CN110787194A - Phellinus igniarius paste and preparation method thereof - Google Patents
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Abstract
The invention relates to a phellinus igniarius paste and a preparation method thereof, wherein the method comprises the following steps: (1) pulverizing mature Phellinus linteus fruiting body, sieving, adding sucrose and water, and preparing culture medium; (2) performing primary fermentation, and inoculating cellulose degrading bacteria; (3) secondary fermentation, inoculating yeast; grinding the pulp, and performing low-temperature ultrahigh-pressure treatment; (4) performing ultrafiltration on the mixed solution obtained in the step (3) to remove salt, small molecular impurities and pigments to obtain a concentrated solution; (5) carrying out low-temperature ultrahigh-pressure sterilization on the obtained concentrated solution; (6) freeze drying; (7) pressing into solid to obtain Phellinus Linteus paste. The present disclosure provides a new phellinus igniarius product, and enriches the variety of phellinus igniarius products. All the process steps of the method are completed at a lower temperature, and the activity of the effective ingredients is retained to the maximum extent.
Description
Technical Field
The present disclosure relates to a phellinus igniarius paste and a preparation method thereof.
Background
The information in this background section is only for enhancement of understanding of the general background of the disclosure and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art that is already known to a person of ordinary skill in the art.
The traditional Chinese medicine paste is a thick semi-fluid preparation which is prepared by decocting the traditional Chinese medicines with water, removing residues, concentrating and adding sugar or refined honey, and is also called soft extract. Although it has the advantages of high drug concentration, small volume, etc., the preparation process has high temperature step, easily inactivates heat-sensitive substances, and the finished product is easily polluted by microorganisms, and needs to be sealed and added with preservative.
At present, the reported phellinus igniarius products mainly comprise phellinus igniarius vinegar beverage, phellinus igniarius health oral liquid, phellinus igniarius yellow wine and the like, and a phellinus igniarius paste product taking phellinus igniarius sporocarp as a main raw material and a corresponding preparation method thereof are not reported yet.
Disclosure of Invention
In view of the above background technologies, the present disclosure provides a phellinus linteus paste prepared by a low temperature technology, which solves the problem that a thermosensitive substance is easily inactivated in the traditional paste preparation process, and also solves the problem that a finished product is easily contaminated by microorganisms, etc.
Specifically, the following technical scheme is adopted in the disclosure:
in a first aspect of the present disclosure, there is provided a method for preparing phellinus linteus paste, which comprises the steps of:
(1) pulverizing mature Phellinus linteus fruiting body, sieving, adding sucrose and water, and preparing culture medium;
(2) primary fermentation: inoculating cellulose degrading bacteria into the culture medium in the step (1), and fermenting for a certain time; after the fermentation is finished, grinding the mixture into thick liquid; after pulping, carrying out low-temperature ultrahigh-pressure sterilization;
(3) and (3) secondary fermentation: inoculating yeast into the slurry obtained in the step (2), and fermenting for a certain time; after fermentation is finished, carrying out solid-liquid separation to obtain fermentation liquor;
(4) performing ultrafiltration on the mixed solution obtained in the step (3) to remove salt, small molecular impurities and pigments to obtain a concentrated solution;
(5) low-temperature ultrahigh pressure: carrying out low-temperature ultrahigh-pressure sterilization on the obtained concentrated solution;
(6) freeze drying;
(7) pressing into solid to obtain Phellinus Linteus paste.
In the step (1), preferably, the species of Phellinus linteus include Phellinus linteus, Phellinus baumii and Phellinus igniarius, etc. In one or more embodiments of the present disclosure, the Phellinus linteus is Phellinus igniarius, and dried Phellinus linteus fruiting body with water content of 1.5 w/w% or less is used.
In the step (1), preferably, the powder is sieved by a sieve of 100-120 meshes. The grinding granularity is moderate, which is beneficial to the utilization of strains.
In the step (1), preferably, the addition amount of the sieved phellinus linteus powder, sucrose and water is (3-6) kg: (0.5-1.5) kg: (5-8) L.
Based on the characteristics of high-content crude fiber, complex components and hard texture of the phellinus igniarius sporocarp, the effective active ingredients in the phellinus igniarius sporocarp are not easy to release, the invention adopts cellulose degrading bacteria which secrete cellulase, hemicellulase, pectinase and the like by utilizing cane sugar and nitrogen sources contained in the sporocarp per se to efficiently degrade the crude fiber and cell walls in the phellinus igniarius sporocarp, so that the crude fiber is degraded into low-molecular carbohydrate and the like, the cell walls can be broken to release the active ingredients such as intracellular polysaccharide and the like, and the cellulose degrading hard bacteria can break the structure of the phellinus igniarius sporocarp to ensure that the effective active ingredients are easier to dissolve out.
