CN104045735A - Method for efficiently extracting full-component grateloupia filicina extract by utilizing two-enzyme method - Google Patents
Method for efficiently extracting full-component grateloupia filicina extract by utilizing two-enzyme method Download PDFInfo
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Abstract
The invention provides a method for efficiently extracting full-component grateloupia filicina extract by utilizing a two-enzyme method. The method comprises the following steps: adding water into grateloupia filicina, and uniformly stirring to form a backing material; regulating the pH to 6.5-7.5 by adding anhydrous sodium carbonate; weighing agarase which is 0.25-0.39 percent of the weight of the backing material, dissolving with water, and performing enzymolysis at 20-35 DEG C for 7-9 hours in the pH value regulated backing material; adding lactic acid and potassium sorbate into the enzymatic hydrolysate, uniformly stirring, and controlling the pH to be 4.8-5.2; weighing cellulase which is 0.17-0.22 percent of the weight of the backing material, dissolving with water, and performing enzymolysis at 45-55 DEG C for 11-14 hours in the previous liquid; centrifuging or performing pump filtration after the reaction is completed to obtain a clear liquid; and adding anhydrous sodium carbonate into the clear liquid to regulate the pH to be 6.5-7.5, heating to perform sterilization and enzyme deactivation to obtain a liquid which is the full-component grateloupia filicina extract. According to the method, the purity of the full-component grateloupia filicina extract can be guaranteed, and the extraction rate of effective components of grateloupia filicina can be effectively improved.
Description
Technical field
The present invention relates to the extracting method of marine alga, particularly a kind of method with the full composition Grateloupia filicina (Wulf.) of double-enzyme method high efficiency extraction extracting solution.
Background technology
China is in the world main marine alga producing country at present, gathers in the crops every year various marine algas (by dry product) more than 1,500,000 tons.These marine algas, in order to produce alginates and ester, agar, carrageenin, are widely used in the industries such as processed food, foodstuff additive, medicine, Chemicals.Research shows: the active substance of marine alga, there are Sargassum polysaccharides, alga oligosaccharide, small-molecular peptides and various VITAMIN, trace element, there is the blood circulation that improves brain, strengthen myocardial function, the effect of preventing cardiovascular disease, especially hypertension, cerebral thrombosis etc. is had to special efficacy, can strengthen body immunity, the auxiliary tumour that suppresses simultaneously.Traditional marine alga deep processing method, be all extract certain single component as alginates and ester, agar, carrageenin, Sargassum polysaccharides etc. be main, do not have the report of seeing that full composition extracts, other does not have the effective constituent of fully decomposing and extracting to run off can to cause like this marine alga.
Existing marine alga processing technology has: (1) prior art 1: in periodical < < guangdong agricultural science > >, the extraction process of the banded Grateloupia filicina (Wulf.) Crude polysaccharides of < < is optimized > > by single factor experiment and L
9(3
3) orthogonal test technique that hot water extraction is extracted to banded Grateloupia filicina (Wulf.) polysaccharide is optimized.Preferred version is solid-liquid ratio 1: 70,95 ℃ of leaching temperatures, leaching time 4.5 h, alcohol precipitation concentration 95%.With this understanding, banded Grateloupia filicina (Wulf.) polysaccharide extract rate is 20.20%, and yield is 67.3%; (2) prior art 2: in periodical < < food and machinery > >, the Study of optimization > > of < < ultrasonic-assisted extraction Grateloupia filicina (Wulf.) polysaccharide adopts single factor experiment and L
9(3
3) orthogonal test research temperature, supersound process time, extraction time and the impact of ultrasonic power on polysaccharide extract rate.Optimised process is that ultrasonic extraction temperature 70 C, supersound process time 15min, ultrasonic power are 450W, extract 3 times; Polysaccharide extract rate is 18.83%, and yield is 62.75%; (3) prior art 3: the manufacture method > > (patent announcement number: CN101012286B) of patent of invention < < water soluble seaweed powder, the preparation of employing enzymolysis process, take the prozyme that cellulase and proteolytic enzyme is main component, part by weight is 50: 1～1: 1, enzymatic hydrolysis condition is pH 3～10,35 ℃～65 ℃ of temperature, the enzyme that goes out filters, and getting filter cake, to add concentration be 20% Na
2cO
3solution is to all dissolving, then gained filtrate regulates pH value to neutrality after mixing by filter cake lysate and while filtering, dry powder process.
