CN107936129B - Preparation process of full-series molecular weight tremella polysaccharide - Google Patents
Preparation process of full-series molecular weight tremella polysaccharide Download PDFInfo
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Abstract
The invention discloses a preparation process of a full series of molecular weight tremella polysaccharides, which comprises the working procedures of extraction, impurity removal and decoloration, coarse filtration, fine filtration, concentration, precipitation, suction filtration and drying, and is characterized in that: the extraction process comprises two extraction steps of steam pressure-maintaining boiling extraction and high-efficiency enzyme activity promoter combined complex enzyme extraction, and the method has the beneficial effects that: the polyoxypropylene polyoxyethylene glyceryl ether added in the steam pressure-maintaining boiling extraction effectively solves the problem of foaming, and meanwhile has a remarkable promoting effect on the enzymolysis efficiency of subsequent acid protease, cellulase and pectinase.
Description
The technical field is as follows:
the invention belongs to the technical field of cosmetic material production equipment, and particularly relates to a preparation process of a full-series molecular weight tremella polysaccharide.
Background art:
tremella fuciformis (Tremellla fuciformis), also known as tremella fuciformis, tremella sanguinea and Chrysanthemum morifolium, is a colloid edible fungus and a medicinal fungus with high economic value and rich nutrition.
The tremella polysaccharide, also called tremella heteropolysaccharide, is also considered as a plant-derived hyaluronic acid, has a molecular weight of more than 150 ten thousand, and is the only plant-derived humectant with a molecular weight of more than 100 ten thousand in the cosmetic raw material world so far. The tremella polysaccharide has high-efficiency moisturizing capability, has certain capability of removing superoxide radicals and hydroxyl radicals, and can prevent skin aging, increase skin cell activity, promote skin regeneration, delay skin aging and repair photodamaged skin by inhibiting peroxidation of cell lipid. In addition, the skin-friendly oil has good skin-friendly property and higher viscosity, has obvious lubrication feeling when being added into a skin care product for smearing, and feels moist, dry, not tight and smooth after being used. The molecular weight of the tremella polysaccharide has great influence on the viscosity of the aqueous solution of the tremella polysaccharide, the larger the molecular weight is, the higher the viscosity of the aqueous solution is, the tremella polysaccharide with different molecular weights is applied to cosmetics, the tremella polysaccharide has different functions, and the tremella polysaccharide with high molecular weight (100-160 ten thousand Da) can form a compact protective film on the surface of skin, so that long-acting moisture preservation can be realized; the tremella polysaccharide with medium and low molecular weight (1-100 ten thousand Da) has good moisture retention and lubricity, and can stabilize emulsification and reduce the addition of emulsifier. The Tremella polysaccharide with ultra-low molecular weight (3000-1 ten thousand Da) can be absorbed through skin, and has effects of deep moisturizing, antiaging and repairing after sunburn. The antioxidant and anti-aging activities of tremella polysaccharides with different molecular weights are different. At present, more tremella polysaccharides sold in the market are in food grade, the number of cosmetic grades is small, the problems of low purity, low light transmittance of solution and the like exist, and the tremella polysaccharides with high purity and different molecular weight sections, which can be applied to the cosmetic industry, are greatly demanded at home and abroad.
At present, the production process of the tremella polysaccharide mainly adopts the working procedures of extraction, decoloration, purification, concentration, precipitation, dehydration, suction filtration, drying and the like;
the conventional extraction method of the tremella polysaccharide comprises 4 methods of hot water extraction, alkaline extraction, enzymatic extraction, ultrasonic-assisted extraction and the like, pectinase and 2 enzymes are mostly adopted in the report of the enzymatic extraction of the tremella polysaccharide, for example, a national invention patent with the patent number of 201410845693.4 discloses a method for purifying the edible fungus polysaccharide by an enzyme pretreatment-microwave-assisted extraction-chitosan flocculation technology, a compound enzyme system is utilized, the enzymes are selected from cellulase, pectinase or a mixture of the cellulase and the pectinase, but a simple compound enzyme system is used for extraction, the efficiency is low, no high-efficiency enzyme activity promoter is reported, particularly, part of cell structures are damaged in a steam pressure-maintaining boiling extraction method, part of proteins are denatured and dissolved out, acid protease is added for enzymolysis of glycoprotein capable of cracking part of cell walls of tremella cell walls, lipoprotein of cell membranes and part of intracellular proteins, the enzymolysis of the intercellular substance of pectinase and the enzymolysis of cellulose cell walls are, the technical scheme of the patent cannot effectively solve the problems.