In the step (2), the cellulose-degrading bacteria are complex bacteria or single bacteria, and are preferably complex bacteria in view of the degrading effect of crude phellinus linteus fibers. The inventor screens and optimizes the species and proportion of the composite bacteria aiming at the phellinus igniarius, and the obtained composite bacteria are a mixture of Bacillus subtilis, Bacillus licheniformis and Aspergillus carbonarius.
Wherein, the bacillus subtilis, the bacillus licheniformis and the aspergillus carbonarius are all strains for producing cellulase and/or hemicellulase and/or pectinase, and can be obtained by conventional commercial routes. In one or more embodiments of the present disclosure, bacillus subtilis is numbered cic 10088, bacillus licheniformis is numbered cic 10831, and aspergillus carbonarius is numbered cic 41254.
In the step (2), preferably, the inoculation amount of the cellulose-degrading bacteria is 1-1.5% (1 is inoculated per 100g of the culture medium)About 1.5mL of bacterial solution), 1mL of bacterial solution contains the following strains: the number of the bacillus subtilis, the bacillus licheniformis and the aspergillus carbonarius is (1-2) multiplied by 10 respectively10cfu、(2～4)×1010cfu and (1-2). times.1010cfu。
In the step (2), the fermentation conditions are as follows: the fermentation time is 2-3 days, the fermentation temperature is normal temperature or room temperature (preferably 25-35 ℃), and stirring or shake culture is carried out. The fermentation time is 2-3 days, so that the effective active ingredients in the phellinus igniarius sporocarp are fully dissolved, and the content of soluble solids is increased most obviously.
In the step (2), a low-temperature ultrahigh-pressure technology is adopted, so that cellulose degrading bacteria and active enzymes can be inactivated, the dissolution of active ingredients in fermentation liquor solids can be further promoted, and the problem that high-temperature sensitive active ingredients such as phellinus igniarius polysaccharides are easy to inactivate is solved by adopting the low-temperature ultrahigh-pressure technology. Preferably, the low temperature ultra high pressure conditions: the ultrahigh pressure is 400 MPa-500 MPa, the ultrahigh pressure time is 10 min-20 min, and the ultrahigh temperature is 25-50 ℃. Tests prove that the three requirements can be well met by adopting the process conditions.
In the test process, the prepared product has a bad flavor (green and astringent woody flavor) only by adopting cellulose degrading bacteria for fermentation, and yeast fermentation is carried out in order to optimize the flavor of the product and further increase nutrient substances.
In the step (3), preferably, the yeast is isolated to the surface of wild phellinus linteus fruiting body or the surrounding environment or is a purchased saccharomyces cerevisiae.
In the step (3), the yeast fermentation is natural fermentation at normal temperature, the fermentation time is 8-24 h, and the preferred temperature is 25-42 ℃.
In the step (4), the range of the cut-off molecular weight of the ultrafiltration membrane is 10000-15000 Da, and the content of effective active ingredients can be improved by selecting the cut-off molecular weight in the range through determination.
In the step (5), the low-temperature ultrahigh-pressure condition is as follows: the ultrahigh pressure is 400 MPa-500 MPa, the ultrahigh pressure time is 10 min-20 min, and the ultrahigh temperature is 25-50 ℃.
In the step (6), freeze drying is carried out for 16-18 hours, so that the water content of the product is less than or equal to 5 w/w%.
And (7) pressing the mixture into granules or blocky solids.
In a second aspect of the disclosure, the phellinus igniarius paste prepared by the method is characterized in that: the product is in solid form, the content of crude polysaccharide of Phellinus Linteus is not less than 30%, the content of total flavonoids is not less than 5%, the content of triterpenes is not less than 2%, and the water content is not more than 5%.
In a third aspect of the disclosure, an application of the phellinus linteus paste in preparing an anti-tumor or anti-oxidation medicine or health-care product is provided.
Compared with the related technology known by the inventor, one technical scheme of the present disclosure has the following beneficial effects:
(1) the present disclosure provides a new phellinus igniarius product, and enriches the variety of phellinus igniarius products.
(2) All the process steps of the method are completed at a lower temperature, and the activity of the effective ingredients is retained to the maximum extent.
(3) The method adopts a secondary fermentation process, so that the content of effective active ingredients in the final product is high, and the product has good flavor.
