CN110710569A - Phellinus igniarius active lactobacillus beverage and preparation method thereof - Google Patents

Phellinus igniarius active lactobacillus beverage and preparation method thereof Download PDF

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CN110710569A
CN110710569A CN201910978667.1A CN201910978667A CN110710569A CN 110710569 A CN110710569 A CN 110710569A CN 201910978667 A CN201910978667 A CN 201910978667A CN 110710569 A CN110710569 A CN 110710569A
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bacteria
phellinus
cfu
phellinus igniarius
fermentation
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CN110710569B (en
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陈安徽
邵颖
秦杰
张爱文
朱文静
岳明宇
郭红伟
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Chen Anhui
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Xuzhou University of Technology
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Abstract

The invention relates to a phellinus igniarius active lactobacillus beverage and a preparation method thereof, wherein the method comprises the following steps: (1) pulverizing Phellinus linteus fruiting body, and preparing culture medium; (2) inoculating cellulose degrading bacteria into the culture medium in the step (1) and fermenting; (3) carrying out low-temperature ultrahigh pressure on the fermentation liquor obtained in the step (2), and carrying out solid-liquid separation after the fermentation liquor is finished to obtain supernatant for later use; (4) adding milk powder into the supernatant, stirring uniformly, inoculating lactobacillus, and fermenting for a certain time; (5) blending; (6) homogenizing the mixed solution in the step (5); homogenizing to obtain Phellinus Linteus active lactobacillus beverage. The phellinus igniarius active lactobacillus beverage prepared by the method disclosed by the invention is high in active bacteria content and active ingredients, and has high nutritional value and health care value.

Description

Phellinus igniarius active lactobacillus beverage and preparation method thereof
Technical Field
The present invention relates to a phellinus igniarius active lactobacillus beverage 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.
Phellinus genus is Basidiomycetes, Polyporales, Polyporaceae, Phellinus genus (Phellinus), and is also called Morriscus, Tree chicken, Hoffonia simplicifolia, Phellinus linteus, Phellinus igniarius, Phellinus linteus, and Prunus mume. Modern medical research finds that phellinus igniarius has various pharmacological actions and has obvious curative effects in the aspects of antibiosis, immunoregulation, sweat gland secretion inhibition, tumor resistance, fibrosis resistance, inflammation diminishing, pain relieving, oxidation resistance and the like.
At present, the reported phellinus igniarius beverages mainly comprise phellinus igniarius vinegar beverages, phellinus igniarius hypha black tea fungus beverages, phellinus igniarius health-care beverages and the like, and a phellinus igniarius lactic acid beverage 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 background technologies, the present disclosure provides a preparation method of a phellinus igniarius active lactobacillus beverage, and the phellinus igniarius active lactobacillus beverage prepared by the method of the present disclosure has high active bacteria content and active ingredients, and has a good application prospect.
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 a phellinus linteus active lactic acid bacteria beverage, the method comprising the steps of:
(1) pulverizing Phellinus linteus fruiting body, sieving, adding sucrose and water, and preparing culture medium;
(2) inoculating cellulose degrading bacteria into the culture medium in the step (1), and fermenting for a certain time;
(3) carrying out low-temperature ultrahigh pressure on the fermentation liquor obtained in the step (2), carrying out solid-liquid separation after the fermentation liquor is finished, and removing mycelium and other solid impurities to obtain supernatant for later use;
(4) adding milk powder into the supernatant, stirring uniformly, inoculating lactobacillus, and fermenting for a certain time;
(5) after fermentation is finished, adding a stabilizer and a sweetening agent into the fermentation liquor, and uniformly mixing;
(6) homogenizing the mixed solution in the step (5); homogenizing to obtain Phellinus Linteus active lactobacillus beverage.
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) kg: (5-8) L.
Based on the characteristics of high content of crude fiber, complex components and hard texture of phellinus igniarius sporocarp, active ingredients such as polysaccharide, flavone and triterpenes in the phellinus igniarius sporocarp are not easy to extract efficiently, the inventor adopts cellulose degrading bacteria which utilize cane sugar and nitrogen sources contained in the sporocarp to secrete cellulase, hemicellulase, pectinase and the like to degrade the crude fiber and cell walls in the phellinus igniarius sporocarp efficiently, so that the crude fiber is degraded into glucose and the like which can be utilized by paenibacillus polymyxa, the cell walls can be broken to release various active ingredients in cells, the effect is realized, the cellulose degrading bacteria can break the hard structure of the phellinus igniarius sporocarp, various active ingredients are dissolved out more easily, and the dissolution rate of various active ingredients is improved.
