Ginger resistant starch as well as preparation method and application thereof in regulating intestinal flora

Abstract

According to the scheme, B-type seed crystal ginger resistant starch is introduced into a 3-type ginger resistant starch system, the endogenous B-type seed crystal is high in induction capacity and can serve as a crystal nucleus to accelerate the recrystallization rate, and the yield of the ginger resistant starch is increased. A-type ginger starch molecular chains are secondarily nucleated on the surfaces of B-type seed crystals during retrogradation, so that growth of a crystal layer structure is promoted, the thickness of a crystal layer is increased, formation and arrangement of a retrogradation starch double-helix structure can be promoted while the crystal nucleus effect is achieved, growth of starch crystal grains and formation of perfect crystals are promoted, the total amount of the crystal grains is increased, and the retrogradation rate is increased. The cell structure is more compact, so that the content of resistant starch is increased; according to the scheme, a cold and hot table precise temperature control technology is used for temperature control, so that the ordered structure of starch particles is more easily damaged, the dissolution of amylose is enhanced, and the retrogradation of unstable amylose and amylopectin in the subsequent cooling process is accelerated; the ginger resistant starch can be utilized by intestinal microorganisms.

Description

Technical Field

[0001] The present invention relates to the technical field of food processing, and particularly to ginger resistant starch, a preparation method thereof, and an application thereof in regulating the intestinal flora. Background Art

[0002] Ginger (Zingiber officinale Roscoe) is a perennial herb belonging to the Zingiberaceae family and the Zingiber genus, and is one of the Chinese medicinal materials that can be used both as medicine and food in China. The starch content in ginger accounts for 40-70% of the dry weight and is the main carbohydrate in the rhizome. Research shows that the natural resistant starch (type 2 resistant starch) in ginger starch accounts for more than 70%, and the amylose content accounts for 25-45% of the starch. It is easy to age and retrograde during the processing to generate type 3 resistant starch.

[0003] Resistant starch is a kind of dietary fiber with prebiotic effects, which cannot be directly digested and absorbed in the small intestine, can be fermented and utilized in the large intestine, and affects human health by regulating the composition of the microbiome through the interaction with the intestinal microbiome. Type 3 resistant starch is mainly formed during food production or processing, and has good structural and functional properties. The type 3 ginger resistant starch of the present invention uses a precise temperature control technology with a hot and cold stage, so that the crystallinity of ginger starch is improved during the formation of resistant starch, and at the same time the RS content increases. The invention patent "A Resistant Starch and a Preparation Method Thereof" (publication number CN116355976A) uses three physical field treatments such as extrusion treatment, ultrasonic treatment, and microwave treatment to prepare resistant starch, and obtains resistant starch with a relatively high degree of order and a relatively dense surface, but does not directly prepare resistant starch by heat temperature treatment. In addition, the digestion, absorption, metabolism and entry into the blood of resistant starch in the human body are relatively slow, and it can improve diseases such as diabetes and hypertension by reducing the reactions of blood sugar and insulin and reducing related blood lipid indexes such as cholesterol in the serum. The type 3 ginger resistant starch in the present invention has prebiotic effects, can mediate bile acid metabolism by regulating the intestinal flora in the large intestine, and the change of the bile acid pool in turn regulates the composition of the intestinal flora, so as to achieve the dual regulation of bile acid and intestinal flora. The invention patent "A Heat-resistant Resistant Starch and a Preparation Method Thereof" (publication number CN116693705A) prepares small particle heat-resistant high-crystalline starch, making it have higher thermal stability, relative crystallinity and RS content, but does not study the prebiotic effects of resistant starch; in the invention patent "A Highly Prebiotic Lotus Seed Resistant Starch Prepared by Enzymolysis Synergistic Pressure Heat Treatment, a Preparation Method Thereof and an Application Thereof" (publication number CN117106833A), the yield of lotus seed resistant starch is increased, and the prepared lotus seed resistant starch has significant in vitro prebiotic effects, while this invention does not verify the in vivo prebiotic effects through animal experiments.

[0004] The present invention uses a precise temperature control technology of a hot and cold stage to promote the gelatinization degree of ginger resistant starch at a continuous and stable temperature; endogenous B-type crystal seeds are introduced during starch retrogradation, and the retrogradation temperature is precisely regulated. The endogenous B-type crystal seeds promote the growth of ginger starch platelets, making the crystal cell structure denser, promoting the formation and arrangement of the double helix structure of retrograded starch, promoting the growth of starch grains and the formation of perfect crystals, thereby promoting the growth of the crystalline structure, further increasing the content of resistant starch. The stable retrogradation temperature further induces starch recrystallization. While the crystallization degree of the obtained ginger resistant starch is increased, it can have the effect of regulating the intestinal flora. Summary of the Invention

[0005] In view of this, the purpose of the present invention is to provide a ginger resistant starch that is convenient to prepare, reliable and stable in efficacy application, and its preparation method and application in regulating the intestinal flora.

