TWI750817B - Preparation method of tremella composition and tremella composition - Google Patents

Abstract

A preparation method of tremella composition and tremella composition. Its technical characteristics are the use of a white fungus, a dietary fiber, a high protein, and a nucleic acid solution, which can enhance the cell repair function and maintain a sense of satiety at the same time, and its manufacturing method is through The specific temperature and pressure cooking conditions provide high light transmittance, and allow direct visual inspection of the solvent stability during testing.

Description

Preparation method of tremella composition and tremella composition

The present invention is related to Tremella nutritional supplements, especially a preparation method of Tremella composition and Tremella composition.

With the improvement of health awareness, people often take fungi or dietary fiber foods to promote intestinal health. In addition to improving physical fitness, such as lowering cholesterol, lowering high blood pressure, improving allergies, enhancing innate immunity, etc. The addition of fiber achieves satiety. At present, most of the common fungi-containing beverages on the market are fungi-based beverages preserved in low temperature acidity, such as yeast beverages, black fungus beverages, etc., or medicinal fungi beverages, such as Cordyceps sinensis beverages, Antrodia cinnamomea beverages, and Poria beverages. In the above-mentioned fungal drinks, there is no dietary fiber added, so it is difficult to achieve the effect of satiating or replacing food. Ganoderma lucidum, Antrodia cinnamomea, Poria, Poria, Cordyceps sinensis enhance immunity, but lack of increasing satiety.

In order to increase the sense of satiety, Patent No. I353222 discloses a baked product of cold weather powder and probiotics, which uses flour, the raw material of the baked product, as the main raw material for satiety. The patent No. I692361 uses Xiangshanzhi, the patent No. I679983 uses Poria, and the patent No. I453025 uses Ganoderma lucidum.

Also, Tremella is popular in China and Southeast Asia for its nutritional value and attractive flavor. Tremella is a typical dimorphic fungus with The ability to switch between the cellular yeast phase and the multicellular filamentous phase. Dimorphism provides a model for studying eukaryotic morphogenesis and cell differentiation, which is directly related to the invasion of pathogenic dimorphic fungal hosts. It is known that the research on the two forms of Tremella is focused on environmental factors and differences in cell wall composition, and the extraction of RNA is the premise of molecular biology research. Tremella contains tough cell walls and high levels of polysaccharides that can precipitate or bind with RNA. Standard RNA isolation kits, such as Trizol reagent (developed by Invitrogen, California, USA) and RNAiso plus (developed by Dalian Shubao Bioengineering Co., Ltd., China) are used to isolate RNA from samples, but they are not effective for RNA isolation. Among the Tremella polysaccharides, it is less conducive to the purpose of manufacturing quality control.

In addition, dietary fiber can generally be divided into soluble dietary fiber and insoluble fiber. Soluble dietary fiber refers to pectin, vegetable gum, viscose, etc., or insoluble fiber includes cellulose, hemicellulose, lignin, etc. Generally, dietary fiber can be consumed by adding water and heating. However, due to poor light transmittance, dietary fiber is not transparent and has a poor appearance. Although Tremella also contains about 70% dietary fiber, which can help gastrointestinal motility and reduce fat absorption, it must be thoroughly extracted and adjusted to a specific pH value, followed by centrifugation and precipitation to effectively extract water-soluble dietary fiber, which is inconvenient for mass production.

Before formulating the nutritional composition, the ion concentration and solubility must be calculated to further calculate the pH value:

Solubility = X grams / 100 grams of water

The pH value is a measure of the hydrogen ion's activity [H+] in a solution, the higher the [H+], the more acidic it is. In the state of pure water at 25 degrees Celsius, the dissociation constant is:

Kw=[H+][OH-]=10^(-14) The pH value of the solution is less than 7 is acidic, and the pH value is more than 7 is alkaline, and the pH value will change due to temperature. The conventional calculation method of pH is as follows:

pH=-log10[ionic molarity]

In addition, there are many common sources of high protein, including milk protein powder, milk protein concentrate, milk protein isolate, skim milk solids, skim milk, skim milk powder, etc. Corresponding to different sample processing methods. Although the liquid fermentation product of medicinal fungus Ganoderma lucidum has been used to improve fatty liver, the fungal silk of Tremella has not yet been used to inhibit inflammation and reduce antioxidants.

In addition, the currently common black fungus drink is a homogenous polysaccharide, and its biological classification is Agaric fungus. The main nutrients are iron and potassium. The ear slices can only increase the amount of sugar on one side, which is different from white fungus. Tremella is a A weak acid polysaccharide, the main components are vitamin B and trace elements.