The steam pressure-maintaining boiling extraction method is a new extraction method, can promote the dissolution of polysaccharide, denatures protein, is beneficial to subsequent enzymolysis treatment and removal of foreign protein, is beneficial to shortening the extraction time and improving the extraction rate, and a national invention patent with the patent number of 201611128959.9 discloses a compound algal polysaccharide blood fat-reducing oral liquid and a preparation method thereof, wherein the algal polysaccharide is obtained by utilizing high-pressure steam extraction, but because part of protein is dissolved in the high-pressure steam extraction process, foam is easy to generate, and the patent does not solve the problems;
the membrane distillation technology is a novel membrane separation technology, and takes the steam pressure difference at two sides of a hydrophobic membrane as a mass transfer driving force, and utilizes the characteristics of the hydrophobic membrane that only air permeability and water impermeability are realized to achieve the purpose of concentrating the feed liquid, so that the feed liquid can be concentrated to the maximum extent, the using amount of ethanol for precipitation can be greatly saved after concentration, and the cost can be saved. At present, the membrane distillation technology is mainly applied to the aspects of sea water desalination, high-concentration brackish water desalination and the like, and can also be applied to the concentration and purification of aqueous solution, but at present, only the membrane distillation is mentioned in the high-purity extraction method of tremella polysaccharide with the application number of 201710712040.2 in the preparation field of tremella polysaccharide, but the patent only mentions the adoption of the method and does not relate to all conditions required by the process and the like. The tremella polysaccharide is a high molecular polymer, the molecular weight distribution range is wide, and the conditions required by tremella polysaccharide with different molecular weights have certain specificity. In order to increase the membrane filtration speed, a filter membrane with a larger membrane pore size can be considered, but excessive expansion of the membrane pore size may cause material to be pumped from the feed liquid side to the vacuum side, so that material leakage brings additional problems, how to balance the membrane pore size and the membrane filtration speed, and how to increase the distillation speed by some means is the focus of research.
In conclusion, the simple and efficient preparation process of the tremella polysaccharide with high purity, which is developed by the new technical and novel method, has important practical significance and application value.
The invention content is as follows:
in order to solve the problems and overcome the defects of the prior art, the invention provides a preparation process of a full series of tremella polysaccharides with high extraction rate and good product quality, and can effectively solve the problems of low extraction rate and poor product quality.
The specific technical scheme for solving the technical problems comprises the following steps: the preparation process of the full-series molecular weight tremella polysaccharide comprises the working procedures of extraction, impurity removal and decoloration, coarse filtration, fine filtration, concentration, precipitation, suction filtration and drying, and is characterized in that: the extraction process comprises steam pressure-maintaining boiling extraction and high-efficiency enzyme activity promoter combined complex enzyme extraction;
the steam pressure maintaining boiling extraction comprises the following steps: putting the whole tremella into an extraction pot, adding 0.02-0.8% of polyoxypropylene polyoxyethylene glycerol ether, heating by a steam jacket, introducing steam into the pot after the temperature of the material is raised to 95 ℃, keeping the material liquid in a boiling state, keeping the pressure of the pot at 0.01-0.1MPa, keeping the temperature of the material liquid at 110-121 ℃, keeping the pressure for boiling extraction for 5min-4h, and after the reaction is finished, crushing the material liquid by a colloid mill wet method, and controlling the particle size of solid suspended matters to be 10-100 mu m to obtain a primary extracting solution;
the extraction of the high-efficiency enzyme activity promoter combined with the complex enzyme comprises the following steps: adding 1-4% of sodium acetate and 0.1-0.3% of magnesium sulfate into the primary extract obtained by steam pressure-maintaining boiling extraction, wherein the complex enzyme is a mixture of two or more of acid protease, pectinase or cellulase, the enzymolysis temperature is 40-60 ℃, the pH value is 2.5-5.0, the enzymolysis time is 2-6h, and the dosage of the enzyme is 0.1-3% of the dry weight of the tremella fuciformis, so as to obtain an enzymolysis treatment solution;