[0006] In order to achieve the above technical objectives, the technical solutions adopted by the present invention are as follows:

[0007] A processing method of ginger resistant starch, which includes:

[0008] S01. Using ginger as a raw material, preprocessing it to make ginger starch;

[0009] S02. Mixing ginger starch with water, then performing gelatinization and adding an acid solution for hydrolysis treatment, and then washing and drying the hydrolysis product to obtain B-type crystal seed resistant starch;

[0010] S03. Mixing ginger starch with water, then performing gelatinization treatment, and then adding B-type crystal seed resistant starch to mix with it. After cooling and retrogradation treatment, ginger retrograded starch is prepared;

[0011] S04. Performing drying treatment on the ginger retrograded starch and making it into a starch homogenate, then adjusting its pH to a preset value, and then performing enzymatic hydrolysis treatment on it according to preset conditions. After centrifugation, drying, pulverization and sieving of the enzymatic hydrolysis product, ginger resistant starch is obtained.

[0012] As a possible implementation manner, further, in S01 of this solution, the preprocessing of ginger to make ginger starch includes:

[0013] Using fresh ginger as a raw material, peeling and slicing it, adding 2 times the mass of water to it, then performing crushing treatment, and then sieving the crushed material through a 100-mesh sieve. After standing the filtrate under the sieve for 12 h, the precipitate obtained by standing is taken, and the precipitate is washed with distilled water, and then washed with 95% ethanol until the washing liquid is transparent to remove impurities in the precipitate. Then, the washed precipitate is placed in an oven at 45 °C and dried until the moisture content is less than 8.00% to obtain ginger starch;

[0014] Among them, the impurities include one or more of lipids, proteins, and pigments.

[0015] As a possible implementation, further, the present solution S02 includes:

[0016] Mix ginger starch and deionized water in a ratio of 3:7 (w / v) and stir well to obtain a uniformly dispersed starch milk. Then place it in a water bath at 100 °C for gelatinization for 30 min, then transfer it to a storage at 4 °C for 24 h, then take it out and add a citric acid aqueous solution with an acid concentration of 3.6% for hydrolysis. The hydrolysis temperature is 40 °C and the hydrolysis time is 12 h. Finally, wash the hydrolysis product with deionized water until the pH of the washing solution is 6.5, and then dry the washed product at 105 °C to obtain B-type seed resistant starch.

[0017] As a possible implementation, further, the present solution S03 includes:

[0018] Mix ginger starch and water in a ratio of 3:7 (w / v) and stir evenly at room temperature, then heat it up to the gelatinization temperature of 95 - 105 °C at a heating rate of 0.5 - 1.5 °C / min for gelatinization for 10 - 15 min;

[0019] Cool the gelatinized ginger starch at a cooling rate of 0.5 - 1.5 °C / min to 45 °C, then add B-type seed resistant starch in a ratio of 5 - 10% (w / v) based on the dry mass of the starch and mix. Then cool the mixture to 2 - 6 °C at a cooling rate of 0.5 - 1.5 °C / min and retrograde for 20 - 28 h to obtain ginger retrograded starch.

[0020] As a better implementation option, preferably, in the present solution S03, the temperature of the material is raised and lowered through a hot and cold stage device.

[0021] As a better implementation option, preferably, the present solution S04 includes:

[0022] After drying the ginger retrograded starch at 45 °C for 12 h, then make it into a starch homogenate with a mass concentration of 6%, then add citric acid - disodium hydrogen phosphate to adjust its pH to 6.0, and then add α-amylase and amyloglucosidase in sequence according to preset conditions for enzymatic hydrolysis. After the enzymatic hydrolysis product is centrifuged, dried at 45 °C, pulverized, and sieved, ginger resistant starch is obtained.

[0023] As a better implementation option, preferably, in the present solution S04, the enzymatic hydrolysis treatment includes:

[0024] Add α - amylase to the starch homogenate at 500 U / g, enzymatically hydrolyze it at 45 °C for 2 h, then adjust its pH to 4.5 with citric acid, and then add amyloglucosidase at 5000 U / L and carry out enzymatic hydrolysis at 60 °C for 1 h;

[0025] Among them, after the enzymatic hydrolysis product is centrifuged, dried at 45 °C, pulverized and sieved through a 100 - mesh sieve, ginger resistant starch is obtained.

[0026] Based on the above, this solution also proposes a kind of ginger resistant starch, which is prepared by the above - mentioned processing method.

[0027] Based on the above, this solution also proposes the application of the above - mentioned ginger resistant starch in the preparation of products for regulating intestinal flora - mediated bile acid metabolism.

[0028] Based on the above, this solution also proposes the application of the above - mentioned ginger resistant starch in the preparation of products for regulating intestinal flora.

[0029] Adopting the above - mentioned technical solution, compared with the prior art, the beneficial effects of the present invention are as follows:

[0030] (1) This solution introduces the pre - prepared B - type seeded ginger resistant starch into the type 3 ginger resistant starch system. Based on the strong induction ability of the endogenous B - type seed, it can be used as a crystal nucleus to accelerate the recrystallization rate, enabling the A - type ginger starch molecular chains to nucleate again on the surface of the B - type seed during retrogradation, thereby promoting the growth of the crystal layer structure and increasing the thickness of the crystal layer. While playing the role of the crystal nucleus, it can promote the formation and arrangement of the double - helix structure of retrograded starch, and promote the growth of starch grains and the formation of perfect crystals, increasing the total amount of grains, making the unit cell structure more dense, and thus increasing the content of resistant starch.