Referring to Fraser CG, Jennings HJ, Moyna P. (1973) Structural analysis of an acidic polysaccharide from Tremella mesenterica NRRL Y-6158, the main structure of Tremella polysaccharide is shown in Figure 1. In addition, some related researchers (such as Proma Khondka) have also revealed that polysaccharides can be extracted from Tremella in large quantities through the precipitation of the chemical reagent lithium chloride (LiCl). When extracting or extracting polysaccharides, not only the amount of extraction, but also the following factors will be considered in the manufacturing process of good saccharide materials: (1) the moisture of the sample; (2) the amount of nitrogen compounds and proteins; (3) ) mineral content by dry ashing method; (4) starch content; (5) sugar properties measured by gas chromatography.

The main purpose of the present invention is to provide a preparation method of a Tremella composition and a Tremella composition, which have the function of repairing human cells and make drinking more satiety.

The reason is that, in order to achieve the aforementioned purpose, the present invention provides a method for preparing a Tremella composition, comprising a Tremella dehydration step, dehydrating a fresh Tremella; a first grinding step, grinding the dehydrated Tremella; a first Two grinding steps, grinding a dietary fiber; a first mixing step, adding if disturbed starch to the ground dietary fiber to mix evenly; a second mixing step, adding the ground white fungus and a high protein to the dietary fiber; A decoction step, heating the mixed white fungus, high protein and dietary fiber for a predetermined time; a pH adjustment step, adjusting the pH value of the white fungus, high protein and dietary fiber after heating to obtain a white fungus composition; and A detection step is to detect the human cell repairing effect of the Tremella composition.

Preferably, after the second mixing step, a third mixing step is further included, which is to add a nucleic acid solution to the mixed white fungus, high protein and dietary fiber.

Preferably, in the first grinding step, 3 to 10 pieces of dried white fungus are taken out, each piece of white fungus has a diameter of about 10 to 15 cm, ground for 15 to 20 minutes to form powder, and sieved; the second grinding step When grinding dietary fiber, add MES/TRIS buffer solution, adjust the pH value to 8.2, add amylase and heat at 65-95 degrees Celsius, then centrifuge, heat and dry to obtain water-soluble Dietary fiber; the composition ratio of each component in the second mixing step is as follows: the weight percent of the white fungus is between 15~30%, the weight percent of the dietary fiber is between 10~20%, the weight percent of the high protein is 40% or less; in the third mixing step, add water to cover the white fungus, the dietary fiber and the high protein; the cooking step is at a pressure of 0.6 MPa and a heating time of 65 to 95 degrees Celsius for 20 minutes; the adjusted acid-base The pH value of Tremella is fixed between 5.68-5.83 in the value step, and the pH value of the whole solution is adjusted between 7.05 and 7.45 through whey protein.

Preferably, according to the method described in claim 3, in the step of dehydrating Tremella, the tremella is heated between 65 and 95 degrees Celsius to produce polysaccharides with a molecular weight between 9,000 and 10,000 Da.

Preferably, the method according to claim 3, wherein the weight percentage (wt%) of the white fungus, dietary fiber, high protein and nucleic acid solution is 8:8:3:2.

In addition, the present invention further provides a white fungus composition prepared by the aforementioned method, comprising a white fungus in powder form with a particle size between 1.6 microns and 25 microns; a dietary fiber, which is extracted from vegetable gum or algae and a high protein, the high protein being whey protein.

Preferably, the tremella composition according to claim 6, further comprises a nucleic acid solution.

Preferably, the white fungus composition according to claim 7, wherein the nucleic acid solution is DNA extracted from white fungus. protein, RNA. Protein and vitamin group, the weight ratio of various components is: DNA protein is 0-40%, RNA protein is White is 0-40%, and vitamins are less than 10%.

Preferably, the tremella composition according to claim 6, wherein the dietary fiber is one of plant gums such as tremella polysaccharide fermentation, glucose fermentation or sucrose fermentation.

Preferably, the white fungus composition according to claim 7, wherein the overall pH value of the white fungus, the dietary fiber, the high protein and the nucleic acid solution is between 7.05 and 7.45.

110: Tremella dehydration step

120: First grinding step

130: Second grinding step

140: First mixing step

150: Second mixing step

160: Third mixing step

170: Cooking Step

180: Reconcile pH steps

190: Detection steps

Figure 1 shows the main structural diagram of the known Tremella polysaccharide.

