CN116114867B - Anti-saccharification pagodatree flower bud powder and preparation method and application thereof - Google Patents
Anti-saccharification pagodatree flower bud powder and preparation method and application thereof Download PDFInfo
- Publication number
- CN116114867B CN116114867B CN202211480228.6A CN202211480228A CN116114867B CN 116114867 B CN116114867 B CN 116114867B CN 202211480228 A CN202211480228 A CN 202211480228A CN 116114867 B CN116114867 B CN 116114867B
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- China
- Prior art keywords
- flower bud
- rice flour
- pagodatree flower
- enzyme
- saccharification
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Classifications
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- 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
-
- 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
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/06—Enzymes
-
- 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
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/065—Microorganisms
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- 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
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- 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
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/175—Amino acids
-
- 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)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a preparation method of anti-saccharification sophora japonica rice flour, which comprises the following steps: (1) The method comprises the steps of obtaining pagodatree flower bud raw materials, cleaning, placing the cleaned pagodatree flower bud raw materials into a fermentation box, fermenting by spraying 0.5-4wt% of fermentation liquor, drying and crushing to obtain fermented pagodatree flower powder, wherein the fermentation liquor consists of aspergillus niger and bifidobacterium; (2) Adding complex enzyme into the fermented locust rice flour for enzymolysis reaction; (3) Performing enzyme deactivation treatment on the fermented sophora japonica rice flour after the enzymolysis reaction to obtain an enzyme deactivation mixture; (4) And mixing the enzyme-deactivated mixture with amino acid, and drying to obtain the finished product of the pagodatree flower bud powder. Correspondingly, the invention also provides the sophora japonica rice flour prepared by the preparation method and application of the sophora japonica rice flour in preparing anti-saccharification food or health care products. The invention can better improve the functional components in the pagodatree flower bud, has simple preparation process and good anti-saccharification effect, and is beneficial to human body absorption.
Description
Technical Field
The invention relates to the technical field of high-value utilization of edible plant raw materials, in particular to anti-saccharification pagodatree flower bud powder and a preparation method and application thereof.
Background
The glycosylated end products (AGEs) are the end products of non-enzymatic browning reactions of reducing sugars and amino acid groups in proteins. AGEs are growing and accumulating in human tissue with age, causing physiological aging of the skin. The glycation cross-linking between the molecules of the aging skin tissue promotes the accelerated aging of the skin and gradually loses elasticity, and the skin becomes stiff and dull. After collagen in the dermis is saccharified, the elasticity of collagen fibers is reduced, so that the skin is loose due to the loss of skin tenseness, and wrinkles are formed. Dicarbonyl compounds, such as Glyoxal (GO), methylglyoxal (MGO), etc., are important precursors for AGEs, and are responsible for saccharification. In recent years, anti-saccharification foods and health care products are continuously appeared, but the defects that the anti-saccharification effect is not ideal, the anti-saccharification active ingredients are difficult to exert the functions and the like are overcome. Therefore, it is highly demanded to develop a food having high safety, no irritation and remarkable anti-glycation effect.
The prior research discovers that the pagodatree flower bud contains various bioactive substances, mainly comprising rutin, triterpenoid saponin, betulin, pagodatree flower diol, glucose, gluconic acid aldehyde and the like, and the functional active ingredients have wide application potential in the fields of medical care products, cosmetic skin care, non-staple food products, daily necessities and the like. Rutin and quercetin are main active ingredients of flos Sophorae Immaturus and its product with effects of resisting sugar and free radical. Therefore, the proper treatment process is explored, the metabolism, absorption and bioavailability of the active ingredients in the pagodatree flower bud are improved to the greatest extent, and the exertion of the bioactivity of the pagodatree flower bud becomes the key of the deep processing research of the pagodatree flower bud.
Disclosure of Invention
The first technical problem to be solved by the invention is to provide the preparation method of the anti-saccharification sophora japonica rice flour, which effectively improves the effective components in the sophora japonica, has simple preparation process and good anti-saccharification effect, and can inhibit the activity of MGO and GO.
The second technical problem to be solved by the invention is to provide the anti-saccharification pagodatree flower bud powder and the application thereof, and the pagodatree flower bud powder has good taste, easy absorption, high safety and good anti-saccharification effect.
