CN112438404B - Application of radix scrophulariae polysaccharide - Google Patents
Application of radix scrophulariae polysaccharide Download PDFInfo
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- CN112438404B CN112438404B CN202011340019.2A CN202011340019A CN112438404B CN 112438404 B CN112438404 B CN 112438404B CN 202011340019 A CN202011340019 A CN 202011340019A CN 112438404 B CN112438404 B CN 112438404B
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- flavescent sophora
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/20—Reducing nutritive value; Dietetic products with reduced nutritive value
- A23L33/21—Addition of substantially indigestible substances, e.g. dietary fibres
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/13—Fermented milk preparations; Treatment using microorganisms or enzymes using additives
- A23C9/1307—Milk products or derivatives; Fruit or vegetable juices; Sugars, sugar alcohols, sweeteners; Oligosaccharides; Organic acids or salts thereof or acidifying agents; Flavours, dyes or pigments; Inert or aerosol gases; Carbonation methods
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Medicines Containing Plant Substances (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention discloses application of a flavescent sophora root polysaccharide, and relates to the technical field of functional food additives and nutritional health products. The flavescent sophora polysaccharide serving as the prebiotic raw material is derived from flavescent sophora, has rich raw material sources, low production cost and simple preparation process, has a prebiotic effect, can promote the growth of probiotics, can resist the hydrolysis of gastrointestinal fluid, and can reach the intestinal tract to be utilized by the probiotics in the intestinal tract, thereby improving the intestinal flora and promoting the health of human bodies.
Description
Technical Field
The invention relates to the technical field of functional food additives and nutritional health products, in particular to application of flavescent sophora root polysaccharide.
Background
Probiotics are important intestinal microorganisms of a human body, can improve intestinal immunity or non-immune barrier, activate endogenous bacterial metabolism, keep balance of intestinal microecology, and can also play a certain role in regulating metabolic diseases such as hypertension, diabetes and the like by regulating organism metabolism.
In order to promote the colonization of the intestinal tract by the probiotics, the balance of the intestinal flora is regulated, and the prebiotics play an important role therein. Prebiotics refer to organic substances that are not digested and absorbed by the host but which selectively promote the metabolism and proliferation of probiotics in the body, thereby improving the health of the host.
Common prebiotics are fructo-oligosaccharides (FOS), galacto-oligosaccharides, inulin, polydextrose, isomalto-oligosaccharides, resistant dextrins, resistant starches, mannooligosaccharides, trehalose, xylooligosaccharides, stachyose, chitosan, lactoketose, palatinose, gentian oligospermins and soy oligosaccharides, etc., which can be consumed alone or added to foods enriched with probiotics to promote the growth and reproduction of the probiotics. Among these functional oligosaccharides are the most predominant and most studied class of prebiotics.
The yellow ginseng is a plant of the genus Miquel (SpHallerocarpus gracilis) of the family Umbelliferae, and is known as Huang, wild carrot, fruit (Qinghai), small She Shangong radish (Hebei varian), daqu (Sichuan Deg) and the like, and is similar to ginseng, golden in whole body, and has long utilization history as a traditional Chinese medicine because of medicinal effects and higher nutritional value. The radix scrophulariae polysaccharide is one of main components of radix scrophulariae, and researches on the radix scrophulariae polysaccharide are mainly focused on an extraction process and a detection method, and the researches on functional characteristics of the radix scrophulariae polysaccharide are less. According to the existing data, the functions of the yellow ginseng polysaccharide are mainly reflected in the aspects of immunity enhancement, oxidation resistance, blood sugar reduction, fatigue resistance, nitrite removal and the like, but the development of the yellow ginseng polysaccharide as a prebiotic is not reported yet.
Disclosure of Invention
Therefore, the invention provides application of the flavescent sophora root polysaccharide to solve the problems that the flavescent sophora root polysaccharide has high nutritive value but is not reported yet when being developed as a prebiotic.
In order to achieve the above object, the present invention provides the following technical solutions:
according to a first aspect of the present invention, the use of a flavescent sophora polysaccharide as a prebiotic in functional food additives and nutritional health products.
Further, the flavescent sophora root polysaccharide is one or only one of the active ingredients of the functional food additive and the nutritional health care product.
