CN111172208A - Method for preparing 2-monoglyceride type n-3PUFA by enzyme method - Google Patents
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- CN111172208A CN111172208A CN202010167155.XA CN202010167155A CN111172208A CN 111172208 A CN111172208 A CN 111172208A CN 202010167155 A CN202010167155 A CN 202010167155A CN 111172208 A CN111172208 A CN 111172208A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
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Abstract
The invention discloses a method for preparing 2-monoglyceride type n-3PUFA by an enzyme method, belonging to the technical field of separation and application of enzyme. And (2) carrying out an ethylation reaction and purification on the oil containing n-3PUFA to obtain fatty acid ethyl ester containing n-3PUFA, carrying out transesterification reaction under the catalysis of immobilized lipase, carrying out alcoholysis reaction by using the immobilized lipase, and extracting and separating to finally obtain the 2-monoglyceride type n-3 PUFA. The glyceride mixture with high sn-2 n-3PUFA content prepared by enzymatic transesterification is used as a substrate to prepare the 2-monoglyceride type n-3PUFA, so that the loss of the n-3PUFA in the subsequent purification process is effectively avoided, and the recovery rate of the n-3PUFA is greatly improved. Meanwhile, the enzymatic alcoholysis by-product n-3PUFA ethyl ester can be recovered as a reaction substrate for enzymatic transesterification, thereby greatly improving the comprehensive utilization rate of raw materials.
Description
Technical Field
The invention belongs to the technical field of grease processing, and particularly relates to a method for preparing 2-monoglyceride type n-3PUFA by an enzymatic method.
Background
Scientific research shows that n-3PUFA (Polyunsaturated fatty acids, such as EPA, DPA and DHA, etc.) plays an important role in human development and growth, vision and cell membrane fluidity. In addition, n-3PUFA has the functions of correcting arrhythmia, inhibiting platelet aggregation, prolonging blood coagulation time, lowering blood pressure, reducing serum triglyceride and plasma homocysteine level, resisting inflammation, regulating immunity, protecting heart and blood vessels, improving insulin sensitivity, reducing the risk of breast cancer and colorectal cancer and the like. In general, n-3PUFA is mainly present in the form of a free fatty acid type, a fatty acid ester type, a glyceride type, or the like. Studies have shown that monoglyceride type n-3 PUFAs have higher bioavailability than free fatty acid type, triglyceride type, diglyceride type, and ethyl fatty acid type, and among monoglyceride type n-3 PUFAs, 2-monoglyceride type n-3 PUFAs have higher bioavailability than 1-monoglyceride type n-3 PUFAs. Therefore, the research on the production of 2-monoglyceride type n-3PUFA has been the current focus.
The biological enzyme method has attracted extensive attention in recent years due to the advantages of mild reaction conditions, high catalytic specificity, environmental protection and the like. The 2-monoglyceride type n-3PUFA prepared by adopting a biological enzyme method instead of a chemical method can effectively avoid the oxidation and isomerization of the n-3PUFA at high temperature and can effectively avoid the generation of 1-monoglyceride type n-3PUFA by the acyl transfer of the 2-monoglyceride type n-3 PUFA. Enzymatic alcoholysis is currently used to produce 2-monoglyceride type n-3 PUFAs. Zhang et al (Food chem.,2018,250:60-66) prepared 2-monoglyceride type n-3PUFA by enzymatic alcoholysis of tuna oil, the 2-monoglyceride content in the product was higher than 90% and the n-3PUFA content in the product 2-monoglyceride was 81.13%, but the recovery rate of n-3PUFA was only 50%; zhang et al (Bioresource technol.,2018,251:334-340) prepared 2-monoglyceride type n-3PUFA by enzymatic alcoholysis of algal oil, the 2-monoglyceride content of the product was 95%, and the n-3PUFA content of the product 2-monoglyceride was 75.66%, but the recovery rate of n-3PUFA was only about 63%. In a word, when marine fish oil or algae oil is adopted as a substrate to prepare 2-monoglyceride type n-3PUFA by an enzyme method at present, the content of 2-monoglyceride in a product can reach more than 90 percent, the content of n-3PUFA in the 2-monoglyceride can reach more than 80 percent, but the recovery rate of n-3PUFA is lower and is only about 50 percent.