In order to achieve the technical effects, the invention provides a preparation method of anti-saccharification sophorae rice flour, which comprises the following steps:
(1) The method comprises the steps of obtaining pagodatree flower bud raw materials, cleaning, placing the cleaned pagodatree flower bud raw materials into a fermentation box, fermenting by spraying 0.5-4wt% of fermentation liquid, drying and crushing to obtain fermented pagodatree flower powder, wherein the fermentation liquid consists of aspergillus niger and bifidobacterium;
(2) Adding complex enzyme into the fermented locust rice flour for enzymolysis reaction;
(3) Performing enzyme deactivation treatment on the fermented sophora japonica rice flour after the enzymolysis reaction to obtain an enzyme deactivation mixture;
(4) And mixing the enzyme-deactivated mixture with amino acid, and drying to obtain the finished product of the pagodatree flower bud powder.
As an improvement of the scheme, the preparation method of the aspergillus niger comprises the following steps: screening Aspergillus niger strains from the pomelo orchard dead leaves, and inoculating the strains to a liquid culture medium for culture to obtain Aspergillus niger bacterial liquid;
the liquid culture medium comprises the following components: corn starch, bran, bean cake powder, bean dregs, ammonium sulfate, potassium dihydrogen phosphate and calcium chloride.
As an improvement of the scheme, the mass ratio of the aspergillus niger to the bifidobacterium in the step (1) is (5-7): (3-5);
and (3) fermenting the fermentation liquid in the step (1) at the temperature of 35-45 ℃ and the humidity of 60-75% for 5-10 days, and spraying the fermentation liquid every day while fermenting.
As an improvement of the scheme, in the step (2), the adding proportion of the fermented sophorae rice flour to the complex enzyme is (80-90): (10-20);
the complex enzyme comprises one or more of amylase, acid protease, cellulase, glucosidase and pectase.
As an improvement of the scheme, the compound enzyme is a mixture of cellulase, beta-glucosidase and pectase, and the mass ratio of the cellulase to the beta-glucosidase to the pectase is (0.8-1.8): (0.5-1): (0.3-1.5).
As an improvement of the scheme, in the step (2), the temperature of the enzymolysis reaction is 30-50 ℃, the pH is 6-7, and the reaction time is 1-3 h.
As an improvement of the above scheme, the amino acid is arginine.
As an improvement of the above scheme, the addition ratio of the enzyme-inactivating mixture to the amino acid is (90-100): (9-15).
Correspondingly, the invention also provides the anti-saccharification pagodatree flower bud powder prepared by the preparation method and the application of the anti-saccharification pagodatree flower bud powder in preparation of anti-saccharification foods or health care products. The concentration of the pagodatree flower bud powder finished product in the anti-saccharification food or the health care product is 3-5 mug/ml, so that good inhibition effect can be achieved on MGO and GO; the concentration of the pagodatree flower bud powder finished product in the anti-saccharification food or the health care product is 500-1000 mug/ml, and the pagodatree flower bud powder finished product can have good inhibition effect on the formation of AGEs in various saccharide systems such as glucose, fructose, galactose, ribose and the like.
The implementation of the invention has the following beneficial effects:
the anti-saccharification pagodatree flower bud powder disclosed by the invention can better improve the functional components in the pagodatree flower bud, and the prepared product has high purity and strong anti-saccharification capability, and is more beneficial to human body absorption. In addition, the bitter taste of quercetin can be reduced, the taste of the sophorae rice flour is improved, and the absorption capacity is further improved while the sensory quality is not affected.
The anti-saccharification pagodatree flower bud powder disclosed by the invention can realize a better anti-saccharification effect at a lower dosage, and inhibit the formation of glycosylation end products by inhibiting the activity of glycosylation precursor dicarbonyl compounds, wherein the inhibition rate of 3-5 mug/ml pagodatree flower bud powder to MGO can reach more than 75% within 10min, and the inhibition rate of pagodatree flower bud powder with the final concentration of 500-1000 mug/ml in a bovine serum protein glycosylation model can reach more than 60%.
The anti-saccharification pagodatree flower bud powder disclosed by the invention has a wide application range, and is simple in component, convenient to prepare and good in anti-saccharification effect when being used for anti-saccharification foods or health-care products.
Drawings
FIG. 1 is a graph showing the effect of anti-glycation sophorae flour concentration on Glyoxal (GO) activity in example 4 of the present invention;
FIG. 2 shows the effect of anti-glycation sophorae flour concentration on Methylglyoxal (MGO) activity in example 4 of the present invention;
FIG. 3 is a graph showing the effect of anti-glycation sophorae flour concentration on formation of glycation end products (AGEs) in comparative example 5 of the present invention.
Detailed Description
The present invention will be described in further detail below for the purpose of making the objects, technical solutions and advantages of the present invention more apparent.