the impurity removal and decoloration process comprises the following steps: swelling a flocculating agent in a 1% acetic acid solution to obtain a flocculating agent solution, slowly adding the flocculating agent solution into the enzymolysis treatment solution under a stirring state, and carrying out heat preservation flocculation and adsorption for 0.5-2h at the flocculation and adsorption temperature of 40-70 ℃ to obtain a feed liquid after impurity removal and decoloration; the flocculant is chitosan, and the dosage of the chitosan is 0.1-0.5% of the enzymolysis treatment liquid;
the coarse filtration and fine filtration process comprises the following steps: the coarse filtration process comprises the steps of adding 2-10% of an auxiliary agent into the feed liquid after impurity removal and decoloration, wherein the auxiliary agent is one or a mixture of more than two of sodium chloride, sodium acetate or potassium chloride, and filtering the feed liquid by using a 200-mesh filter cloth to remove and extract residual tremella residue and flocculating agent chitosan to obtain a coarse filtrate; the fine filtration process adopts a clarification filter plate with the filtration precision of 5 mu m, and adopts a perlite and diatomite ratio of 1: 1-5 as filter aid, adjusting pH of the feed liquid to 9.0-10.0, and filtering the coarse filtrate to obtain fine filtrate;
the concentration step includes: adjusting the pH of the fine filtrate to 7.0, and concentrating the fine filtrate by vacuum distillation under reduced pressure by using a PTFE hollow fiber membrane with a membrane aperture of 0.45 μm, wherein the temperature of the fine filtrate is kept at 70-95 ℃ during concentration, and the concentration is carried out until the volume is reduced to half of the original volume to obtain a concentrated solution;
the precipitation, suction filtration and drying process comprises the following steps: slowly adding absolute ethyl alcohol into the concentrated solution under the stirring condition for precipitation, wherein the alcohol degree at the ethanol precipitation end point is 58-60 degrees, standing for 1h after precipitation, removing supernatant alcohol, and then adding 1-2.5 times of volume of absolute ethyl alcohol to dehydrate precipitates; carrying out suction filtration on the dehydrated precipitate by using vacuum filtration equipment; and (3) drying the dried precipitate in vacuum drying equipment at the drying temperature of 50-80 ℃ and the vacuum degree of 0.09-0.1MPa to obtain the finished product of the tremella polysaccharide.
Furthermore, in the process of extracting the high-efficiency enzyme activity promoter by combining with the complex enzyme, the enzymolysis sequence of the complex enzyme is set to be a mode of adding enzymolysis at the same time or a mode of carrying out enzymolysis by enzyme sequences with different time periods.
Further, the tremella polysaccharide finished product is of a series of molecular weights, the molecular weight is 160 ten thousand-5000 Da, the color of the product is white powder, the total sugar content is greater than 95%, the content of foreign proteins is less than 0.1%, the light transmittance of 0.1% tremella polysaccharide solution is more than 99.8%, the drying weight loss is less than 8%, and the extraction rate of a target product is 16-21%.
The invention has the beneficial effects that: the polyoxypropylene polyoxyethylene glycerol ether added in the steam pressure-maintaining boiling extraction effectively solves the problem of foaming, and meanwhile, the polyoxypropylene polyoxyethylene glycerol ether is unexpectedly found to have a remarkable promoting effect on the subsequent enzymolysis efficiency of acid protease, cellulase and pectinase, so that the extraction rate of polysaccharide is improved; the high-efficiency enzyme activity promoter is unexpectedly found in the complex enzyme extraction, the extraction rate is different due to the different enzymolysis sequences of the complex enzyme, the molecular weight distribution of the tremella polysaccharide whole series is different due to different boiling extraction process conditions, and the diversity and the flexibility of the product are improved; the unexpected discovery that the auxiliary agent in the rough filtration process can not only improve the state of the precipitated material and make the precipitated material in uniformly dispersed fine particles, but also improve the distillation speed in the next membrane distillation process.