Further, the yellow ginseng polysaccharide is extracted by adopting a complex enzyme method or an ultrasonic wave combined complex enzyme method.
Further, the use includes the use of prebiotics in the production of yogurt or the consumption of prebiotics alone.
According to a second aspect of the present invention, a prebiotic comprises an effective amount of a flavescent sophora polysaccharide.
Further, the effective amount of the flavescent sophora polysaccharide is one or only one of the effective components of the prebiotic.
The invention has the following advantages:
the flavescent sophora polysaccharide serving as the prebiotic raw material is derived from flavescent sophora, has rich raw material sources, low production cost and simple preparation process, has a prebiotic effect, can promote the growth of probiotics, can resist the hydrolysis of gastrointestinal fluid, and can reach the intestinal tract to be utilized by the probiotics in the intestinal tract, thereby improving the intestinal flora and promoting the health of human bodies. The yellow ginseng polysaccharide can be taken alone as a prebiotic or added into foods rich in probiotics to promote the growth and propagation of the probiotics.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below. It will be apparent to those skilled in the art from this disclosure that the drawings described below are merely exemplary and that other embodiments may be derived from the drawings provided without undue effort.
The structures, proportions, sizes, etc. shown in the present specification are shown only for the purposes of illustration and description, and are not intended to limit the scope of the invention, which is defined by the claims, so that any structural modifications, changes in proportions, or adjustments of sizes, which do not affect the efficacy or the achievement of the present invention, should fall within the scope of the invention.
FIG. 1 is a graph showing the effect of different amounts of polysaccharide added to Lactobacillus bulgaricus and Streptococcus thermophilus during fermentation in example 1;
FIG. 2 is a graph showing the effect of different amounts of polysaccharide from yellow ginseng on titrating acidity during fermentation of Lactobacillus bulgaricus and Streptococcus thermophilus in example 1 of the present invention.
FIG. 3 is a graph showing the effect of different amounts of polysaccharide from yellow ginseng on proliferation of Streptococcus thermophilus in example 2 of the present invention;
FIG. 4 is a graph showing the effect of different prebiotics on the proliferation of thermophilic chain balls in example 2 of the present invention;
FIG. 5 is a graph showing comparison of the degree of hydrolysis of the polysaccharide of flavescent sophora root at different pH values for simulating gastric juice of human body in example 3 of the present invention;
FIG. 6 is a graph showing comparison of the degree of hydrolysis of the polysaccharide from Huangshan in the simulated intestinal fluid of human body at different pH values, in example 3 of the present invention, using FOS and inulin as controls.
Detailed Description
Other advantages and advantages of the present invention will become apparent to those skilled in the art from the following detailed description, which, by way of illustration, is to be read in connection with certain specific embodiments, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The yellow ginseng polysaccharide in the following examples was extracted by the following method:
1. pretreatment of yellow ginseng
Cutting appropriate amount of dried Huang Cangen into small pieces, pulverizing in pulverizer until there is no large residue, sieving with 20 mesh sieve, and collecting the sieved radix Ginseng Rubra powder.
2. Ultrasonic wave combined complex enzyme method for extracting polysaccharide of radix scrophulariae
The experiment adopts ultrasonic wave combined complex enzyme method to extract the polysaccharide of the radix scrophulariae, and the specific operation flow is as follows: drying radix Ginseng Rubra powder at 60-70deg.C, adding appropriate amount of distilled water, and dissolving with the ratio of 1:40 (mg/mL), regulating pH of the solution to 5.8 with 12mol/L concentrated hydrochloric acid, adding complex enzyme (papain 0.4% and cellulase 2%), and performing ultrasonic-assisted enzymolysis at 60deg.C for 25min at 152.5W. Centrifuging the enzymolysis solution at 5000r/min for 10min, collecting supernatant, concentrating at 60deg.C under reduced pressure, and adding 4 times of anhydrous ethanol to precipitate overnight. Centrifuging at 8000r/min for 15min the next day, taking out precipitate, pre-freezing the precipitate in a refrigerator at-80deg.C for 12 hr, and vacuum freeze drying for 10-12 hr to obtain radix Ginseng Rubra crude polysaccharide. The crude polysaccharide of the yellow ginseng is extracted and then deproteinized and purified by a papain-Sevag method.