FIG. 2 is a flow chart of a preferred embodiment of the present invention.

FIG. 3 is a cell repair verification diagram of a preferred embodiment of the present invention.

Hereinafter, a preferred embodiment of the present invention is given, and further detailed description is done in conjunction with the drawings as follows:

As shown in Figure 2, a method for preparing a Tremella composition according to a preferred embodiment of the present invention includes a Tremella dehydration step 110, dehydrating a fresh Tremella; a first grinding step 120, grinding the dehydrated Tremella; a The second grinding step 130 is to grind a dietary fiber; a first mixing step 140 is to add the starch to the ground dietary fiber to mix evenly; a second mixing step 150 is to mix the ground white fungus and a high protein adding dietary fiber; a third mixing step 160, adding a nucleic acid solution to the mixed white fungus, high protein and dietary fiber; a decoction step 170, mixing the mixed The white fungus, high protein and dietary fiber are heated for a predetermined time; a pH adjustment step 180 is to adjust the pH value of the white fungus, high protein and dietary fiber after heating to obtain a white fungus composition; and a detection step 190 is to detect the white fungus Human cell repairing effect of the composition.

The white fungus required in the first grinding step 120 must be selected from the dried white fungus pieces, and 3 to 10 pieces can be collected during each grinding, and the best is 5 to 77 pieces; through experiments, it was found that Wei Yu Co., Ltd. The produced sub-large T8 tremella is the best; when it is selected from the tremella chips provided by the company, and at least 10 to 15 cm is selected for grinding for at least 15 to 20 minutes, and sieved, the best polysaccharide extraction can be obtained. quantity. In every 100 grams of sub-large T8 dried white fungus, the particle size of grade A (Mesh less than 8000) can be obtained about 15.0%±0.17; the particle size of grade B (Mesh500-8000) is about 38.0%±0.15; the particle size of grade C (Mesh100-500 ) about 22.0%±0.2; particle size D grade (Mesh greater than 100) about 25.0%±0.75. In order to reduce the manufacturing cost and use the dried Tremella as the material for the extraction of polysaccharides, the particle size, the particle size of B grade (Mesh500-8000) is the best, and the extraction amount of the polysaccharide liquid can be optimized.

To be more specific, Tremella is selected from a factory with a temperature of 23-25 degrees Celsius, a humidity of 85%-90%, a light of 250-450lux, and an oxygen supply for five minutes per hour. Tremella planted on more than 26 days but no more than 26 days, and select 3 to 10 ear pieces, and each piece is about 10 to 15 cm long for harvesting.

After the white fungus was ground, the white fungus powder with different particle size grades was obtained per 100 grams of dried white fungus according to the grade (Table 1).

The different particle size grade Tremella (Table 2) and the polysaccharide extraction amount (Table 3) that can be obtained per 100 grams are as follows:

Reference is made to [Table 3] below.

In one example, the polysaccharide content test was carried out at a solid-liquid ratio of 1:20 through experiments, and it was also found that the content of polysaccharide exhibited by particle size grade B was better than that of particle size grades A, C, and D.

In another example, the polysaccharide content test was conducted with a solid-liquid ratio of 1:40 through experiments, and it was also found that the content of polysaccharide exhibited by particle size grade B was better than that of particle size grades A, C, and D.

In yet another example, the polysaccharide content test was carried out at a solid-liquid ratio of 1:60 through experiments, and it was also found that the polysaccharide content of particle size grade B was better than that of particle size grades A, C, and D.

In conclusion, Tremella granules or Tremella powder of different sieved particle size grades can be properly dried and adjusted for use in nutritional product mixtures. Observing the obtained data, the particle size of grade A and particle size of grade B are similar in the ratio of material to liquid of 1:20 and 1:40. Therefore, when grade of particle size A and grade of particle size B are selected for production, the material-to-liquid ratio should be The optimal operation range is selected from 1:20 and 1:40.

The second grinding step 120 is to grind dietary fiber, and the dietary fiber is added with MES/TRIS buffer solution; the purpose of its use is to facilitate observation and purification without interfering with the activity of the protein. Determine the purification method and the cell sample to be assayed. Adjust to pH 8.2, mix, add amylase, heat at 65-95 degrees Celsius, then centrifuge, heat and dry the water-soluble dietary fiber obtained, and grind it independently. The dietary fiber is one of plant gums such as Tremella polysaccharide fermentation product, glucose fermentation product or sucrose fermentation product.

