CN108948115B - Method for efficiently extracting adenosine from cordyceps militaris - Google Patents
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Abstract
The invention belongs to the technical field of extraction of effective ingredients of cordyceps militaris, and particularly relates to an extraction method for efficiently extracting adenosine from cordyceps militaris. According to the method for extracting adenosine from cordyceps militaris, on the basis of taking water as an extraction solvent in the prior art, a selected amount of composite extraction enzyme comprising laccase or xylanase and cellulase or pectinase is further added for enzymolysis extraction, so that the extraction rate of adenosine from cordyceps militaris is effectively improved; according to the method for extracting adenosine from cordyceps militaris, the dissolution and extraction of adenosine from cordyceps militaris are effectively improved by adding polyethylene glycol in the step of pulping; preferably, in the method, before extraction, the cordyceps militaris sporocarp is dried in a gradient drying mode, and the temperature difference of each stage is utilized, so that the wall breaking effect of the cordyceps militaris sporocarp is further improved, the dissolution of adenosine is effectively promoted, and the extraction rate of adenosine is also improved.
Description
Technical Field
The invention belongs to the technical field of extraction of effective ingredients of cordyceps militaris, and particularly relates to an extraction method for efficiently extracting adenosine from cordyceps militaris.
Background
Cordyceps militaris (L.) Link), also known as Cordyceps militaris, is a model species belonging to the phylum Eumycota, the phylum Ascomycota, the subdivision Ascomycotina, the class of Chaetomium (Sordariomycetes), the subclass Hypocrea (Hypocrea), the order Hypocrea (Hypocrea), the family of Cordyceps (Cordycipitaceae), the genus of Cordyceps (Cordyceps). Cordyceps militaris usually has similar bioactive components to Cordyceps militaris, mainly comprises cordycepin, cordycepic acid, Cordyceps polysaccharide, ergosterol, amino acid, nucleoside substances and the like, has similar efficacy to Cordyceps militaris, and is a good substitute for Cordyceps militaris. Cordyceps militaris is a natural resource and is distributed worldwide, but the quantity of the Cordyceps militaris is small, and most of the Cordyceps militaris depends on artificial cultivation. In recent years, the chemical components and pharmacological activity of the artificial cordyceps are widely researched by human beings, and the results show that the chemical components and pharmacological effects of the artificial cordyceps are basically the same as those of natural cordyceps, wherein the contents of cordyceps polysaccharide and cordycepin are higher than those of the natural cordyceps. A large number of modern pharmacological studies show that the cordyceps militaris not only has special nutritional value, but also has obvious medicinal value.
Adenosine is an endogenous purine nucleotide, belongs to nucleoside substances, is one of active ingredients of cordyceps sinensis, is a direct premise of cordycepin, and has wide physiological activity, and the content of adenosine in cordyceps militaris is more than 0.01 percent, is similar to that of cordyceps sinensis, and is even higher. To date, 11 nucleosides have been isolated and identified from cordyceps and fermentation hyphae, including adenine, adenosine, uracil, uridine, guanine, guanosine, hypoxanthine, inosine, thymine, thymidine, and uridine. Cordyceps adenosine can improve cardiovascular and cerebrovascular blood circulation, prevent arrhythmia, and lower blood pressure; can reduce the release of norepinephrine, and the cordyceps adenosine weakens the excitation effect, namely stimulation, of catecholamine delivery on the heart, thereby protecting the heart; plays an important role in myocardial ischemia pre-adaptation and ischemia reperfusion injury; cordyceps adenosine also has strong platelet aggregation inhibiting effect, is a strong vasoactive substance, and can dilate systemic circulation blood vessels to cause blood pressure to be stabilized at normal low level. Thus, the cordyceps adenosine has better effects of resisting virus and bacteria, preventing and treating cerebral thrombosis and cerebral hemorrhage, inhibiting platelet accumulation and preventing thrombosis. However, both natural cordyceps and artificial cordyceps have very low content of cordyceps adenosine, which causes certain difficulty in extracting cordyceps adenosine. In the prior art, the cordyceps sinensis adenosine is extracted by adopting an enzyme method, but the problem of low extraction efficiency generally exists.
Disclosure of Invention
Therefore, the technical problem to be solved by the invention is to provide a method for efficiently extracting adenosine from cordyceps militaris so as to solve the problem of low extraction rate of adenosine from cordyceps militaris in the prior art.