Existing these extract the technology of Grateloupia filicina (Wulf.) polysaccharide, and as hot water extraction method, alcohol precipitation extraction method and ultrasonic extraction exist polysaccharide extract rate lower, and long-time high temperature and ultrasonication such as may damage to the effective constituent in Grateloupia filicina (Wulf.) at the shortcoming.Another Grateloupia filicina (Wulf.) cell walls main component is Mierocrystalline cellulose and agar-agar, and the combined-enzyme method in prior art 3 adopts cellulase and proteolytic enzyme, produces a large amount of filter cakes after enzymolysis, need to introduce a large amount of Na
2cO
3,k
2cO
3further filter cake is dissolved and purified.Also have the components such as the more difficult extraction halfcystine of existing method, Histidine, proline(Pro), tryptophane, zinc, iron, cause the effective constituent of extraction to run off.Add normal hydrochloric acid, the sulphur acid for adjusting pH of adopting in prior art, hydrochloric acid and sulfuric acid are strong, and corrodibility is large, and adding conditional is difficult to be controlled, difficult operation in implementation process, and extracting method is loaded down with trivial details, and extract is applied to food, cosmetics safety is low.
Summary of the invention
The object of the invention is to for above-mentioned existing problems and deficiency, provide a kind of and can farthest destroy Grateloupia filicina (Wulf.) cell walls, most insolubless are degraded to solvend, make the abundant stripping of active substance in Grateloupia filicina (Wulf.), thereby guarantee the purity and the extraction yield that has improved Grateloupia filicina (Wulf.) effective constituent of full composition Grateloupia filicina (Wulf.) extract, and the easy method with the full composition Grateloupia filicina (Wulf.) of double-enzyme method high efficiency extraction extracting solution of extracting method operation.In addition the extracting solution in the present invention is safe, can be applicable to being applied in food, cosmetic field.
Technical scheme of the present invention is achieved in that
Method with the full composition Grateloupia filicina (Wulf.) of double-enzyme method high efficiency extraction extracting solution of the present invention, is characterized in comprising the following steps:
(1), take the Grateloupia filicina (Wulf.) of aequum, after cleaning, add deionized water, stir, form bed material;
(2), in above-mentioned bed material, add anhydrous sodium carbonate, tune pH to 6.5～7.5;
(3), take the agarase that 0.25%～0.39% bed material is heavy, with adding after deionized water dissolving in the above-mentioned bed material that mixes up pH value, stir; In reactor, carry out enzymolysis for the first time, the temperature of enzymolysis is 20～35 ℃ again, and the time is 7～9h;
(4), in above-mentioned enzymolysis solution, add lactic acid, potassium sorbate, stir, pH is controlled in 4.8～5.2 scopes;
(5), take the cellulase that 0.17%～0.22% bed material is heavy, with adding after deionized water dissolving in step (fours') solution, after stirring, in reactor, carry out enzymolysis for the second time, the temperature of enzymolysis is 45～55 ℃, the time is 11-14h;
(6), reaction finish after, enzymolysis is completed to liquid and carries out centrifugal or suction filtration, obtain clear liquid;
(7), in clear liquid, add anhydrous sodium carbonate to adjust potential of hydrogen, adjust pH to 6.5～7.5 post-heating to carry out sterilization enzyme inactivation (cooling stand-by), gained solution is full composition Grateloupia filicina (Wulf.) extracting solution, the extraction yield of described full composition Grateloupia filicina (Wulf.) extracting solution is 26.32%～27.92%, and yield is 87.73%～93.07%.
Because working condition should be controlled in the suitableeest scope of enzyme, the height of temperature and the size of pH all can affect the activity size of enzyme or cause the inactivation of enzyme, so the optimum temps of agarase is 25 ℃, optimal ph is 7; The optimum temps of cellulase is 50 ℃, and optimal ph is 5.
Because being to use newborn acid for adjusting pH, lactic acid is gentle, can be used for food, makeup simultaneously, easily-controlled reaction conditions, and processing safety is high.
The agarase wherein adopting in above-mentioned steps (three) and step (five) and the vigor of cellulase are 30000 units.The add-on of the middle deionized water of above-mentioned steps () is generally 30-50 times of Grateloupia filicina (Wulf.) amount.In above-mentioned steps (two) add-on of anhydrous sodium carbonate be generally bed material heavy 0.01%～0.016%.In above-mentioned steps (four) in above-mentioned enzymolysis solution the add-on of lactic acid be generally bed material heavy 0.02%～0.032%.In above-mentioned steps (seven) add-on of anhydrous sodium carbonate be generally bed material heavy 0.03%～0.037%.