[0031] (2) This solution uses a hot - cold stage device for auxiliary processing and realizes precise control of the reaction temperature during the preparation of ginger resistant starch through its precise temperature control technology. Among them, both the temperature and time of heat treatment will affect the degree of damage to the starch granule structure and the gelatinization degree of starch. During the gelatinization process, the starch granules absorb water, swell and finally rupture, releasing linear and branched starch molecules to form a viscous paste. The continuous stability of temperature is crucial for the gelatinization process. Too high a temperature may lead to starch degradation, and too low a temperature may result in incomplete gelatinization. Precise temperature control is more likely to destroy the ordered structure of starch granules, enhance the dissolution of linear starch, and thus accelerate the retrogradation of unstable linear and branched starch during the subsequent cooling process. On the basis of introducing endogenous B - type seeds during retrogradation, a stable retrogradation temperature can further induce the growth of crystal structures. This processing method has a constant heating and cooling rate, which can further promote the increase of crystallinity and the content of resistant starch.

[0032] (3) The ginger resistant starch prepared by this solution can be utilized by intestinal microorganisms, improve the diversity of intestinal flora in the large intestine of obese rats, promote intestinal bile acid metabolism, and exhibit good regulation of intestinal flora. Through experimental demonstration, ginger resistant starch significantly promotes the proliferation of beneficial bacterial genera such as Allobaculum and Bifidobacterium in the large intestine of obese rats, reduces the abundance of potentially harmful bacterial genera such as Helicobacter, Corynebacterium, and Bacillus in the large intestine of rats, secretes bile salt hydrolase, and through the deconjugation pathway, promotes the conversion of conjugated bile acids (such as taurocholic acid TCA and tauro-β-muricholic acid T-β-MCA) in the large intestine to free primary bile acids (such as β-muricholic acid β-MCA and α-muricholic acid α-MCA). The changes in the composition and content of the bile acid pool in turn regulate the intestinal flora composition of obese rats, thereby achieving a two-way regulatory effect between bile acids and intestinal flora. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required for use in the description of the embodiments or the prior art. Obviously, the drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0034] Figure 1 is the swelling degree of ginger starch paste under different treatment conditions;

[0035] Figure 2 is the RS content of ginger resistant starch under different treatment conditions;

[0036] Figure 3 is the RS content of ginger resistant starch in Examples 1-3 and the control group;

[0037] Figure 4 Effect of ginger resistant starch on the content of primary bile acids in the large intestine of obese rats;

[0038] Figure 5 Heatmap of the intestinal flora community in the large intestine of obese rats treated with ginger resistant starch. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0039] The present invention will be further described in detail below with reference to the drawings and embodiments. It should be particularly noted that the following embodiments are only used to illustrate the present invention, but do not limit the scope of the present invention. Similarly, the following embodiments are only some embodiments of the present invention rather than all embodiments. All other embodiments obtained by those of ordinary skill in the art without creative efforts fall within the scope of protection of the present invention.

[0040] Example 1

[0041] A processing method of ginger resistant starch in this example includes:

[0042] (1) Preparation of ginger starch:

[0043] Using fresh ginger as raw material, after peeling and slicing, add 2 times the mass of water, crush with a juicer, pass through a 100-mesh sieve, take the filtrate and let it stand for precipitation for 12 h, and then take the precipitate. Wash the precipitate three times with distilled water and then with 95% ethanol to remove impurities such as lipids, proteins, and pigments until the ethanol extract is colorless and transparent. Then place it in an oven at 45 °C and dry until the moisture content is less than 8.00% to obtain ginger starch;

[0044] (2) Preparation of B-type seed resistant starch:

[0045] Add deionized water according to the ratio of ginger starch to water of 3:7 (w / v) and stir well. Gelatinize the uniformly dispersed starch milk in a water bath at 100 °C for 30 min, then transfer it to a storage condition at 4 °C for 24 h. Add the starch to an aqueous solution of citric acid with an acid concentration of 3.6% for hydrolysis. The hydrolysis temperature is 40 °C and the hydrolysis time is 12 h. Wash the hydrolysis product repeatedly with deionized water until the pH is 6.5, and dry it at 105 °C to obtain B-type seed resistant starch;

[0046] (3) Precision gelatinization technology on a hot and cold stage:

[0047] Accurately weigh ginger starch and add water to a beaker. The ratio of starch to water is 3:7. Stir evenly at room temperature and then place it on a hot and cold stage for heating and gelatinization. The heating rate is 0.5 °C / min, the gelatinization temperature is 95 °C, and the gelatinization time is 10 min;

[0048] (4) Introduce B-type seed resistant starch after cooling:

[0049] After gelatinization of ginger starch, set the temperature of the hot and cold stage to decrease to 45 °C, the cooling rate is 0.5 °C / min, and add B-type seed resistant starch with a mass of 5% (w / w) of the starch dry basis;