The specific implementation mode is as follows:
in the description of the invention, specific details are given only to enable a full understanding of the embodiments of the invention, but it should be understood by those skilled in the art that the invention is not limited to these details for the implementation. In other instances, well-known structures and functions have not been described or shown in detail to avoid obscuring the points of the embodiments of the invention. The specific meanings of the above terms in the present invention can be understood in specific cases to those skilled in the art.
The specific implementation mode of the invention is as follows:
example 1
Selecting 3kg of mildew-free whole tremella raw materials for each group, putting the raw materials into an extraction pot, adding 0.2% of defoaming agent and enzymolysis accelerator polyoxypropylene polyoxyethylene glycerol ether, introducing a steam jacket for heating and raising the temperature, introducing steam into a tank for boiling extraction after the temperature is raised to 95 ℃, maintaining the temperature of feed liquid for boiling extraction at 110-121 ℃, and extracting the tremella raw materials for each group under the tank pressure and the pressure-maintaining boiling extraction time as shown in the following table 1:
after the material liquid after boiling extraction is discharged, wet grinding is carried out by adopting a colloid mill, and the particle size of the solid suspended matter after grinding is 80 mu m. Extracting the feed liquid after wet grinding by adopting a high-efficiency enzyme activity promoter and combining with a complex enzyme, adding 2% of sodium acetate and 0.2% of magnesium sulfate into the feed liquid to serve as an enzymolysis promoter, wherein the complex enzyme is acid protease, pectinase and cellulase, the enzymolysis temperature is 50 ℃, the pH value is 3.5, the enzymolysis time is 3 hours, and the dosage of the enzyme is 1% of the dry weight of the tremella. After enzymolysis, deproteinizing and decolorizing by adopting a chitosan flocculation method, wherein the dosage of a flocculating agent chitosan is 0.1 percent of the extraction liquid, slowly adding a flocculating agent solution under the stirring state, and carrying out heat preservation flocculation and adsorption for 1 hour at the flocculation and adsorption temperature of 40 ℃. And (3) performing coarse filtration and deslagging after flocculation, adding 8% of sodium chloride into the feed liquid to improve the filtration performance of the feed liquid and contribute to the improvement of the state of precipitates in the subsequent alcohol precipitation process, filtering the feed liquid by adopting 200-mesh filter cloth to remove and extract residual tremella residues and flocculating agent chitosan, and separately collecting filter residues for later use. And after rough filtration, plate-and-frame fine filtration is adopted, the pH of the feed liquid is adjusted to 9.5, a clarifying filter plate with the filtering precision of 5 mu m is adopted, perlite and diatomite which are mixed according to the proportion of 1:1 are adopted as filter aids, the pH of the feed liquid after fine filtration is adjusted to 7.0, and the feed liquid is concentrated by adopting a PTFE hollow fiber membrane with the membrane aperture of 0.45 mu m to carry out vacuum membrane distillation, wherein the temperature of the feed liquid is kept at 80 ℃ during concentration, and the feed liquid is concentrated until the volume of the feed liquid is reduced to half of the original volume. Slowly adding anhydrous ethanol into the concentrated feed liquid under stirring for precipitation, wherein the alcohol degree at the ethanol precipitation end point is 60 degrees, standing for 1h after precipitation, removing supernatant alcohol, and dehydrating the precipitate with 1.5 times of volume of anhydrous ethanol. And (4) after dehydration, using vacuum filtration equipment to filter and dry the dehydrated precipitate. And (3) drying the dried precipitate in vacuum drying equipment at 60 ℃ under the vacuum degree of 0.09-0.1MPa for 6 h. The prepared tremella polysaccharide products are all white powder with good color, and other indexes of the products are shown in table 1:
TABLE 1 preparation of Tremella polysaccharide products of different molecular weights under different boiling extraction conditions
As can be seen from Table 1: different boiling extraction process conditions have little influence on the total sugar content, the light transmittance, the chromaticity b value and the extraction rate of the product, have larger influence on the molecular weight of the product, and the tremella polysaccharides with different molecular weights have different biological activities and purposes, so that the distribution of the tremella polysaccharides with the full series of molecular weights is increased by the different boiling extraction process conditions, and the application range of the product is enlarged.