Disclosure of Invention
In order to solve the above-mentioned drawbacks of the prior art, it is an object of the present invention to provide a process for producing 2-monoglyceride type n-3PUFA by an enzymatic method, which improves the recovery rate of n-3PUFA and the overall utilization rate of raw materials.
The invention is realized by the following technical scheme:
an enzymatic process for the preparation of 2-monoglyceride type n-3PUFA comprising the steps of:
step 1: carrying out an ethylation reaction on the oil containing n-3PUFA and absolute ethyl alcohol under the catalysis of an alkaline catalyst, and purifying a reaction product to obtain fatty acid ethyl ester containing n-3 PUFA;
step 2: taking glycerol and the n-3 PUFA-containing fatty acid ethyl ester obtained in the step 1 as substrates, carrying out transesterification reaction on the substrates by adopting immobilized lipase, and centrifuging to collect an upper oil phase;
and step 3: and (3) taking the upper oil phase collected in the step (2) as a substrate, carrying out alcoholysis reaction by using immobilized lipase, and then extracting and separating to obtain the 2-monoglyceride type n-3 PUFA.
Preferably, in the step 1, NaOH is used as an alkaline catalyst, and the mass of the NaOH is 0.2-0.4% of that of the oil containing n-3 PUFA; the molar ratio of the ethanol to the n-3 PUFA-containing oil is 6-9: 1.
Preferably, in the step 1, the temperature of the ethylation reaction is 70-75 ℃, and the reaction time is 1.5-2 h.
Preferably, in step 1, the purification is a four-stage molecular distillation purification, wherein the four stages have feed flow rates of 3mL/min, 2mL/min, 1mL/min and 0.5mL/min, evaporation surface temperatures of 110 deg.C, 140 deg.C, 150 deg.C and 160 deg.C, and operating pressures of 30Pa, 15Pa, 5Pa and 1Pa, respectively.
Preferably, the immobilized lipase used in step 2 and step 3 is novozyme 435 or Lipozyme 435.
Preferably, in the step 2, the molar ratio of the n-3 PUFA-containing fatty acid ethyl ester to the glycerol is 3:1, the enzyme adding amount is 2-5% of the total mass of the substrate, the reaction temperature is 55-65 ℃, and the reaction time is 12-24 hours.
Preferably, in the step 3, in the alcoholysis reaction, the alcohol is ethanol, the mass ratio of the upper oil phase to the ethanol is 1: 2.5-3, the enzyme is added in an amount of 6-10 wt% of the total mass of the substrate, the reaction temperature is 25-30 ℃, and the reaction time is 1.5-3 hours.
Preferably, in step 3, the specific steps of extraction are as follows: adding n-hexane into the alcoholysis reaction product, adding ethanol, and washing with n-hexane for several times.
Further preferably, the volume fraction of ethanol is 85%.
Preferably, in step 3, the separation is performed by rotary evaporation or reduced pressure distillation.
Compared with the prior art, the invention has the following beneficial technical effects:
the invention discloses a method for preparing 2-monoglyceride type n-3PUFA by an enzyme method, which comprises the steps of carrying out ethylation reaction and purification on n-3 PUFA-containing grease to obtain fatty acid ethyl ester containing n-3PUFA, carrying out transesterification reaction by using immobilized lipase, carrying out alcoholysis reaction by using immobilized lipase, extracting and separating to finally obtain the 2-monoglyceride type n-3 PUFA. Since the distribution of n-3PUFA in the sn-2 position of the triglyceride skeleton is generally 65% or less, in order to obtain 2-monoglyceride type n-3PUFA having a high n-3PUFA content, it is necessary to further increase the n-3PUFA content in 2-monoglyceride by low-temperature crystallization or molecular distillation purification of 2-monoglyceride separated from the enzymatic alcoholysis product, and the n-3PUFA content in 2-monoglyceride has been increased to some extent by low-temperature crystallization and molecular distillation purification methods which have been generally employed so far, but when the n-3PUFA content in 2-monoglyceride has been increased by both methods, the n-3PUFA loss is large, and the final recovery rate of n-3PUFA is only about 50%. When the glyceride mixture rich in n-3PUFA is prepared by enzymatic transesterification, the n-3PUFA can be uniformly distributed at the sn-1, sn-2 and sn-3 positions of the triglyceride skeleton, so that the glyceride mixture with high n-3PUFA content at the sn-2 position obtained by enzymatic transesterification is used as a substrate to prepare the 2-monoglyceride type n-3PUFA, the loss of the n-3PUFA in the subsequent purification process is effectively avoided, and the recovery rate of the n-3PUFA is greatly improved. Meanwhile, the enzymatic alcoholysis by-product n-3PUFA ethyl ester can be recovered as a reaction substrate for enzymatic transesterification, thereby greatly improving the comprehensive utilization rate of raw materials.