In order to solve the technical problems, the method for efficiently extracting adenosine from cordyceps militaris comprises the following steps:
(1) drying Cordyceps militaris fruiting body, and adding water to make into Cordyceps militaris pulp for use;
(2) adding compound extraction enzyme into the obtained Cordyceps militaris pulp, and extracting to obtain Cordyceps adenosine-containing extract.
In the step (2), the compound extraction enzyme comprises laccase or xylanase and cellulase or pectinase.
Preferably, the complex extraction enzyme comprises:
the mass ratio is 1-2: 1-3 laccase and cellulase; alternatively, the first and second electrodes may be,
the mass ratio is 1-2: 2-4 laccase and pectinase; alternatively, the first and second electrodes may be,
the mass ratio is 2-3: 1-2 xylanase and cellulase; alternatively, the first and second electrodes may be,
the mass ratio is 2-3: 3-4 xylanase and pectinase.
Preferably, the adding amount of the compound extracting enzyme accounts for 3-8 wt% of the cordyceps militaris sporocarp.
Preferably, in the step of pulping, the ratio of the cordyceps militaris sporocarp to water is 1: 20-30 g/mL.
Preferably, the pulping step further comprises the step of adding polyethylene glycol into water for pulping.
More preferably, the polyethylene glycol comprises polyethylene glycol 400.
Preferably, in the pulping step, the mass concentration of the polyethylene glycol aqueous solution is controlled to be 10-20 wt%.
Preferably, the step (1) further comprises the step of performing gradient drying on the cordyceps militaris sporocarp at 40-80 ℃.
Specifically, the gradient drying step specifically includes: controlling the initial drying temperature of the cordyceps militaris sporocarp to be 40-50 ℃, and heating and drying the cordyceps militaris sporocarp at the temperature of 5 ℃/10min until the weight of the cordyceps militaris sporocarp is constant, and controlling the temperature of the drying step to be not more than 80 ℃.
According to the method for extracting adenosine from cordyceps militaris, on the basis of taking water as an extraction solvent in the prior art, a selected amount of composite extraction enzyme comprising laccase or xylanase and cellulase or pectinase is further added for enzymolysis extraction, so that the extraction rate of adenosine from cordyceps militaris is effectively improved; according to the method for extracting adenosine from cordyceps militaris, the dissolution and extraction of adenosine from cordyceps militaris are effectively improved by adding polyethylene glycol in the step of pulping; preferably, in the method, before extraction, the cordyceps militaris sporocarp is dried in a gradient drying mode, and the temperature difference of each stage is utilized, so that the wall breaking effect of the cordyceps militaris sporocarp is further improved, the dissolution of adenosine is effectively promoted, and the extraction rate of adenosine is also improved.
Detailed Description
Example 1
The method for efficiently extracting adenosine from cordyceps militaris comprises the following steps:
(1) drying Cordyceps militaris fruiting body at 50 deg.C to constant temperature, collecting dried Cordyceps militaris fruiting body 1kg, adding water 20L, and conventionally pulping to obtain Cordyceps militaris pulp;
(2) adding 30g of compound extraction enzyme into the obtained cordyceps militaris pulp for extraction for 2h, wherein the compound extraction enzyme comprises the following components in percentage by mass: 3 laccase and cellulase; obtaining the extract containing the cordyceps sinensis adenosine.
Example 2
The method for efficiently extracting adenosine from cordyceps militaris comprises the following steps:
(1) drying Cordyceps militaris fruiting body at 50 deg.C to constant temperature, collecting dried Cordyceps militaris fruiting body 1kg, adding water 30L, and conventionally pulping to obtain Cordyceps militaris pulp;
(2) adding 80g of compound extraction enzyme into the obtained cordyceps militaris pulp for extraction for 2h, wherein the compound extraction enzyme comprises the following components in percentage by mass: 1 laccase and cellulase; obtaining the extract containing the cordyceps sinensis adenosine.
Example 3
The method for efficiently extracting adenosine from cordyceps militaris comprises the following steps:
(1) drying Cordyceps militaris fruiting body at 50 deg.C to constant temperature, collecting dried Cordyceps militaris fruiting body 1kg, adding water 25L, and conventionally pulping to obtain Cordyceps militaris pulp;
(2) adding 50g of compound extraction enzyme into the obtained cordyceps militaris pulp for extraction for 2 hours, wherein the compound extraction enzyme comprises the following components in a mass ratio of 1.5: 2 laccase and cellulase; obtaining the extract containing the cordyceps sinensis adenosine.
