阅读说明：本技术 一种黄酮发酵物及其制备方法和在血糖调节中的应用 (Flavone fermentation product, preparation method thereof and application thereof in blood sugar regulation ) 是由 刘飞 王锐 陈勉 马文靖 袁丹丹 郑亚军 陈磊 李倩 张晓元 张艳艳 于 2020-07-03 设计创作，主要内容包括：本发明提供一种黄酮发酵物及其制备方法和在血糖调节中的应用,涉及医药技术领域。所述发酵方法包括：在灭菌豆皮中接种保加利亚乳杆菌和嗜热链球菌进行一阶静态发酵,然后接种马克思克鲁维酵母通气搅拌进行二阶发酵。本发明的方法原料价格低廉易得,工艺步骤少,获得的发酵物与大豆蛋白、胶原肽和可溶性膳食纤维以特定比例制备成组合物后具有较好的降空腹血糖功效。(The invention provides a flavone ferment, a preparation method thereof and application thereof in blood sugar regulation, relating to the technical field of medicines. The fermentation method comprises the following steps: inoculating lactobacillus bulgaricus and streptococcus thermophilus in the sterilized soybean hulls for first-order static fermentation, and then inoculating kluyveromyces marxianus for ventilation stirring for second-order fermentation. The method has the advantages of low price and easy obtainment of raw materials, few process steps, and better fasting blood sugar reducing effect after the obtained fermentation product, the soybean protein, the collagen peptide and the soluble dietary fiber are prepared into a composition according to a specific proportion.)

1. A method of fermenting a fermentation of flavones, the method comprising: inoculating lactobacillus bulgaricus and streptococcus thermophilus in a sterilized beancurd skin culture medium for first-order static fermentation, and then inoculating kluyveromyces marxianus for second-order fermentation by aeration stirring.

2. The fermentation method according to claim 1, wherein the testa Phaseoli is selected from one or more of semen Phaseoli, semen Phaseoli Radiati, semen Sojae Atricolor, semen Phaseoli vulgaris, semen Phaseoli Radiati and semen glycines.

3. The fermentation method according to claim 1, wherein the conditions for sterilizing the bean curd skin are: sterilizing at 60-130 deg.C for 10-40 min.

4. The fermentation process of claim 1, wherein the amount of lactobacillus bulgaricus and streptococcus thermophilus inoculated is 2-5%;

preferably, the ratio of Lactobacillus bulgaricus to Streptococcus thermophilus is 1-9: 3.

5. The fermentation process of claim 1, wherein the first-order static fermentation is carried out at a temperature of 38-44 ℃ for a fermentation time of 5-24 hours.

6. The fermentation method of claim 1, wherein kluyveromyces marxianus is inoculated when the pH of the first-order fermentation is reduced to 4-5;

preferably, the inoculation amount of the kluyveromyces marxianus is 1-3%;

preferably, the temperature of the second-order fermentation is 25-45 ℃, preferably 33-35 ℃, and the fermentation time is 48-96 h.

7. A fermentation product obtained by fermentation according to the fermentation method of any one of claims 1 to 6.

8. A composition comprising the fermentate of claim 7.

9. The composition of claim 8, further comprising soy protein, collagen peptide, and soluble dietary fiber;

Preferably, the mixing ratio of the fermentation product, the soy protein, the collagen peptide and the soluble dietary fiber is: 10-30 parts of fermentation product, 40-90 parts of soybean protein, 1-8 parts of collagen peptide and 0.5-15 parts of soluble dietary fiber;

preferably, the soluble dietary fiber is selected from one or more of polydextrose, resistant dextrin, pectin, konjac flour, fructo-oligosaccharide, inulin and xylo-oligosaccharide.

10. Use of the fermentation product of claim 7 or the composition of claim 8 or 9 for the preparation of a medicament, food or health product for the prevention and/or treatment of diabetes.

Technical Field

The invention relates to the technical field of medicines, in particular to a flavone ferment, a preparation method thereof and application thereof in blood sugar regulation.

Background

The information in this background section is only for enhancement of understanding of the general background of the invention and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art that is already known to a person of ordinary skill in the art.