The invention provides a preparation method of anti-saccharification sophora japonica rice flour, which comprises the following steps:
(1) And (3) obtaining pagodatree flower bud raw materials, cleaning, placing the cleaned pagodatree flower bud raw materials into a fermentation box, fermenting by spraying 0.5-4wt% of fermentation liquid, drying and crushing to obtain fermented pagodatree flower powder, wherein the fermentation liquid consists of aspergillus niger and bifidobacterium.
The fermentation liquor adopts the complex formulation of aspergillus niger and bifidobacterium, wherein the aspergillus niger can decompose macromolecular sugar into monosaccharide and oligosaccharide through fermentation, and organic acid, vitamin, biological enzyme, growth factor and the like are generated, so that the nutrition level and digestibility of the fermented pagodatree flower bud powder are improved; the bifidobacterium is beneficial to the health of human bodies, can enhance the immune function in gastrointestinal tracts, increase the absorption and digestion of nutrient substances and balance the dysbacteriosis in human bodies.
Wherein the mass ratio of aspergillus niger to bifidobacterium in the fermentation liquor is preferably (5-7): (3-5); more preferably (5 to 6): (3-4).
The sophora flower powder is fermented by aspergillus niger and bifidobacterium before enzymolysis, the content of convertible components in the sophora flower powder is improved by fermentation, and the better absorption of the human body on the active components in the sophora flower powder is promoted.
Preferably, the fermentation process is carried out for 5-10 days under the conditions that the temperature is 35-45 ℃ and the humidity is 60-75%, and the fermentation liquid is sprayed every day while fermentation is carried out, so that the fermentation effect is better improved.
Further, the preparation method of aspergillus niger comprises the following steps: screening Aspergillus niger strains from the pomelo orchard dead leaves, and inoculating the strains to a liquid culture medium for culture to obtain Aspergillus niger bacterial liquid; the liquid medium comprises the following components: corn starch, bran, bean cake powder, bean dregs, ammonium sulfate, potassium dihydrogen phosphate and calcium chloride.
The aspergillus niger obtained by the method has rich raw material sources, can realize full utilization of biomass resources, can be used for large-scale production, and can be used for fermentation in cooperation with bifidobacteria to improve the content of active ingredients with anti-saccharification effect in the sophora japonica rice flour.
Specifically, the preparation process of the aspergillus niger used in the invention comprises the following steps: selecting a single colony from aspergillus niger screened from the dead leaves of the grapefruit orchard, inoculating the single colony into a liquid culture medium, and shake-culturing the single colony for 3-6 days at 25-35 ℃ and 150-200 rpm to obtain aspergillus niger bacterial liquid;
wherein, the liquid culture medium comprises the following raw materials in percentage by weight: corn starch 2-6%, bran 3-8%, bean cake powder 2-6%, bean dregs 6-15%, ammonium sulfate 0.5-1.5%, monopotassium phosphate 0.1-0.5%, calcium chloride 0.2-0.7%, and water the balance, wherein the pH is natural, and steam pressure sterilization is carried out for 20min under 0.1 Mpa.
(2) Adding complex enzyme into the fermented sophora japonica rice flour obtained in the step (1) to carry out enzymolysis reaction;
wherein, the adding proportion of the fermented locust rice flour and the complex enzyme is (80-90): (10-20); the complex enzyme comprises one or more of amylase, acid protease, cellulase, glucosidase and pectase.
Preferably, the complex enzyme is a mixture of cellulase, beta-glucosidase and pectinase.
The cellulase mainly comprises endo-and exo-beta-glucanase, beta-glucosidase and the like, and can decompose cellulose into polysaccharide or monosaccharide protein or RNA. Pectic enzymes are capable of degrading pectin, and typically include protopectic enzymes, pectate methyl hydrolase, and the like, and the combined action of the multiple enzyme complexes results in complete pectic decomposition. Beta-glucosidase may be involved in the metabolism of cellulose and in a variety of physiological and biochemical pathways.
The complex enzyme is utilized to destroy the tissue structure of the pagodatree flower bud, especially the situation that the components of the pagodatree flower bud cannot be fully exerted due to the wrapping of the components such as cellulose, pectin and the like can be avoided, the effective components in the pagodatree flower bud are fully exposed, the effective components in the pagodatree flower bud are enzymatically decomposed into carbohydrate substances which are easier to absorb, and the synergistic fermentation process and the subsequent operation steps are facilitated, so that the absorption capacity of the effective components in the pagodatree flower bud is improved.
Wherein, the mass ratio of the cellulase to the beta-glucosidase to the pectase is (0.8-1.8): (0.5-1): (0.3-1.5).
