CN113817077A - Extraction method of caulerpa lentillifera polysaccharide - Google Patents
Extraction method of caulerpa lentillifera polysaccharide Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
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Abstract
The invention discloses a method for extracting caulerpa lentillifera polysaccharide, which comprises the steps of washing the caulerpa lentillifera with seawater and distilled water in sequence, drying and crushing to obtain caulerpa lentillifera powder; adding the caulerpa lentillifera powder into ultrapure water and papain, extracting for 2-4 h, inactivating enzyme under the condition of a boiling water bath, and then performing centrifugal separation to obtain supernatant and algae residues; concentrating the supernatant under negative pressure, adding ethanol for alcohol precipitation, centrifuging, and removing the upper layer of ethanol to obtain precipitate A; simultaneously, extracting the algae residue according to the same steps to obtain a precipitate B, combining the precipitate A and the precipitate B, completely dissolving the precipitate A and the precipitate B in ultrapure water, and freeze-drying to obtain the caulerpine lentinan. The method can improve the extraction rate of the caulerpa lentillifera polysaccharide, and has the advantages of easy preparation, low cost, no pollution, high yield and the like.
Description
Technical Field
The invention belongs to the technical field of extraction of long-stem grape bracken polysaccharide, and particularly relates to an extraction method of long-stem grape bracken polysaccharide.
Background
The Caulerpa longipedicularis is a genus of green algae, commonly known as sea grape (acalerpa lentillifera) native to the lucidus, the shape of the erect branch of the Caulerpa lentillifera is similar to that of the grape, the algal body can be divided into three parts, namely a stolon, an erect branch and a filiform rhizoid, and the main axis of the erect branch is covered by small spherical branches. The whole algal body is a multinucleated cell, and although a plurality of nuclei exist, no cell wall is arranged between the nuclei. The caulerpa longipedicularis is an edible green alga rich in nutrition, and contains various amino acids, polysaccharides and vitamins required by a human body, omega-6 fatty acids such as oleic acid, arachidonic acid and the like, wherein the content of essential amino acids/non-essential amino acids (EAA/NEAA) in the caulerpa longipedicularis is 54.36-69.75%, the content of essential amino acids/total amino acids (EAA/TAA) is 35.22-48.19%, the content of total phenols is 2.04mg/g, the content of flavonoids compounds is 4.93mg/g, the content of vitamin E is 0.22mg/kg-0.31 mg/kg, the content of vitamin C is 3.47mg/kg-5.03 mg/kg, the content of crude fibers is 1.91-12.98%, the content of crude fat is 0.81-4.40%, the content of protein is 9.22-19.38%, and the content of ash is 24.00-56.08%; the botryococcus longipediculus polysaccharide is mainly sulfated polysaccharide with the content of 34.00-64.97%, and in the monosaccharide composition, the contents of mannose, galactose and xylose are the highest, namely 28.38-48.26%, 28.47-49.93% and 13.32-59.8% respectively. Researches show that the Caulerpa lentillifera polysaccharide has alpha-glucosidase inhibition effect, antioxidant effect and immunoregulation effect, and the Caulerpa lentillifera has immunostimulation activity on macrophages and has wide application value and prospect. However, when the polysaccharide of the caulerpa lentillifera is separated and extracted at present, the extraction methods of a water extraction and alcohol precipitation method and an alkali extraction method are adopted, so that the problems of low polysaccharide yield, high cost, complex preparation process and the like exist, the production efficiency is low, the product quality is poor, and the application and popularization of the caulerpa lentillifera polysaccharide are hindered.
Disclosure of Invention
Aiming at the defects, the invention discloses a method for extracting the caulerpa lentillifera polysaccharide, which improves the extraction rate of the caulerpa lentillifera polysaccharide and has the advantages of easy preparation, low cost, no pollution, high yield and the like.
