AU2021104239A4

AU2021104239A4 - Effects of Ganoderma lucidum fermentation on the main chemical components in mulberry leaves α- Effects of glucosidase inhibitors on their properties

Info

Publication number: AU2021104239A4
Application number: AU2021104239A
Authority: AU
Inventors: Tianyi WU
Original assignee: Individual
Current assignee: Individual
Priority date: 2021-07-16
Filing date: 2021-07-16
Publication date: 2021-09-16
Anticipated expiration: 2029-07-16

Abstract

In recent years, as a global chronic disease, the incidence rate of diabetes has been increasing rapidly, and it has become the third serious threat to human health after cancer and cardiovascular and cerebrovascular diseases. Traditional diabetes drugs are expensive and have great side effects on the body. Therefore, it is safe and inexpensive to develop new effective multi targets from natural Chinese herbal medicine a- Glucosidase inhibitors have become a hot topic in recent years. Mulberry leaf, as a traditional Chinese medicine in China, is used as a traditional Chinese medicine to control diabetes. It contains a variety of functional components of hypoglycemia. The mechanism of hypoglycemic effect in mulberry leaves is characterized by multi-component, multi-target, multi-channel and multi effect, which has attracted the attention of scholars at home and abroad. However, the research on mulberry leaves is almost based on its dry leaves, and few researches on fresh medicine. The project applied microbial fermentation engineering to the biological processing of fresh mulberry leaves, in order to enrich the active components of hypoglycemic and improve its efficacy, which has important theoretical and applied research value. 1

Description

TITLE Effects of Ganoderma lucidum fermentation on the main chemical components in mulberry leaves a- Effects of glucosidase inhibitors on their properties

FIELD OF THE INVENTION This paper proposes a goal detection network of end-to-end multi-scale feature fusion because of the tiny pedestrian target and blocking in pedestrian detection.

BACKGROUND OF THE INVENTION

Current situation of diabetes and its drug treatment Diabetes mellitus (also known as "diabetes" in traditional Chinese Medicine) is a series of metabolic disorders, such as sugar, fat, protein, water, electrolyte and so on, caused by the absolute or relative deficiency of insulin secretion or the decrease of insulin biological effect caused by genetic and environmental factors. Diabetes is divided into type I diabetes (insulin dependent) and type II diabetes (non insulin dependent). Type I diabetes is usually caused by autoimmune system defects, virus infection, genetic factors and so on. Patients need to rely on insulin injection to survive, which may lead to the occurrence of type I diabetes. Type II diabetes is the main type of diabetes. Its pathogenesis is very complex. It is affected by the interaction of genetic factors and environmental factors, insulin resistance and islets p-Cell dysfunction is closely related to obesity. It is generally believed that it is caused by the relative deficiency of insulin and tissue insensitivity to insulin. Now the most important drugs for diabetes: insulin, biguanides, sulfonylureas and insulin a-Glucosidase inhibitors, etc. Traditional hypoglycemic drugs such as insulin, biguanides and sulfonylureas can reduce blood glucose for a short time, but they often have some side effects on gastrointestinal function, and may make the body dependent. In contrast, the hypoglycemic substances obtained from natural plants usually have the characteristics of mild and long-lasting effects, small side effects, and integrated action mechanism of multi targets, multi components and multi effects

Hypoglycemic components of plants and their hypoglycemic effects Mulberry leaves are cold in nature, sweet and bitter in taste, and have the functions of dispersing wind heat, clearing lung and moistening dryness, clearing liver and improving eyesight. Modem studies have shown that mulberry leaves are rich in a variety of active ingredients and show a variety of biological activities. The clinical results showed that mulberry leaves had the effects of lowering blood sugar, blood lipid and anti inflammation. Mulberry leaves contain flavonoids, alkaloids and polysaccharides.

Plant polysaccharides Plant polysaccharide is a kind of biological macromolecule widely existing in plant roots, stems and leaves. It is composed of many monosaccharide molecules through glycosidic bond polymerization, dehydration,A polyhydroxy polymer containing ketone group or aldehyde group is formed. It has good hypoglycemic function, mainly including: promoting insulin secretion and improving insulin resistance; Regulate the activities of related enzymes; Improve the metabolic disorder of the body; It can promote the synthesis of liver glycogen and inhibit gluconeogenesis.