Example 2
In order to more intuitively show the process advantages of the composite enzymolysis, the novel process technology of the invention for the customized preparation of the high-purity tremella polysaccharide with a full series of molecular weights, different enzymes, different enzymolysis and different enzymolysis promoters are used for the preparation of the tremella polysaccharide, and the grouping and experimental results are shown in the following table 2 and comprise the following steps:
selecting 3kg of mildew-free whole tremella raw material for each group, putting the raw material into an extraction pot, wherein the material-water ratio is 1:70, 0.2% of defoaming agent and enzymolysis accelerant polyoxypropylene polyoxyethylene glycerol ether are added or not added, heating by introducing a steam jacket, introducing steam into a tank for boiling extraction after the temperature is raised to 95 ℃, keeping the temperature of the material liquid for boiling extraction at 110-121 ℃, keeping the tank pressure of each group at 0.05MPa, and keeping the pressure for boiling extraction for 1 h; after the material liquid after boiling extraction is discharged, wet grinding is carried out by adopting a colloid mill, and the particle size of the solid suspended matter after grinding is 40 mu m. And carrying out enzymolysis and extraction on the feed liquid crushed by the wet method by adopting a high-efficiency enzyme activity promoter combined with a complex enzyme according to the grouping shown in the table 1, wherein the enzymolysis promoter is 2% of sodium acetate or 0.02% of magnesium sulfate or the combination of the sodium acetate and the magnesium sulfate, the complex enzyme is three enzymes of acid protease, pectinase and cellulase, the enzymolysis temperature is 45 ℃, the pH value is 4.0, the enzymolysis time is 4 hours, and the dosage of the enzyme is 1.5% of the dry weight of the tremella fuciformis. After enzymolysis, deproteinizing and decolorizing by adopting a chitosan flocculation method, wherein the dosage of a flocculating agent chitosan is 0.2 percent of the extraction liquid, slowly adding a flocculating agent solution under the stirring state, and carrying out heat preservation flocculation and adsorption for 1 hour at the flocculation and adsorption temperature of 50 ℃. And (3) performing coarse filtration and deslagging after flocculation, adding 6% of sodium chloride into the feed liquid to improve the filtration performance of the feed liquid and contribute to the improvement of the state of precipitates in the subsequent alcohol precipitation process, filtering the feed liquid by adopting 200-mesh filter cloth to remove and extract residual tremella residues and flocculating agent chitosan, and separately collecting filter residues for later use. After rough filtration, plate-frame fine filtration is adopted, the pH of the feed liquid is adjusted to 9.8, a clarifying filter plate with the filtering precision of 5 mu m is adopted, and perlite and diatomite which are mixed according to the ratio of 1:1 are adopted as filter aids. Adjusting the pH value of the finely filtered feed liquid to 7.0, and concentrating the feed liquid by adopting a PTFE hollow fiber membrane with a membrane aperture of 0.45 mu m to perform vacuum membrane distillation, wherein the temperature of the feed liquid is kept at 75 ℃ during concentration, and the feed liquid is concentrated until the volume of the feed liquid is reduced to half of the original volume. Slowly adding anhydrous ethanol into the concentrated feed liquid under stirring for precipitation, wherein the alcohol degree at the ethanol precipitation end point is 60 degrees, standing for 1h after precipitation, removing supernatant alcohol, and dehydrating the precipitate with 1.5 times of volume of anhydrous ethanol. And (4) after dehydration, using vacuum filtration equipment to filter and dry the dehydrated precipitate. And (3) drying the dried precipitate in vacuum drying equipment at 60 ℃ under the vacuum degree of 0.09-0.1MPa for 6 h. The prepared tremella polysaccharide products are all white powder with good color, and other indexes of the products are shown in table 2:
TABLE 2 Effect of different enzymatic conditions on the Process
As can be seen from Table 2: the extraction rate is improved from 12.2% to 17.7% by comparing the group 1 and the group 10; from a comparison of groups 4 and 9: when the enzyme promoter is added, the extraction rate is improved from 20.1 percent to 21.4 percent compared with the simultaneous addition of the enzyme preparation in sequence; from groups 9 and 10, it can be seen that: when the enzyme preparation is added in sequence, the extraction rate is improved from 17.7% to 21.4% by adding the enzyme promoter compared with the extraction rate without adding the enzyme promoter, so the addition sequence of the compound enzyme system and the addition of the enzymolysis promoter have great beneficial effects on the extraction rate.