[0050] (5) Precision retrogradation technology on a hot and cold stage

[0051] Precisely cool the temperature of the hot and cold stage to 2 °C, the cooling rate is 0.5 °C / min, and retrograde for 20 h to obtain ginger retrograded starch;

[0052] (6) Drying, enzymatic hydrolysis, and purification of retrograded starch

[0053] After drying ginger reactivated starch at 45 °C for 12 h, a 6% starch slurry was prepared, the pH was adjusted to 6.0 with citric acid - disodium hydrogen phosphate, α - amylase was added at 500 U / g, and enzymatic hydrolysis was carried out at 45 °C for 2 h; then the pH was adjusted to 4.5 with citric acid, and amyloglucosidase was added at 5000 U / L, and enzymatic hydrolysis was carried out at 60 °C for 1 h. After enzymatic hydrolysis, the starch precipitate was centrifuged, dried at 45 °C, pulverized, and passed through a 100 - mesh sieve to obtain ginger resistant starch.

[0054] Example 2

[0055] A processing method of ginger resistant starch in this example includes:

[0056] (1) Preparation of ginger starch:

[0057] Using fresh ginger as raw material, after peeling and slicing, add 2 times the mass of water, crush with a juicer, pass through a 100 - mesh sieve, take the filtrate and let it stand for precipitation for 12 h, then take the precipitate. Wash the precipitate three times with distilled water and then with 95% ethanol to remove impurities such as lipids, proteins, and pigments until the ethanol extract is colorless and transparent. Then place it in an oven at 45 °C and dry until the moisture content is less than 8.00% to obtain ginger starch;

[0058] (2) Preparation of B - type seed resistant starch:

[0059] Add deionized water according to the ratio of ginger starch to water of 3:7 (w / v) and stir well. Gelatinize the uniformly dispersed starch milk in a water bath at 100 °C for 30 min, then transfer it to a 4 °C storage for 24 h. Hydrolyze the starch in an aqueous solution of citric acid with an acid concentration of 3.6%. The hydrolysis temperature is 40 °C and the hydrolysis time is 12 h. Wash the hydrolysis product repeatedly with deionized water until the pH is 6.5, and dry it at 105 °C to obtain B - type seed resistant starch;

[0060] (3) Precise gelatinization technology of hot - cold stage:

[0061] Accurately weigh ginger starch into a beaker and add water. The ratio of starch to water is 3:7. Stir evenly at room temperature and then place it on the hot - cold stage for heating and gelatinization. The heating rate is 1 °C / min, the gelatinization temperature is 100 °C, and the gelatinization time is 12.5 min;

[0062] (4) Introduce B - type seed resistant starch after cooling:

[0063] After gelatinization of ginger starch, set the temperature of the hot - cold stage to decrease to 45 °C, the cooling rate is 1 °C / min, and add B - type seed resistant starch accounting for 7.5% (w / w) of the dry starch mass;

[0064] (5) Precise retrogradation technology of hot - cold stage

[0065] The temperature of the hot and cold stage is accurately cooled to 4 °C, the cooling rate is 1 °C / min, and it is retrograded for 24 h to obtain ginger retrograded starch;

[0066] (6) Drying, enzymolysis and purification of retrograded starch

[0067] After the ginger retrograded starch is dried at 45 °C for 12 h, a 6% starch slurry is prepared, the pH is adjusted to 6.0 with citric acid - disodium hydrogen phosphate, α - amylase is added at 500 U / g, and enzymolysis is carried out at 45 °C for 2 h; then the pH is adjusted to 4.5 with citric acid, and amyloglucosidase is added at 5000 U / L, and enzymolysis is carried out at 60 °C for 1 h. After enzymolysis, the starch precipitate is centrifuged, dried at 45 °C, pulverized, and then passed through a 100 - mesh sieve to obtain ginger resistant starch.

[0068] Example 3

[0069] A processing method of ginger resistant starch in this example includes:

[0070] (1) Preparation of ginger starch:

[0071] Using fresh ginger as raw material, after peeling and slicing, add 2 times the mass of water, crush with a juicer, pass through a 100 - mesh sieve, let the filtrate stand and precipitate for 12 h, and then take the precipitate. Wash the precipitate three times with distilled water and then with 95% ethanol to remove impurities such as lipids, proteins, and pigments until the ethanol extract is colorless and transparent. Then place it in an oven at 45 °C and dry until the moisture content is less than 8.00% to obtain ginger starch;

[0072] (2) Preparation of B - type seed resistant starch:

[0073] Add deionized water according to the ratio of ginger starch to water of 3:7 (w / v) and stir well. Gelatinize the uniformly dispersed starch milk in a 100 °C water bath for 30 min, then transfer it to be stored at 4 °C for 24 h. Add the starch to an aqueous solution of citric acid with an acid concentration of 3.6% for hydrolysis, the hydrolysis temperature is 40 °C, and the hydrolysis time is 12 h. Wash the hydrolysis product repeatedly with deionized water until the pH is 6.5, and dry it at 105 °C to obtain B - type seed resistant starch;

[0074] (3) Precision gelatinization technology of hot and cold stage:

[0075] Accurately weigh ginger starch into a beaker and add water. The ratio of starch to water is 3:7. After stirring evenly at room temperature, place it on the hot and cold stage for heating and gelatinization. The heating rate is 1.5 °C / min, the gelatinization temperature is 105 °C, and the gelatinization time is 15 min;

[0076] (4) Introducing B - type seed resistant starch after cooling

[0077] After the ginger starch is gelatinized, the temperature of the hot stage is set to decrease to 45 °C at a cooling rate of 1.5 °C / min, and B-type crystalline resistant starch accounting for 10% (w / w) of the dry starch mass is added.