(4) Because the product of the present disclosure is in a solid form, it is easier to store and has an extended shelf life compared to the prior art of semi-solid fluid form pastes.
Drawings
The accompanying drawings, which are incorporated in and constitute a part of this disclosure, illustrate embodiments of the disclosure and, together with the description, serve to explain the disclosure and not to limit the disclosure.
Fig. 1 shows a phellinus linteus paste prepared in example 1 of the present disclosure.
Fig. 2 is a phellinus linteus liquid brewed in example 1 of the present disclosure.
Detailed Description
It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of example embodiments according to the present disclosure. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of the stated features, steps, operations, and/or combinations thereof, unless the context clearly indicates otherwise.
In order to make the technical solutions of the present disclosure more clearly understood by those skilled in the art, the technical solutions of the present disclosure will be described in detail below with reference to specific embodiments.
Example 1
A preparation method of phellinus igniarius paste comprises the following steps:
(1) pulverizing mature dried Phellinus igniarius fruiting body, grinding, sieving with 100 mesh sieve, adding sucrose and water, and mixing well to prepare culture medium; wherein, the adding amount of the phellinus igniarius powder, the cane sugar and the water is 5 kg: 2 kg: 8L of the compound;
(2) primary fermentation: the adopted cellulose degrading bacteria are composite strains consisting of bacillus subtilis CICC 10088, bacillus licheniformis CICC10831 and aspergillus carbonarius CICC 41254;
inoculating cellulose-degrading bacteria into the culture medium in the step (1), and fermenting for 48 hours at room temperature and about 30 ℃ in a shaking table manner, wherein the rotation speed of the shaking table is 150 rpm;
wherein the inoculation amount of the cellulose-degrading bacteria is that 1.5mL of cellulose-degrading bacteria solution is inoculated per 100g of culture medium, and each mL of the cellulose-degrading bacteria solution contains 2 multiplied by 1010cfu Bacillus subtilis CICC 10088, 2 × 1010cfu Bacillus licheniformis CICC10831 and 1 × 1010cfu aspergillus carbonarius CICC 41254;
grinding the mixture into pulp after the fermentation is finished, so as to break the cell wall again and further release the effective active ingredients in the cells;
performing low-temperature ultrahigh-pressure treatment after grinding, wherein the pressure is 400MPa, the time is 15min, and the temperature is 30 ℃;
(3) and (3) secondary fermentation: activating Saccharomyces cerevisiae CICC 1839 with brown sugar water, and then adding into the slurry obtained in step (2) for natural normal temperature fermentation, wherein the inoculation amount is as follows: 5mL of the bacterial suspension was inoculated per 100mL of the slurry, and 1X 10 of the bacterial suspension was contained in each mL of the bacterial suspension10The fermentation time of cfu yeast is 12 hours;
after fermentation, centrifuging to obtain supernatant and precipitate; washing the precipitate, and combining the washing liquid into the supernatant;
(4) performing ultrafiltration desalination and concentration on the mixed solution obtained in the step (3), wherein the cut-off molecular weight range of an ultrafiltration membrane is 12000Da, so as to obtain a concentrated solution;
(5) low-temperature ultrahigh pressure: sterilizing the obtained concentrated solution at low temperature and ultrahigh pressure, wherein the pressure is 400MPa, the time is 15min, and the temperature is 30 ℃;
(6) freeze drying in a freeze drier overnight;
(7) pressing the powder into block solid with a briquetting machine under 15MPa to obtain Phellinus linteus paste, and further packaging with food packaging paper.
According to the detection by the conventional method, the content of crude polysaccharide substances is 35.3 percent (namely 353mg/1g), the content of total flavonoids substances is 6.5 percent (namely 65mg/1g), the content of triterpenoid substances is 3.3 percent (namely 33mg/1g), the water content is less than or equal to 5w/w percent, and the product photo is shown in figure 1 and has dark brown-yellow color.
The eating method comprises the following steps: soaking in warm water below 60 deg.C, stirring or shaking to make Phellinus Linteus paste be soaked, and become clear transparent yellow liquid after soaking completely, with remarkable fragrance of Phellinus Linteus as shown in FIG. 2.