Example 1: effect of Huangshen polysaccharide on lactic acid bacteria acidogenesis
1. Experimental method
1.1 preparation of fermented milk
Sterilizing fresh milk at 85deg.C for 15min, cooling to 43deg.C, inoculating Lactobacillus bulgaricus and Streptococcus thermophilus into sterilized milk at appropriate ratio (w/v), adding Ginseng radix Rubri polysaccharide into fresh milk at appropriate ratio (w/v) of 0.3%,0.6%,0.9%,1.2%,1.5%, and 2.0%, stirring, and fermenting to curd at 43deg.C.
1.2 determination of pH
During the fermentation at 43 ℃, the pH values of each group were measured every 0.5h, each group of samples was measured 3 times in parallel and averaged.
1.3 determination of titrated acidity
The measurement of titrating acidity is carried out synchronously with the measurement of pH value, and the specific operation is as follows: 10mL of milk samples are taken every 0.5h in the fermentation process, placed in a 250mL conical flask, diluted by 2 times of deionized water, 100 mu L of phenolphthalein ethanol solution with the volume fraction of 0.5% is added dropwise, and then 0.1mol/LNaOH solution is used for titration until pink, and 30s does not fade as an end point. The titrated acidity of the sample, here denoted Ji Ernie degrees (°t), is obtained by multiplying the milliliters of NaOH solution consumed by 10. Each group of samples was measured 3 times in parallel and averaged.
2. Experimental results
2.1 Effect of Huangshen polysaccharide on lactic acid bacteria acidogenesis
As can be seen from fig. 1 and 2, the flavescent sophora root polysaccharide can promote the acid production of lactobacillus bulgaricus and streptococcus thermophilus, and improve the fermentation efficiency. The lactobacillus bulgaricus and streptococcus thermophilus are basic fermentation agents of all the yogurt, when the adding amount of the flavescent sophora root polysaccharide is 0.9%, the lactobacillus bulgaricus and the streptococcus thermophilus can ferment for 2 hours to ensure that the titrated acidity of the fermented milk is higher than 70 (° T), and the pH of the fermented milk can be lower than 4.5 after 3 hours of fermentation, so that the acid production rate (p < 0.05) of the lactobacillus bulgaricus and the streptococcus thermophilus is obviously improved, and the production efficiency can be improved.
Example 2: proliferation research of streptococcus thermophilus, lactobacillus plantarum and lactobacillus rhamnosus by using flavescent sophora polysaccharide
1. Experimental method
1.1 Effect of flavescent Ginseng polysaccharide on proliferation of Streptococcus thermophilus
Sterilizing fresh milk at 85deg.C for 15min, cooling to 43deg.C, inoculating Streptococcus thermophilus into sterilized milk at a ratio of 0.3%,0.6%,0.9%,1.2%,1.5%, and 2.0% (w/v), respectively, adding radix Ginseng Rubra polysaccharide into fresh milk, stirring, and fermenting to curd at 43deg.C.
The M17 medium and MRS medium cooled to 60℃are poured into about 15mL plates, and the plates are rotated to mix well until they cool and solidify. After milk coagulation of fresh cows, each group of samples was diluted to different gradients according to the method of national standard GB4789.35-2016, 3 consecutive appropriate dilutions were selected, 100. Mu.L of sample homogenate was drawn from each dilution, added to a petri dish, coated uniformly with a coating rod, then incubated for 72 h.+ -. 2h at 36.+ -. 1 ℃ in an aerobic environment, and counting of Streptococcus thermophilus was performed on plates of M17 medium.
1.2 Effect of Huangshan polysaccharide on proliferation of Lactobacillus plantarum and Lactobacillus rhamnosus
Taking FOS and inulin as positive control, pasteurizing fresh cow milk for 15min, placing on a clean bench, cooling to 37deg.C by a thermometer, and inoculating 0.6% (w/w) Lactobacillus plantarum/Lactobacillus rhamnosus respectively. The polysaccharide concentration was set to 0%,1%,1.5%,2%,2.5%,3%,4%, the polysaccharide content in the yellow ginseng polysaccharide was about 60%, and the polysaccharide amounts in FOS and inulin were about 90%, and in this experiment, the polysaccharide was added by concentration calculation according to the polysaccharide amount actually contained. After fully stirring, the milk is fermented in a constant temperature box at 37 ℃.