The present invention is owing to adopting cellulase and agarase to be used in conjunction with the method for Grateloupia filicina (Wulf.) being carried out to full composition extraction, synergy and the promoter action full composition Grateloupia filicina (Wulf.) extract extracted by these two kinds of enzymes, can farthest destroy Grateloupia filicina (Wulf.) cell walls, most insolubless (are mainly to agar-agar, carrageenin etc.) be degraded to solvend, make the abundant stripping of active substance in Grateloupia filicina (Wulf.) (as the halfcystine with the more difficult extraction of other method, Histidine, proline(Pro), tryptophane, zinc, the extraction yield of the active substances such as iron is greatly enhanced), farthest retain the effective constituent of Grateloupia filicina (Wulf.), improved the extraction yield of full composition Grateloupia filicina (Wulf.) extract.This is because the main component of Grateloupia filicina (Wulf.) cell walls is agar-agar and Mierocrystalline cellulose, agarase can destroy the cell walls of Grateloupia filicina (Wulf.), be conducive to polysaccharide and other composition stripping in cell content, and can reduce the viscosity of Grateloupia filicina (Wulf.) liquid, being conducive to cellulase is combined with bed material, thereby reduce the addition of cellulase, improve the extraction yield of Grateloupia filicina (Wulf.) effective constituent.The effects such as that the agar-agar degradation product oligosaccharides simultaneously producing in enzymolysis process also has is antiviral, antitumor, strengthening immunity.Because the present invention also adopts edible lactic acid, replace hydrochloric acid and sulfuric acid, lactic acid is gentle, can be used for food, makeup, easily-controlled reaction conditions, and processing safety is high.Enzyme digestion reaction in the technology of the present invention is abundant, gentle, extraction yield is high, insolubles filter cake is few, do not need to add again sodium carbonate dissolving filter cake, can save labour's material resources, and working method is simple and easy, guarantee the purity of full composition Grateloupia filicina (Wulf.) extract, these technique effects are that existing Grateloupia filicina (Wulf.) extracting method is beyond one's reach simultaneously, creative high.
Embodiment
Method with the full composition Grateloupia filicina (Wulf.) of double-enzyme method high efficiency extraction extracting solution of the present invention, is characterized in comprising the following steps:
(1), take the Grateloupia filicina (Wulf.) of aequum, after cleaning (wash, guarantee without sandstone, earth or other impurity), add deionized water, stir, form bed material;
(2), in above-mentioned bed material, add anhydrous sodium carbonate, in the scope of tune pH to 6～8;
(3), take the agarase that 0.25%～0.39% bed material is heavy, with adding after deionized water dissolving in the bed material of the above-mentioned pH of mixing up value, stir; In reactor, carry out enzymolysis for the first time, the temperature of enzymolysis is 20～35 ℃ again, and the time is 7～9h;
(4), in above-mentioned enzymolysis solution, add lactic acid, potassium sorbate, stir, pH is controlled in 4.8～5.2 scopes;
(5), take the cellulase that 0.17%～0.22% bed material is heavy, with adding after deionized water dissolving in step (fours') solution, after stirring, in reactor, carry out enzymolysis for the second time, the temperature of enzymolysis is 45～55 ℃, the time is 11-14h;
(6), reaction finish after, enzymolysis is completed to liquid and carries out centrifugal or suction filtration, obtain clear liquid;
(7), in clear liquid, add anhydrous sodium carbonate to adjust potential of hydrogen, adjust pH to 6.5～7.5 post-heating to carry out sterilization enzyme inactivation (can cool stand-by), gained solution is full composition Grateloupia filicina (Wulf.) extracting solution, the extraction yield of described full composition Grateloupia filicina (Wulf.) extracting solution is 26.32%～27.92%, and yield is 87.73%～93.07%.
The present invention is in the process of extracting, synergy and the promoter action that can to full composition Grateloupia filicina (Wulf.) extract, extract by cellulase and these two kinds of enzymes of agarase, can farthest destroy Grateloupia filicina (Wulf.) cell walls, most insolubless (being mainly agar-agar, carrageenin etc.) are degraded to solvend, make the abundant stripping of active substance in Grateloupia filicina (Wulf.), farthest retain the effective constituent of Grateloupia filicina (Wulf.), improved the extraction yield of full composition Grateloupia filicina (Wulf.) extract.