Flavonoids Flavonoids are a kind of polyphenols widely existing in nature. Its structure is often connected with phenolic hydroxyl, methylamino, methyl, isopentenyl and other functional groups. Flavonoids in insulin sensitive tissues regulate carbohydrate digestion, insulin secretion, insulin signaling pathway and glucose uptake through various intracellular signaling pathways, thus showing good hypoglycemic and improving insulin resistance.

alkaloid Alkaloids are a class of basic organic compounds containing nitrogen. Most of them have complex ring structure, and most of them contain nitrogen in the ring, which has significant bioactivity and is one of the important effective components in medicinal plants. Alkaloids in mulberry leaves is a special deoxygenistein (DNJ) a- The inhibition of glucosidase activity can reduce the peak value of postprandial blood glucose and inhibit the absorption of sugar by small intestine. Objective to establish the characterization method of total flavonoids, total alkaloids and total polysaccharides in fresh mulberry leaves; Probiotics (yeast, lactic acid bacteria, etc.), medicinal fungi (Ganoderma lucidum, Eurotium coronatum, etc.), edible fungi (Pleurotus eryngii, coriolopsis strogii, etc.) were selected to screen the excellent fermentation strains of fresh mulberry leaves; With the help of modem analysis technology, the differences of hypoglycemic components in fresh mulberry leaves before and after fermentation were revealed a- The inhibition rate of glucosidase was used to characterize the hypoglycemic effect of fresh mulberry leaves before and after fermentation; Finally, the structure-activity relationship between the hypoglycemic effect and the composition and structure changes of Hypoglycemic Active Substances in fresh mulberry leaves during microbial transformation was elucidated. Natural hypoglycemic components At present, the main drugs for diabetes are insulin, biguanides, sulfonylureas Glucose inhibitors, thiazolidinediones, etc. Although these drugs can reduce blood glucose in a short time, they often have some side effects, especially on the gastrointestinal tract, and long-term use of the body will produce dependence. In contrast, the hypoglycemic substances extracted from natural plants usually have the characteristics of mild and long-lasting effects, small side effects, and the integrated action mechanism of multi-target, multi component and multi effect.

Research and application of fresh Chinese Medicine Mulberry leaves contain a large number of medicinal active components, macromolecular compounds and trace elements beneficial to human body [2]. In clinical practice, mulberry leaf has long been used as a drug to control diabetes (also known as diabetes). At present, the research on mulberry leaves is based on the dry products of mulberry leaves. There are no studies on the changes of active components and pharmacodynamics of fresh mulberry leaves before and after fermentation. The main reasons are that we do not know enough about the material structure and chemical composition of fresh medicine, and the concentration of active ingredients in fresh medicine is low. In addition, in terms of current science and technology, the cost of cultivation, transportation and storage of fresh medicine is high, the application method of fresh medicine, the backward preservation technology of medicinal materials, and the constraints of its own deterioration and failure, It brings challenges to the research of fresh Chinese medicine. In this project, fresh mulberry leaves are used as raw materials, and on the basis of the existing research on dry preparations of traditional Chinese medicine, a method for analyzing the physical and chemical properties of fresh medicines is established to analyze the differences between dry and fresh traditional Chinese medicine, so as to provide a theoretical basis for the development of fresh preparations of traditional Chinese medicine.

Fermentation processing technology of medicinal plants Traditional fermentation technology mainly uses natural inoculation and multi strain fermentation. The bacteria are fungi and bacteria in natural environment. There are many kinds of microorganisms added in the fermentation process, but the number of bacteria that play a role is limited. In general, the traditional fermentation method is also mixed with other strains, such as,In the process of fermentation, the nutrients in the effective components will be greatly reduced, and the fermentation efficiency will be reduced sharply, and the fermentation products will be easily polluted. Different from traditional Chinese herbal medicine processing, it combines Chinese herbal fermentation with bioengineering and microbiology. With the development of real-time fluorescence quantitative PCR and RNA and the in-depth development of microbial diversity research, it can now be based on the changes of bacterial colony and its composition structure in the process of edible and same source plants, The source and species of medicinal plant fermentation strains are more abundant by using modem fermentation technology to screen out the beneficial bacteria, such as 16sma and 26srdna sequence analysis. The core of solid-state fermentation of mulberry leaves by Ganoderma lucidum is that Ganoderma lucidum can proliferate by using the nutrients in mulberry leaves. The mycelium will produce various enzymes during the process of proliferation. Some components of mulberry leaves will decompose and combine with each other under the action of these metabolic enzymes, and form new active chemical components.