Optimally, the mass ratio of the cellulase to the beta-glucosidase to the pectase is 1:0.5:1.
when the mass ratio of the cellulase to the beta-glucosidase to the pectase is (0.8-1.8): (0.5-1): (0.3-1.5), the synergistic effect of the complex enzyme can be well realized, and under the synergistic effect of the complex enzyme, the fermented locust flour after enzymolysis treatment has high purity of the functional components and strong anti-saccharification capability, and is more beneficial to human body absorption.
Preferably, the temperature of the enzymolysis reaction is 30-50 ℃, the pH is 6-7, and the reaction time is 1-3 h.
More preferably, the temperature of the enzymolysis reaction is 40 ℃, the pH is 6.5, and the reaction time is 2 hours.
The enzymolysis conditions are selected to be suitable for the temperature, pH and time of the complex enzyme reaction, so that the activity of the complex enzyme is optimal, the enzymolysis of the complex enzyme can be better performed, and the obtained enzymatic locust flour has more active ingredients.
(3) Performing enzyme deactivation treatment on the mixture obtained in the step (2) to obtain an enzyme-deactivated mixture;
in order to ensure the preservation quality of the sophorae rice flour, the sophorae rice flour is subjected to enzyme deactivation treatment after enzymolysis, wherein the enzyme deactivation treatment can be conventional roasting, normal pressure cooking, high-pressure cooking, far infrared treatment or microwave treatment and the like, and the heat treatment enzyme deactivation method can also increase the flavor of the sophorae rice flour.
(4) Mixing the enzyme-deactivated mixture obtained in the step (3) with amino acid, and drying to obtain the finished product of the pagodatree flower bud powder.
The amino acid is preferably arginine; the addition ratio of the enzyme-inactivating mixture to the amino acid is (90-100): (9-15).
The addition of arginine is favorable for improving the taste of the pagodatree flower powder, and the content of the quercetin with bitter taste is increased after the pagodatree flower powder is fermented and hydrolyzed, so that the content of active ingredients is improved, and meanwhile, the adverse effect is caused on the whole flavor of the pagodatree flower powder, so that the whole taste of the pagodatree flower powder is improved, the bitter taste is reduced, meanwhile, the nutritional ingredients are increased, meanwhile, the water solubility and the fat solubility of flavonoid ingredients are very small, and the human body is difficult to absorb, so that the bioavailability is very low, and the pagodatree flower powder after enzymolysis treatment and arginine are combined to prepare mixed powder, so that the solubility of the quercetin in water can be increased, and the absorptivity of nutrient substances is improved.
Correspondingly, the invention also provides the anti-saccharification pagodatree flower bud powder prepared by the preparation method and the application of the anti-saccharification pagodatree flower bud powder in preparing anti-saccharification food or health care products.
The invention is further illustrated by the following specific examples.
Example 1
(1) Selecting a single colony from the aspergillus niger obtained by screening, inoculating the single colony into a liquid culture medium, and carrying out shake culture for 4 days at 30 ℃ and 180rpm to obtain aspergillus niger bacterial liquid; wherein, the liquid culture medium comprises the following raw materials in percentage by weight: corn starch 4%, bran 5%, bean cake powder 4%, bean dreg 10%, ammonium sulfate 1%, monopotassium phosphate 0.3%, calcium chloride 0.5%, and water the balance, wherein the pH is natural, and steam pressure of 0.1Mpa is used for sterilization for 20min.
(2) The method comprises the steps of obtaining pagodatree flower bud raw materials, cleaning, placing the cleaned pagodatree flower bud raw materials into a fermentation box, fermenting for 6 days at the temperature of 40 ℃ and the humidity of 65% by spraying 3wt% of fermentation liquid, spraying the fermentation liquid every day while fermenting, wherein the fermentation liquid comprises aspergillus niger and bifidobacterium according to the mass ratio of 3:2, drying and crushing the pagodatree flower bud after fermentation is completed to obtain fermented pagodatree flower powder;
(3) Adding 1kg of complex enzyme into 9kg of fermented locust rice flour, wherein the complex enzyme is a mixture of cellulase, beta-glucosidase and pectase, and the mass ratio of the cellulase to the beta-glucosidase to the pectase is 1:0.5:1, carrying out enzymolysis reaction in a liquid system with the temperature of 40 ℃ and the pH value of=6.5, and obtaining a mixture after 2 hours.
(4) And (3) carrying out enzyme deactivation treatment on the mixture obtained in the step (3) to obtain an enzyme-deactivated mixture.
(5) Mixing the enzyme-inactivating mixture obtained in the step (4) with arginine 10:1, mixing and drying to obtain the finished product of the pagodatree flower bud powder.