Wherein the optimal ph of agarase is 7; The optimal ph of cellulase is 5.In above-mentioned steps (three) and step (five), the general agarase adopting and the vigor of cellulase are 30000 units.The add-on of the middle deionized water of above-mentioned steps () is generally 30-50 times of Grateloupia filicina (Wulf.) amount.Preferably 35 times.
For further improving extraction yield of the present invention and yield, in above-mentioned steps (two) add-on of anhydrous sodium carbonate be generally bed material heavy 0.01%～0.016%.In above-mentioned steps (seven) add-on of anhydrous sodium carbonate be generally bed material heavy 0.03%～0.037%.In above-mentioned steps (four) in above-mentioned enzymolysis solution the add-on of lactic acid be generally bed material heavy 0.02%～0.032%.
The Main Function of above-mentioned potassium sorbate is as sanitas, optionally adds.
The present invention can farthest destroy Grateloupia filicina (Wulf.) cell walls, most insolubless are degraded to solvend, make the abundant stripping of active substance in Grateloupia filicina (Wulf.), thereby guarantee the purity and the extraction yield that has improved Grateloupia filicina (Wulf.) effective constituent of full composition Grateloupia filicina (Wulf.) extract, and extracting method operation is simple and easy, extracting solution is safe, can extensively be applicable to being applied in food, cosmetic field.
below in conjunction with specific embodiment, the present invention is done to the detailed description of this step:
embodiment mono-:take the Grateloupia filicina (Wulf.) of 2kg, add 70kg deionized water, stir evenly, add 7.2g anhydrous sodium carbonate, adjust pH to 7.The agarase that adds 200g, stirs evenly, 25 ℃ of heating 8h in reactor.The lactic acid that adds 144g potassium sorbate, 23mL, stirs evenly, and adjusts pH to 5.0.The cellulase that adds 122g, stirs evenly, 50 ℃ of heating 12h in reactor.After reaction finishes, by enzymolysis solution centrifugal (slag is 576g).In centrifugal clear liquid, add 22g anhydrous sodium carbonate, adjust pH to 7, be heated to 100 ℃ of sterilization enzyme inactivations, gained solution is full composition Grateloupia filicina (Wulf.) extracting solution, cools stand-by.The extraction yield of full composition Grateloupia filicina (Wulf.) extracting solution is 27.74%, and yield is 92.47%.
embodiment bis-:take the Grateloupia filicina (Wulf.) of 2kg, add 70kg deionized water, stir evenly, add 8.4g anhydrous sodium carbonate, adjust pH to 7.5.The agarase that adds 240g, stirs evenly, 25 ℃ of heating 8h in reactor.The lactic acid that adds 144g potassium sorbate, 25mL, stirs evenly, and adjusts pH to 5.1.The cellulase that adds 140g, stirs evenly, 52 ℃ of heating 12h in reactor.After reaction finishes, by enzymolysis solution centrifugal (slag is 920g).In centrifugal clear liquid, add 20g anhydrous sodium carbonate, adjust pH to 7, be heated to 100 ℃ of sterilization enzyme inactivations, gained solution is full composition Grateloupia filicina (Wulf.) extracting solution, cools stand-by.The extraction yield of full composition Grateloupia filicina (Wulf.) extracting solution is 26.83%, and yield is 89.43%.
embodiment tri-:take the Grateloupia filicina (Wulf.) of 2kg, add 70kg deionized water, stir evenly, add 7.2g anhydrous sodium carbonate, adjust pH to 7.The agarase that adds 144g, stirs evenly, 25 ℃ of heating 8h in reactor.The lactic acid that adds 144mL potassium sorbate, 23mL, stirs evenly, and adjusts pH to 5.0.The cellulase that adds 72g, stirs evenly, 50 ℃ of heating 12h in reactor.After reaction finishes, by enzymolysis solution centrifugal (slag is 1728g).In centrifugal clear liquid, add 22g anhydrous sodium carbonate, adjust pH to 7, be heated to 100 ℃ of sterilization enzyme inactivations, gained solution is full composition Grateloupia filicina (Wulf.) extracting solution, cools stand-by.The extraction yield of full composition Grateloupia filicina (Wulf.) extracting solution is 24.53%, and yield is 81.77%.