Example 4
The method for efficiently extracting adenosine from cordyceps militaris comprises the following steps:
(1) drying Cordyceps militaris fruiting body at 50 deg.C to constant temperature, collecting dried Cordyceps militaris fruiting body 1kg, adding water 20L, and conventionally pulping to obtain Cordyceps militaris pulp;
(2) adding 30g of compound extraction enzyme into the obtained cordyceps militaris pulp for extraction for 2h, wherein the compound extraction enzyme comprises the following components in percentage by mass: 4 laccase and pectinase; obtaining the extract containing the cordyceps sinensis adenosine.
Example 5
The method for efficiently extracting adenosine from cordyceps militaris comprises the following steps:
(1) drying Cordyceps militaris fruiting body at 50 deg.C to constant temperature, collecting dried Cordyceps militaris fruiting body 1kg, adding water 30L, and conventionally pulping to obtain Cordyceps militaris pulp;
(2) adding 80g of compound extraction enzyme into the obtained cordyceps militaris pulp for extraction for 2h, wherein the compound extraction enzyme comprises the following components in percentage by mass: 2 laccase and pectinase; obtaining the extract containing the cordyceps sinensis adenosine.
Example 6
The method for efficiently extracting adenosine from cordyceps militaris comprises the following steps:
(1) drying Cordyceps militaris fruiting body at 50 deg.C to constant temperature, collecting dried Cordyceps militaris fruiting body 1kg, adding water 25L, and conventionally pulping to obtain Cordyceps militaris pulp;
(2) adding 50g of compound extraction enzyme into the obtained cordyceps militaris pulp for extraction for 2 hours, wherein the compound extraction enzyme comprises the following components in a mass ratio of 1.5: 3 laccase and pectinase; obtaining the extract containing the cordyceps sinensis adenosine.
Example 7
The method for efficiently extracting adenosine from cordyceps militaris comprises the following steps:
(1) drying Cordyceps militaris fruiting body at 50 deg.C to constant temperature, collecting dried Cordyceps militaris fruiting body 1kg, adding water 20L, and conventionally pulping to obtain Cordyceps militaris pulp;
(2) adding 30g of compound extraction enzyme into the obtained cordyceps militaris pulp for extraction for 2h, wherein the compound extraction enzyme comprises the following components in percentage by mass: 2 and a cellulase; obtaining the extract containing the cordyceps sinensis adenosine.
Example 8
The method for efficiently extracting adenosine from cordyceps militaris comprises the following steps:
(1) drying Cordyceps militaris fruiting body at 50 deg.C to constant temperature, collecting dried Cordyceps militaris fruiting body 1kg, adding water 30L, and conventionally pulping to obtain Cordyceps militaris pulp;
(2) adding 80g of compound extraction enzyme into the obtained cordyceps militaris pulp for extraction for 2 hours, wherein the compound extraction enzyme comprises the following components in percentage by mass: 1, xylanase and cellulase; obtaining the extract containing the cordyceps sinensis adenosine.
Example 9
The method for efficiently extracting adenosine from cordyceps militaris comprises the following steps:
(1) drying Cordyceps militaris fruiting body at 50 deg.C to constant temperature, collecting dried Cordyceps militaris fruiting body 1kg, adding water 25L, and conventionally pulping to obtain Cordyceps militaris pulp;
(2) adding 50g of compound extraction enzyme into the obtained cordyceps militaris pulp for extraction for 2 hours, wherein the compound extraction enzyme comprises the following components in a mass ratio of 2.5: 1.5 xylanase and cellulase; obtaining the extract containing the cordyceps sinensis adenosine.
Example 10
The method for efficiently extracting adenosine from cordyceps militaris comprises the following steps:
(1) drying Cordyceps militaris fruiting body at 50 deg.C to constant temperature, collecting dried Cordyceps militaris fruiting body 1kg, adding water 20L, and conventionally pulping to obtain Cordyceps militaris pulp;
(2) adding 30g of compound extraction enzyme into the obtained cordyceps militaris pulp for extraction for 2h, wherein the compound extraction enzyme comprises the following components in percentage by mass: 4 xylanase and pectinase; obtaining the extract containing the cordyceps sinensis adenosine.