Dietary fiber is not ground together with Tremella because the grinding stress (Grinding Stress) of dietary fiber and Tremella is different. With reference to [Table 4], when the feeding speed is 50~100cm/sec, 500RPM enters the grinding machine, and after the screening of the holes, the average value of 25 tests is calculated, and the Mesh 500-8000 weight percentage of the gained Mesh 500-8000 residues is observed, and the white fungus is known. Only 5.9% of the grinding residue was measured within 10 seconds, and 8.6% of the dietary fiber treated as above. Observing the average residue measurement per second, it can be known that the residual percentage of dietary fiber is higher than that of white fungus, and its residual pressure is about 41-45% higher than that of white fungus, so dietary fiber is higher than that of white fungus. The ears are more resistant to grinding.

The first mixing step 140 is to add dietary fiber to interfere with starch; its purpose is to convert the fluid prepared by oxidation, acid hydrolysis and enzymatic hydrolysis, so as to promote its physical modification including facilitating subsequent processing, shaping, shearing and thermal inhibition. After the first mixing step 140, the second mixing step 150 is continued, wherein the weight percentage of white fungus is between 15 and 30%, the weight percentage of the dietary fiber is between 10 and 20%, and the high protein is between 10 and 20% by weight. The weight percentage is below 40%. In the third mixing step 160, the nucleic acid solution is added to the container containing Tremella, dietary fiber, and high protein, and water is added to cover, and the weight percentage (wt%) of the Tremella, dietary fiber, high protein, and nucleic acid solution is 8. :8:3:2, the nucleic acid solution is DNA extracted from Tremella. protein, RNA. For protein and vitamin groups, the weight ratio of various components is: DNA protein is 0-40%, RNA protein is 0-40%, and vitamins are less than 10%.

Referring to [Table 5], the third mixing step 160 is followed by a steaming Cooking step 170, the cooking step is to stabilize the suspension of Tremella granules and dietary fiber in the nucleic acid solution, and to make the overall light transmittance of the solvent good. The main light transmittance effect can be selected at a pressure of 0.6~0.9MPa, and the cooking step 170 is performed at a heating time between 65 and 95 degrees Celsius for 20 minutes; the best result of the cooking step is a pressure of 0.6 MPa and a temperature between 65 and 95 degrees Celsius. The heating time is 20 minutes, and the white fungus is heated between 65 and 95 degrees Celsius to produce polysaccharides with molecular weights between 9000 and 10000 Da. The light transmission at 388 nm is carried out through Shimadzu's UV-3600 spectrometer, as shown in [Table] 5].

In the step 180 of adjusting the pH value, except that the white fungus is fixed at a pH value of 5.68-5.83, the whole solvent is adjusted between 7.05 and 7.45 through whey protein.

The detection step 190 is as follows:

Referring to [FIG. 3A-3B], the Tremella fuciformis prepared in order to prove the present invention The composition has the function of cell repair, and the experiment was conducted by entrusting Dr. Yang Jiaxin from the Medical Research Department of the Hospital Affiliated to China Medical University to conduct an experiment for the detection of cell wound healing activity. In the experimental method, the wound healing activity of the human skin keratinocyte strain (HaCaT) was detected by the IncuCyte Kinetic Live Cell Imaging System with the particle size specially ground. After HaCaT cells were added with 100 and 200 μg/mL of the composition, the changes of cell healing were observed every 30 minutes, and images were taken at the 0th hour, the 6th hour, and the 12th hour to measure the state of cell growth. In the control group, no additives were added after the skin cells were scraped out, while the experimental group 1 was treated with the tremella extract of the present invention at a concentration of 100 μg/mL, and the Tremella tremella extract at a concentration of 200 μg/mL. to be processed. After 12 hours of uninterrupted observation, it can significantly promote the human skin keratinocyte line HaCaT cells and wound healing. Apparently, the tremella composition prepared by the present invention has the effect of repairing wound healing of human skin keratinocytes.

During extraction, the polysaccharide can be extracted with different particle-size Tremella powder through different material-liquid ratios (1:20, 1:40, 1:60). The extracted amount of polysaccharide was detected by phenol sulfur method. According to the results of our company's development experiments, the polysaccharide content of Tremella and the particle size of Tremella have differences in output. The finer tremella particle size produces more polysaccharides, and the coarser tremella particle size produces less polysaccharide.