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), 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, which is enough to fully dissolve active ingredients such as polysaccharide in phellinus igniarius sporocarp, the content of soluble solid matters is high, and lactobacillus can quickly grow by utilizing nutrient substances in the culture medium. If the fermentation time is shorter than 2 days, the dissolution rate of active ingredients such as phellinus linteus polysaccharide is low; the fermentation time is longer than 3 days, the nutrient medium is insufficient, the nutrient components such as polysaccharide of phellinus igniarius sporocarp and the like need to be consumed, and the total content of the active components is reduced.
In the step (2), the inoculation amount of the cellulose degrading bacteria is 1-1.5% (1-1.5 mL of bacterial liquid is inoculated per 100g of culture medium), and the bacterial strains contained in 1mL of bacterial liquid are as follows: 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 (3), 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 step (3), preferably, the solid-liquid separation is performed by centrifugation under the following conditions: centrifuging at 4000-5000 rpm for 5-15 min.
In the step (4), preferably, the addition amount of the milk powder is as follows: 1L of supernatant: 10-40 g of milk powder.
In the step (4), the lactic acid bacteria are composite bacteria or single bacteria, and preferably composite bacteria in terms of fermentation effect and efficiency. Aiming at specific phellinus igniarius fermentation liquor, the selected composite bacteria are streptococcus thermophilus and Lactobacillus acidophilus. Experiments prove that the composite bacteria have high fermentation efficiency and good fermentation flavor.
Wherein said Streptococcus thermophilus and Lactobacillus acidophilus are commercially available in a conventional manner.
In the step (4), preferably, the inoculation amount of the lactic acid bacteria is 1-3% (1-3 mL of lactic acid bacteria liquid is inoculated per 100mL of fermentation medium), and the strains contained in 1mL of the lactic acid bacteria liquid are as follows: streptococcus thermophilus and lactobacillus acidophilus ═ (2-4) × 1010cfu:(1~2)×1010cfu。
In the step (4), preferably, the fermentation condition is 37-42 ℃ for 4-6 h.
In the step (5), preferably, the fermentation broth: a stabilizer: the adding proportion of the sweetening agent is 100 mL: (0.1-1.5) g: (0.01-1) g.
In order to effectively prevent the phenomena of elutriation and precipitation, even water-milk stratification, the inventor selects the xanthan gum and the locust bean gum as the stabilizing agents through screening and optimization, wherein the mass ratio of the xanthan gum to the locust bean gum is (1-2) to (1-2).
The sweetener in the present disclosure is not particularly limited and includes aspartame, acesulfame potassium, sucrose, various sugar alcohols, and the like.
In the step (6), preferably, the homogenization conditions are as follows: the pressure is 10-15 MPa, the temperature is 40-50 ℃, and the time is 5-15 min.
In a second aspect of the present disclosure, there is provided a phellinus linteus active lactobacillus beverage prepared by the above method, which is characterized in that: the Phellinus Linteus active lactobacillus beverage is light yellow, sour and sweet, and has good flavor, and viable count of lactobacillus is not less than 1 × 109cfu/mL, the content of soluble solids is more than or equal to 12 percent, the pH value is 4-5, the content of protein is more than or equal to 1.0 percent, the content of phellinus igniarius crude polysaccharide is more than or equal to 3 percent, the content of total flavone is more than or equal to 0.5 percent, and the content of triterpenes is more than or equal to 0.3 percent.
Compared with the related technology known by the inventor, one technical scheme of the present disclosure has the following beneficial effects:
(1) the method adopts cellulose degrading bacteria to degrade the phellinus igniarius sporocarp, improves the dissolution rate of active ingredients of the phellinus igniarius sporocarp, and also provides a carbon source for lactic acid bacteria.
(2) The fermentation process of the method does not need to adjust pH, and the production efficiency is high.
(3) The phellinus igniarius active lactobacillus beverage prepared by the method disclosed by the invention is high in active bacteria content and active ingredients, and has high nutritional value and health care value.