Furthermore, NaOH is used as the alkaline catalyst, the mass of the NaOH is 0.2-0.4% of that of the n-3 PUFA-containing oil, the molar ratio of ethanol to the n-3 PUFA-containing oil is 6-9: 1, and the ethyl esterification of the n-3 PUFA-containing oil to the n-3 PUFA-containing fatty acid ethyl ester can be effectively promoted.
Furthermore, the temperature of the ethylation reaction is 70-75 ℃, the reaction time is 1.5-2 h, and the n-3PUFA is prevented from being oxidized as far as possible while high ethylation is realized.
Furthermore, the purification is four-stage molecular distillation purification, so that the fatty acid ethyl ester containing n-3PUFA can be effectively recovered, and the n-3PUFA can be prevented from being oxidized; the fatty acid ethyl ester containing n-3PUFA can be recovered with high recovery rate by setting parameters of each stage.
Furthermore, the immobilized lipase used in the step 2 and the step 3 is Novozym 435 or Lipozyme435, which can realize the high-efficiency transesterification of the n-3 PUFA-containing fatty acid ethyl ester to generate the n-3 PUFA-containing glyceride and ensure the specific conversion of the n-3 PUFA-containing glyceride to generate the 2-glycerol monoester type n-3 PUFA.
Furthermore, in the step 2, the molar ratio of the n-3 PUFA-containing fatty acid ethyl ester to the glycerol is 3:1, the enzyme is added in an amount of 2-5 wt% of the total mass of the substrate, the reaction temperature is 55-65 ℃, and the reaction time is 12-24 h, so that the conversion of the n-3 PUFA-containing fatty acid ethyl ester into the n-3 PUFA-containing glyceride can be effectively promoted.
Furthermore, in the alcoholysis reaction, the used alcohol is ethanol, the mass ratio of the upper oil phase to the ethanol is 1: 2.5-3, the enzyme is added in an amount of 6-10 wt% of the total mass of the substrate, the reaction temperature is 25-30 ℃, and the reaction time is 1.5-3 h, so that the 2-monoglyceride type n-3PUFA can be generated through efficient and thorough conversion of the glyceride containing the n-3 PUFA.
Further, in the extraction, n-hexane is added into the alcoholysis reaction product, ethanol is added, and then n-hexane is used for washing for a plurality of times to completely remove the residual non-polar compounds such as fatty acid ethyl ester, diglyceride and triglyceride in the ethanol phase.
Furthermore, the volume fraction of ethanol used is 85%, which increases the solubility of polar compounds such as monoglycerides and reduces the solubility of non-polar compounds such as fatty acid ethyl esters, diglycerides and triglycerides.
Furthermore, the separation is carried out by rotary evaporation or reduced pressure distillation, so that the 2-monoglyceride type n-3PUFA can be effectively prevented from being subjected to acyl transfer and the n-3PUFA can be effectively prevented from being oxidized.
Detailed Description
The present invention will now be described in further detail with reference to specific examples, which are intended to be illustrative, but not limiting, of the invention.
The oil containing n-3PUFA is marine fish oil, seal oil or their mixture containing EPA, DPA, DHA, and has acid value lower than 1mg KOH/g.