Example 3
In order to more intuitively show the technological advantages of the steam pressure-maintaining boiling extraction, 3kg of mildew-free whole tremella raw materials are selected for each group, the raw materials are put into an extraction pot, the material-water ratio is 1:40, 0.2% of defoaming agent is added or not added according to the grouping of the following table 3, a steam jacket is firstly introduced for heating and temperature rise, steam is introduced into the pot for boiling extraction after the temperature rises to 95 ℃, the temperature of the feed liquid for boiling extraction is maintained at 110-121 ℃, the pot pressure of each group is 0.06MPa, and the pressure-maintaining boiling extraction time is 1 h; after the material liquid after boiling extraction is discharged, wet grinding is carried out by adopting a colloid mill, and the particle size of the solid suspended matter after grinding is 50 mu m. And respectively adding 2% of sodium acetate and 0.02% of magnesium sulfate into the feed liquid obtained after wet grinding to serve as enzymolysis promoters, and performing complex enzyme enzymolysis and extraction by using three enzymes of acid protease, pectinase and cellulase at 45 ℃ and pH4.0 for 4h, wherein the enzyme consumption is 1% of the dry weight of the tremella. After enzymolysis, deproteinizing and decolorizing by adopting a chitosan flocculation method, wherein the dosage of a flocculating agent chitosan is 0.3 percent of the extraction liquid, slowly adding a flocculating agent solution under the stirring state, and carrying out heat preservation flocculation and adsorption for 1 hour at the flocculation and adsorption temperature of 50 ℃. And (3) performing coarse filtration and deslagging after flocculation, adding 8% of sodium chloride into the feed liquid to improve the membrane distillation rate, filtering the feed liquid by adopting 200-mesh filter cloth to remove and extract residual tremella slag and flocculating agent chitosan, and separately collecting filter residues for later use. After rough filtration, plate-frame fine filtration is adopted, the pH of the feed liquid is adjusted to 9.8, a clarifying filter plate with the filtering precision of 5 mu m is adopted, and perlite and diatomite which are mixed according to the ratio of 1:2 are adopted as filter aids. Adjusting the pH value of the finely filtered feed liquid to 7.0, and concentrating the feed liquid by adopting a PTFE hollow fiber membrane with a membrane aperture of 0.45 mu m to perform vacuum membrane distillation, wherein the temperature of the feed liquid is kept at 75 ℃ during concentration, and the feed liquid is concentrated until the volume of the feed liquid is reduced to half of the original volume. Slowly adding anhydrous ethanol into the concentrated feed liquid under stirring for precipitation, wherein the alcohol degree at the ethanol precipitation end point is 60 degrees, standing for 1h after precipitation, removing supernatant alcohol, and dehydrating the precipitate with 1.5 times of volume of anhydrous ethanol. And (4) after dehydration, using vacuum filtration equipment to filter and dry the dehydrated precipitate. And (3) drying the dried precipitate in vacuum drying equipment at 60 ℃ under the vacuum degree of 0.09-0.1MPa for 6 h. The prepared tremella polysaccharide products are all white powder with good color, and the foreign protein indexes and extraction rates of the products are shown in the following table:
TABLE 3 steam pressure-maintaining boiling extraction contrast test
As can be seen from Table 3: compared with other defoaming agents, the polyoxypropylene polyoxyethylene glycerol ether has a remarkable promoting effect on the enzymolysis efficiency of subsequent acid protease, cellulase and pectinase, so that the enzymolysis efficiency of the complex enzyme and the extraction rate of the tremella polysaccharide are improved.
