reaction for 2 hours, and then 0.5 g of neutral protease is added for reaction for 2 hours; then heating to 90 deg.C, reacting at constant temperature for half an hour to inactivate enzyme, and filtering. And concentrating the supernatant under reduced pressure at the vacuum degree of 0.06-0.10 Mpa and the temperature of 40-65 ℃, and freeze-drying to obtain solid ginseng zymolyte dry powder. The yield thereof was found to be 42.8%.

Example 3 preparation of ginseng zymolyte of the present invention

Crushing 100 g of American ginseng to 60-80 meshes, adding 1000 ml of water, keeping the temperature at 30-65 ℃, adding 1 g of cellulase and 2 g of amylase, and reacting for 3 hours; then, under the same temperature, the pH value is kept constant to 8, and 0.5 g of alkaline protease and 0.2 g of neutral protease are added to react for 2 hours under constant temperature stirring; then heating to 90 deg.C, reacting at constant temperature for half an hour to inactivate enzyme, and filtering. And concentrating the supernatant under reduced pressure at the vacuum degree of 0.06-0.10 Mpa and the temperature of 40-65 ℃, and freeze-drying to obtain solid ginseng zymolyte dry powder. The yield thereof was found to be 53.6%.

Example 4 Industrial production of ginseng zymolyte of the present invention

Crushing 100 kg of ginseng to be dried in the sun to 60-80 meshes, adding 1000 kg of water, keeping the temperature at 30-65 ℃, adding 1 kg of cellulase and 2 kg of amylase, and reacting for 3 hours; then, under the same temperature, 2N sodium hydroxide is used for adjusting the pH value to be 8, 500 g of alkaline protease and 200 g of neutral protease are added under constant temperature stirring for reaction for 2 hours; then heating to 90 deg.C, reacting at constant temperature for half an hour to inactivate enzyme, and filtering. Concentrating the supernatant under reduced pressure at 40-65 deg.C under 0.06-0.10 Mpa, and spray drying (inlet temperature 190 deg.C and outlet temperature 95 deg.C) to obtain solid Ginseng radix zymolyte dry powder with yield of 55%.

Comparative example 1 preparation of ginseng zymolyte

The alkaline protease was replaced with the acidic protease in the same manner as in example 1, and the pH was adjusted to 4, and the obtained enzymatic hydrolysate was filtered. And concentrating the supernatant under reduced pressure at the vacuum degree of 0.06-0.10 Mpa and the temperature of 40-65 ℃, and freeze-drying to obtain solid ginseng zymolyte dry powder. To obtain solid ginseng zymolyte dry powder. The yield thereof was found to be 23.6%.

Comparative example 2 preparation of ginseng zymolyte

Crushing 100 g of ginseng sun-dried ginseng to 60-80 meshes, adding 1000 ml of water, keeping the temperature at 30-65 ℃, adding 2 g of cellulase and 1 g of amylase, and reacting for 3 hours; then, under the same temperature, the pH value is kept constant to 3, and 0.5 g of acid protease is added under constant temperature stirring for reaction for 3 hours; then heating to 90 deg.C, reacting at constant temperature for half an hour to inactivate enzyme, and filtering. And concentrating the supernatant under reduced pressure at the vacuum degree of 0.06-0.10 Mpa and the temperature of 40-65 ℃, and freeze-drying to obtain solid ginseng zymolyte dry powder. The yield thereof was found to be 31.5%.

The advantageous effects of the present invention are further described below by way of test examples.

Test example 1 characterization of the composition of matter of the dry powder of the enzymatic hydrolysate

The contents of polysaccharide, flavone, total nitrogen, protein and total saponin in the zymolyte dry powder of the previous examples were measured, and the results are shown in table 1.