Example 2
(1) Selecting a single colony from the aspergillus niger obtained by screening, inoculating the single colony into a liquid culture medium, and carrying out shake culture for 3 days at 35 ℃ and 190rpm to obtain aspergillus niger bacterial liquid; wherein, the liquid culture medium comprises the following raw materials in percentage by weight: corn starch 4.5%, bran 4%, bean cake powder 3.5%, bean dreg 11%, ammonium sulfate 1%, monopotassium phosphate 0.3%, calcium chloride 0.5%, and water in balance, and sterilizing for 20min under natural pH and steam pressure of 0.1 Mpa.
(2) The pagodatree flower bud raw material is obtained and cleaned, the pagodatree flower bud raw material after cleaning is placed in a fermentation box, fermentation is carried out for 7 days at the temperature of 35 ℃ and the humidity of 65% by spraying 3.5wt% of fermentation liquid, the fermentation liquid is sprayed every day while fermentation, and the fermentation liquid is prepared from aspergillus niger and bifidobacterium according to the mass ratio of 7:5, drying and crushing the pagodatree flower bud after fermentation is completed to obtain fermented pagodatree flower powder;
(3) Adding 1kg of complex enzyme into 9kg of fermented locust rice flour, wherein the complex enzyme is a mixture of cellulase, beta-glucosidase and pectase, and the mass ratio of the cellulase to the beta-glucosidase to the pectase is 1.5:0.8:1, carrying out enzymolysis reaction in a liquid system with the temperature of 40 ℃ and the pH value of=6.5, and obtaining a mixture after 2 hours.
(4) And (3) carrying out enzyme deactivation treatment on the mixture obtained in the step (3) to obtain an enzyme-deactivated mixture.
(5) Mixing the enzyme-inactivating mixture obtained in the step (4) with arginine 9:1, mixing and drying to obtain the finished product of the pagodatree flower bud powder.
Example 3
(1) Selecting a single colony from the aspergillus niger obtained by screening, inoculating the single colony into a liquid culture medium, and carrying out shake culture for 5 days at 25 ℃ and 160rpm to obtain aspergillus niger bacterial liquid; wherein, the liquid culture medium comprises the following raw materials in percentage by weight: corn starch 5%, bran 5%, bean cake powder 4%, bean dregs 9%, ammonium sulfate 1%, monopotassium phosphate 0.3%, calcium chloride 0.5%, and water the balance, wherein the pH is natural, and steam pressure of 0.1Mpa is used for sterilization for 20min.
(2) The method comprises the steps of obtaining pagodatree flower bud raw materials, cleaning, placing the cleaned pagodatree flower bud raw materials into a fermentation box, fermenting for 5 days at the temperature of 35 ℃ and the humidity of 65% by spraying 3wt% of fermentation liquid, spraying the fermentation liquid every day while fermenting, wherein the fermentation liquid comprises aspergillus niger and bifidobacterium according to the mass ratio of 1:1, drying and crushing pagodatree flower bud after fermentation to obtain fermented pagodatree rice flour;
(3) Adding 1kg of complex enzyme into 8kg of fermented locust rice flour, wherein the complex enzyme is a mixture of cellulase, beta-glucosidase and pectase, and the mass ratio of the cellulase to the beta-glucosidase to the pectase is 1:0.5: and (3) performing enzymolysis reaction in a liquid system with the pH of=6.5 at the temperature of 40 ℃ for 2 hours to obtain a mixture.
(4) And (3) carrying out enzyme deactivation treatment on the mixture obtained in the step (3) to obtain an enzyme-deactivated mixture.
(5) Mixing the enzyme-inactivating mixture obtained in the step (4) with arginine 10:1, mixing and drying to obtain the finished product of the pagodatree flower bud powder.
Comparative example 1
(1) Selecting a single colony from the aspergillus niger obtained by screening, inoculating the single colony into a liquid culture medium, and carrying out shake culture for 4 days at 30 ℃ and 180rpm to obtain aspergillus niger bacterial liquid; wherein, the liquid culture medium comprises the following raw materials in percentage by weight: corn starch 4%, bran 5%, bean cake powder 4%, bean dreg 10%, ammonium sulfate 1%, monopotassium phosphate 0.3%, calcium chloride 0.5%, and water the balance, wherein the pH is natural, and steam pressure of 0.1Mpa is used for sterilization for 20min.
(2) The method comprises the steps of obtaining pagodatree flower bud raw materials, cleaning, placing the cleaned pagodatree flower bud raw materials into a fermentation box, fermenting for 6 days at the temperature of 40 ℃ and the humidity of 65% by spraying 3wt% of fermentation liquid, spraying the fermentation liquid every day while fermenting, wherein the fermentation liquid comprises aspergillus niger and bifidobacterium according to the mass ratio of 3:2, drying and crushing the pagodatree flower bud after fermentation is completed to obtain fermented pagodatree flower powder;
(3) Mixing the fermented locust rice flour obtained in the step (2) with arginine 10:1, mixing and drying to obtain the finished product of the pagodatree flower bud powder.