Example 4
In order to more intuitively show the process advantages of the membrane distillation technology of the invention, a novel process technology for the customizable preparation of a full-series of high-purity tremella polysaccharides with molecular weights, a comparative example for preparing the tremella polysaccharides under improved membrane distillation process conditions, and grouping and experimental results are shown in table 3 below, and the method comprises the following steps:
selecting 3kg of mildew-free whole tremella raw material for each group, putting the raw material into an extraction pot, adding 0.2% of defoaming agent and enzymolysis accelerant polyoxypropylene polyoxyethylene glycerol ether at a material-water ratio of 1:40, introducing steam into a steam jacket for heating and raising the temperature, introducing steam into a tank for boiling extraction after the temperature is raised to 95 ℃, keeping the temperature of feed liquid for boiling extraction at 110-121 ℃, keeping the tank pressure of each group at 0.06MPa, and keeping the pressure for boiling extraction for 1 h; after the material liquid after boiling extraction is discharged, wet grinding is carried out by adopting a colloid mill, and the particle size of the solid suspended matter after grinding is 50 mu m. And respectively adding 2% of sodium acetate and 0.02% of magnesium sulfate into the feed liquid obtained after wet grinding to serve as enzymolysis promoters, and performing complex enzyme enzymolysis and extraction by using three enzymes of acid protease, pectinase and cellulase at 45 ℃ and pH4.0 for 4h, wherein the enzyme consumption is 1% of the dry weight of the tremella. After enzymolysis, deproteinizing and decolorizing by adopting a chitosan flocculation method, wherein the dosage of a flocculating agent chitosan is 0.3 percent of the extraction liquid, slowly adding a flocculating agent solution under the stirring state, and carrying out heat preservation flocculation and adsorption for 1 hour at the flocculation and adsorption temperature of 50 ℃. And (3) performing coarse filtration and deslagging after flocculation, adding or not adding 8% of sodium chloride into the feed liquid according to the grouping shown in the table 3 so as to improve the membrane distillation rate, filtering the feed liquid by adopting 200-mesh filter cloth to remove and extract residual tremella residues and flocculating agent chitosan, and separately collecting filter residues for later use. After rough filtration, plate-frame fine filtration is adopted, the pH of the feed liquid is adjusted to 9.8, a clarifying filter plate with the filtering precision of 5 mu m is adopted, and perlite and diatomite which are mixed according to the ratio of 1:2 are adopted as filter aids. And (3) adjusting the pH value of the feed liquid after fine filtration to 7.0, concentrating the feed liquid by adopting PTFE hollow fiber membranes with different membrane apertures to perform vacuum membrane distillation according to the grouping shown in the table 3, and concentrating the feed liquid to half of the original volume according to the grouping shown in the table 3. Slowly adding anhydrous ethanol into the concentrated feed liquid under stirring for precipitation, wherein the alcohol degree at the ethanol precipitation end point is 60 degrees, standing for 1h after precipitation, removing supernatant alcohol, and dehydrating the precipitate with 1.5 times of volume of anhydrous ethanol. And (4) after dehydration, using vacuum filtration equipment to filter and dry the dehydrated precipitate. And (3) drying the dried precipitate in vacuum drying equipment at 60 ℃ under the vacuum degree of 0.09-0.1MPa for 6 h. The prepared tremella polysaccharide products of each group are all white powder with good color, and the intermediate detection indexes of each group are shown in table 4:
TABLE 4 comparative experimental data for membrane distillation technology
As can be seen from Table 4, the auxiliary agent in the rough filtration process improves the state of the precipitated material and increases the distillation speed of the membrane distillation process.
In summary, the following steps: the polyoxypropylene polyoxyethylene glycerol ether added in the steam pressure-maintaining boiling extraction effectively solves the problem of foaming, and meanwhile, the polyoxypropylene polyoxyethylene glycerol ether is unexpectedly found to have a remarkable promoting effect on the subsequent enzymolysis efficiency of acid protease, cellulase and pectinase, so that the enzymolysis efficiency of complex enzyme and the extraction rate of tremella polysaccharide are improved; the high-efficiency enzyme activity promoter is discovered unexpectedly in the complex enzyme extraction, the difference of the extraction rate caused by the difference of the enzymolysis sequence of the complex enzyme is discovered unexpectedly, and the different boiling extraction process conditions can cause the different molecular weight distributions of the whole series of tremella polysaccharides, thereby increasing the diversity and the flexibility of the product; the auxiliary agent in the rough filtration process can improve the state of the precipitated material to ensure that the precipitated material is in uniformly dispersed fine particles, and also plays a role in improving the distillation speed in the next membrane distillation process.