(4) The phellinus igniarius active lactobacillus beverage obtained by the method is light yellow, and the phellinus igniarius has aromatic flavor.
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.
Figure 1 is a graph of the degradation effect of different samples 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.
In the examples of the present disclosure, both lactic acid bacteria were purchased from wanfang organisms.
Example 1 investigation of the Effect of different cellulose-degrading bacteria on fermentation
Treatment 1: the cellulose degrading bacteria are Bacillus subtilis CICC10088 and Bacillus licheniformis CICC10831, and each mL of the bacteria liquid contains 2 × 1010cfu Bacillus subtilis CICC10088, 3 × 1010cfu Bacillus licheniformis CICC 10831.
And (3) treatment 2: the cellulose-degrading bacteria are Bacillus licheniformis CICC10831 and Aspergillus carbonarius CICC41254, and each mL of the bacteria solution contains 2 × 1010cfu Bacillus licheniformis CICC10831, 3 × 1010cfu aspergillus carbonarius CICC 41254.
And (3) treatment: the cellulose-degrading bacteria are Bacillus subtilis CICC10088 and Aspergillus carbonarius CICC41254, and each mL of the bacteria liquid contains 2 × 1010cfu Bacillus subtilis CICC10088, 3 × 1010cfu aspergillus carbonarius CICC 41254.
And (4) treatment: the cellulose-degrading bacteria are Trichoderma reesei, Pseudomonas sp and Bacillus subtilis CICC10088 producing cellulase and/or hemicellulase and/or pectinase, and each mL of the bacteria liquid contains 1 × 1010cfu Trichoderma reesei, 2X 1010Pseudomonas cfu and 2X 1010cfu Bacillus subtilis CICC 10088.
And (4) treatment 5: the adopted cellulose degrading bacteria are composite strains consisting of bacillus subtilis CICC10088, bacillus licheniformis CICC10831 and aspergillus carbonarius CICC 41254; 2X 10 of bacterial suspension per mL10cfu Bacillus subtilis CICC10088, 2 × 1010cfu Bacillus licheniformis CICC10831 and 1 × 1010cfu aspergillus carbonarius CICC 41254.
The treatment 1-5 is carried out according to the following steps: (1) pulverizing dried Phellinus igniarius fruiting body, grinding, sieving with 100 mesh sieve, adding sucrose and water, and mixing to obtain culture medium; wherein, the adding amount of the phellinus igniarius powder, the sucrose and the water is 4 kg: 1 kg: 5L.
(2) 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 liquid is inoculated to every 100g of culture medium.
(3) And after fermentation is finished, centrifuging at 5000rpm for 10min to obtain a supernatant, and recording the supernatant after 1-5 treatment as a sample 1, a sample 2, a sample 3, a sample 4 and a sample 5 in sequence. The soluble solids content of each sample was determined in triplicate and averaged. The test results are shown in FIG. 1.
As can be seen from figure 1, aiming at a specific culture medium containing phellinus igniarius substances, different cellulose degrading bacteria have larger difference in degrading effect, in sum, a composite strain consisting of bacillus subtilis CICC10088, bacillus licheniformis CICC10831 and aspergillus carbonarius CICC41254 is selected as the cellulose degrading bacteria with the best degrading effect, and the content of soluble solid matters is higher, so that the method is used as a basis for fermenting lactic acid bacteria and obtaining beverages containing more active ingredients.
Example 2
A preparation method of phellinus igniarius active lactobacillus beverage comprises the following steps:
(1) pulverizing dried Phellinus igniarius fruiting body, grinding, sieving with 100 mesh sieve, adding sucrose and water, and mixing to obtain culture medium; wherein, the adding amount of the phellinus igniarius powder, the cane sugar and the water is 3 kg: 1 kg: 4L.
(2) The adopted cellulose degrading bacteria are composite strains consisting of bacillus subtilis CICC10088, 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 every 100g of culture medium is inoculated with 2mL of cellulose-degrading bacteria liquid, and every mL of the cellulose-degrading bacteria liquid contains 2 multiplied by 1010cfu Bacillus subtilis CICC10088, 2 × 1010cfu Bacillus licheniformis CICC10831 and 1 × 1010cfu aspergillus carbonarius CICC 41254.