The invention is realized by adopting the following technical scheme:
a method for extracting Caulerpa lentillifera polysaccharide comprises the following steps:
(1) washing caulerpa lentillifera with seawater, washing with distilled water for 3 times, drying in an oven at 50 deg.C, pulverizing, sieving with 50 mesh sieve to obtain caulerpa lentillifera powder, and drying and storing;
(2) adding ultrapure water into caulerpa longipedica powder, uniformly mixing to obtain a mixed solution A, wherein the ratio of the mass of the caulerpa longipedica powder to the volume of the ultrapure water is 1g (40-50) ml, then adjusting the pH of the mixed solution A to 6, adding papain, and extracting at 40-60 ℃ for 2-4 h to obtain a mixed solution B, wherein the adding amount of the papain is 1-2.5% of the mass of the mixed solution A;
(3) inactivating enzyme of the mixed liquor B obtained in the step (2) for 5min under the condition of boiling water bath, centrifuging for 15min under the condition that the centrifugal force is 3500Xg, and separating to obtain supernatant and algae residue; placing the supernatant in a rotary evaporator, concentrating under negative pressure at 50 ℃ until the concentration is 1g/ml to obtain a concentrated extracting solution, naturally cooling the concentrated extracting solution to room temperature, and adding 95% ethanol by volume to obtain a mixed solution C, wherein the volume of the ethanol added is 5 times of the volume of the concentrated extracting solution; precipitating the mixed solution C at 4 deg.C for 12h, centrifuging at 3500Xg for 15min, removing the upper layer ethanol, and collecting precipitate A;
(4) and (3) sequentially extracting the algae residues obtained in the step (3) according to the step (2) and the step (3) to obtain a precipitate B, combining the precipitate A and the precipitate B, completely dissolving the precipitate A and the precipitate B in ultrapure water to obtain a mixed solution D, and freeze-drying the mixed solution D for 36 hours to obtain the caulerpine lentinan.
Further, in the step (2), the pH is adjusted to 6 by adding hydrochloric acid having a mass concentration of 10% or a sodium hydroxide solution having a mass concentration of 5% to the mixed solution a.
Further, in the extraction process of the step (2), the mixed solution A is stirred every 30min for 2min each time.
Compared with the prior art, the technical scheme has the following beneficial effects:
aiming at the physiological structure and the nutritional composition of the caulerpa lentillifera, the invention adopts papain to carry out enzymolysis, scientifically controls various factor parameters such as the feed-liquid ratio, the extraction temperature, the extraction time, the addition of the papain, the total extraction frequency and the like which influence the extraction rate in the extraction process, and combines an optimized alcohol precipitation process to form a complete set of complete extraction method of the caulerpa lentillifera polysaccharide, thereby improving the extraction rate of the caulerpa lentillifera polysaccharide and leading the extraction rate of the caulerpa lentillifera polysaccharide to reach more than 40 percent.
Drawings
FIG. 1 is a graph showing the results obtained in step (2) of the one-factor test of Experimental example 1.
FIG. 2 is a graph showing the results obtained in step (3) of the one-factor test of Experimental example 1.
FIG. 3 is a graph showing the results obtained in step (4) in the one-factor test of Experimental example 1.
FIG. 4 is a graph showing the results obtained in step (5) in the one-factor test of Experimental example 1.
FIG. 5 is a graph showing the results obtained in step (6) in the one-factor test of Experimental example 1.
FIG. 6 shows the design and results of Box-Behnken test for polysaccharide extraction from Botrytis longipedicularis in Experimental example 2.
FIG. 7 is the results of analysis of the variance and regression coefficient of polysaccharide yields in the response surface quadratic model of Experimental example 2.
FIG. 8 is a graph showing the effect of feed-to-liquid ratio and extraction time on the extraction yield of Caulerpa lentinan in Experimental example 2.
FIG. 9 is a graph showing the effect of feed-to-liquid ratio and extraction temperature on the extraction rate of Caulerpa lentinan in Experimental example 2.
FIG. 10 is a graph showing the effect of extraction time and extraction temperature on the extraction rate of Caulerpa lentinan in Experimental example 2.
Detailed Description
The invention is further illustrated by the following examples, which are not to be construed as limiting the invention thereto. The specific experimental conditions and methods not indicated in the following examples are generally conventional means well known to those skilled in the art.