In this embodiment, the addition minimizing because of agarase and cellulase, causes cell walls to destroy not exclusively, and insolubles increases, and extraction yield declines.
embodiment tetra-:take the Grateloupia filicina (Wulf.) of 2kg, add 70kg deionized water, stir evenly, add 7.2g anhydrous sodium carbonate, adjust pH to 8.0.The agarase that adds 200g, stirs evenly, 45 ℃ of heating 8h in reactor.The lactic acid that adds 144g potassium sorbate, 23mL, stirs evenly, and adjusts pH to 6.0.The cellulase that adds 122g, stirs evenly, 65 ℃ of heating 12h in reactor.After reaction finishes, by enzymolysis solution centrifugal (slag is 6840g).In centrifugal clear liquid, add 22g anhydrous sodium carbonate, adjust pH to 7, be heated to 100 ℃ of sterilization enzyme inactivations, gained solution is Grateloupia filicina (Wulf.) extracting solution, cools stand-by.The extraction yield of Grateloupia filicina (Wulf.) extracting solution is 19.88%, and yield is 66.27%.
The reason that in this embodiment, the extraction yield of each component is low is: reaction conditions is outside the optimum condition of agarase and cellulase, and enzymic activity declines or inactivation, causes extraction yield to decline.
embodiment five: take the Grateloupia filicina (Wulf.) of 2kg, add 70kg deionized water, the lactic acid of 144g potassium sorbate, 23mL, stirs evenly, and adjusts pH to 4.8.The cellulase that adds 122g, stirs evenly, 50 ℃ of heating 12h in reactor.After reaction finishes, by enzymolysis solution centrifugal (slag is 2448g).In centrifugal clear liquid, add 22g anhydrous sodium carbonate, adjust pH to 7, be heated to 100 ℃ of sterilization enzyme inactivations, gained solution is Grateloupia filicina (Wulf.) extracting solution, cools stand-by.The extraction yield of Grateloupia filicina (Wulf.) extracting solution is 24.32%, and yield is 81.07%.
The present embodiment is because not adding agarase, and cell walls destroys not exclusively, causes extraction yield to decline.
implement six:take the Grateloupia filicina (Wulf.) of 2kg, add 70kg deionized water, stir evenly, add 6.5g anhydrous sodium carbonate, adjust pH to 6.5.The proteolytic enzyme that adds 200g, stirs evenly, 25 ℃ of heating 8h in reactor.The lactic acid that adds 144g potassium sorbate, 21mL, stirs evenly, and adjusts pH to 5.0.The cellulase that adds 122g, stirs evenly, 50 ℃ of heating 12h in reactor.After reaction finishes, by enzymolysis solution centrifugal (slag is 6344g).In centrifugal clear liquid, add 22g anhydrous sodium carbonate, adjust pH to 7, be heated to 100 ℃ of sterilization enzyme inactivations, gained solution is Grateloupia filicina (Wulf.) extracting solution, cools stand-by.The extraction yield of Grateloupia filicina (Wulf.) extracting solution is 20.24%, and yield is 67.45%.
This embodiment is owing to adopting other double-enzyme method to extract Grateloupia filicina (Wulf.), causes that slag is many, extraction yield is low.