(3) After fermentation, filling the fermentation liquor in a sealed polyvinylidene bag for low-temperature ultrahigh-pressure treatment, wherein the ultrahigh-pressure is 450MPa, the time is 10min, and the temperature is 30 ℃;
after the low-temperature ultrahigh pressure is finished, performing centrifugal separation, wherein the centrifugal conditions are as follows: centrifuging at 5000rpm for 10min to remove solid and insoluble substances including mycelium to obtain supernatant.
(4) The supernatant was diluted to 1L: 20g of supernatant: adding milk powder into the milk powder according to the proportion, uniformly stirring to prepare a fermentation medium, and then adding the milk powder into the fermentation medium according to the proportion of 100 mL: 1.5mL of the lactobacillus liquid is inoculated, and the 1mL of the lactobacillus liquid contains the following strains: streptococcus thermophilus 4X 1010cfu, Lactobacillus acidophilus 1X 1010cfu, anaerobic fermentation at 40 ℃ for 6h after inoculation.
(5) After the fermentation of the lactic acid bacteria is finished, adding 100mL of: 0.2 g: 0.03g ═ broth: a stabilizer: uniformly mixing the stabilizer and the acesulfame potassium in the acesulfame potassium proportion;
wherein the stabilizer is xanthan gum and locust bean gum with the mass ratio of 1:1.2, and the stabilizer is fully dissolved and uniformly mixed by using a proper amount of warm water before adding.
(6) Homogenizing the mixed solution in the step (5), wherein the homogenizing condition is as follows: the pressure is 15MPa, the temperature is 45 ℃, and the time is 8 min.
(7) And filling the homogenized mixed solution to obtain the phellinus igniarius active lactobacillus beverage.
The obtained Phellinus Linteus active lactobacillus beverage is light yellow, has good fragrance, sour and sweet taste, and has viable count of lactobacillus not less than 1 × 109cfu/mL, the content of soluble solids is 12.5%, the pH value is 4.3, the content of protein is 1.3%, the content of crude polysaccharide of phellinus igniarius is 3.9%, the content of total flavonoids is 0.61%, and the content of triterpenoids is 0.32%.
Example 3
A preparation method of phellinus igniarius active lactobacillus beverage comprises the following steps:
(1) pulverizing dried Phellinus igniarius fruiting body, grinding, sieving with 100 mesh sieve, adding sucrose and water, and mixing to obtain culture medium; wherein, the adding amount of the phellinus igniarius powder, the sucrose and the water is 4 kg: 1 kg: 5L.
(2) The adopted cellulose degrading bacteria are composite strains consisting of bacillus subtilis CICC10088, 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 CICC10088, 2 × 1010cfu Bacillus licheniformis CICC10831 and 1 × 1010cfu aspergillus carbonarius CICC 41254.
(3) After fermentation, filling the fermentation liquor in a sealed polyvinylidene bag for low-temperature ultrahigh-pressure treatment, wherein the ultrahigh-pressure is 400MPa, the time is 15min, and the temperature is 30 ℃;
after the low-temperature ultrahigh pressure is finished, performing centrifugal separation, wherein the centrifugal conditions are as follows: centrifuging at 5000rpm for 10min to remove solid and insoluble substances including mycelium to obtain supernatant.
(4) The supernatant was diluted to 1L: 25g of supernatant: adding milk powder into the milk powder according to the proportion, uniformly stirring to prepare a fermentation medium, and then adding the milk powder into the fermentation medium according to the proportion of 100 mL: inoculating lactobacillus liquid in a proportion of 2mL, wherein the lactobacillus liquid in 1mL contains the following strains: streptococcus thermophilus 4X 1010cfu, Lactobacillus acidophilus 1X 1010cfu, anaerobic fermentation at 40 ℃ for 8h after inoculation.
(5) After the fermentation of the lactic acid bacteria is finished, adding 100mL of: 0.2 g: 0.02g ═ broth: a stabilizer: uniformly mixing the stabilizer with aspartame according to the proportion of aspartame;
wherein the stabilizer is xanthan gum and locust bean gum with the mass ratio of 1:1, and a proper amount of warm water is adopted to fully dissolve and uniformly mix the stabilizer before adding.
(6) Homogenizing the mixed solution in the step (5), wherein the homogenizing condition is as follows: the pressure is 12MPa, the temperature is 405 ℃, and the time is 10 min.
(7) And filling the homogenized mixed solution to obtain the phellinus igniarius active lactobacillus beverage.