Example 1:
a method for extracting Caulerpa lentillifera polysaccharide comprises the following steps:
(1) washing caulerpa lentillifera with seawater, washing with distilled water for 3 times, drying in an oven at 50 deg.C, pulverizing, sieving with 50 mesh sieve to obtain caulerpa lentillifera powder, and drying and storing;
(2) adding ultrapure water into caulerpa longipedica powder, uniformly mixing to obtain a mixed solution A, wherein the ratio of the mass of the caulerpa longipedica powder to the volume of the ultrapure water is 1g:45ml, adding 10% hydrochloric acid or 5% sodium hydroxide solution into the mixed solution A to adjust the pH value to 6, adding papain, extracting at 60 ℃ for 3 hours, stirring the mixed solution A every 30 minutes for 2 minutes every time to obtain a mixed solution B, wherein the adding amount of the papain is 2% of the mass of the mixed solution A;
(3) inactivating enzyme of the mixed liquor B obtained in the step (2) for 5min under the condition of boiling water bath, centrifuging for 15min under the condition that the centrifugal force is 3500Xg, and separating to obtain supernatant and algae residue; placing the supernatant in a rotary evaporator, concentrating under negative pressure at 50 ℃ until the concentration is 1g/ml to obtain a concentrated extracting solution, naturally cooling the concentrated extracting solution to room temperature, and adding 95% ethanol by volume to obtain a mixed solution C, wherein the volume of the ethanol added is 5 times of the volume of the concentrated extracting solution; precipitating the mixed solution C at 4 deg.C for 12h, centrifuging at 3500Xg for 15min, removing the upper layer ethanol, and collecting precipitate A;
(4) and (3) sequentially extracting the algae residues obtained in the step (3) according to the step (2) and the step (3) to obtain a precipitate B, combining the precipitate A and the precipitate B, completely dissolving the precipitate A and the precipitate B in ultrapure water to obtain a mixed solution D, and freeze-drying the mixed solution D for 36 hours to obtain the caulerpine lentinan.
Example 2:
a method for extracting Caulerpa lentillifera polysaccharide comprises the following steps:
(1) washing caulerpa lentillifera with seawater, washing with distilled water for 3 times, drying in an oven at 50 deg.C, pulverizing, sieving with 50 mesh sieve to obtain caulerpa lentillifera powder, and drying and storing;
(2) adding ultrapure water into caulerpa longipedica powder, uniformly mixing to obtain a mixed solution A, wherein the mass ratio of the caulerpa longipedica powder to the volume of the ultrapure water is 1g:50ml, adding 10% hydrochloric acid or 5% sodium hydroxide solution into the mixed solution A to adjust the pH value to 6, adding papain, extracting at 60 ℃ for 4 hours, stirring the mixed solution A every 30 minutes for 2 minutes every time to obtain a mixed solution B, wherein the adding amount of the papain is 2.5% of the mass of the mixed solution A;
(3) inactivating enzyme of the mixed liquor B obtained in the step (2) for 5min under the condition of boiling water bath, centrifuging for 15min under the condition that the centrifugal force is 3500Xg, and separating to obtain supernatant and algae residue; placing the supernatant in a rotary evaporator, concentrating under negative pressure at 50 ℃ until the concentration is 1g/ml to obtain a concentrated extracting solution, naturally cooling the concentrated extracting solution to room temperature, and adding 95% ethanol by volume to obtain a mixed solution C, wherein the volume of the ethanol added is 5 times of the volume of the concentrated extracting solution; precipitating the mixed solution C at 4 deg.C for 12h, centrifuging at 3500Xg for 15min, removing the upper layer ethanol, and collecting precipitate A;
(4) and (3) sequentially extracting the algae residues obtained in the step (3) according to the step (2) and the step (3) to obtain a precipitate B, combining the precipitate A and the precipitate B, completely dissolving the precipitate A and the precipitate B in ultrapure water to obtain a mixed solution D, and freeze-drying the mixed solution D for 36 hours to obtain the caulerpine lentinan.
Example 3:
a method for extracting Caulerpa lentillifera polysaccharide comprises the following steps:
(1) washing caulerpa lentillifera with seawater, washing with distilled water for 3 times, drying in an oven at 50 deg.C, pulverizing, sieving with 50 mesh sieve to obtain caulerpa lentillifera powder, and drying and storing;
(2) adding ultrapure water into caulerpa longipedica powder, uniformly mixing to obtain a mixed solution A, wherein the ratio of the mass of the caulerpa longipedica powder to the volume of the ultrapure water is 1g:40ml, adding 10% hydrochloric acid or 5% sodium hydroxide solution into the mixed solution A to adjust the pH value to 6, adding papain, extracting at 50 ℃ for 3 hours, stirring the mixed solution A every 30 minutes for 2 minutes every time to obtain a mixed solution B, wherein the adding amount of the papain is 2% of the mass of the mixed solution A;
(3) inactivating enzyme of the mixed solution B obtained in the step (2) for 5min in boiling water under the condition of boiling water bath, centrifuging for 15min under the condition that the centrifugal force is 3500Xg, and separating to obtain supernatant and algae residue; placing the supernatant in a rotary evaporator, concentrating under negative pressure at 50 ℃ until the concentration is 1g/ml to obtain a concentrated extracting solution, naturally cooling the concentrated extracting solution to room temperature, and adding 95% ethanol by volume to obtain a mixed solution C, wherein the volume of the ethanol added is 5 times of the volume of the concentrated extracting solution; precipitating the mixed solution C at 4 deg.C for 12h, centrifuging at 3500Xg for 15min, removing the upper layer ethanol, and collecting precipitate A;
(4) and (3) sequentially extracting the algae residues obtained in the step (3) according to the step (2) and the step (3) to obtain a precipitate B, combining the precipitate A and the precipitate B, completely dissolving the precipitate A and the precipitate B in ultrapure water to obtain a mixed solution D, and freeze-drying the mixed solution D for 36 hours to obtain the caulerpine lentinan.