Ways of plant fermentation Traditional Chinese medicine fermentation technology mainly includes liquid deep fermentation and solid fermentation. In the production process, due to the high technological requirements of submerged fermentation, this process will reduce the conversion rate of effective components of traditional Chinese medicine, improve the production cost, and consume a lot of medicinal materials resources. The current expenditure of this research is similar to that of the laboratory. Nowadays, the traditional solid-state fermentation method is still used in traditional Chinese medicine fermentation, so this experiment uses the solid-state fermentation technology. In most cases, solid-state fermentation is a biological reaction process in which there is no or almost no free water and one or more microorganisms ferment in water-insoluble solid substrate with certain humidity. Compared with liquid fermentation, solid-state fermentation has the following advantages: (1)after complex processing procedures, energy saving, fermentation process is more environmentally friendly and green. (2)Simulation of natural growth environment, maintain the growth state of microorganisms. (3)The post-treatment is simple, no need to extract and dry directly, so as to keep the active ingredients in the drug to the maximum extent.

SUMMARY OF THE INVENTION Materials and methods Raw materials and strains Ganoderma lucidum was purchased from CICC (No. 14029), and mulberry leaf was purchased from Kuqa, Xinjiang.

Chemicals and reagents Corn flour (Henan Feitian Agricultural Development Co., Ltd.); Wheat bran (Henan Feitian Agricultural Development Co., Ltd.) rutin standard (Yuanye Biology),Technology Co., Ltd.); Ethanol (Shanghai Aladdin Biochemical Technology Co., Ltd.); Acetone (Shanghai Aladdin Biochemical Technology Co., Ltd.); 4-hydroxypiperidinol (Sinopharm Chemical Reagent Co., Ltd.);

Saturated Wright's salt solution (Sinopharm Chemical Reagent Co., Ltd.); Hydrochloric acid (Sinopharm Chemical Reagent Co., Ltd.), PBS solution (Sinopharm Chemical Reagent Co., Ltd.); Acarbose (Sinopharm Chemical Reagent Co., Ltd.); DNJ (Shanghai Aladdin Biochemical Technology Co., Ltd.); PNPG (Sinopharm Chemical Reagent Co., Ltd.); Glucosamine standard solution (Sinopharm Chemical Reagent Co., Ltd.); Sulfuric acid (Sinopharm Chemical Reagent Co., Ltd.); Sodium hydroxide (Sinopharm Chemical Reagent Co., Ltd.); Acetylacetone (Sinopharm Chemical Reagent Co., Ltd.); Eurlich reagent (Sinopharm Chemical Reagent Co., Ltd.); Sodium carbonate (Sinopharm Chemical Reagent Co., Ltd.); The water is ultrapure water (self made), and the other reagents are analytically pure.

Solid state fermentation of mulberry leaves Weigh 10g fresh mulberry leaves, add corn flour and bran, then add distilled water, adjust the final solid-liquid ratio of medium to 1:4; Stir evenly under natural pH, and sterilize in 120 °C with high pressure steam sterilizer 0 C sterilization for 20 min. After cooling, 2 ml of Ganoderma lucidum seed solution was inoculated, and then 30 ml of Ganoderma lucidum seed solution was placed 0 The control group was not inoculated with microorganisms. On day 0, respectively; 2d ; 4d ; 6d ; 8d ; Ten days later, samples were taken to determine the indicators (including biomass, total flavonoids content, total polysaccharides, total alkaloids content), and three parallel time points were set at each time point.

Determination of biomass Draw the standard curve of glucosamine: weigh an appropriate amount of glucosamine standard accurately after prepare a series of solutions. Then measure the absorption value at 520nm with spectrophotometer. Taking the concentration as the abscissa and the absorption value as the ordinate, the standard curve of glucosamine content was drawn. The membrane culture method was used: a layer of sterile cellophane was laid on the wort plate, 0.5ml spore suspension prepared from the inclined plane was absorbed and laid on the membrane. It was incubated for 7 days under 30 0 C , and then cultured for 14 days after decreased to 25 0 C, The biofilm was peeled off from the cellophane for 80 days 0 C drying is pure bacteria. Weigh 0.3g dry bacteria precisely, add 2 ml of 60% h2so4,25 0 Soak for 24h at room temperature, dilute to Imol / L H2SO4, place in 250ml flask, 9.8 ml x After cooling, neutralize with Imol / L NaOH to pH7, and fix the volume to 100ml. Take 0.5ml (blank is 0.5ml distilled water) and 1ml acetylacetone reagent (1.5ml acetylacetone + 50ml 1.25mol/1 Na2CO3), 90 0 After heating for 1 h at room temperature and cooling in cold water, 1 mleurlich reagent was added and colorimetric determination was carried out at 520 nm.