Example 11
The method for efficiently extracting adenosine from cordyceps militaris comprises the following steps:
(1) drying Cordyceps militaris fruiting body at 50 deg.C to constant temperature, collecting dried Cordyceps militaris fruiting body 1kg, adding water 30L, and conventionally pulping to obtain Cordyceps militaris pulp;
(2) adding 80g of compound extraction enzyme into the obtained cordyceps militaris pulp for extraction for 2 hours, wherein the compound extraction enzyme comprises the following components in percentage by mass: 3 xylanase and pectinase; obtaining the extract containing the cordyceps sinensis adenosine.
Example 12
The method for efficiently extracting adenosine from cordyceps militaris comprises the following steps:
(1) drying Cordyceps militaris fruiting body at 50 deg.C to constant temperature, collecting dried Cordyceps militaris fruiting body 1kg, adding water 25L, and conventionally pulping to obtain Cordyceps militaris pulp;
(2) adding 50g of compound extraction enzyme into the obtained cordyceps militaris pulp for extraction for 2 hours, wherein the compound extraction enzyme comprises the following components in a mass ratio of 2.5: 3.5 xylanase and pectinase; obtaining the extract containing the cordyceps sinensis adenosine.
Example 13
The method for efficiently extracting adenosine from cordyceps militaris is the same as that in example 3, and the difference is only that the step of pulping is to adopt polyethylene glycol 400 aqueous solution with the mass concentration of 10 wt% to carry out pulping.
Example 14
The method for efficiently extracting adenosine from cordyceps militaris is the same as that in example 6, and the difference is only that the step of pulping is to adopt polyethylene glycol 400 aqueous solution with mass concentration of 20 wt% to carry out pulping.
Example 15
The method for efficiently extracting adenosine from cordyceps militaris is the same as that in example 9, and the difference is only that the step of pulping is to adopt polyethylene glycol 400 aqueous solution with mass concentration of 15 wt% to carry out pulping.
Example 16
The method for efficiently extracting adenosine from cordyceps militaris is the same as that in example 12, and the difference is only that the step of pulping is to adopt polyethylene glycol 400 aqueous solution with mass concentration of 15 wt% to carry out pulping.
Example 17
The method for efficiently extracting adenosine from cordyceps militaris is the same as that in example 13, and the difference is that the step of drying cordyceps militaris sporocarp is a gradient drying process, and the method specifically comprises the following steps: controlling the initial drying temperature of the cordyceps militaris sporocarp to be 40 ℃, and performing heating and drying by a program of 5 ℃/10min until the cordyceps militaris sporocarp is constant in weight, and controlling the temperature of the drying step to be not more than 80 ℃.
Example 18
The method for efficiently extracting adenosine from cordyceps militaris is the same as that in the embodiment 14, and the difference is that the step of drying the cordyceps militaris sporocarp is a gradient drying process, and the method specifically comprises the following steps of: controlling the initial drying temperature of the cordyceps militaris sporocarp to be 50 ℃, and heating and drying the cordyceps militaris sporocarp at the temperature of 5 ℃/10min until the weight of the cordyceps militaris sporocarp is constant, wherein the temperature in the drying step is controlled not to exceed 80 ℃.
Example 19
The method for efficiently extracting adenosine from cordyceps militaris is the same as that in example 15, and the difference is that the step of drying cordyceps militaris sporocarp is a gradient drying process, and the method specifically comprises the following steps: controlling the initial drying temperature of the cordyceps militaris sporocarp to be 45 ℃, and performing heating and drying by a program of 5 ℃/10min until the cordyceps militaris sporocarp is constant in weight, and controlling the temperature of the drying step to be not more than 80 ℃.
Example 20
The method for efficiently extracting adenosine from cordyceps militaris is the same as that in example 16, and the difference is only that the step of drying cordyceps militaris sporocarp is a gradient drying process, and the method specifically comprises the following steps: controlling the initial drying temperature of the cordyceps militaris sporocarp to be 45 ℃, and performing heating and drying by a program of 5 ℃/10min until the cordyceps militaris sporocarp is constant in weight, and controlling the temperature of the drying step to be not more than 80 ℃.
Comparative example 1
The method for extracting adenosine from cordyceps militaris adopts the method in the prior art, namely, the method directly adds the same amount of water to extract the adenosine.
Comparative example 2
The method for extracting adenosine from cordyceps militaris adopts the method in the prior art, namely, 60 wt% of ethanol solution with the same amount is directly added for pulping and extracting.