Example 4:
a method for extracting Caulerpa lentillifera polysaccharide comprises the following steps:
(1) washing caulerpa lentillifera with seawater, washing with distilled water for 3 times, drying in an oven at 50 deg.C, pulverizing, sieving with 50 mesh sieve to obtain caulerpa lentillifera powder, and drying and storing;
(2) adding ultrapure water into caulerpa longipedica powder, uniformly mixing to obtain a mixed solution A, wherein the ratio of the mass of the caulerpa longipedica powder to the volume of the ultrapure water is 1g:40ml, adding 10% hydrochloric acid or 5% sodium hydroxide solution into the mixed solution A to adjust the pH value to 6, adding papain, extracting at 40 ℃ for 2 hours, stirring the mixed solution A every 30 minutes for 2 minutes every time to obtain a mixed solution B, wherein the adding amount of the papain is 1% of the mass of the mixed solution A;
(3) inactivating enzyme of the mixed liquor B obtained in the step (2) for 5min under the condition of boiling water bath, centrifuging for 15min under the condition that the centrifugal force is 3500Xg, and separating to obtain supernatant and algae residue; placing the supernatant in a rotary evaporator, concentrating under negative pressure at 50 ℃ until the concentration is 1g/ml to obtain a concentrated extracting solution, naturally cooling the concentrated extracting solution to room temperature, and adding 95% ethanol by volume to obtain a mixed solution C, wherein the volume of the ethanol added is 5 times of the volume of the concentrated extracting solution; precipitating the mixed solution C at 4 deg.C for 12h, centrifuging at 3500Xg for 15min, removing the upper layer ethanol, and collecting precipitate A;
(4) and (3) sequentially extracting the algae residues obtained in the step (3) according to the step (2) and the step (3) to obtain a precipitate B, combining the precipitate A and the precipitate B, completely dissolving the precipitate A and the precipitate B in ultrapure water to obtain a mixed solution D, and freeze-drying the mixed solution D for 36 hours to obtain the caulerpine lentinan.
Example 5:
a method for extracting Caulerpa lentillifera polysaccharide comprises the following steps:
(1) washing caulerpa lentillifera with seawater, washing with distilled water for 3 times, drying in an oven at 50 deg.C, pulverizing, sieving with 50 mesh sieve to obtain caulerpa lentillifera powder, and drying and storing;
(2) adding ultrapure water into caulerpa longipedica powder, uniformly mixing to obtain a mixed solution A, wherein the mass ratio of the caulerpa longipedica powder to the volume of the ultrapure water is 1g:45ml, adding 10% hydrochloric acid or 5% sodium hydroxide solution into the mixed solution A to adjust the pH value to 6, adding papain, extracting at 50 ℃ for 4 hours, stirring the mixed solution A every 30 minutes for 2 minutes every time to obtain a mixed solution B, wherein the adding amount of the papain is 2.5% of the mass of the mixed solution A;
(3) inactivating enzyme of the mixed liquor B obtained in the step (2) for 5min under the condition of boiling water bath, centrifuging for 15min under the condition that the centrifugal force is 3500Xg, and separating to obtain supernatant and algae residue; placing the supernatant in a rotary evaporator, concentrating under negative pressure at 50 ℃ until the concentration is 1g/ml to obtain a concentrated extracting solution, naturally cooling the concentrated extracting solution to room temperature, and adding 95% ethanol by volume to obtain a mixed solution C, wherein the volume of the ethanol added is 5 times of the volume of the concentrated extracting solution; precipitating the mixed solution C at 4 deg.C for 12h, centrifuging at 3500Xg for 15min, removing the upper layer ethanol, and collecting precipitate A;
(4) and (3) sequentially extracting the algae residues obtained in the step (3) according to the step (2) and the step (3) to obtain a precipitate B, combining the precipitate A and the precipitate B, completely dissolving the precipitate A and the precipitate B in ultrapure water to obtain a mixed solution D, and freeze-drying the mixed solution D for 36 hours to obtain the caulerpine lentinan.