Determination of flavonoids Draw the standard curve of flavonoids: prepare 0.2mg/ml rutin solution, accurately measure 0, 1, 2, 3, 4 and 5ml of the reference solution, put them into ml volumetric flask, add 0.3ml of 5% sodium nitrite solution, shake and stand for 6minutes 0 Add 0.3ml of 10% aluminum nitrate solution, shake and stand for 6minutes, add 4ml of 4% sodium hydroxide solution, dilute with 70% ethanol to 10ml scale, shake and stand for minutes, The absorbance was measured at 510nm. The standard curve was drawn with absorbance (a) as ordinate and reference concentration (c) as abscissa. The regression equation was y = 13.9x-0.0577 (R2 = 0.9913). Preparation of sample extract: 4.Og mulberry leaves were accurately weighed in a 50ml centrifuge tube, and 40 ml of 70% ethanol solution was added 50°C water bath ultrasonic extraction for 40 minutes, filtration, collection of supernatant, sediment and then repeat the above steps to extract once, combined with supernatant, the sample extract was obtained. 2 ml of mulberry leaf extract was precisely measured and put into a ml measuring flask. 0.3 ml of 5% sodium nitrite solution was added to each flask. The mixture was shaken well and left standing for 6 min; Add 0.3ml of % aluminum nitrate solution, shake up, stand for 6minutes, then add 4ml of 4% sodium hydroxide solution, dilute with 60% ethanol, set volume scale, shakeup, stand for minutes, take 60% ethanol solution as blank, measure the absorbance at 51Onm, calculate the content of total flavonoids.

Determination of polysaccharide content Draw polysaccharide standard curve: prepare 1mg / ml glucose standard solution, take 0, 0.2, 0.4, 0.6, 0.8, 1.ml glucose standard solution respectively, add water to 1ml, add dns1.5ml, and develop color in boiling water bath for minutes. After cooling, add water to 10ml, measure the absorbance at 550nm, and draw the standard curve with the absorbance (a) as the ordinate and the reference concentration (c) as the abscissa. Regression equation: y = 0.7251x 0.0326 (R2 = 0.9956) preparation of sample extract: accurately weigh 4.Og of mulberry leaves to be tested in a 50ml centrifuge tube, add 30ml distilled water, mix well and mix for 80 minutes water bath ultrasound 20 minutes, filter, collect supernatant, sediment. Then repeat the above steps to extract once, combined with supernatant to obtain sample extract.Hydrolysis of the extract: precisely transfer 4ml of mulberry leaf sample solution, put it into a triangular conical flask, add 2ml of 6mol / L HC solution (currently prepared), seal it, heat it on a boiling water bath for 30minutes, take it out to cool it down to room temperature. Adjust the pH value to 8.0 with 6mol / L NaOH solution, fix the volume to 20ml with distilled water and shake up. Take 1.Oml of different grades of mulberry leaves polysaccharide hydrolysate samples, put them into a tube with plug, add water to 2.Oml, add 1.5ml DNS reagent, shake them up, heat them in boiling water bath for minutes, take them out and cool them fast, then use 2.Oml distilled water as blank,measure the absorbance at 550nm with ultraviolet visible spectrophotometer.