Be below each embodiment extracting solution Analysis of primary nutritive compositions table (measuring the mg/ml of unit of component)
| ? | Embodiment mono- | Embodiment bis- | Embodiment tri- | Embodiment tetra- | Embodiment five | Embodiment six |
| Protein | 6.98 | 6.72 | 6.12 | 4.72 | 5.36 | 5.08 |
| Crude fat | 0.73 | 0.66 | 0.59 | 0.45 | 0.56 | 0.53 |
| Polysaccharide | 7.65 | 7.24 | 6.56 | 5.28 | 6.33 | 5.68 |
| Aspartic acid (Asp) | 0.564 | 0.503 | 0.495 | 0.395 | 0.480 | 0.411 |
| Threonine (Thr) | 0.254 | 0.244 | 0.232 | 0.185 | 0.216 | 0.192 |
| Serine (Ser) | 0.287 | 0.245 | 0.237 | 0.189 | 0.225 | 0.210 |
| L-glutamic acid (Glu) | 1.123 | 1.085 | 1.033 | 0.826 | 1.002 | 0.830 |
| Glycine (Gly) | 0.284 | 0.257 | 0.245 | 0.191 | 0.244 | 0.206 |
| L-Ala (Ala) | 0.481 | 0.463 | 0.422 | 0.301 | 0.401 | 0.350 |
| Half Guang ammonia (Cys) | 0.045 | 0.032 | 0.021 | 0.00 | 0.020 | 0.00 |
| α-amino-isovaleric acid (Val) | 0.473 | 0.455 | 0.421 | 0.278 | 0.391 | 0.348 |
| Methionine(Met) (Met) | 0.287 | 0.256 | 0.251 | 0.205 | 0.253 | 0.217 |
| Different bright ammonia (Ile) | 0.275 | 0.232 | 0.227 | 0.162 | 0.247 | 0.443 |
| Leucine (Leu) | 0.392 | 0.352 | 0.345 | 0.275 | 0.335 | 0.289 |
| Tyrosine (Tyr) | 0.194 | 0.181 | 0.172 | 0.136 | 0.155 | 0.134 |
| Phenylalanine (Phe) | 0.192 | 0.178 | 0.175 | 0.140 | 0.171 | 0.145 |
| Methionin (Lys) | 0.246 | 0.232 | 0.203 | 0.159 | 0.202 | 0.185 |
| Histidine (His) | 0.059 | 0.056 | 0.054 | 0.042 | 0.0524 | 0.00 |
| Arginine (Arg) | 0.858 | 0.820 | 0.755 | 0.511 | 0.722 | 0.620 |
| Proline(Pro) (Pro) | 0.089 | 0.084 | 0.082 | 0.060 | 0.078 | 0.00 |
| Tryptophane (Trp) | 0.076 | 0.065 | 0.045 | 0.046 | 0.058 | 0.00 |
| Calcium (Ca) | 0.325 | 0.302 | 0.295 | 0.245 | 0.285 | 0.231 |
| Magnesium (Mg) | 0.178 | 0.165 | 0.154 | 0.122 | 0.145 | 0.128 |
| Iron (Fe) | 0.038 | 0.034 | 0.031 | 0.023 | 0.030 | 0.00 |
| Zinc (Zn) | 0.026 | 0.018 | 0.014 | 0.00 | 0.012 | 0.00 |
| The extraction yield of Grateloupia filicina (Wulf.) extracting solution | 27.74% | 26.83% | 24.53% | 19.88% | 24.32% | 20.24% |
| Yield | 92.47% | 89.43% | 81.77% | 66.27% | 81.07% | 67.45% |
By embodiment mono-and embodiment tetra-, can be found out, embodiment tetra-does not add agarase, and its extraction yield is significantly less than embodiment mono-, and the weight of centrifugal slag is many, so the composition of extracting solution is comprehensive not enough.By enforcement one and embodiment five, can be found out, what in embodiment five, add is other pair of enzyme, and its extraction yield is very low, and extracting solution composition is incomplete, and a lot of components are 0.Separately from above-mentioned list, can obviously find out, adopt extracting method of the present invention (embodiment mono-and embodiment bis-), in its extracting solution, very multi-component content all will be higher than other embodiment, especially the extraction yield of the active substance such as polysaccharide, halfcystine, Histidine, proline(Pro), tryptophane, zinc, iron is apparently higher than other embodiment, and extraction yield of the present invention and yield are high more a lot of than other embodiment.
Although the present invention describes with reference to specific embodiment, this description not meaning that is construed as limiting the present invention.With reference to description of the invention, other of the disclosed embodiments change, and all can expect for those skilled in the art, and this variation should belong in affiliated claim limited range.