TABLE 1 content of major components of the enzymatic hydrolysate (in weight percent)

	Polysaccharides	Flavone	Total nitrogen	Protein content	Total saponin content
						Example 1	54.14％	0.80％	2.45％	15.31％	10.18％
Example 2	76.86％	1.04％	2.49％	15.57％	6.75％
						Example 3	57.16％	0.87％	2.34％	14.63％	5.85％
Comparative example 1	68.16％	0.80％	1.97％	12.33％	5.28％
						Comparative example 2	16.53％	0.97％	1.57％	9.82％	3.86％

Wherein, the HPLC peak diagrams of the saponin detection of comparative example 1 and example 1 are shown in FIG. 1 and FIG. 2, respectively, and the corresponding statistics of various saponins are shown in Table 2 and Table 3, respectively.

TABLE 2 comparative example 1 saponin substance HPLC detection values

TABLE 3 HPLC-DETECTION PEAK PATTERN OF SAPONIN IN DRY POWDER OF THE zymolyte obtained in example 1

From the comparison results, the method of the invention uses a specific enzymolysis process, which not only can improve the yield of the zymolyte, but also can improve the content of the saponin in the zymolyte.

Test example 2 molecular weight characterization of ginseng zymolyte of example 1

The number average molecular weight, weight average molecular weight, Z average molecular weight, and Z +1 average molecular weight of the ginseng zymolyte in example 1 were measured. The detection spectrum is shown in FIG. 3. The relative molecular weight results are shown in table 4.

Table 4 example 1 zymolyte molecular weight characterization

Experimental example 3 application experiment of ginseng zymolyte in anti-fatigue population

The crowd data: selection of 10 subjects, age: the age of the male is 30-55 years, and the male and the female are 5 cases respectively. The testees are easy to fatigue, sleepiness is achieved within 1 hour of driving, afternoon nap is needed, otherwise, the work cannot concentrate on the work.

The subject takes the sun-dried ginseng powder continuously for 10 days; after 20 days, the zymolyte of example 4 is taken for 10 consecutive days; after another 20 days, the enzymatic hydrolysate of comparative example 1 was administered for 10 consecutive days.

The taking method comprises the following steps: the preparation is taken orally once a day, 1 g each time, dissolved in warm water. It is administered in the morning on an empty stomach.

As shown in Table 5, the fatigue-preventing effect of example 4 was the best.

TABLE 5 anti-fatigue test of ginseng zymolyte

And (4) conclusion: the ginseng zymolyte has good anti-fatigue effect, and the effect is better than that of the untreated ginseng powder and ginseng zymolyte produced by other processes.

Test example 4 application experiment of ginseng zymolyte in invigorating kidney and strengthening yang

1. Method of producing a composite material

(1) Experimental group

In total 8 cases, 45-65 years old, male, no morning bobble had been for about 1 year.

The ginseng zymolyte and the fish scale collagen polypeptide are compounded at a ratio of 1: 9(w/w), 10 g of the compound is taken once a day on an empty stomach. After taking for 15 days, the effect is observed.

The ginseng zymolyte is prepared by the process of example 4.

(2) Control group

In total 4 cases, 55-60 years old, male, no morning bobble had been for about 1 year.

The ginseng zymolyte and the fish scale collagen polypeptide are compounded at a ratio of 1: 9(w/w), 10 g of the compound is taken once a day on an empty stomach. After taking for 15 days, the effect is observed.

The ginseng zymolyte is prepared by the process of comparative example 1.

The fish scale collagen polypeptide of the experimental group and the control group is prepared by taking tilapia scales as raw materials, wherein the weight percentage of hydroxylysine is more than 1.3%; the weight average molecular weight is 800-2400; the hydroxyproline accounts for more than 7.3 percent by weight;

the fish scale collagen polypeptide can be prepared according to the method disclosed by the patent publication No. CN104223115B, and specifically comprises the following steps:

a. taking 100 parts by weight of tilapia scales as a raw material, adding 800-1200 parts by weight of water, heating to 80-90 ℃, and keeping the temperature for 20-60 minutes;

b. then cooling to 30-65 ℃, adjusting the pH value to 8-9, adding 1-8 parts of alkaline protease, and degrading for 0.5-12 hours at constant temperature;

c. adding 1-8 parts of neutral protease at the constant temperature of 45-50 ℃, and stirring for reaction for 0.5-12 hours;

d. then heating to 75-90 ℃, and preserving the heat for 15-30 minutes to inactivate the enzyme;

e. adding flocculant, flocculating to precipitate impurities, filtering, and spray drying the filtrate.