The detection shows that the obtained phellinus igniarius active lactobacillus beverage is light yellow, the phellinus igniarius fungus has obvious fragrance and is sour, sweet and delicious, and the number of live lactobacillus is more than or equal to 1 multiplied by 109cfu/mL, a soluble solid content of 12% (i.e., 12g/100g), a pH of 4.5, a protein content of 1.2% (i.e., 1.2g/100g), a phellinus linteus polysaccharide content of 3.4% (i.e., 3.4g/100g), a flavone content of 0.54% (i.e., 0.54g/100g), and a triterpene content of 0.30% (i.e., 0.30g/100 g).
Example 4
A preparation method of phellinus igniarius active lactobacillus beverage comprises the following steps:
(1) pulverizing dried Phellinus igniarius fruiting body, grinding, sieving with 100 mesh sieve, adding sucrose and water, and mixing to obtain culture medium; wherein, the adding amount of the phellinus igniarius powder, the cane sugar and the water is 5 kg: 1 kg: 8L.
(2) The adopted cellulose degrading bacteria are composite strains consisting of bacillus subtilis CICC10088, 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 every 100g of culture medium is inoculated with 2mL of cellulose-degrading bacteria liquid, and every mL of the cellulose-degrading bacteria liquid contains 2 multiplied by 1010cfu Bacillus subtilis CICC10088, 2 × 1010cfu Bacillus licheniformis CICC10831 and 1 × 1010cfu aspergillus carbonarius CICC 41254.
(3) After fermentation, filling the fermentation liquor in a sealed polyvinylidene bag for low-temperature ultrahigh-pressure treatment, wherein the ultrahigh-pressure is 500MPa, the time is 15min, and the temperature is 30 ℃;
after the low-temperature ultrahigh pressure is finished, performing centrifugal separation, wherein the centrifugal conditions are as follows: centrifuging at 5000rpm for 10min to remove solid and insoluble substances including mycelium to obtain supernatant.
(4) The supernatant was diluted to 1L: 40g of supernatant: adding milk powder into milk powder at a certain proportion, stirring well to obtain fermentation culture medium, and then mixing according to the above-mentioned proportionAccording to 100mL of fermentation medium: inoculating lactobacillus liquid in a proportion of 2mL, wherein the lactobacillus liquid in 1mL contains the following strains: streptococcus thermophilus 4X 1010cfu, Lactobacillus acidophilus 1X 1010cfu, anaerobic fermentation at 40 ℃ for 4.5h after inoculation.
(5) After the fermentation of the lactic acid bacteria is finished, adding 100mL of: 0.2 g: 0.03g ═ broth: a stabilizer: uniformly mixing the stabilizer and the acesulfame potassium in the acesulfame potassium proportion;
wherein the stabilizer is xanthan gum and locust bean gum with the mass ratio of 1.5:1, and the stabilizer is fully dissolved and uniformly mixed by using a proper amount of warm water before adding.
(6) Homogenizing the mixed solution in the step (5), wherein the homogenizing condition is as follows: the pressure is 12MPa, the temperature is 45 ℃, and the time is 10 min.
(7) And filling the homogenized mixed solution to obtain the phellinus igniarius active lactobacillus beverage.
The detection shows that the obtained phellinus igniarius active lactobacillus beverage is light yellow, the phellinus igniarius fungus has obvious fragrance and is sour, sweet and delicious, and the number of live lactobacillus is more than or equal to 1 multiplied by 109cfu/mL, the content of soluble solids is 13%, the pH value is 4.8, the content of protein is 1.1%, the content of phellinus linteus polysaccharide is 3.7%, the content of flavone is 0.58%, and the content of triterpenes is 0.38%.
Experimental example 1
A preparation method of phellinus igniarius lactic acid beverage comprises the following steps:
(1) crushing dried Phellinus igniarius sporocarp, grinding, sieving by a 100-mesh sieve, and adding water to mix uniformly, wherein the adding amount of Phellinus igniarius powder and water is 4 kg: 10L.
(2) The pH of the mixed solution in step (1) was adjusted to 4.5, and then cellulase (activity: 10U/mg) was added thereto, cellulase: the amount of the mixture added was 1 g: 1L, enzymolysis temperature of 45 ℃ and enzymolysis time of 4 h.