Example 2
A preparation method of phellinus igniarius paste comprises the following steps:
(1) pulverizing mature dried Phellinus igniarius fruiting body, grinding, sieving with 80 mesh sieve, adding sucrose and water, and mixing well to prepare culture medium; wherein, the adding amount of the phellinus igniarius powder, the sucrose and the water is 4 kg: 1 kg: 5L;
(2) primary fermentation: the adopted cellulose degrading bacteria are composite strains consisting of bacillus subtilis CICC 10088, bacillus licheniformis CICC10831 and aspergillus carbonarius CICC 41254;
inoculating cellulose-degrading bacteria into the culture medium in the step (1), fermenting for 60 hours at room temperature and about 30 ℃, and performing shake culture at a shaking table rotating speed of 150 rpm;
wherein the inoculation amount of the cellulose-degrading bacteria is that 1.5mL of cellulose-degrading bacteria solution is inoculated per 100g of culture medium, and each mL of the cellulose-degrading bacteria solution contains 2 multiplied by 1010cfu Bacillus subtilis CICC 10088, 2 × 1010cfu Bacillus licheniformis CICC10831 and 1 × 1010cfu aspergillus carbonarius CICC 41254;
grinding the mixture into thick liquid after the fermentation is finished, and then carrying out low-temperature ultrahigh-pressure treatment under the pressure of 450MPa for 10min at the temperature of 30 ℃;
(3) and (3) secondary fermentation: activating Saccharomyces cerevisiae CICC 1839 with brown sugar water, and then adding into the slurry obtained in step (2) for natural normal temperature fermentation, wherein the inoculation amount is as follows: inoculating 4mL of bacterial liquid per 100mL of the slurry, wherein each mL of the bacterial liquid contains 1X 1010The fermentation time of cfu yeast is 15 h;
after fermentation, centrifuging to obtain supernatant and precipitate; washing the precipitate, and combining the washing liquid into the supernatant;
(4) performing ultrafiltration desalination and concentration on the mixed solution obtained in the step (3), wherein the cut-off molecular weight range of an ultrafiltration membrane is 10000Da, so as to obtain a concentrated solution;
(5) low-temperature ultrahigh pressure: sterilizing the obtained concentrated solution at low temperature and ultrahigh pressure, wherein the pressure is 450MPa, the time is 10min, and the temperature is 30 ℃;
(6) freeze drying in a freeze drier overnight;
(7) pressing the powder into block-shaped solid by a briquetting machine under the pressure of 15MPa to obtain the phellinus igniarius paste, wherein the content of crude polysaccharide substances is 34.1 percent, the content of total flavonoids is 6.2 percent, the content of triterpenes substances is 3.0 percent, and the water content is less than or equal to 5w/w percent.
Example 3
(1) Pulverizing mature dried Phellinus igniarius fruiting body, grinding, sieving with 100 mesh sieve, adding sucrose and water, and mixing well to prepare culture medium; wherein, the adding amount of the phellinus igniarius powder, the cane sugar and the water is 3 kg: 2 kg: 5L;
(2) primary fermentation: the adopted cellulose degrading bacteria are composite strains consisting of bacillus subtilis CICC 10088, bacillus licheniformis CICC10831 and aspergillus carbonarius CICC 41254;
inoculating cellulose-degrading bacteria into the culture medium in the step (1), fermenting for 72 hours at room temperature and about 30 ℃, and performing shake culture at a shaking table rotating speed of 150 rpm;
wherein the inoculation amount of the cellulose-degrading bacteria is that 1.5mL of cellulose-degrading bacteria solution is inoculated per 100g of culture medium, and each mL of the cellulose-degrading bacteria solution contains 2 multiplied by 1010cfu Bacillus subtilis CICC 10088, 2 × 1010cfu Bacillus licheniformis CICC10831 and 1 × 1010cfu aspergillus carbonarius CICC 41254;
grinding the mixture into pulp after the fermentation is finished, so as to break the cell wall again and further release the effective active ingredients in the cells;
performing low-temperature ultrahigh-pressure treatment after grinding, wherein the pressure is 500MPa, the time is 10min, and the temperature is 30 ℃;
(3) and (3) secondary fermentation: activating Saccharomyces cerevisiae CICC 1839 with brown sugar water, and then adding into the slurry obtained in step (2) for natural normal temperature fermentation, wherein the inoculation amount is as follows: inoculating 3mL of bacterial liquid per 100mL of the slurry, wherein each mL of the bacterial liquid contains 1X 1010The fermentation time of the cfu yeast is 24 hours;
after fermentation, centrifuging to obtain supernatant and precipitate; washing the precipitate, and combining the washing liquid into the supernatant;
(4) performing ultrafiltration desalination and concentration on the mixed solution obtained in the step (3), wherein the cut-off molecular weight range of an ultrafiltration membrane is 15000Da, so as to obtain a concentrated solution;
(5) low-temperature ultrahigh pressure: sterilizing the obtained concentrated solution at low temperature and ultrahigh pressure, wherein the pressure is 500MPa, the time is 10min, and the temperature is 30 ℃;
(6) freeze drying in a freeze drier overnight;
(7) pressing the powder into block-shaped solid by a briquetting machine under the pressure of 15MPa to obtain the phellinus igniarius paste, wherein the content of crude polysaccharide substances is 33.0 percent, the content of total flavonoids is 5.7 percent, the content of triterpenes substances is 2.9 percent, and the water content is less than or equal to 5w/w percent.