[0078] (5) Precision retrogradation technology of the hot stage

[0079] The temperature of the hot stage is precisely decreased to 6 °C at a cooling rate of 1.5 °C / min, and ginger retrograded starch is obtained after retrogradation for 28 h.

[0080] (6) Drying, enzymatic hydrolysis and purification of the retrograded starch

[0081] After the ginger retrograded starch is dried at 45 °C for 12 h, a 6% starch slurry is prepared, the pH is adjusted to 6.0 with citric acid-sodium hydrogen phosphate, α-amylase is added at 500 U / g, and enzymatic hydrolysis is carried out at 45 °C for 2 h; then the pH is adjusted to 4.5 with citric acid, amyloglucosidase is added at 5000 U / L, and enzymatic hydrolysis is carried out at 60 °C for 1 h. After enzymatic hydrolysis, the starch precipitate is centrifuged, dried at 45 °C, pulverized, and passed through a 100-mesh sieve to obtain ginger resistant starch.

[0082] Control Example 1

[0083] The processing method of a type 3 ginger resistant starch in this example includes:

[0084] (1) Preparation of ginger starch:

[0085] Using fresh ginger as raw material, after peeling and slicing, add 2 times the mass of water, crush with a juicer, pass through a 100-mesh sieve, take the filtrate and let it stand for precipitation for 12 h, and then take the precipitate. Wash the precipitate three times with distilled water and then with 95% ethanol to remove impurities such as lipids, proteins, and pigments until the ethanol extract is colorless and transparent. Then place it in an oven at 45 °C and dry until the moisture content is less than 8.00% to obtain ginger starch.

[0086] (2) Gelatinization and retrogradation treatment

[0087] Accurately weigh ginger starch and add water to a beaker. The ratio of starch to water is 3:7. Stir evenly at room temperature and then gelatinize in a 100 °C water bath for 15 min. After cooling to room temperature, store it in a 4 °C refrigerator for 24 h to obtain ginger retrograded starch.

[0088] (3) After drying the ginger retrograded starch at 45 °C for 12 h, a 6% starch slurry was prepared, the pH was adjusted to 6.0 with citric acid - disodium hydrogen phosphate, α - amylase was added at 500 U / g, and enzymatic hydrolysis was carried out at 45 °C for 2 h; then the pH was adjusted to 4.5 with citric acid, and glucoamylase was added at 5000 U / L, and enzymatic hydrolysis was carried out at 60 °C for 1 h. After enzymatic hydrolysis, the starch precipitate was centrifuged, dried at 45 °C, pulverized, and passed through a 100 - mesh sieve to obtain ginger resistant starch.

[0089] Control Example 2

[0090] The processing method of a type 3 ginger resistant starch in this example includes:

[0091] (1) Preparation of ginger starch:

[0092] Using fresh ginger as raw material, after peeling and slicing, 2 times the mass of water was added, crushed with a juicer, passed through a 100 - mesh sieve, the filtrate was allowed to stand and precipitate for 12 h, and then the precipitate was taken. The precipitate was washed three times with distilled water and then with 95% ethanol to remove impurities such as lipids, proteins, and pigments until the ethanol extract was colorless and transparent. Then it was placed in an oven at 45 °C and dried until the moisture content was less than 8.00% to obtain ginger starch;

[0093] (2) Gelatinization and retrogradation treatment

[0094] Accurately weigh ginger starch into a beaker and add water. The ratio of starch to water is 3:7. After stirring evenly at room temperature, it was placed on a hot - cold stage for heating gelatinization. The heating rate was 1.5 °C / min, the gelatinization temperature was 100 °C, and the gelatinization time was 15 min;

[0095] After gelatinization of ginger starch, the temperature of the hot - cold stage was set to decrease to room temperature, the cooling rate was 1.5 °C / min, and finally it was stored in a 4 °C refrigerator for 24 h to obtain ginger retrograded starch;

[0096] (3) After drying the ginger retrograded starch at 45 °C for 12 h, a 6% starch slurry was prepared, the pH was adjusted to 6.0 with citric acid - disodium hydrogen phosphate, α - amylase was added at 500 U / g, and enzymatic hydrolysis was carried out at 45 °C for 2 h; then the pH was adjusted to 4.5 with citric acid, and glucoamylase was added at 5000 U / L, and enzymatic hydrolysis was carried out at 60 °C for 1 h. After enzymatic hydrolysis, the starch precipitate was centrifuged, dried at 45 °C, pulverized, and passed through a 100 - mesh sieve to obtain ginger resistant starch.