2. Results

In the experimental group: morning bobs appear on the second day after 1 case is taken, morning bobs appear on the second day after 2 cases are taken for one week, and morning bobs appear on the second day after 5 cases are taken for 10 days.

In the control group: after one week, morning puffs appeared in 1 person, and in other 3 cases, mental condition was improved, but no morning puffs appeared.

3. Conclusion

The zymolyte of the invention has good effects of invigorating the kidney and strengthening yang.

In conclusion, the product prepared by the preparation process of the ginseng zymolyte has high yield, rich enzymolysis active substances, good effects of resisting fatigue, tonifying kidney and strengthening yang, and good application prospect.

Claims (10)

1. A ginseng enzymolysis method is characterized by comprising the following steps:

1) pulverizing 100 weight parts of dry root of Panax plant, adding 500-2000 weight parts of water, adding 1-8 weight parts of carbohydrase at 30-65 deg.C, and reacting for 0.5-12 hr;

2) simultaneously adding 0.1-8 parts by weight of alkaline protease and 0.1-8 parts by weight of neutral protease to react for 0.5-12 hours at the same temperature, or adding 0.1-8 parts by weight of alkaline protease to react for 0.5-12 hours, and then adding 0.1-8 parts by weight of neutral protease to react for 0.5-12 hours;

3) terminating the enzyme reaction of step 2);

step 2) maintaining the pH value of 6-9.

2. The ginseng enzymatic hydrolysis method according to claim 1, wherein: the Panax plants in the step 1) are as follows: ginseng (Panax ginseng), american ginseng (Panax quinquefolius), pseudoginseng (Panax pseudoginseng), or Panax notoginseng (Panax zingiberensis);

and/or the reaction time of step 1) is 3 hours;

and/or, the carbohydrases of step 1) are amylases and cellulases; preferably, the amylase in step 1) is used in an amount of 1-2 parts by weight, and the cellulase is used in an amount of 0.5-2 parts by weight.

3. The ginseng enzymatic hydrolysis method according to claim 1 or 2, wherein: the pH value in step 2) is 8.

4. The ginseng enzymatic hydrolysis method according to claim 1 or 2, wherein: 0.5-3 parts by weight of alkaline protease in the step 2); and/or the reaction time is 2-4 h.

5. The ginseng enzymatic hydrolysis method according to claim 1 or 2, wherein: 0.2 to 0.5 weight portion of neutral protease in the step 3); and/or the reaction time is 2 h.

6. The ginseng enzymatic hydrolysis method according to claim 1 or 2, wherein: the step 3) is followed by the following steps:

4) filtering to obtain supernatant, concentrating under reduced pressure, and freeze drying to obtain dry powder.

7. A ginseng zymolyte is characterized in that: the zymolyte is prepared by the method of any one of claims 1 to 8.

8. The ginseng zymolyte of claim 7, wherein: the number average molecular weight of the ginseng zymolyte is 346;

and/or a number average molecular weight of 274;

and/or, Z average molecular weight is 515;

and/or, Z +1 has an average molecular weight of 878.

9. Use of the ginseng zymolyte of claim 7 or 8 in preparing a nutritional food, a health product or a medicament for resisting fatigue, tonifying kidney and strengthening yang.

10. Use according to claim 9, characterized in that: the nutritional food, the health product or the medicine also contains fish scale collagen;

preferably, the fish scale collagen is prepared by taking tilapia scales as raw materials, wherein the weight percentage of hydroxylysine is more than 1.3%; the weight average molecular weight is 800-2400; the hydroxyproline accounts for more than 7.3 percent by weight;

preferably, the weight ratio of the ginseng zymolyte to the fish scale collagen is 1: 9.

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