Taking out the whole fermented milk, placing on a clean workbench for dilution, adding 0.1ml fermented milk into 0.9ml sterile NaCl solution for continuous dilution, wherein the dilution multiple is 10 5 Plating after dilution, selecting MRS culture medium for plating, making 3 groups of plates in parallel, culturing in an incubator at 37 ℃ for 48 hours after plating, and counting colonies.
2. Experimental results and discussion
2.1 Streptococcus thermophilus counts
As can be seen from fig. 3, when the addition amount of the flavescent sophora root polysaccharide is between 0.0% and 0.6%, the colony count of streptococcus thermophilus increases with the increase of the addition amount of the flavescent sophora root polysaccharide, and when the addition amount of the flavescent sophora root polysaccharide is more than 0.6%, the colony count of streptococcus thermophilus gradually decreases with the increase of the addition amount, and after 1.5%, the colony count gradually becomes stable.
As can be seen from FIG. 4, the same amount of flavescent holothurian polysaccharide has the best effect on promoting the proliferation of Streptococcus thermophilus, and then FOS, inulin and blank have no statistical difference.
2.2 Lactobacillus plantarum and Lactobacillus rhamnosus counts
TABLE 1 influence of different prebiotics on proliferation of Lactobacillus plantarum and Lactobacillus rhamnosus
Data are presented as mean ± standard deviation
The same letters indicate no difference
Through the data analysis, the flavescent sophora root polysaccharide has the effect of promoting the proliferation of lactobacillus plantarum>Inulin>FOS, radix Ginseng Rubra polysaccharide has maximum proliferation effect on Lactobacillus plantarum at concentration of 1%, and is (1.81+ -0.06) ×10 8 cfu/ml, inulin and FOS were maximized at 3%, respectively (4.11.+ -. 0.06). Times.10 7 cfu/ml，(55.33±0.03)×10 7 cfu/ml。
For promoting proliferation of lactobacillus rhamnosus, radix Ginseng Rubra polysaccharide>Fos=inulin, and the proliferation effect of the flavescent sophora root polysaccharide on lactobacillus plantarum is maximum at the concentration of 1%, which is (2.12+/-0.06) ×10 8 cfu/ml, inulin and FOS were maximized at 3%, respectively (8.93.+ -. 0.03). Times.10 7 cfu/ml，(9.11±0.06)×10 7 cfu/ml。
It can be seen from this that the flavescent ginseng polysaccharide is superior to inulin and FOS in comparison of the proliferation of lactobacillus plantarum and lactobacillus rhamnosus.
Example 3: simulation of gastrointestinal solution digestion experiments
1. The experimental method comprises the following steps:
1g of the yellow ginseng polysaccharide was dissolved in 100ml of distilled water to prepare a 1% (w/v) solution.
1.1 gastric juice simulation experiment
FOS and inulin were used as positive controls.
The formula of simulated gastric juice: naCl 8g, KCl 0.2g, na 2 HPO 4 ·2H 2 08.25g,NaHPO 4 14.35g，CaCl 2 ·2H 2 O 0.1g,MgCl 2 ·6H 2 O0.18 g was dissolved in distilled water to prepare 1000ml of a solution. The pH of the solution was adjusted to 1,2,3,4,5 with 5mol/L hydrochloric acid, respectively. 5ml of each pH solution is respectively taken and added with 5ml of polysaccharide solution, then the mixture is reacted on a shaking table at 37 ℃ and 130r/min for 6 hours, the movement condition of human stomach is simulated, the total sugar content and the reducing sugar content are respectively measured at 0h,1h,2h,4h and 6h, the total sugar is measured by a phenol-sulfuric acid method, and the reducing sugar is measured by a DNS method. The degree of hydrolysis was calculated according to the following formula:
degree of hydrolysis (100%) =hydrolyzed reducing sugar/(total sugar-initial reducing sugar) ×100%.
A comparison graph of the degree of hydrolysis of the simulated human gastrointestinal solution at different pH values for the yellow ginseng polysaccharide in comparison with FOS and inulin is shown in FIG. 5.
1.2 simulation of intestinal juice experiments
FOS and inulin were used as positive controls.