Experimental example 1:
and (3) drawing a polysaccharide standard curve:
accurately weighing 10mg of standard glucose, and putting the standard glucose into a 100ml volumetric flask for constant volume; taking a 7-support glass test tube, respectively adding 0ml, 0.1ml, 0.2ml, 0.4ml, 0.6ml, 0.8ml and 1.0m of glucose solution, adding ultrapure water to make up the volume to 1.0ml, fully and uniformly mixing, quickly placing in an ice bath for cooling for 5min, taking out, adding a prepared 0.2% anthrone sulfuric acid solution, shaking uniformly, immediately placing in a boiling water bath for boiling for 10min, taking out, cooling to room temperature, and measuring the OD value at the wavelength of 492 nm; drawing a standard curve by taking the standard glucose content (microgram) as a horizontal coordinate and the light absorption value as a vertical coordinate to obtain a linear regression equation: y =0.0034x +0.0841, R2= 0.9992.
Determination of polysaccharide extraction rate of Caulerpa lentillifera:
dissolving the cauliflower within ultrapure water, adding ultrapure water for dilution, accurately sucking 1ml of the solution, placing the solution into a glass test tube with a plug (three times for each sample), rapidly placing the glass test tube in an ice bath for cooling for 5min, taking out the sample, adding a prepared 0.2% anthrone sulfuric acid solution, shaking up, immediately placing the sample in a boiling water bath for boiling for 10min, taking out the sample, cooling to room temperature, and measuring the OD value at the wavelength of 492 nm; calculating the polysaccharide concentration according to the standard curve, and calculating the polysaccharide extraction rate in the caulerpa lentillifera sample according to the following formula:
extraction rate of Caulerpa lentillifera polysaccharide = (polysaccharide concentration x to-be-measured volume x dilution multiple/mass of polysaccharide sample) × polysaccharide yield x 100%
Single factor test:
(1) adding 10g of Caulerpa longipediculosa powder into 200ml of ultrapure water to obtain mixed solution A, adding papain with a mass of 2.5% of that of the mixed solution A, extracting at 60 deg.C for 2h with pH of 6, wherein the mixed solution A is stirred every 30min for 2min each time; inactivating enzyme for 5min under boiling water bath condition after extraction, centrifuging for 15min under centrifugal force of 3500Xg, and collecting supernatant and algae residue; placing the supernatant in a rotary evaporator, concentrating under negative pressure at 50 ℃ to obtain 10ml of concentrated extract, adding 50ml of ethanol with the volume fraction of 95%, then carrying out alcohol precipitation at 4 ℃ for 12h, then centrifuging for 15min under the condition that the centrifugal force is 3500Xg, removing the upper layer of ethanol, collecting the precipitate, dissolving the precipitate in 50ml of ultrapure water, carrying out freeze drying for 36h, collecting the caulerpa lentinan, and weighing.
(2) According to the method in the step (1), 10g of caulerpa longipedicellata powder is respectively added into 100ml, 200ml, 300ml, 400ml and 500ml of ultrapure water to obtain mixed liquid A, the caulerpa longipedicularis polysaccharide is prepared and weighed, then the polysaccharide concentration is calculated according to the method, the polysaccharide extraction rate in the caulerpa longipedicularis sample is calculated according to the formula, and the specific result is shown in figure 1.
As can be seen from fig. 1, when the feed-liquid ratio is too low, the caulerpa lentillifera powder is added into water to make the solution in a rice paste shape, the solution is nearly saturated, and the space for dissolving out the polysaccharide becomes smaller, so that the extraction rate of the polysaccharide is lower, when the feed-liquid ratio is 1:40, the extraction rate of the caulerpa lentillifera polysaccharide reaches the maximum, and then the extraction rate is reduced along with the increase of the feed-liquid ratio, because when the feed-liquid ratio is too high, the impurity content is increased, so that the extraction rate of the polysaccharide is reduced.
(3) According to the method in the step (1), 10g of caulerpa lentillifera powder is added with 200ml of ultrapure water to obtain a mixed solution A, papain with the mass being 2.5% of the mass of the mixed solution A is added, extraction is carried out for 2 hours at the temperatures of 30 ℃, 40 ℃, 50 ℃, 60 ℃ and 70 ℃ respectively, caulerpa lentillifera polysaccharide is prepared and weighed, then the polysaccharide concentration is calculated according to the method, the polysaccharide extraction rate in a caulerpa lentillifera sample is calculated according to the formula, and the specific result is shown in figure 2.