Determination of alkaloid content Draw the standard curve of alkaloid: accurately absorb 5ml of 0.4mol/1 mother solution of 4-hydroxypiperidinol standard, add 45ml of 0.05mol/1 hydrochloric acid to dilute it to 0.04mol/1 standard solution, take 0.1, 0.2, 0.4, 0.6, 0.8, 1ml of diluted standard solution respectively when making the standard curve. Add 0.05mol/1 hydrochloric acid solution to prepare 2ml of 0.002, 0.004, 0.008, 0.012, 0.016, 0.02mol/1 solution for standby. Suck 2ml standard substance into ml centrifuge tube, add 3ml of 0.02g/ml Wright's salt solution, and take ice bath for 2hours. After centrifugation at 8000 rpm / min for 10 minutes, the supernatant was skimmed off; and ethyl acetate was added into the centrifugation tube. After centrifugation at 8000 rpm / min for 10 minutes, the precipitate was retained and dissolved in 10 ml of 70% acetone solution. The absorbance of the solution was measured at 523 nm. The standard curve was drawn with absorbance (a) as ordinate and reference concentration (c) as abscissa. Regression equation: y = 22.4555x-0.0196 (R2 = 0.9973). Preparation of sample extract: accurately weigh 5g mulberry leaves to be tested and put them into a 50ml centrifuge tube, add 50ml 25% ethanol-0.05mol/1 hydrochloric acid solution, mix well and mix for 30 minutes water bath ultrasound for 20 minutes, filter and collect the supernatant to obtain the sample extract. The sample extract was concentrated and evaporated to dryness with rotary evaporator, dissolved with 0.05mol/1 hydrochloric acid and diluted to 4ml to obtain the total alkaloids of mulberry leaves to be determined solution. 2ml of the solution to be determined was centrifuged at 8000rpm / min for 10 minutes. The supernatant was put into a 10ml centrifuge tube. 3ml of 0.02g/ml Reynold's salt solution was added and ice bath was used for 2hours. After centrifugation at 8000 rpm / min for 10 minutes, the supernatant was skimmed off, and ethyl acetate was added into the centrifugation tube. After centrifugation at 8000 rpm / min for 10 minutes, the precipitate was retained and dissolved in 10 ml of 70% acetone solution. The absorbance of the solution was measured at 523 nm.

Inhibitors for a- Analysis of inhibition rate of glucosidase The experiment was carried out on 96 well plate. There were four groups in the experiment: blank group (substrate), control group (substrate + enzyme solution), sample blank group (substrate + sample), sample group (substrate +

sample + enzyme solution). First of all, 80% was added into 96 well plate p L (blank group, b), 70 p L (control group C), 60 p L (sample blank group, sb), 80 p L (sample group, s) PBS solution with pH = 6.8; Then, 20 samples were added to the sample group and the sample blank group (sb) p L the inhibitor to be tested (the sample is each extract.The control substance is acarbose, DNJ and rutin); Then added 10 samples to the sample group (s) and the control group (b) p L lU/mL a- Glucosidase; Mix well, 37 0 C. oven activation for 15 minutes; Add another 20 p LIOmmol/LpNPG ; After mixing 37 0 C. oven reaction for 30 minutes; Finally, add 100 p The absorbance of 1Imol / L Na2CO3 solution was determined at 405nm. Inhibitors for a- The inhibition rate of glucosidase was calculated as follows:

Inhibition rate%=AC-AB)-(AS-ASB) (AC-AB) X 100%

DESCRIPTION OF DRAWINGS

1. Fig.1 shows the growth curve of Ganoderma lucidum on dry and fresh mulberry leaves; 2. Fig. 2 shows the polysaccharide content of Ganoderma lucidum fermented on dry and fresh mulberry leaves; 3. Fig. 3 shows the alkaloid content of Ganoderma lucidum fermented on dry and fresh mulberry leaves.

DESCRIPTION OF PREFERRED EMBODIMENT Results and analysis Determination of biomass It can be seen from Fig. 1 that Ganoderma lucidum grows rapidly and logarithmically from day 0 to 8 of fermentation; From the 8th day to the 12th day, the growth rate was gradually stable and reached the peak on the 12th day; After 12 days, the biomass of Ganoderma lucidum began to decline. On day 0 2, the biomass of Ganoderma lucidum cultured in fresh mulberry leaves and dry mulberry leaves was almost the same; From the second day of fermentation, the biomass and growth rate of Ganoderma lucidum cultivated with fresh mulberry leaves were higher than those cultivated with dry mulberry leaves. After the 12th day of fermentation, the biomass of Ganoderma lucidum cultivated with fresh mulberry leaves tended to be the same.

Determination of polysaccharide content From Fig. 2, the fermentation of mulberry leaves by Ganoderma lucidum, the content of polysaccharide generally presents a downward trend. On day 0, the polysaccharide content in fresh mulberry leaf medium (224.18 mg / g) was much higher than that in dry mulberry leaf medium (113.48 mg / g). However, it decreased to 22.08 mg / G on day 10. While the polysaccharide content in dry mulberry leaf medium decreased to 46.76 mg / g. There's a possibility that with time-varying, polysaccharides were decomposed by microorganisms, which reduced the content of polysaccharides, and microorganisms may be easier to use polysaccharides in fresh mulberry leaves.