Comparative example 2
(1) Crushing the dried pagodatree flower bud to obtain pagodatree rice flour;
(2) Adding 1kg of complex enzyme into 9kg of locust rice flour, wherein the complex enzyme is a mixture of cellulase, beta-glucosidase and pectase, and the mass ratio of the cellulase, the beta-glucosidase and the pectase is 1:0.5:1, carrying out enzymolysis reaction in a liquid system with the temperature of 40 ℃ and the pH value of=6.5, and obtaining a mixture after 2 hours.
(3) And (3) carrying out enzyme deactivation treatment on the mixture obtained in the step (2) to obtain an enzyme-deactivated mixture.
(4) Mixing the enzyme-inactivating mixture obtained in the step (3) with arginine 10:1, mixing and drying to obtain the finished product of the pagodatree flower bud powder.
Comparative example 3
(1) Selecting a single colony from the aspergillus niger obtained by screening, inoculating the single colony into a liquid culture medium, and carrying out shake culture for 4 days at 30 ℃ and 180rpm to obtain aspergillus niger bacterial liquid; wherein, the liquid culture medium comprises the following raw materials in percentage by weight: corn starch 4%, bran 5%, bean cake powder 4%, bean dreg 10%, ammonium sulfate 1%, monopotassium phosphate 0.3%, calcium chloride 0.5%, and water the balance, wherein the pH is natural, and steam pressure of 0.1Mpa is used for sterilization for 20min.
(2) The pagodatree flower bud raw material is obtained and cleaned, the pagodatree flower bud raw material after cleaning is placed in a fermentation box, fermentation is carried out for 6 days at the temperature of 40 ℃ and the humidity of 65% by spraying 3wt% of fermentation liquid, the fermentation liquid is sprayed every day while fermentation, and the fermentation liquid is prepared from aspergillus niger and bifidobacterium according to the mass ratio of 6:1, drying and crushing pagodatree flower bud after fermentation to obtain fermented pagodatree rice flour;
(3) Adding 1kg of complex enzyme into 9kg of fermented locust rice flour, wherein the complex enzyme is a mixture of cellulase, beta-glucosidase and pectase, and the mass ratio of the cellulase to the beta-glucosidase to the pectase is 1:1:2, performing enzymolysis reaction in a liquid system with the temperature of 40 ℃ and the pH value of=6.5, and obtaining a mixture after 2 hours.
(4) And (3) carrying out enzyme deactivation treatment on the mixture obtained in the step (3) to obtain an enzyme-deactivated mixture.
(5) Mixing the enzyme-inactivating mixture obtained in the step (4) with arginine 10:1, mixing and drying to obtain the finished product of the pagodatree flower bud powder.
The pagodatree flower bud powders prepared in examples 1 to 3 and comparative examples 1 to 3 were tested for the effect of pagodatree flower bud powder on the activity of Methylglyoxal (MGO) and Glyoxal (GO).
The testing method comprises the following steps: in a 15ml centrifuge tube, adding 5ml of 1mM MGO and GO sample standard solution, adding 5ml of 5 mug/ml of methanol preparation solution of sophorae rice flour, uniformly mixing, vibrating and reacting in a shaking table at 37 ℃ for 50 r/min, taking out 1ml of sample solution at 60min, adding 10ul of acetic acid to terminate the reaction, and freezing at-80 ℃ to obtain three samples. And measuring the content of MGO and GO of different samples, and calculating the inhibition rate of the sophorae flour on the MGO and the GO.
The test results are shown in table 1:
TABLE 1 Effect of Sophora flour on the activity of Methylglyoxal (MGO) and Glyoxal (GO)
| Group of | MGO inhibition rate | Inhibition of GO |
| Example 1 | 80.13％ | 49.86％ |
| Example 2 | 79.66％ | 51.67％ |
| Example 3 | 79.74％ | 49.91％ |
| Comparative example 1 | 58.40％ | 43.06％ |
| Comparative example 2 | 53.91％ | 31.67％ |
| Comparative example 3 | 61.78％ | 44.27％ |
As can be seen from Table 1, the sophorae rice flour of examples 1-3 can significantly improve the inhibition of the activity of MGO and GO, and research shows that compared with GO, the reaction activity of MGO and bovine serum albumin glycosylation is high and fast, AGEs generated by MGO is far higher than AGEs generated by GO, the inhibition rate of the sophorae rice flour of examples 1-3 on the MGO can reach 80%, the inhibition rate of the sophorae rice flour of examples 1-3 on the MGO is significantly higher than that of comparative examples 1-3, and the inhibition rate of the sophorae rice flour on the GO is also higher than that of comparative examples 1-3. In conclusion, the sophorae rice flour provided by the invention can obviously improve the inhibition rate of glycosylated precursor dicarbonyl compounds, so that the effect of inhibiting the generation of AGEs is achieved.