(3) After enzymolysis, filling the enzymolysis liquid in a sealed polyvinylidene bag for low-temperature ultrahigh-pressure treatment, wherein the ultrahigh-pressure is 400MPa, the time is 15min, and the temperature is 30 ℃;
after the low-temperature ultrahigh pressure is finished, performing centrifugal separation, wherein the centrifugal conditions are as follows: centrifuging at 5000rpm for 10min, removing solid, and collecting supernatant.
(4) The supernatant was diluted to 1L: 25g of supernatant: adding milk powder into the milk powder according to the proportion, uniformly stirring to prepare a fermentation medium, and then adding the milk powder into the fermentation medium according to the proportion of 100 mL: inoculating lactobacillus liquid in a proportion of 2mL, wherein the lactobacillus liquid in 1mL contains the following strains: streptococcus thermophilus 4X 1010cfu, Lactobacillus acidophilus 1X 1010cfu, anaerobic fermentation at 40 ℃ for 8h after inoculation.
(5) After the fermentation of the lactic acid bacteria is finished, adding 100mL of: 0.2 g: 0.02g ═ broth: a stabilizer: uniformly mixing the stabilizer with aspartame according to the proportion of aspartame;
wherein the stabilizer is xanthan gum and locust bean gum with the mass ratio of 1:1, and a proper amount of warm water is adopted to fully dissolve and uniformly mix the stabilizer before adding.
(6) Homogenizing the mixed solution in the step (5), wherein the homogenizing condition is as follows: the pressure is 12MPa, the temperature is 405 ℃, and the time is 10 min.
(7) And (4) after homogenizing, obtaining the lactobacillus beverage.
The obtained lactobacillus beverage is light yellow, soluble solid content is 11%, pH is 4.4, and viable count is about 1 × 108cfu/mL, protein content 1.1%, phellinus igniarius crude polysaccharide content 1.6%, total flavone content 0.22%, and triterpenes content 0.11%. Compared with examples 2-4, the nutrition content is lower, and the difference is obvious.
Experimental example 2
A preparation method of phellinus igniarius lactic acid beverage comprises the following steps:
(1) pulverizing dried Phellinus igniarius fruiting body, grinding, sieving with 100 mesh sieve, adding sucrose and water, and mixing to obtain culture medium; wherein, the adding amount of the phellinus igniarius powder, the cane sugar and the water is 3 kg: 1 kg: 4L.
(2) The adopted cellulose degrading bacteria are composite strains consisting of bacillus subtilis CICC10088, 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 every 100g of culture medium is inoculated with 2mL of cellulose-degrading bacteria liquid, and every mL of the cellulose-degrading bacteria liquid contains 2 multiplied by 1010cfu Bacillus subtilis CICC10088, 2 × 1010cfu Bacillus licheniformis CICC10831 and 1 × 1010cfu aspergillus carbonarius CICC 41254.
(3) After fermentation, performing centrifugal separation, wherein the centrifugal conditions are as follows: centrifuging at 5000rpm for 10min to remove solid and insoluble substances including mycelium to obtain supernatant, and sterilizing at high temperature and inactivating enzyme.
(4) The supernatant was diluted to 1L: 20g of supernatant: adding milk powder into the milk powder according to the proportion, uniformly stirring to prepare a fermentation medium, and then adding the milk powder into the fermentation medium according to the proportion of 100 mL: 1.5mL of the lactobacillus liquid is inoculated, and the 1mL of the lactobacillus liquid contains the following strains: streptococcus thermophilus 4X 1010cfu, Lactobacillus acidophilus 1X 1010cfu, anaerobic fermentation at 40 ℃ for 6h after inoculation.
(5) After the fermentation of the lactic acid bacteria is finished, adding 100mL of: 0.2 g: 0.03g ═ broth: a stabilizer: uniformly mixing the stabilizer and the acesulfame potassium in the acesulfame potassium proportion;
wherein the stabilizer is xanthan gum and locust bean gum with the mass ratio of 1:1.2, and the stabilizer is fully dissolved and uniformly mixed by using a proper amount of warm water before adding.
(6) Homogenizing the mixed solution in the step (5), wherein the homogenizing condition is as follows: the pressure is 15MPa, the temperature is 45 ℃, and the time is 8 min.
(7) And filling the homogenized mixed solution to obtain the phellinus igniarius active lactobacillus beverage.