[0097] Control Example 3

[0098] The processing method of a type 3 ginger resistant starch in this example includes:

[0099] (1) Preparation of ginger starch:

[0100] Using fresh ginger as raw material, after peeling and slicing, add water with a mass twice that of the ginger, crush it with a juicer, sieve through a 100-mesh sieve, take the filtrate and let it stand for precipitation for 12 h, and then take the precipitate. Wash the precipitate three times with distilled water and then with 95% ethanol to remove impurities such as lipids, proteins, and pigments until the ethanol extract is colorless and transparent. Then place it in an oven at 45 °C and dry it until the moisture content is less than 8.00% to obtain ginger starch;

[0101] (2) Add deionized water according to the ratio of ginger starch to water of 3:7 (w / v) and stir well. Gelatinize the uniformly dispersed starch milk in a water bath at 100 °C for 30 min, then transfer it to a storage condition at 4 °C for 24 h. Add the starch to an aqueous solution of citric acid with an acid concentration of 3.6% for hydrolysis. The hydrolysis temperature is 40 °C and the hydrolysis time is 12 h. Wash the hydrolysis product repeatedly with deionized water until the pH is 6.5, and dry it at 105 °C to obtain B-type crystalline resistant starch;

[0102] (3) Gelatinization and retrogradation treatment

[0103] Accurately weigh ginger starch into a beaker and add water. The ratio of starch to water is 3:7. Stir evenly at room temperature and then gelatinize it in a water bath at 100 °C for 15 min. Then cool it. When the temperature drops to 45 °C, add B-type crystalline resistant starch (prepared in step (2)) with a mass of 5% (w / w) of the dry starch basis; finally, when it reaches room temperature, place it in a refrigerator at 4 °C for storage for 24 h to obtain ginger retrograded starch;

[0104] (4) After drying the ginger retrograded starch at 45 °C for 12 h, make a 6% starch slurry, adjust the pH to 6.0 with citric acid-sodium hydrogen phosphate, add α-amylase at 500 U / g, and enzymatically hydrolyze it at 45 °C for 2 h; then adjust the pH to 4.5 with citric acid, add amyloglucosidase at 5000 U / L, and enzymatically hydrolyze it at 60 °C for 1 h. After enzymatic hydrolysis, the starch precipitate is centrifuged, dried at 45 °C, pulverized, and sieved through a 100-mesh sieve to obtain ginger resistant starch.

[0105] Control Example 4

[0106] A processing method of type 3 ginger resistant starch in this example includes:

[0107] (1) Preparation of ginger starch:

[0108] Using fresh ginger as raw material, after peeling and slicing, add water with a mass twice that of the ginger, crush it with a juicer, sieve through a 100-mesh sieve, take the filtrate and let it stand for precipitation for 12 h, and then take the precipitate. Wash the precipitate three times with distilled water and then with 95% ethanol to remove impurities such as lipids, proteins, and pigments until the ethanol extract is colorless and transparent. Then place it in an oven at 45 °C and dry it until the moisture content is less than 8.00% to obtain ginger starch;

[0109] (2) Add deionized water according to the ratio of ginger starch to water of 3:7 (w / v) and stir well. Gelatinize the uniformly dispersed starch milk in a water bath at 100 °C for 30 min, then transfer it to a storage at 4 °C for 24 h. Add the starch to an aqueous solution of citric acid with an acid concentration of 3.6% for hydrolysis. The hydrolysis temperature is 40 °C and the hydrolysis time is 12 h. Wash the hydrolysis product repeatedly with deionized water until the pH is 6.5, and dry it at 105 °C to obtain B-type seed resistant starch;

[0110] (3) Gelatinization and retrogradation treatment

[0111] Accurately weigh ginger starch into a beaker and add water. The ratio of starch to water is 3:7. Stir evenly at room temperature and then place it on a hot and cold stage for heating and gelatinization. The heating rate is 1.5 °C / min, the gelatinization temperature is 105 °C, and the gelatinization time is 15 min;

[0112] After the ginger starch is gelatinized, set the temperature of the hot and cold stage to decrease to 45 °C, and the cooling rate is 1.5 °C / min. Add B-type seed resistant starch (prepared in step (2)) accounting for 5% (w / w) of the dry basis mass of the starch;

[0113] Precisely cool the temperature of the hot and cold stage to room temperature, and the cooling rate is 1.5 °C / min. Finally, store it in a 4 °C refrigerator for 24 h to obtain ginger retrograded starch;

[0114] (4) After drying the ginger retrograded starch at 45 °C for 12 h, make it into a 6% starch slurry, adjust the pH to 6.0 with citric acid - disodium hydrogen phosphate, add α-amylase at 500 U / g, and enzymatically hydrolyze it at 45 °C for 2 h; then adjust the pH to 4.5 with citric acid, add amyloglucosidase at 5000 U / L, and enzymatically hydrolyze it at 60 °C for 1 h. After enzymatic hydrolysis, the starch precipitate is centrifuged, dried at 45 °C, pulverized, and then passed through a 100-mesh sieve to obtain ginger resistant starch.