Example 4
Effect of Sophora flower bud powder concentration on the activity of Methylglyoxal (MGO) and Glyoxal (GO)
Adding 5ml of 1mM MGO and GO sample standard solution into a 15ml centrifuge tube, adding 5ml of pagodatree flower bud powder methanol preparation solution with different concentrations into the centrifuge tube, wherein the pagodatree flower powder is prepared by the example 1, the concentration of the pagodatree flower powder is respectively 1 mug/ml, 3 mug/ml and 5 mug/ml, mixing the sample standard solution and the pagodatree flower bud powder methanol preparation solution uniformly, then carrying out shaking reaction in a shaking table at 37 ℃ and 50 revolutions/min, taking out 1ml of sample solution at 0, 10, 30, 60, 120 and 240min respectively, adding 10ul of acetic acid to terminate the reaction, and freezing and storing each sample at the temperature of-80 ℃, and carrying out three parallel experiments. And (3) measuring the content of MGO and GO of different samples, and calculating the inhibition rate of the sophorae rice flour to the MGO and GO, wherein the results are shown in figures 1 and 2.
Example 5
Effect of pagodatree flower bud powder concentration on formation of glycosylated end products (AGEs)
Dissolving Bovine Serum Albumin (BSA) in phosphate buffer solution with the concentration of 0.2mM and the pH of 7.4, adding 0.2ml of mixed solution of penicillin and streptomycin and 2ml of 4.5mg/ml BSA diluent into a 10ml sample tube, adding diluted glucose, fructose, galactose and ribose solution till the saccharide concentration is 10mM, adding pagodatree flower bud powder solutions with different concentrations into the four saccharide solution system, wherein the pagodatree flower bud powder is prepared in the embodiment 1, adding phosphate balanced normal saline (PBS) to 6ml, enabling the final concentration of the pagodatree flower powder to be 5 mug/ml, 50 mug/ml, 250 mug/ml and 1000 mug/ml respectively, heating in a water bath at 37 ℃, and freezing and storing at the temperature of minus 20 ℃ after 720 hours. The relative fluorescence values were measured and the inhibition ratios were calculated, and the results are shown in FIG. 3.
As can be seen from fig. 1 and fig. 2, the flos Sophorae Immaturus powder prepared by the method has good effect of inhibiting the MGO and the GO. In the process of the reaction from 0min to 10min, the inhibition rate of the pagodatree flower bud powder to the MGO and the GO is rapidly increased, the inhibition rate of the pagodatree flower bud powder to the MGO is slowly increased and gradually becomes gentle along with the continuous transition of the reaction time, when the reaction is carried out for 10min, the inhibition rate of the pagodatree flower bud powder with 1 mug/ml to the MGO is close to 60%, the low-concentration pagodatree flower bud powder can achieve a good inhibition effect on the MGO in a short time, and when the reaction is carried out for 60min, the inhibition rate of the pagodatree flower bud powder with 5 mug/ml to the GO can also reach more than 50%. Because AGEs generated by MGO are far higher than AGEs generated by GO, the high inhibition rate of MGO can achieve a good effect of inhibiting the generation of AGEs. Therefore, the sophorae rice flour provided by the invention can have a good inhibition effect on MGO and GO in a relatively low concentration (the concentration is 3-5 mug/ml) and a relatively short time (60 min).
As can be seen from FIG. 3, the sophorae rice flour prepared by the method has an inhibition effect on AGEs generated by different saccharides and BSA, especially glucose, and the inhibition rate on the AGEs is obviously increased along with the increase of the concentration of the added sophorae rice flour. When the concentration of the pagodatree flower bud powder is 500 mug/ml, the inhibiting rate of the pagodatree flower bud powder on AGEs generated by glucose, fructose, galactose, ribose and BSA is more than 50%, and especially the inhibiting rate of the pagodatree flower bud powder on the AGEs generated by glucose can reach more than 60%; when the concentration of the pagodatree flower bud powder is 1000 mug/ml, the inhibiting rate of the pagodatree flower bud powder on AGEs generated by glucose, fructose, galactose, ribose and BSA is more than 70%, and especially the inhibiting rate of the pagodatree flower bud powder on the AGEs generated by glucose can reach more than 80%. Therefore, the pagodatree flower bud powder provided by the invention can play a good role in inhibiting AGEs generated by various saccharides such as glucose, fructose, galactose, ribose and the like and BSA under the condition that the final concentration is 500-1000 mug/ml.