Because temperature can affect the activity of papain, and thus the extraction rate of polysaccharides. As can be seen from FIG. 2, when the temperature is less than 50 ℃, the extraction rate of Caulerpa lentillifera polysaccharide increases with the increase of the temperature, because the collision of the enzyme with the substrate is aggravated by the increase of the temperature, and the probability of the enzyme contacting with the substrate increases, thereby increasing the extraction rate of polysaccharide. When the temperature reaches 50 ℃, the activity of the papain is maximum, and the extraction rate of the polysaccharide reaches a peak. After the temperature is higher than 50 ℃, the enzyme structure is damaged at high temperature, the enzyme activity is reduced along with the increase of the temperature, and the polysaccharide extraction rate is also gradually reduced.
(4) According to the method in the step (1), 10g of caulerpa longipedicellata powder is added with 200ml of ultrapure water to obtain a mixed solution A, papain with the mass being 2.5% of that of the mixed solution A is added, extraction is carried out at 60 ℃, the extraction time is 1h, 2h, 3h, 4h and 5h respectively, caulerpa longipedicularis polysaccharide is prepared and weighed, then the polysaccharide concentration is calculated according to the method, the polysaccharide extraction rate in a caulerpa longipedicularis sample is calculated according to the formula, and the specific result is shown in figure 3.
As can be seen from FIG. 3, the extraction rate of Caulerpa lentillifera polysaccharide increases with the increase of the extraction time, but the extraction rate of polysaccharide decreases after the extraction time exceeds 3 hours, because the extraction time is too long, which results in the change of polysaccharide structure or the cleavage of glycosidic bond.
(5) According to the method in the step (1), 10g of caulerpa lentillifera powder is added with 200ml of ultrapure water to obtain mixed liquid A, papain with the mass of 0.5%, 1.0%, 1.5%, 2.0% and 2.5% of the mixed liquid A is respectively added to prepare and weigh caulerpa lentillifera polysaccharide, then the polysaccharide concentration is calculated according to the method, the polysaccharide extraction rate in a caulerpa lentillifera sample is calculated according to the formula, and the specific result is shown in figure 4.
As can be seen from FIG. 4, the extraction rate of Caulerpa lentillifera polysaccharide increases with the increase of the addition amount of papain, and after the addition amount reaches 2%, the extraction rate of polysaccharide tends to be stable, because when the enzyme amount reaches saturation after increasing to a certain concentration, no excess substrate has the opportunity to react with the enzyme. Therefore, in order to save cost, the optimal papain addition amount is determined to be 2%.
(6) After the caulerpa lentillifera polysaccharide is extracted according to the method in the step (1), the collected algal residues are repeatedly extracted according to the same method for multiple times, wherein the extraction times are respectively 0 time, 1 time, 2 times and 3 times, the total extraction times are respectively 1 time, 2 times, 3 times and 4 times, the all extracted caulerpa lentillifera polysaccharide is combined, the polysaccharide concentration is calculated according to the method, the polysaccharide extraction rate in the caulerpa lentillifera sample is calculated according to the formula, and the specific result is shown in fig. 5.
As can be seen from FIG. 5, when the total extraction frequency is increased to 2 times, the extraction rate of Caulerpa lentillifera polysaccharide is significantly increased. When the total extraction times are more than 2 times, the polysaccharide extraction rate is slightly reduced and basically kept unchanged. Therefore, in order to save cost and improve efficiency, the number of extractions was determined to be 2.
Experimental example 2:
response surface test:
(1) according to the results of the one-factor test, the amount of immobilized papain added was 2% of the mass of the mixed solution a and the total extraction times were 2 times, according to the method described in example 1. The effect of feed-to-solution ratio (ratio of Botrytis longituba powder to ultrapure water), extraction temperature and extraction time on the extraction rate of Botrytis longituba polysaccharide was then examined using Design-Eepert.V8.0.6 software, the experimental Design and results are shown in FIG. 6.
(2) Establishing a regression equation and variance analysis:
and (3) performing multiple regression fitting on the data in the graph 6 by using Design-Expert 8.0.6 software to obtain a quadratic regression equation: y = +41.63+4.97 a + 0.15B + 0.74C +0.27 a B +0.77 a C + 0.32B C-6.02 a2-3.87*B2-2.35*C2. Wherein Y is the extraction rate of Caulerpa lentillifera polysaccharide, and A, B and C respectively represent the feed-liquid ratio, extraction time and extraction temperature.