[0115] Comparative test

[0116] Define the ginger resistant starch prepared in Comparative Examples 1 - 4 as Sample A, Sample B, Sample C, and Sample D in sequence; conduct crystallinity measurement, swelling degree measurement, and resistant starch (RS) content measurement on the obtained ginger resistant starch, and the results are shown in Table 1 and Figure 1 、 Figure 2 。

[0117] Table 1

[0118]

[0119] Note: In the table, lowercase letters in the same column represent significant differences in data of different samples, p < 0.05

[0120] Among them, it can be seen from the data of each sample in Table 1 that, compared with sample A, the proportion of microcrystalline regions, the proportion of subcrystalline regions, and the crystallinity of samples B-D increase, while the proportion of amorphous regions decreases. From Figure 1 the data of each sample, it can be seen that, compared with the gelatinized starch in the preparation process of sample A, the swelling degree of the gelatinized starch in the process of making sample B increases, indicating that the precise gelatinization technology of the hot and cold stage can promote the degree of starch gelatinization. From Figure 2 the data of each sample, it can be seen that, compared with sample A, the RS content of samples B-D increases, and the resistant starch content of sample D is the highest.

[0121] The ginger resistant starch prepared in Comparative Example 1 (defined as sample A) was subjected to crystallinity measurement, swelling degree measurement, and resistant starch (RS) content measurement with the ginger resistant starch prepared in Examples 1-3. The results are shown in Table 2 and Figure 3 .

[0122] Table 2

[0123]

[0124] Note: In the table, lowercase letters in the same column represent significant differences in the data of different examples, p < 0.05

[0125] Combined with Table 2 and Figure 3 the data of each sample, it can be seen that, compared with sample A, the proportion of microcrystalline regions, the proportion of subcrystalline regions, and the crystallinity of Examples 1-3 increase, the proportion of amorphous regions decreases, and at the same time, the RS content increases significantly. In order to verify the effects of ginger resistant starch with the function of regulating intestinal flora, the following tests were carried out.

[0126] Study on the function of regulating intestinal flora

[0127] The grouping and administration methods of animals are as follows: 84 SPF-grade male SD rats were adaptively fed for 1 week, during which they had free access to water and food. They were randomly divided into 2 groups according to body weight. One group of 12 was given a basal diet, and one group of 72 was given a high-fat diet (high-fat diet formula: 89.25% of ordinary feed, 5.00% of egg yolk powder, 5.00% of lard, 0.50% of cholesterol, 0.25% of pig bile salt. The ordinary feed meets the nutritional components of the experimental animal compound feed in GB 14924.3-2010. Purchased from Fuzhou Wu's Animal Experiment Co., Ltd.). After 2 weeks of feeding, the 3 rats with the highest and lowest body weights in the basal diet group were respectively eliminated, and the remaining 6 were the normal control group (Normal control: NC), and they continued to be fed with the ordinary feed for 4 weeks, and were intragastrically administered 2 mL / 100 g·bw of normal saline every day. After 2 weeks of high-fat diet feeding, 18 obese-sensitive rats with higher body weights and 18 obese-resistant rats with lower body weights were eliminated, and the remaining 36 rats were randomly divided into 6 groups, with 6 rats in each group.

[0128] Hyperlipidemic model group (HM): Fed with a high-fat diet for 4 weeks, and intragastrically administered with 2 mL / 100 g·bw of normal saline daily.

[0129] Positive control group (PC): Continued to be fed with a high-fat diet for 4 weeks, and simultaneously given 4 mg / kg of simvastatin (simvastatin 10 mg / tablet, dissolved them in 50 mL of distilled water to obtain a 0.2 mg / mL suspension), and the intragastric administration dose was 2.0 mL / 100 g·bw, once a day, for a total of 4 weeks.

[0130] Using the product ginger resistant starch prepared in Example 3 as the RS3 type ginger resistant starch group (GRS3): Continued to be fed with a high-fat diet for 4 weeks, and simultaneously intragastrically administered with a GRS3 resistant starch suspension (0.125 g / mL), and the intragastric administration dose was 0.25 g / 100 g·bw, once a day, for a total of 4 weeks.

[0131] High amylose maize starch group (HMS): Continued to be fed with a high-fat diet for 4 weeks, and simultaneously intragastrically administered with a high amylose maize starch suspension (0.125 g / mL), and the intragastric administration dose was 0.25 g / 100 g·bw, once a day, for a total of 4 weeks; among them, the high amylose maize starch suspension was purchased from a commercially available product, and its source was Yingxin Laboratory, product number: YX0688.

[0132] As Figure 4 shown, compared with the obesity model group, type 3 ginger resistant starch significantly promoted the proliferation of beneficial bacterial genera such as Allobaculum and Bifidobacterium in the large intestine of obese rats, and reduced the abundances of potentially harmful bacterial genera such as Helicobacter, Corynebacterium, and Bacillus in the large intestine of rats.

[0133] As Figure 5 shown, compared with the obesity model group, type 3 ginger resistant starch could significantly reduce the contents of taurocholic acid (TCA) and tauro-β-muricholic acid (T-β-MCA) in the large intestine of obese rats, and could increase the contents of β-muricholic acid (β-MCA) and α-muricholic acid (α-MCA) in the large intestine of rats. The type 3 ginger resistant starch group promoted the conversion of conjugated bile acids taurocholic acid (TCA) and tauro-β-muricholic acid (T-β-MCA) in the large intestine into free primary bile acids β-muricholic acid (β-MCA) and α-muricholic acid (α-MCA). The changes in the composition and content of the bile acid pool in turn regulated the intestinal flora composition of obese rats, thus achieving a two-way regulatory effect between bile acids and intestinal flora.