Comparative example 3
The method for extracting adenosine from cordyceps militaris adopts the method in the prior art, namely, cellulase with the same quantity as the compound extracting enzyme is added for enzymolysis extraction after the step of adding water for pulping.
Comparative example 4
The method for extracting adenosine from cordyceps militaris adopts the method in the prior art, namely, pectinase with the same amount as the compound extracting enzyme is added for enzymolysis extraction after the step of adding water for pulping.
Comparative example 5
The method for extracting adenosine from cordyceps militaris adopts the method in the prior art, namely adding protein with the same amount as the compound extracting enzyme for enzymolysis extraction after the step of adding water for pulping.
Examples of the experiments
The extract liquids collected in the above examples 1-20 and comparative examples 1-5 were respectively used for detecting the content of cordyceps adenosine, and the content of cordyceps adenosine in the extract liquid was determined by the conventional HPLC method in the prior art, i.e. the detection method described in chinese patent CN102558265A was used to calculate the extraction amount of cordyceps adenosine in the extract liquid relative to the extraction efficiency of cordyceps militaris fruiting bodies, and the results are recorded in table 1 below.
TABLE 1 Cordyceps militaris adenosine extraction Rate results
From the data in the table, the method for extracting adenosine from cordyceps militaris further adds a selected amount of composite extracting enzyme comprising laccase or xylanase and cellulase or pectinase for enzymolysis extraction on the basis of taking water as an extraction solvent in the prior art, so that the extraction rate of adenosine from cordyceps militaris is effectively improved; according to the method for extracting adenosine from cordyceps militaris, the dissolution and extraction of adenosine from cordyceps militaris are effectively improved by adding polyethylene glycol in the step of pulping; preferably, in the method, before extraction, the cordyceps militaris sporocarp is dried in a gradient drying mode, and the temperature difference of each stage is utilized, so that the wall breaking effect of the cordyceps militaris sporocarp is further improved, the dissolution of adenosine is effectively promoted, and the extraction rate of adenosine is also improved.
It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications therefrom are within the scope of the invention.
Claims (5)
1. A method for extracting adenosine from Cordyceps militaris is characterized by comprising the following steps:
(1) gradient drying fruiting body of Cordyceps militaris at 40-80 deg.C, adding water solution containing polyethylene glycol, and pulping to obtain Cordyceps militaris pulp;
the gradient drying step specifically comprises: controlling the initial drying temperature of the cordyceps militaris sporocarp to be 40-50 ℃, and heating and drying the cordyceps militaris sporocarp at the temperature of 5 ℃/10min until the weight of the cordyceps militaris sporocarp is constant, wherein the temperature in the drying step is controlled not to exceed 80 ℃;
(2) adding compound extraction enzyme into the obtained Cordyceps militaris pulp, and extracting for 2h to obtain extract containing adenosine of Cordyceps militaris; the adding amount of the compound extracting enzyme accounts for 3-8 wt% of the cordyceps militaris sporocarp;
the compound extraction enzyme is composed of laccase or xylanase and cellulase or pectinase.
2. The method for extracting adenosine from cordyceps militaris, according to claim 1, wherein the compound extraction enzyme comprises the following components in percentage by mass:
the mass ratio is 1-2: 1-3 laccase and cellulase; alternatively, the first and second electrodes may be,
the mass ratio is 1-2: 2-4 laccase and pectinase; alternatively, the first and second electrodes may be,
the mass ratio is 2-3: 1-2 xylanase and cellulase; alternatively, the first and second electrodes may be,
the mass ratio is 2-3: 3-4 xylanase and pectinase.
3. The method for extracting adenosine from cordyceps militaris, according to claim 2, wherein in the step of pulping, the ratio of the cordyceps militaris fruit body to water is 1: 20-30 g/mL.
4. The method for extracting adenosine from Cordyceps militaris as claimed in any one of claims 1 to 3, wherein the polyethylene glycol comprises polyethylene glycol 400.
5. The method for extracting adenosine from Cordyceps militaris (L.) Link, as claimed in claim 4, wherein in the step of pulping, the mass concentration of the aqueous solution of polyethylene glycol is controlled to be 10-20 wt%.
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| CN113480587B (en) * | 2021-05-20 | 2024-05-03 | 湖南合健生物科技有限公司 | Method for efficiently extracting N6- (2-hydroxyethyl) adenosine from cordyceps sobolifera fruiting bodies |
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