The simulated intestinal juice formula comprises: naCl 5.4g,KCl 0.65g,CaCl 2 0.33g of the electrolyte was dissolved in distilled water to prepare 1000ml of an intestinal electrolyte solution. 7g/100ml trypsin solution: 7g trypsin was dissolved in 100ml PBS solution with pH=7.2. 100ml of intestinal electrolyte solution was taken, 100ml of trypsin solution was added, and 1mol/LNaHCO was used 3 The pH of the solution was adjusted to 7.2. 30ml of simulated intestinal juice is taken and added with pig bile salt, so that the final concentration of the bile salt in the solution is 0.05%,0.15% and 0.3% respectively. Adding 5ml of gastric juice obtained by the above reaction for 6 hours into 5ml of bile salt intestinal juice with different concentrations, immediately using 1mol/LNaHCO 3 The pH of the solution was adjusted to 7.2. Then the reaction is carried out on a shaking table at 37 ℃ for 6 hours at 45r/min, the total sugar content and the reducing sugar content are respectively measured at 0h,1h,2h,4h and 6h, the total sugar is measured by a phenol-sulfuric acid method, and the reducing sugar is measured by a DNS method. The degree of hydrolysis was calculated according to the following formula:
degree of hydrolysis (100%) =hydrolyzed reducing sugar/(total sugar-initial reducing sugar) ×100%.
A comparison graph of the degree of hydrolysis of the flavescent sophora polysaccharide in simulated human intestinal fluid at different pH values using FOS and inulin as controls is shown in FIG. 6.
2. Analysis of results
According to data analysis, the hydrolysis degree of 5 pH values has a remarkable difference (P < 0.05) in the range of 1,2,3,4 and 5 pH values of simulated gastric juice, the hydrolysis degree is FOS > inulin > flavescent sophora polysaccharide, and the pH value range of human gastric juice is 1-2, so that the resistance of the flavescent sophora polysaccharide to gastric juice hydrolysis is greater than that of FOS and inulin.
In 0.05%,0.15% and 0.3% of bile salt intestinal juice, the hydrolysis degree has a remarkable difference (P < 0.05) through SPSS analysis, the hydrolysis degree is FOS > inulin > flavescent sophora root polysaccharide, in simulated human intestinal juice, the bile salt concentration is 0.1% -0.3%, and the resistance of the flavescent sophora root polysaccharide to intestinal juice hydrolysis is stronger than that of FOS and inulin.
In conclusion, the flavescent holothurian polysaccharide can withstand hydrolysis of simulated gastrointestinal solution, and has higher gastric juice hydrolysis resistance than inulin and fructo-oligosaccharides (FOS), which means that the flavescent holothurian polysaccharide can reach the intestinal tract more directly than the flavescent holothurian polysaccharide and can be utilized by intestinal probiotics.
While the invention has been described in detail in the foregoing general description and specific examples, it will be apparent to those skilled in the art that modifications and improvements can be made thereto. Accordingly, such modifications or improvements may be made without departing from the spirit of the invention and are intended to be within the scope of the invention as claimed.
Claims (1)
1. The application of the flavescent sophora root polysaccharide as a prebiotic in the preparation of functional food additives and nutritional health care products is characterized in that the preparation method of the flavescent sophora root polysaccharide is as follows:
drying radix Ginseng Rubra powder at 60-70deg.C, adding appropriate amount of distilled water for dissolving, and mixing with the feed liquid with a ratio of mg/mL of 1:40, regulating the pH value of the solution to 5.8 by using 12mol/L concentrated hydrochloric acid, and adding complex enzyme, wherein the complex enzyme consists of papain and cellulase, the addition amount of the papain is 0.4%, the addition amount of the cellulase is 2%, and simultaneously, the enzymolysis is assisted by 152.5W of ultrasound, the enzymolysis temperature is 61 ℃, and the enzymolysis time is 25min;
centrifuging the enzymolysis solution at 5000r/min for 10min, collecting supernatant, concentrating under reduced pressure at 60deg.C, and adding 4 times volume of absolute ethanol to precipitate overnight;
centrifuging at 8000r/min for 15min the next day, taking out precipitate, pre-freezing the precipitate in a refrigerator at-80deg.C for 12 hr, and vacuum freeze drying for 10-12 hr to obtain radix Ginseng Rubra crude polysaccharide;
the crude polysaccharide of the yellow ginseng is extracted and then deproteinized and purified by a papain-Sevag method.
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