The detection shows that the obtained Phellinus Linteus active lactobacillus beverage is light yellow, the content of soluble solids is 10%, and the number of viable bacteria is about 1 × 109cfu/mL, pH 4.4, protein content 1.0%, crude polysaccharide content of Phellinus linteus 2.3%, total flavone content 0.20%, and crude triterpene content 0.23%. Compared with the examples 2-4, the active ingredients of the obtained beverage contain no low-temperature ultrahigh-pressure technologyThe amount is low, and the difference is obvious.
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. A preparation method of phellinus igniarius active lactobacillus beverage is characterized by comprising the following steps:
(1) pulverizing Phellinus linteus fruiting body, sieving, adding sucrose and water, and preparing culture medium;
(2) inoculating cellulose degrading bacteria into the culture medium in the step (1), and fermenting for a certain time;
(3) carrying out low-temperature ultrahigh pressure on the fermentation liquor obtained in the step (2), carrying out solid-liquid separation after the fermentation liquor is finished, and removing mycelium and other solid impurities to obtain supernatant for later use;
(4) adding milk powder into the supernatant, stirring uniformly, inoculating lactobacillus, and fermenting for a certain time;
(5) after fermentation is finished, adding a stabilizer and a sweetening agent into the fermentation liquor, and uniformly mixing;
(6) homogenizing the mixed solution in the step (5); homogenizing to obtain Phellinus Linteus active lactobacillus beverage.
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, sieving the mixture by a sieve of 100-120 meshes;
preferably, the addition amount of the sieved phellinus igniarius powder, sucrose and water is (3-6) kg: (0.5-1) 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 complex bacteria;
preferably, the complex bacteria are a mixture of Bacillus subtilis, Bacillus licheniformis and Aspergillus carbonarius.
4. The method of claim 1, wherein in step (2), the fermentation conditions are: fermenting for 2-3 days at normal temperature or room temperature, and stirring or shake culturing;
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。
5. The method as set forth in claim 1, wherein in the step (3), the low-temperature ultrahigh-pressure condition: the ultrahigh pressure is 400MPa to 500MPa, the ultrahigh pressure time is 10min to 20min, and the ultrahigh temperature is 25 ℃ to 50 ℃;
preferably, the solid-liquid separation adopts centrifugal treatment, and the centrifugal conditions are as follows: centrifuging at 4000-5000 rpm for 5-15 min.
6. The method as claimed in claim 1, wherein in the step (4), the added amount of the powdered milk is: 1L of supernatant: 10-40 g of milk powder;
preferably, the lactic acid bacteria are compound bacteria or single bacteria, and preferably compound bacteria;
further preferably, the complex bacteria are Streptococcus thermophilus and Lactobacillus acidophilus.
7. The method according to claim 1, wherein in the step (4), the amount of the inoculated lactic acid bacteria is 1 to 3%, and 1mL of the bacterial solution contains the following species: streptococcus thermophilus and lactobacillus acidophilus ═ (2-4) × 1010cfu:(1~2)×1010cfu;
Preferably, the fermentation condition is 37-42 ℃ for 4-6 h.
8. The method of claim 1, wherein in step (5), the fermentation broth: a stabilizer: the adding proportion of the sweetening agent is 100 mL: (0.1-1.5) g: (0.01-1) g;
preferably, the stabilizer is xanthan gum and locust bean gum, and the mass ratio of the xanthan gum to the locust bean gum is (1-2) to (1-2).
9. The method according to claim 1, wherein in the step (6), the homogenization conditions are: the pressure is 10-15 MPa, the temperature is 40-50 ℃, and the time is 5-15 min.
10. The phellinus igniarius active lactic acid bacteria beverage prepared by the method of any one of claims 1 to 9, which is characterized in that the phellinus igniarius active lactic acid bacteria beverage is light yellow, sour and sweet and delicious, the flavor of phellinus igniarius is obvious, and the number of viable lactic acid bacteria is more than or equal to 1 x 109cfu/mL, the content of soluble solids is more than or equal to 12 percent, the pH value is 4-5, the content of protein is more than or equal to 1.0 percent, the content of phellinus linteus polysaccharide is more than or equal to 3 percent, the content of flavone is more than or equal to 0.5 percent, and the content of triterpenes is more than or equal to 0.3 percent.
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