Claims (6)
1. by a method for the full composition Grateloupia filicina (Wulf.) of double-enzyme method high efficiency extraction extracting solution, it is characterized in that comprising the following steps:
(1), take the Grateloupia filicina (Wulf.) of aequum, after cleaning, add deionized water, stir, form bed material;
(2), in above-mentioned bed material, add anhydrous sodium carbonate, tune pH to 6.5～7.5;
(3), take the agarase that 0.25%～0.39% bed material is heavy, with adding after deionized water dissolving in the bed material of the above-mentioned pH of mixing up value, stir; In reactor, carry out enzymolysis for the first time, the temperature of enzymolysis is 20～35 ℃ again, and the time is 7～9h;
(4), in above-mentioned enzymolysis solution, add lactic acid, potassium sorbate, stir, and pH be controlled in 4.8～5.2 scopes;
(5), take the cellulase that 0.17%～0.22% bed material is heavy, with adding after deionized water dissolving in step (fours') solution, after stirring, in reactor, carry out enzymolysis for the second time, the temperature of enzymolysis is 45～55 ℃, the time is 11-14h;
(6), reaction finish after, enzymolysis is completed to liquid and carries out centrifugal or suction filtration, obtain clear liquid;
(7), in clear liquid, add anhydrous sodium carbonate to adjust potential of hydrogen, adjust pH to 6.5～7.5 post-heating to carry out sterilization enzyme inactivation, gained solution is full composition Grateloupia filicina (Wulf.) extracting solution, and the extraction yield of described full composition Grateloupia filicina (Wulf.) extracting solution is 26.32%～27.92%, and yield is 87.73%～93.07%.
2. the method with the full composition Grateloupia filicina (Wulf.) of double-enzyme method high efficiency extraction extracting solution according to claim 1, is characterized in that the agarase of employing in above-mentioned steps (three) and step (five) and the vigor of cellulase are 30000 units.
3. the method with double-enzyme method high efficiency extraction full composition Grateloupia filicina (Wulf.) extracting solution according to claim 1, the add-on that it is characterized in that deionized water in above-mentioned steps () be Grateloupia filicina (Wulf.) amount 30-50 doubly.
4. the method with double-enzyme method high efficiency extraction full composition Grateloupia filicina (Wulf.) extracting solution according to claim 1, the add-on that it is characterized in that anhydrous sodium carbonate in above-mentioned steps (two) be bed material heavy 0.01%～0.016%.
5. the method with double-enzyme method high efficiency extraction full composition Grateloupia filicina (Wulf.) extracting solution according to claim 1, it is characterized in that in above-mentioned steps (four) add-on of lactic acid in above-mentioned enzymolysis solution be bed material heavy 0.02%～0.032%.
6. the method with double-enzyme method high efficiency extraction full composition Grateloupia filicina (Wulf.) extracting solution according to claim 1, the add-on that it is characterized in that anhydrous sodium carbonate in above-mentioned steps (seven) be bed material heavy 0.03%～0.037%.
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| CN104529570A (en) * | 2014-12-10 | 2015-04-22 | 济南航晨生物科技有限公司 | Novel method for extracting seaweed intermediate by enzyme process |
| CN109096409A (en) * | 2018-10-12 | 2018-12-28 | 国家海洋局第海洋研究所 | Weak solidifying, the ease of solubility Asia polysaccharide extraction of grateloupia filicina of one kind and its application |
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| CN1810842A (en) * | 2005-01-25 | 2006-08-02 | 上海中医药大学 | Longleaf Grateloupia acuminata polysaccharide extract and its prepn and use |
| CN101077356A (en) * | 2006-05-24 | 2007-11-28 | 上海中医药大学 | Application for polysaccharide extraction of grateloupia filicina in preparing antitumor medicine and other medicines |
| CN101932715A (en) * | 2007-02-26 | 2010-12-29 | 韩国生产技术研究院 | Method of producing biofuel using sea algae |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1810842A (en) * | 2005-01-25 | 2006-08-02 | 上海中医药大学 | Longleaf Grateloupia acuminata polysaccharide extract and its prepn and use |
| CN101077356A (en) * | 2006-05-24 | 2007-11-28 | 上海中医药大学 | Application for polysaccharide extraction of grateloupia filicina in preparing antitumor medicine and other medicines |
| CN101932715A (en) * | 2007-02-26 | 2010-12-29 | 韩国生产技术研究院 | Method of producing biofuel using sea algae |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104529570A (en) * | 2014-12-10 | 2015-04-22 | 济南航晨生物科技有限公司 | Novel method for extracting seaweed intermediate by enzyme process |
| CN109096409A (en) * | 2018-10-12 | 2018-12-28 | 国家海洋局第海洋研究所 | Weak solidifying, the ease of solubility Asia polysaccharide extraction of grateloupia filicina of one kind and its application |
| CN109096409B (en) * | 2018-10-12 | 2021-08-13 | 自然资源部第一海洋研究所 | Weakly-coagulated and easily-soluble Asiatica fragrans polysaccharide extract and application thereof |
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| CN104045735B (en) | 2016-03-30 |
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