Example 1
(1) Accurately weighing 1Kg of anchovy oil, placing the anchovy oil into a 5L conical flask with a plug, adding 385mL of absolute ethanol dissolved with 3g of NaOH, reacting at 75 ℃ for 1.5h, centrifugally separating an upper layer solution, distilling and recovering ethanol, and washing with water to remove NaOH to obtain n-3 PUFA-containing ethyl ester; subsequently, ethyl ester containing n-3PUFA was purified by four-stage molecular distillation to obtain 280g of n-3PUFA ethyl ester having an n-3PUFA content of 90.36%; (2) adding 25g of glycerol into the ethyl ester rich in n-3PUFA as a substrate, uniformly stirring at 60 ℃, adding 6.1g of Novozym 435, starting reaction, reacting for 24 hours, and centrifuging to collect an upper oil phase (270 g); (3) adding 810g of absolute ethyl alcohol into the oil phase serving as a substrate, stirring and mixing the mixture evenly at 30 ℃, adding 86.4g of Novozym 435, timing after the reaction, filtering to remove the Novozym 435 after the reaction is carried out for 3 hours, distilling to remove the ethyl alcohol, adding 2.8L of 85% ethyl alcohol and 2.8L of n-hexane into the reaction product, extracting and separating 2-monoglyceride type n-3PUFA, collecting the obtained ethyl alcohol phase, washing the ethyl alcohol phase with the n-hexane for 2 times, and carrying out rotary evaporation to remove the ethyl alcohol and the water to obtain the product. The molecular distillation light phase obtained in the step (1) can be continuously used for molecular distillation to recover the n-3PUFA ethyl ester in the light phase, and the steps are repeated; after recovering n-hexane from the n-hexane phase, the resulting n-3 PUFA-rich ethyl ester can be used again in step (2). Finally, the 2-monoglyceride content of the product was 93.45%, the n-3PUFA content of the 2-monoglyceride was 90.11%, and the recovery yield of n-3PUFA was 86.18%.
Example 2
(1) Accurately weighing 1Kg of tuna oil, placing the tuna oil into a 5L conical flask with a plug, adding 578mL of absolute ethanol dissolved with 2g of NaOH, reacting for 2h at 70 ℃, centrifugally separating the upper solution, distilling and recovering ethanol, and washing with water to remove NaOH to obtain n-3 PUFA-containing ethyl ester; subsequently, the ethyl ester containing n-3PUFA was purified by four-stage molecular distillation to obtain 286g of n-3PUFA ethyl ester having an n-3PUFA content of 90.78%; (2) adding 25g of glycerol into the ethyl ester rich in the n-3PUFA as a substrate, uniformly stirring at 65 ℃, adding 15.5g of Lipozyme435, starting reaction timing, reacting for 12 hours, centrifuging and collecting an upper oil phase (275 g); (3) adding 688g of absolute ethyl alcohol into the oil phase serving as a substrate, stirring and mixing the mixture evenly at 25 ℃, adding 96.3g of Lipozyme435, timing after the reaction, filtering to remove the Lipozyme435 and distilling to remove the ethyl alcohol after the reaction is carried out for 2.5 hours, adding 1.45L of 85% ethyl alcohol and 1.45L of n-hexane into the reaction product, extracting and separating 2-glycerol monoester type n-3PUFA, washing the collected ethyl alcohol phase with the n-hexane for 2 times, and carrying out rotary evaporation to remove the ethyl alcohol and the water to obtain the product. The molecular distillation light phase obtained in the step (1) can be continuously used for molecular distillation to recover the n-3PUFA ethyl ester in the light phase, and the steps are repeated; after recovering n-hexane from the n-hexane phase, the resulting n-3 PUFA-rich ethyl ester can be used again in step (2). Finally, the 2-monoglyceride content of the product was 93.89%, the n-3PUFA content in 2-monoglyceride was 90.34%, and the recovery rate of n-3PUFA was 85.23%.
Example 3
(1) Accurately weighing 1Kg of anchovy oil, placing in a 5L conical flask with a plug, adding 481mL of absolute ethanol dissolved with 4g of NaOH, reacting at 70 ℃ for 1.5h, centrifuging the upper solution, distilling to recover ethanol, and washing with water to remove NaOH to obtain n-3 PUFA-containing ethyl ester; subsequently, the ethyl ester containing n-3PUFA was purified by four-stage molecular distillation to obtain 278g of n-3PUFA ethyl ester with n-3PUFA content of 90.44%; (2) adding 25g of glycerol into the ethyl ester rich in n-3PUFA as a substrate, uniformly stirring at 55 ℃, adding 9.1g of Novozym 435, starting reaction, reacting for 24 hours, and centrifuging to collect an upper oil phase (271 g); (3) adding 680g of absolute ethyl alcohol into the oil phase serving as a substrate, stirring and mixing the mixture evenly at 25 ℃, adding 57g of Lipozyme435, timing when the reaction starts, filtering to remove the Lipozyme435 after the reaction is carried out for 3 hours, distilling to remove the ethyl alcohol, adding 2.8L of 85% ethyl alcohol and 2.8L of n-hexane into the reaction product, extracting and separating 2-glycerol monoester type n-3PUFA, washing the collected ethyl alcohol phase with the n-hexane for 2 times, and carrying out rotary evaporation to remove the ethyl alcohol and the water to obtain the product. The molecular distillation light phase obtained in the step (1) can be continuously used for molecular distillation to recover the n-3PUFA ethyl ester in the light phase, and the steps are repeated; after recovering n-hexane from the n-hexane phase, the resulting n-3 PUFA-rich ethyl ester can be used again in step (2). Finally, the 2-monoglyceride content of the product was 93.21%, the n-3PUFA content of the 2-monoglyceride was 90.15%, and the recovery yield of n-3PUFA was 85.66%.