[0134] The above are only some embodiments of the present invention, and thus do not limit the protection scope of the present invention. Any equivalent device or equivalent process transformation made by using the content of the specification and drawings of the present invention, or directly or indirectly applied in other related technical fields, shall similarly be included in the patent protection scope of the present invention.

Claims

1. A method for processing ginger resistant starch, characterized in that: It includes: S01, using ginger as raw material, pre-treating it to prepare ginger starch; S02, mixing ginger starch with water, then gelatinizing and adding acid solution for hydrolysis, and then washing and drying the hydrolyzate to obtain B-type seed resistant starch; S03, mixing ginger starch with water, and then gelatinizing it, and then adding B-type seed resistant starch to mix with it, and after cooling and retrogradation treatment, preparing ginger retrograded starch; S04, drying the retrograded ginger starch and preparing it into starch homogenate, adjusting its pH to a preset value, and then subjecting it to enzymolysis treatment under preset conditions, and obtaining ginger resistant starch after the enzymolysis product is centrifuged, dried, crushed and sieved.

2. The method for processing ginger resistant starch according to claim 1, characterized in that: In S01, pre-treating ginger to prepare ginger starch includes: Fresh ginger is used as a raw material, peeled and sliced, and then water twice its weight is added, and then the ginger is crushed, and then the crushed ginger is passed through a 100-mesh sieve, and the filtrate under the sieve is taken and allowed to stand for 12 hours, and then the precipitate obtained by standing is removed, and the precipitate is washed with distilled water, and then washed with 95% ethanol until the washing liquid becomes transparent to remove impurities in the precipitate, and then the washed precipitate is placed in a 45° C. oven and dried until the moisture content is less than 8.00%, so as to obtain ginger starch; Wherein, the impurities include one or more of lipids, proteins and pigments.

3. The method for processing ginger resistant starch according to claim 1, characterized in that: S02 includes: Ginger starch and deionized water are mixed in a ratio of 3:7 (w / v) and stirred thoroughly to obtain a uniformly dispersed starch emulsion, which is then placed in a 100°C water bath for gelatinization for 30 minutes, then transferred to a 4°C temperature environment for storage for 24 hours, then taken out and hydrolyzed by adding a 3.6% citric acid aqueous solution at a hydrolysis temperature of 40°C and a hydrolysis time of 12 hours. Finally, the hydrolyzate is washed with deionized water until the pH of the washing liquid is 6.5, and then the washing is dried at 105°C to obtain B-type seed resistant starch.

4. The method for processing ginger resistant starch according to claim 1, characterized in that: S03 includes: Ginger starch and water are mixed in a ratio of 3:7 (w / v) and stirred evenly at room temperature, and then heated to a gelatinization temperature of 95-105°C at a heating rate of 0.5-1.5°C / min for gelatinization treatment for 10-15 minutes; The gelatinized ginger starch is cooled to 45°C at a cooling rate of 0.5-1.5°C / min, and then B-type seed resistant starch is added in a ratio of 5-10% (w / v) of the dry basis mass of starch, and then the mixture is cooled to 2-6°C at a cooling rate of 0.5-1.5°C / min, and retrograded for 20-28h to obtain retrograded ginger starch.

5. The method for processing ginger resistant starch according to claim 4, characterized in that: In S03, the material is subjected to a heating and cooling process by means of a hot and cold stage device.

6. The method for processing ginger resistant starch according to any one of claims 1 to 5, characterized in that: S04 includes: The ginger retrograded starch was dried at 45°C for 12 hours, and then made into a 6% mass concentration starch homogenate. Then, citric acid-disodium hydrogen phosphate was added to adjust the pH to 6.

0. Then, α-amylase and amyloglucosidase were added in sequence according to preset conditions for enzymatic hydrolysis. The enzymatic hydrolysis product was centrifuged, dried at 45°C, crushed and sieved to obtain ginger resistant starch.

7. The method for processing ginger resistant starch according to claim 6, characterized in that: In S04, the enzymatic treatment includes: 500 U / g of α-amylase was added to the starch slurry, and the slurry was subjected to enzymatic hydrolysis at 45°C for 2 h. The pH was then adjusted to 4.5 with citric acid, and then 5000 U / L of amyloglucosidase was added, and the slurry was subjected to enzymatic hydrolysis at 60°C for 1 h. The enzymatic hydrolysis product was centrifuged, dried at 45°C, crushed and sieved through a 100-mesh sieve to obtain ginger resistant starch.

8. A ginger resistant starch, characterized in that: The invention is prepared by the processing method according to any one of claims 1 to 7.

9. Use of the ginger resistant starch as claimed in claim 8 in preparing a product for regulating intestinal flora-mediated bile acid metabolism.

10. Use of the ginger resistant starch according to claim 8 in preparing a product for regulating intestinal flora.

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