To test the effectiveness of the regression model of the Botrytis longipedicularis polysaccharide extraction process, the model was analyzed for variance, and the results are shown in FIG. 7. Regression modelp<0.0001, indicating that the regression model has statistical significance. Correction factor R2(Adj) =0.9568, which shows that the experimental error is small and the accuracy is high. Model determination coefficient R2=0.9811, prediction correlation coefficient R2(Pred) =0.9506, where the values are close, indicating that the deviation is within a reasonable range, the actual measurement value and the predicted value have a good degree of fitting, and the two have a high correlation. Missimilitude termp=0.9530>0.05, the results were not significant, indicating that the selected model fits well to the test. The coefficient of variation, C.V%, was 3.16%, reflecting the higher stability and confidence of the model. The signal-to-noise ratio (Adeq precision) was 17.222, which is greater than the threshold of 4, indicating a high accuracy of the model. The indexes show that the fitting regression equation can be used for predicting and analyzing the optimal extraction process of the caulerpa lentillifera polysaccharide.
The independent variable primary term A and secondary term A in the regression model2、B2、C2Has significant influence on the extraction rate of polysaccharide from Caulerpa lentillifera (A)p<0.05). From the F value, the influence degrees of the three factors on the extraction rate of the polysaccharide from the caulerpa lentillifera are shown in the following sequence: the ratio of material to liquid is more than the extraction temperature and more than the extraction time.
(3) Interaction between factors
According to a regression equation, a 3D response surface graph and a contour graph are drawn by using Design-Expert 8.0.6 software, the relationship between the interaction among factors and the extraction rate of the caulerpa longipedicla polysaccharide is analyzed, and the result is shown in figures 8-10. The degree of steepness of the response surface graph can reflect the influence degree of a single factor in the extraction system on the polysaccharide extraction rate, and the steeper the response surface graph, the greater the influence of the factor on the polysaccharide extraction rate is, and the smaller the influence is. The distance between the contour lines is also an important basis for judging whether the influence of a certain factor on the polysaccharide extraction rate is obvious, and the denser contour lines indicate that the influence degree of the factor on the polysaccharide extraction rate is larger, otherwise, the influence degree is smaller. Furthermore, the closer the contour is to a circle, the less significant the interaction between the two factors, whereas the closer to an ellipse the more significant it is.
Therefore, in the experiment, three groups of contour maps AB, AC and BC are all close to a circle, which shows that the two factors of the feed-liquid ratio and the extraction time, the feed-liquid ratio and the extraction temperature, and the extraction time and the extraction temperature have no significant influence, and the result is consistent with the result of the variance analysis. According to fig. 8 and 9, it can be seen that the response surface graph of the feed-liquid ratio is curved greatly, and the contour lines of the feed-liquid ratio axis are also denser, which indicates that the feed-liquid ratio has a greater influence on the extraction rate of the caulerpa lentinan, and conforms to the result of the analysis of variance.
(4) Response surface test scheme optimization and result verification
Partial derivative analysis is carried out on the regression model equation by using Design-Expert 8.0.6 software, and the result shows that the optimal extraction process of the caulerpa lentillifera polysaccharide is as follows: the ratio of material to liquid is 1:44.77, the extraction temperature is 60 ℃, the extraction time is 3.08h, the addition amount of the fixed papain is 2 percent of the mass of the mixed liquid A, the total extraction times are 2 times, and the maximum theoretical extraction rate of the caulerpa lentillifera polysaccharide under the condition is 41.4068 percent. In consideration of the operability of the experiment and production, the optimization conditions were adjusted to: the material-liquid ratio is 1: 45, the extraction temperature is 60 ℃, the extraction time is 3 hours, the addition amount of the immobilized papain is 2 percent of the mass of the mixed solution A, and the total extraction times are 2 times. Under the condition, the method described in example 1 is used for 3 parallel tests, the actual extraction rate of the botryococcus longipes polysaccharide is (40.2253 + 0.89)%, and the difference is only 1.18 from the predicted value, which shows that the model can better simulate the extraction process of the botryococcus longipes polysaccharide and predict the extraction rate.
Furthermore, it should be understood that although the present description refers to embodiments, not every embodiment may contain only a single embodiment, and such description is for clarity only, and those skilled in the art should integrate the description, and the embodiments may be combined as appropriate to form other embodiments understood by those skilled in the art.