Determination of alkaloid content From Fig. 3, the alkaloid content in mulberry leaves first increased and then decreased with the increase of fermentation time. The content of alkaloids in fresh mulberry leaf medium was higher than that in dry mulberry leaf medium. On the 8th day of fermentation, the alkaloid content reached the maximum, fresh mulberry leaf medium (12.41 mg / g), dry mulberry leaf medium (8.23 mg / g).

Determination of flavonoids From Fig. 4, with the fermentation time, the flavonoid content in mulberry leaves generally increased first and then decreased. The content of flavonoids in fresh mulberry leaves reached the maximum on the 8th day of fermentation (12.41 mg / g). Dry mulberry leaves reached the maximum on the 6th day of fermentation (8.23 mg / g). Bewteen them, the content of flavonoids in fresh mulberry leaf culture medium was generally higher than that in dry mulberry leaf culture medium. However, it took less time for the content of flavonoids in dry mulberry leaf culture medium to reach the maximum value.

Inhibition of active components in mulberry leaves by Ganoderma lucidum fermentation a- Effect of glucosidase on the activity of glucosidase There was a negative correlation between fermentation time and polysaccharide content in mulberry leaf medium, and a positive correlation between fermentation time and flavonoids and alkaloids content at first, and then a negative correlation. Then, 5 UG / ml flavonoids and alkaloids were prepared, and the effects of alkaloids and flavonoids on fermentation time were determined a- The inhibition efficiency of glucosidase.

From Fig. 5, 5 UG / ml alkaloid has a significant effect on the growth of mice which proves a- The inhibition rate of glucosidase first increased and then decreased as the fermentation time increases. It reached the maximum value of 50.732% on the 8th day, and then the inhibition rate began to decline. The inhibition rate increased rapidly on day 0-2 and 4-6, and tended to be flat on day 2-4. From Fig. 6, 5 UG / ml flavone is the best for the treatment a- With the increase of fermentation time, the inhibition rate of glucosidase tended to be gentle at first and then decreased. It reached the maximum value of 19.072% on the fourth day and then decreased gradually. It can be seen that under the same concentration, the alkaloids have a significant effect on the growth of mice a The inhibitory effect of glucosidase was better than that of flavonoids, and the maximum inhibitory rate appeared later than that of flavonoids.

Conclusion and prospect

By measuring the biomass changes of fresh and dry mulberry leaves fermented by Ganoderma lucidum at different fermentation time, and the content changes of polysaccharides, flavonoids, alkaloids and other active substances so as to found that the fresh mulberry leaves medium was more suitable for the growth of Ganoderma lucidum than the dry mulberry leaves medium. The contents of polysaccharides, flavonoids and alkaloids in fresh mulberry leaf medium were higher than those in dry mulberry leaf medium. With the development of fermentation, the content of polysaccharides in fresh and dry mulberry leaves decreased. Also, the content of flavonoids and alkaloids increased first and then decreased, indicating that the fermentation time was negatively correlated with the content of polysaccharides, and positively correlated with the content of flavonoids and alkaloids first and then negatively correlated. The effect of alkaloids on Fermentation a- The inhibitory effect of glucosidase was obviously enhanced, and flavone was the most effective a- The inhibition of glucosidase changed unsignificant. This study provides a theoretical and experimental basis not only for the current clinical trials and laboratory research of traditional Chinese medicine, but also the research and development of functional foods, the prevention and treatment of diabetes, hyperglycemia and secondary diseases of blood lipids. Future research directions should reduce the toxic and side effects and dependence in the process of drug treatment to the greatest extent; improve the utilization rate of active ingredients in mulberry leaves, also including study the preservation method and transportation way of fresh Chinese medicine as soon as possible, so as to realize the reasonable allocation of resources and reduce waste.

Claims

CLAIM

1. Effects of Ganoderma lucidum fermentation on the main chemical components in mulberry leaves a- Effects of glucosidase inhibitors on their properties, characterized in that, including: The project applied microbial fermentation engineering to the biological processing of fresh mulberry leaves, in order to enrich the active components of hypoglycemic and improve its efficacy, which has important theoretical and applied research value.

Figure 2 Figure 1

Figure 3