In addition, different particle size grades of Tremella can be obtained by heating water at different temperatures to obtain polysaccharides, and the relevant solubility can be calculated [Table 6]:

Use and handling of dietary fiber:

Selecting vegetable soluble dietary fiber as the application of the present invention can appropriately select soluble fiber, such as pectin, vegetable gum, viscose, or select insoluble fiber, such as cellulose, hemicellulose, lignin. Insoluble fiber is selected when preparing the powder composition; soluble fiber is selected as the material for preparing the solvent.

Tremella itself also has dietary fiber, but the method of adding it is adopted because the crude fiber of Tremella is not much during production. For example, in the form of meal replacement, insoluble fiber is added to the product, so that when the human body takes it, the gastric acid secreted by the human body can be digested for a longer time. If the product is made to take polysaccharide as the main purpose of ingesting nutrients, soluble fiber or no soluble fiber can be added.

When Tremella is used as dietary fiber, MES/TRIS buffer solution is added, and the pH value is adjusted to 8.2. After mixing, adding amylase and heating at 65-95 degrees Celsius, and then centrifuging, heating and drying to obtain a water-soluble meal made of Tremella. fiber.

Use and handling of high protein:

The present invention uses whey protein, and the main reason for mixing Tremella extract is that the polysaccharide of Tremella extract can reduce bitterness, and at the same time, it contains all the essential amino acids that help the growth of human tissues, through a small amount of fat and carbohydrates in the form It can be used for users with lactose discomfort as their source of nutrition. In addition, whey protein may contain immunoglobulins, lactoperoxidase and lactoferrin, so whey protein may provide advantages over soy protein.

110: Tremella dehydration step

120: First grinding step

130: Second grinding step

140: First mixing step

150: Second mixing step

160: Third mixing step

170: Cooking Step

180: Steps for the pH value of the slip and the blending solvent

190: Detection steps

Claims (8)

A preparation method of white fungus composition, comprising a dehydration step of white fungus, dehydrating a fresh white fungus; a first grinding step, grinding the dehydrated white fungus; a second grinding step, grinding a dietary fiber; A mixing step, adding if disturbing starch to the ground dietary fiber to mix evenly; a second mixing step, adding the ground white fungus and a high protein to the dietary fiber; a third mixing step, in the mixed white fungus, A nucleic acid solution is added to the high protein and dietary fiber; a decoction step is to heat the previously mixed white fungus, high protein and dietary fiber at a pressure of 0.6 MPa and 65 to 95 degrees Celsius for 20 minutes; a pH adjustment step to adjust The above-mentioned pH value of white fungus, high protein and dietary fiber after heating is to fix the pH value of white fungus between 5.68-5.83, and adjust the pH value of the whole solvent between 7.05 and 7.45 through whey protein to obtain white fungus. combination. The method according to claim 1, wherein, in the first grinding step, 3 to 10 pieces of dried white fungus are taken out, each piece of white fungus is about 10 to 15 cm in diameter, ground for 15 to 20 minutes to form a powder, and the sieve; in the second grinding step, MES/TRIS buffer solution is added when the dietary fiber is ground, and the pH value is adjusted to 8.2, then amylase is added to heat at 65-95 degrees Celsius, and water-soluble meals are obtained after centrifugation, heating and drying fiber; the composition ratio of each component in the second mixing step is as follows: the weight percent of this white fungus is between 15~30%, the weight percent of this dietary fiber is between 10~20%, the weight percent of this high protein is 40% % or less; in the third mixing step, water is added to cover the white fungus, the dietary fiber and the high protein. The method of claim 2, wherein the tremella dehydration step Among them, Tremella is heated between 65 and 95 degrees Celsius to produce polysaccharides with molecular weights between 9,000 and 10,000 Da. The method of claim 2, wherein the weight percentage (wt%) of the white fungus, dietary fiber, high protein and nucleic acid solution is 8:8:3:2. A tremella composition prepared by the method of claim 1, comprising a tremella in powder form with a particle size of between 1.6 microns and 25 microns; a dietary fiber, which is vegetable gum or algae extract; And a high protein, the high protein is whey protein. The Tremella composition as claimed in claim 5, wherein the nucleic acid solution is DNA extracted from Tremella. protein, RNA. For protein and vitamin groups, the weight ratio of various components is: DNA protein is 0-40%, RNA protein is 0-40%, and vitamins are less than 10%. The Tremella composition according to claim 5, wherein the dietary fiber is one of vegetable gums such as Tremella polysaccharide fermentation, glucose fermentation or sucrose fermentation. The white fungus composition according to claim 5, wherein the overall pH value of the white fungus, the dietary fiber, the high protein and the nucleic acid solution is between 7.05 and 7.45.

Images