Claims (3)
1. A method for extracting the caulerpa lentillifera polysaccharide is characterized by comprising the following steps: the method comprises the following steps:
(1) washing caulerpa lentillifera with seawater, washing with distilled water for 3 times, drying in an oven at 50 deg.C, pulverizing, sieving with 50 mesh sieve to obtain caulerpa lentillifera powder, and drying and storing;
(2) adding ultrapure water into caulerpa longipedica powder, uniformly mixing to obtain a mixed solution A, wherein the ratio of the mass of the caulerpa longipedica powder to the volume of the ultrapure water is 1g (40-50) ml, then adjusting the pH of the mixed solution A to 6, adding papain, and extracting at 40-60 ℃ for 2-4 h to obtain a mixed solution B, wherein the adding amount of the papain is 1-2.5% of the mass of the mixed solution A;
(3) inactivating enzyme of the mixed liquor B obtained in the step (2) for 5min under the condition of boiling water bath, centrifuging for 15min under the condition that the centrifugal force is 3500Xg, and separating to obtain supernatant and algae residue; placing the supernatant in a rotary evaporator, concentrating under negative pressure at 50 ℃ until the concentration is 1g/ml to obtain a concentrated extracting solution, naturally cooling the concentrated extracting solution to room temperature, and adding 95% ethanol by volume to obtain a mixed solution C, wherein the volume of the ethanol added is 5 times of the volume of the concentrated extracting solution; precipitating the mixed solution C at 4 deg.C for 12h, centrifuging at 3500Xg for 15min, removing the upper layer ethanol, and collecting precipitate A;
(4) and (3) sequentially extracting the algae residues obtained in the step (3) according to the step (2) and the step (3) to obtain a precipitate B, combining the precipitate A and the precipitate B, completely dissolving the precipitate A and the precipitate B in ultrapure water to obtain a mixed solution D, and freeze-drying the mixed solution D for 36 hours to obtain the caulerpine lentinan.
2. The method of extracting Caulerpa lentillifera polysaccharide of claim 1, wherein: in the step (2), the pH is adjusted to 6 by adding 10% by mass hydrochloric acid or 5% by mass sodium hydroxide solution to the mixed solution a.
3. The method of extracting Caulerpa lentillifera polysaccharide of claim 1, wherein: in the extraction process of the step (2), the mixed solution A is stirred every 30min for 2min each time.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104311691A (en) * | 2014-11-11 | 2015-01-28 | 济南凯因生物科技有限公司 | Extraction method of bitter melon polysaccharides |
| CN105646728A (en) * | 2016-03-11 | 2016-06-08 | 江苏农牧科技职业学院 | Honeycomb spleen polysaccharide extract and preparation method thereof |
| CN110256595A (en) * | 2019-06-29 | 2019-09-20 | 深圳市蓝汀鼎执生物科技有限公司 | A kind of extracting method of sea grape polysaccharide |
| CN112279931A (en) * | 2020-10-20 | 2021-01-29 | 中国计量大学 | Method for extracting auricularia auricula polysaccharide by using biological enzyme method |
| CN113274490A (en) * | 2021-05-13 | 2021-08-20 | 华南农业大学 | Preparation method and application of botryococcus longipediculus polysaccharide disease-resistant inducer |
-
2021
- 2021-10-08 CN CN202111169494.2A patent/CN113817077A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104311691A (en) * | 2014-11-11 | 2015-01-28 | 济南凯因生物科技有限公司 | Extraction method of bitter melon polysaccharides |
| CN105646728A (en) * | 2016-03-11 | 2016-06-08 | 江苏农牧科技职业学院 | Honeycomb spleen polysaccharide extract and preparation method thereof |
| CN110256595A (en) * | 2019-06-29 | 2019-09-20 | 深圳市蓝汀鼎执生物科技有限公司 | A kind of extracting method of sea grape polysaccharide |
| CN112279931A (en) * | 2020-10-20 | 2021-01-29 | 中国计量大学 | Method for extracting auricularia auricula polysaccharide by using biological enzyme method |
| CN113274490A (en) * | 2021-05-13 | 2021-08-20 | 华南农业大学 | Preparation method and application of botryococcus longipediculus polysaccharide disease-resistant inducer |
Non-Patent Citations (4)
| Title |
|---|
| 温思萌等: "响应面优化酶法提取茯苓多糖工艺", 《中国食品添加剂》 * |
| 王辉等: "响应面法优化酶提取柿子多糖工艺研究", 《农产品加工》 * |
| 罗云波等: "《食品生物技术导论》", 31 May 2016, 中国农业大学出版社 * |
| 许加超等: "《海藻化学与工艺学》", 30 September 2014, 中国海洋大学出版社 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114276471A (en) * | 2021-12-30 | 2022-04-05 | 江苏海洋大学 | Preparation method and application of caulerpa lentillifera polysaccharide |
| CN114276471B (en) * | 2021-12-30 | 2022-12-06 | 江苏海洋大学 | Application of caulerpa lentillifera polysaccharide |
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