The bean skin (hull) has poor taste and is not easy to digest, thereby affecting the processing and product quality of traditional bean products such as sweetened bean paste, bean curd, soybean oil and the like, and being generally treated as waste after food processing and polluting the environment. The research shows that the beans contain soybean isoflavone components (hereinafter referred to as flavone), wherein the content of the bean skin part is the highest. The flavonoid compounds are plant secondary metabolites with a 2-phenylchromone structure and widely exist in the plant world. Pharmacological activity research of the flavonoid compounds shows that the flavonoid compounds have physiological effects of resisting oxidation, preventing cardiovascular diseases, regulating immunity, resisting fatigue and the like. It usually combines with carbohydrates in plants to form flavonoid glycosides in the form of ligands, and exists in the form of flavonoid aglycones in a small part in the free state. However, the research finds that the aglycosylated flavonoid aglycone is beneficial to the absorption and utilization of the human body. Although plants such as soybean contain beta-glucosidase, the efficiency of obtaining flavonoid aglycone by hydrolysis using beta-glucosidase is only about 20%, and the conversion rate is low. Therefore, for converting flavonoid glycosides into flavonoid aglycones, the current major methods mainly include chemical methods and biological conversion methods. The chemical conversion methods comprise acid hydrolysis and alkali hydrolysis, which can pollute the environment, and the reaction conditions can change the structure and configuration of aglycone violently, which leads to the instability of the product or the disappearance of the drug effect. The biotransformation method is relatively mild and environment-friendly, but most of the strains reported at present are non-edible strains such as aspergillus niger and the like, and are not suitable for food processing. In addition, there are reports of enzymatic conversion, which is suitable for the extraction of flavonoids directly contacted with enzymes, and is not suitable for the treatment of flavonoid glycosides in soybean skins.

Disclosure of Invention

In order to overcome the defects of the prior art, the invention provides a flavone ferment, a preparation method thereof and application thereof in blood sugar regulation, the invention takes soybean hulls which are low in cost and easy to obtain as raw materials, and adopts a secondary inoculation and secondary fermentation mode to ferment the soybean hulls, the fermentation method is simple, easy and easy to control, the fermentation process can fully convert flavone components contained in the soybean hulls from a glucoside form to an aglycone form, the conversion rate is up to more than 90 percent, and soybean hull fibers are continuously acidified, swollen and dispersed in the fermentation process, so that the soybean hull fibers are converted into softened insoluble dietary fibers with high water holding capacity from a compact structure. In addition, the fermented soybean hull can effectively reduce fasting blood sugar and postprandial blood sugar, and when the fermented soybean hull is mixed with components such as soybean protein, collagen peptide and soluble dietary fiber for use, life habit intervention is combined, and the balance of blood sugar can be controlled.

Specifically, the technical scheme of the invention is as follows:

in a first aspect of the present invention, there is provided a method of fermentation of a fermentation product of flavones, the method comprising: lactobacillus delbrueckii subspecies bulgaricus (Lactobacillus delbrueckii subsp. Bulgaricus) and Streptococcus thermophilus (Streptococcus thermophilus) were inoculated into the sterilized soybean hull medium to perform first-order static fermentation, and then Kluyveromyces marxianus was inoculated with aeration stirring to perform second-order fermentation.

In an embodiment of the present invention, the bean skin is selected from whole bean with skin or bean skin of one or more of red bean, mung bean, black bean, flower bean, green bean and soybean, preferably red bean. The soybean hulls are preferably used after being crushed, and the soybean hulls can be sieved by a 100-mesh sieve after being crushed.

The composition of the beancurd skin culture medium is as follows: 7-20% of soybean hull, 0.5-5% of lactose, 0.05-0.5% of sodium citrate, 0.02-0.5% of sodium acetate, 0.5-9% of skimmed milk powder and water, wherein the% is in percentage by weight.

In an embodiment of the present invention, the sterilization conditions of the testa sojae medium are: sterilizing at 60-130 deg.C for 10-40min, preferably 60-70 deg.C for 30 min; 20min at 70-80 ℃; 80-90 deg.C for 15 min; 90-95 deg.C for 10min, preferably 80-90 deg.C for 15 min.

In an embodiment of the invention, the Lactobacillus delbrueckii subsp bulgaricus and Streptococcus thermophilus are inoculated in an amount of 2-5%, preferably 2%.

Wherein the mass ratio of the lactobacillus delbrueckii subspecies bulgaricus to the streptococcus thermophilus is 1-9:3, and the ratio range can be further 1-2: 3. 2-5: 3. 5-9: 3, and the like, and preferably, the mixing ratio of the two bacteria is 1-2: 3, preferably 1: 3. The inventor finds that pH is neutral in the early stage of fermentation, streptococcus thermophilus grows better, and the acid yield is increased, which is a main contributor to the acidity reduction in the stage. When the pH drops to 4.2, the acid production of Streptococcus thermophilus is substantially stopped, and the Lactobacillus delbrueckii subsp.bulgaricus rapidly grows and continues to produce acid, so that the Lactobacillus delbrueckii subsp.bulgaricus and Streptococcus thermophilus have a pH of 1-2: the mixing ratio of 3 is particularly 1:3, which is more favorable for accelerating the acidification of the fermentation liquor and reducing the risk of bacterial contamination.

In an embodiment of the invention, the first-order static fermentation temperature is 38-44 ℃, preferably 40-42 ℃, the fermentation time is 5-24h, and the first-order static fermentation temperature is preferably kept at 40 ℃ for 5 h.

In the embodiment of the invention, the kluyveromyces marxianus is inoculated when the pH value of the first-order fermentation is reduced to 4-5; wherein, the inoculation amount of the kluyveromyces marxianus is 1-3%, and the preferable amount is 1%.