Therefore, the anti-saccharification sophora japonica rice flour can well inhibit the activity of MGO and GO in a short time at a lower dosage, can effectively inhibit the formation of AGEs in various sugar and protein reaction systems, and has a good anti-saccharification effect.
While the foregoing is directed to the preferred embodiments of the present invention, it will be appreciated by those skilled in the art that changes and modifications may be made without departing from the principles of the invention, such changes and modifications are also intended to be within the scope of the invention.
Claims (3)
1. A method for preparing anti-saccharification locust rice flour, which is characterized by comprising the following steps:
(1) Obtaining pagodatree flower bud raw materials, cleaning, placing the cleaned pagodatree flower bud raw materials into a fermentation box, fermenting for 5-10 days at the temperature of 35-45 ℃ and the humidity of 60-75%, spraying fermentation liquor with the weight percentage of 0.5-4% for fermentation every day while fermenting, drying and crushing to obtain fermented pagodatree flower bud rice flour, wherein the fermentation liquor consists of aspergillus niger and bifidobacterium, and the mass ratio of the aspergillus niger to the bifidobacterium is (5-7): (3-5);
(2) Adding complex enzyme into the fermented locust rice flour to carry out enzymolysis reaction, wherein the temperature of the enzymolysis reaction is 30-50 ℃, the pH is 6-7, and the reaction time is 1-3 h; wherein the adding ratio of the fermented locust rice flour to the complex enzyme is (80-90): (10-20); the compound enzyme is a mixture of cellulase, beta-glucosidase and pectase, and the mass ratio of the cellulase to the beta-glucosidase to the pectase is (1-1.5): (0.5 to 0.8): (0.8-1);
(3) Performing enzyme deactivation treatment on the fermented sophora japonica rice flour after the enzymolysis reaction to obtain an enzyme deactivation mixture;
(4) Mixing the enzyme-inactivating mixture with arginine, wherein the adding ratio of the enzyme-inactivating mixture to the arginine is (90-100): (9-15), and drying to obtain the finished product of the pagodatree flower bud powder.
2. The method for preparing anti-saccharification sophorae rice flour according to claim 1, wherein the method for preparing aspergillus niger comprises the following steps: screening Aspergillus niger strains from the pomelo orchard dead leaves, and inoculating the strains to a liquid culture medium for culture to obtain Aspergillus niger bacterial liquid;
the liquid culture medium comprises the following components: corn starch, bran, bean cake powder, bean dregs, ammonium sulfate, potassium dihydrogen phosphate and calcium chloride.
3. Anti-glycation sophorae rice flour prepared by the preparation method of claim 1 or 2.
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| CN113332354A (en) * | 2021-06-25 | 2021-09-03 | 江苏菌钥生命科技发展有限公司 | Anti-saccharification anti-aging fermented composition and preparation method thereof |
| CN113599422A (en) * | 2021-09-07 | 2021-11-05 | 泰州医药城国科化物生物医药科技有限公司 | Composition with anti-sugar effect and application thereof |
| CN114246208A (en) * | 2021-12-29 | 2022-03-29 | 烟台百事特丽特种肥料有限公司 | Method for preparing fruit and vegetable preservative by fermenting and extracting sophora flower buds |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113332354A (en) * | 2021-06-25 | 2021-09-03 | 江苏菌钥生命科技发展有限公司 | Anti-saccharification anti-aging fermented composition and preparation method thereof |
| CN113599422A (en) * | 2021-09-07 | 2021-11-05 | 泰州医药城国科化物生物医药科技有限公司 | Composition with anti-sugar effect and application thereof |
| CN114246208A (en) * | 2021-12-29 | 2022-03-29 | 烟台百事特丽特种肥料有限公司 | Method for preparing fruit and vegetable preservative by fermenting and extracting sophora flower buds |
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| Title |
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| 槐米固态发酵提高槲皮素含量的研究;徐萌萌;沈竞;徐春;王东溯;汪红;孙启玲;;时珍国医国药(第03期);第704页左栏第4段，第1.2.4节、第3节 * |
| 槲皮素抑制蛋白糖基化及DNA损伤;陆敏;李晓明;卢永翎;郑铁松;张丹;吕丽爽;;食品科学(第01期);摘要，第105页左栏第2段 * |
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