Claims (2)
1. A preparation process of a full series of molecular weight tremella polysaccharide comprises the working procedures of extraction, impurity removal and decoloration, coarse filtration, fine filtration, concentration, precipitation, suction filtration and drying, and is characterized in that: the extraction process comprises steam pressure-maintaining boiling extraction and high-efficiency enzyme activity promoter combined complex enzyme extraction;
the steam pressure maintaining boiling extraction comprises the following steps: putting the whole tremella into an extraction pot, adding 0.02-0.8% of polyoxypropylene polyoxyethylene glycerol ether, heating by a steam jacket, introducing steam into the pot after the temperature of the material is raised to 95 ℃, keeping the material liquid in a boiling state, keeping the pressure of the pot at 0.01-0.1MPa, keeping the temperature of the material liquid at 110-121 ℃, keeping the pressure for boiling extraction for 5min-4h, and after the reaction is finished, crushing the material liquid by a colloid mill wet method, and controlling the particle size of solid suspended matters to be 10-100 mu m to obtain a primary extracting solution;
the extraction of the high-efficiency enzyme activity promoter combined with the complex enzyme comprises the following steps: adding 1-4% of sodium acetate and 0.1-0.3% of magnesium sulfate into the primary extract obtained by steam pressure-maintaining boiling extraction, wherein the complex enzyme is a mixture of two or more of acid protease, pectinase or cellulase, the enzymolysis temperature is 40-60 ℃, the pH value is 2.5-5.0, the enzymolysis time is 2-6h, and the dosage of the enzyme is 0.1-3% of the dry weight of the tremella fuciformis, so as to obtain an enzymolysis treatment solution;
the impurity removal and decoloration process comprises the following steps: swelling a flocculating agent in a 1% acetic acid solution to obtain a flocculating agent solution, slowly adding the flocculating agent solution into the enzymolysis treatment solution under a stirring state, and carrying out heat preservation flocculation and adsorption for 0.5-2h at the flocculation and adsorption temperature of 40-70 ℃ to obtain a feed liquid after impurity removal and decoloration; the flocculant is chitosan, and the dosage of the chitosan is 0.1-0.5% of the enzymolysis treatment liquid;
the coarse filtration and fine filtration process comprises the following steps: the coarse filtration process comprises the steps of adding 2-10% of an auxiliary agent into the feed liquid after impurity removal and decoloration, wherein the auxiliary agent is one or a mixture of more than two of sodium chloride, sodium acetate or potassium chloride, and filtering the feed liquid by using a 200-mesh filter cloth to remove and extract residual tremella residue and flocculating agent chitosan to obtain a coarse filtrate; the fine filtration process adopts a clarification filter plate with the filtration precision of 5 mu m, and adopts a perlite and diatomite ratio of 1: 1-5 as filter aid, adjusting pH of the feed liquid to 9.0-10.0, and filtering the coarse filtrate to obtain fine filtrate;
the concentration step includes: adjusting the pH of the fine filtrate to 7.0, and concentrating the fine filtrate by vacuum distillation under reduced pressure by using a PTFE hollow fiber membrane with a membrane aperture of 0.45 μm, wherein the temperature of the fine filtrate is kept at 70-95 ℃ during concentration, and the concentration is carried out until the volume is reduced to half of the original volume to obtain a concentrated solution;
the precipitation, suction filtration and drying process comprises the following steps: slowly adding absolute ethyl alcohol into the concentrated solution under the stirring condition for precipitation, wherein the alcohol degree at the ethanol precipitation end point is 58-60 degrees, standing for 1h after precipitation, removing supernatant alcohol, and then adding 1-2.5 times of volume of absolute ethyl alcohol to dehydrate precipitates; carrying out suction filtration on the dehydrated precipitate by using vacuum filtration equipment; and (3) drying the dried precipitate in vacuum drying equipment at the drying temperature of 50-80 ℃ and the vacuum degree of 0.09-0.1MPa to obtain the finished product of the tremella polysaccharide.
2. The preparation process of the full-range molecular weight tremella polysaccharide as claimed in claim 1, wherein the efficient enzyme activity promoter is combined with a complex enzyme extraction process, and the enzymatic hydrolysis order of the complex enzyme is set to be a simultaneous enzymatic hydrolysis mode or a mode of enzymatic hydrolysis with different enzyme orders at different time intervals.
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