Wherein the second-order fermentation temperature is 25-45 deg.C, preferably 33-35 deg.C, the fermentation time is 48-96 hr, preferably fermentation at 33 deg.C under aeration and stirring for 48 hr.

In an embodiment of the invention, the soy hulls are subjected to secondary inoculation fermentation, wherein the first fermentation is first inoculated with lactobacillus bulgaricus and streptococcus thermophilus, and then static fermentation is performed, wherein the fermentation can convert part of the flavone glycoside and part of acidified soy hull fibers, and the static fermentation has low conversion efficiency of the flavone glycoside, but provides enough time for infiltration and digestion of the soy hulls under the control of probiotics, so as to facilitate the sufficient release of the flavone glycoside and the soy hull fibers. And (2) performing second-order fermentation, when the pH value of the first-order fermentation is reduced to about 4-5 and is suitable for the growth of kluyveromyces marxianus, inoculating kluyveromyces marxianus, starting stirring to perform aerobic fermentation on the soybean hulls, increasing the contact between the kluyveromyces marxianus and loose and dispersed flavonoid glycoside fully released by the first-order fermentation and soybean hull fibers, fully fermenting and converting the flavonoid glycoside which is high in water solubility and difficult to be absorbed and utilized by a human body into flavonoid aglycone which is easy to be absorbed and utilized by the human body, wherein the fermentation conversion rate is more than 90%, and meanwhile, the soybean hull fibers are continuously acidified, swollen and dispersed, so that the soybean hull fibers are converted into softened insoluble dietary fibers with high water holding capacity from a compact structure, thereby being beneficial to intestinal tract movement, improving intestinal tract microecology.

In a second aspect of the present invention, there is provided a fermented product obtained by fermenting soybean hulls according to the fermentation method described in the first aspect above.

The fermentation product contains probiotic bacteria, culture medium components and a soybean hull fermentation product, wherein the soybean hull fermentation product is rich in flavonoid aglycone and loose-dispersed insoluble dietary fibers, and the intestinal microecology and health level of a human body are synergistically improved.

If desired, the flavonoid aglycone and dietary fiber component can be isolated by one skilled in the art for further use.

In a third aspect of the present invention, there is provided a composition comprising the fermented product described in the first aspect above.

In a further embodiment, soy protein, collagen peptide and soluble dietary fiber may also be included in the compositions of the present invention.

Wherein, the mixing proportion of the fermentation product, the soybean protein, the collagen peptide and the soluble dietary fiber is as follows: 10-30 parts of fermentation product, 40-90 parts of soybean protein, 1-8 parts of collagen peptide and 0.5-15 parts of soluble dietary fiber; wherein the soluble dietary fiber is selected from one or more of polydextrose, resistant dextrin, pectin, rhizoma Amorphophalli powder, fructo-oligosaccharide, inulin, and xylo-oligosaccharide. The fermentation product can be dry powder of fermentation liquor, such as obtained by concentrating the fermentation liquor and then spray drying or freeze drying; the fermentation product (fermentation liquor) can also be directly mixed with the soybean protein, the collagen peptide and the soluble dietary fiber powder, the total liquid amount is calculated according to the addition amount of the required solid matter, and then the mixture is uniformly mixed, dried and crushed. Wherein, in a more preferable embodiment, the content of the flavonoid aglycone in the composition is 30-1000 mg/g.

In a fourth aspect of the present invention, there is provided a use of the fermented product of the third aspect or the composition of the fourth aspect in the preparation of a medicament, food or health product for preventing and/or treating diabetes.

Compared with the prior art, the invention has the beneficial effects that:

the invention can efficiently utilize the waste bean curd skin to ferment and generate the dietary fiber and the composite probiotics which are rich in flavonoid aglycone and have high water holding capacity; other nutrient components are compounded, so that the fasting blood glucose can be reduced to a normal level. The raw materials of the invention are cheap and easy to obtain, the process steps are few, the conversion efficiency of the flavonoid aglycone is high, and the water holding capacity of the insoluble dietary fiber is improved.

Detailed Description

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out according to conventional conditions or according to conditions recommended by the manufacturers.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. The reagents or starting materials used in the present invention can be purchased from conventional sources, and unless otherwise specified, the reagents or starting materials used in the present invention can be used in a conventional manner in the art or in accordance with the product specifications. In addition, any methods and materials similar or equivalent to those described herein can be used in the methods of the present invention. The preferred embodiments and materials described herein are intended to be exemplary only.

The Streptococcus thermophilus used in the embodiment of the invention is Streptococcus thermophilus with the strain number CICC 20681; the Lactobacillus bulgaricus is Lactobacillus delbrueckii subsp.Bulgaricus with the strain number of CICC 20361; the used Kluyveromyces marxianus is Kluyveromyces marxianus with the strain number of CICC 32015; the three strains are obtained from the China center for the preservation and management of industrial microbial strains, and the website ishttp://www.china-cicc.org/。