The above embodiments are preferred embodiments of the present disclosure, but the embodiments of the present disclosure are not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present disclosure should be regarded as equivalent replacements within the scope of the present disclosure.
Claims (10)
1. The preparation method of the phellinus igniarius paste is characterized by comprising the following steps of:
(1) pulverizing mature Phellinus linteus fruiting body, sieving, adding sucrose and water, and preparing culture medium;
(2) primary fermentation: inoculating cellulose degrading bacteria into the culture medium in the step (1), and fermenting for a certain time; after the fermentation is finished, grinding the mixture into thick liquid; after pulping, carrying out low-temperature ultrahigh-pressure sterilization;
(3) and (3) secondary fermentation: inoculating yeast into the slurry obtained in the step (2), and fermenting for a certain time; after fermentation is finished, carrying out solid-liquid separation to obtain fermentation liquor;
(4) performing ultrafiltration on the mixed solution obtained in the step (3) to remove salt, small molecular impurities and pigments to obtain a concentrated solution;
(5) low-temperature ultrahigh pressure: carrying out low-temperature ultrahigh-pressure sterilization on the obtained concentrated solution;
(6) freeze drying;
(7) pressing into solid to obtain Phellinus Linteus paste.
2. The method according to claim 1, wherein in the step (1), the Phellinus linteus species include Phellinus linteus, Phellinus baumii and Phellinus igniarius;
preferably, the addition amount of the sieved phellinus igniarius powder, sucrose and water is (3-6) kg: (0.5-1.5) kg: (5-8) L.
3. The method according to claim 1, wherein in the step (2), the cellulose-degrading bacteria are complex bacteria or single bacteria;
preferably, the cellulose degrading bacteria is a mixture of Bacillus subtilis, Bacillus licheniformis and Aspergillus carbonarius;
preferably, the inoculation amount of the cellulose degrading bacteria is 1-1.5%, and 1mL of bacteria solution contains the following strains: bacillus subtilis, Bacillus licheniformis and Aspergillus carbonarius (1-2) x 1010cfu:(2～4)×1010cfu:(1～2)×1010cfu；
Preferably, the fermentation conditions are: fermenting for 2-3 days at normal temperature or room temperature, and stirring or shake culturing;
preferably, the low temperature ultra high pressure conditions: the ultrahigh pressure is 400 MPa-500 MPa, the ultrahigh pressure time is 10 min-20 min, and the ultrahigh temperature is 25-50 ℃.
4. The method according to claim 1, wherein in the step (3), the yeast is isolated from the surface of wild Phellinus linteus fruiting body or the surrounding environment or is obtained from purchased Saccharomyces cerevisiae;
preferably, the yeast fermentation is normal-temperature natural fermentation, and the fermentation time is 8-24 h;
further preferably, the fermentation temperature is 25-42 ℃.
5. The method of claim 1, wherein in step (4), the ultrafiltration membrane has a molecular weight cut-off of 10000-15000 Da.
6. The method as set forth in claim 1, wherein in the step (5), the low-temperature ultrahigh-pressure condition: the ultrahigh pressure is 400 MPa-500 MPa, the ultrahigh pressure time is 10 min-20 min, and the ultrahigh temperature is 25-50 ℃.
7. The method as claimed in claim 1, wherein in the step (6), the product is freeze-dried for 16-18 hours to a water content of 5 w/w% or less.
8. The method as set forth in claim 1, wherein in the step (7), the solid is pressed into granules or blocks.
9. The phellinus igniarius ointment prepared by the method according to any one of claims 1 to 8, which is characterized by being in a solid form, wherein the content of crude phellinus igniarius polysaccharides is not less than 30%, the content of total flavonoids is not less than 5%, the content of triterpenes is not less than 2%, and the water content is not more than 5%.
10. The use of the phellinus linteus paste according to claim 9 in the preparation of an anti-tumor or anti-oxidation medicament or health product.
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| CN111903969A (en) * | 2020-08-10 | 2020-11-10 | 重庆三峡学院 | Pueraria root and impatiens three-treasure paste and production method thereof |
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