Example 4
(1) Accurately weighing 1Kg of tuna oil, placing the tuna oil into a 5L conical flask with a plug, adding 385mL of absolute ethanol dissolved with 3g of NaOH, reacting at 70 ℃ for 1.5h, centrifugally separating the upper solution, distilling to recover ethanol, and washing with water to remove NaOH to obtain n-3 PUFA-containing ethyl ester; subsequently, the ethyl ester containing n-3PUFA was purified by four-stage molecular distillation to obtain 281g of n-3PUFA ethyl ester having an n-3PUFA content of 90.67%; (2) adding 25g of glycerol into the ethyl ester rich in n-3PUFA as a substrate, uniformly stirring at 55 ℃, adding 15.3g of Lipozyme435, starting reaction timing, reacting for 12 hours, centrifuging and collecting an upper oil phase (272 g); (3) adding 816g of absolute ethyl alcohol into the oil phase serving as a substrate, stirring and mixing the mixture evenly at 30 ℃, adding 87g of Novozym 435, timing when the reaction starts, filtering to remove the Novozym 435 after the reaction is carried out for 3 hours, distilling to remove the ethanol, adding 2.8L of 85% ethanol and 2.8L of n-hexane into the reaction product, extracting and separating 2-glycerol monoester type n-3PUFA, washing the collected ethanol phase with the n-hexane for 2 times, and carrying out rotary evaporation to remove the ethanol and the water to obtain the product. The molecular distillation light phase obtained in the step (1) can be continuously used for molecular distillation to recover the n-3PUFA ethyl ester in the light phase, and the steps are repeated; after recovering n-hexane from the n-hexane phase, the resulting n-3 PUFA-rich ethyl ester can be used again in step (2). Finally, the 2-monoglyceride content of the product was 94.67%, the n-3PUFA content of the 2-monoglyceride was 90.52%, and the recovery yield of n-3PUFA was 86.78%.
Claims (10)
1. A method for preparing 2-monoglyceride type n-3PUFA by an enzymatic method is characterized by comprising the following steps:
step 1: carrying out an ethylation reaction on the oil containing n-3PUFA and absolute ethyl alcohol under the catalysis of an alkaline catalyst, and purifying a reaction product to obtain fatty acid ethyl ester containing n-3 PUFA;
step 2: taking glycerol and the n-3 PUFA-containing fatty acid ethyl ester obtained in the step 1 as substrates, carrying out transesterification reaction on the substrates by adopting immobilized lipase, and centrifuging to collect an upper oil phase;
and step 3: and (3) taking the upper oil phase collected in the step (2) as a substrate, carrying out alcoholysis reaction by using immobilized lipase, and then extracting and separating to obtain the 2-monoglyceride type n-3 PUFA.
2. The enzymatic method for producing 2-monoglyceride type n-3PUFA according to claim 1, wherein NaOH is used as the alkaline catalyst in step 1, and the mass of NaOH is 0.2 to 0.4% of that of the n-3 PUFA-containing fat; the molar ratio of the ethanol to the n-3 PUFA-containing oil is 6-9: 1.
3. The enzymatic method for producing 2-monoglyceride type n-3PUFA according to claim 1, wherein the temperature of the ethylation reaction in step 1 is 70 to 75 ℃ and the reaction time is 1.5 to 2 hours.
4. The enzymatic process for producing 2-monoglyceride type n-3PUFA according to claim 1, wherein in step 1, the purification is performed by four-stage molecular distillation wherein the feed flow rates of the four stages are 3mL/min, 2mL/min, 1mL/min and 0.5mL/min, respectively, and the evaporation surface temperatures are 110 ℃, 140 ℃, 150 ℃ and 160 ℃, and the operating pressures are 30Pa, 15Pa, 5Pa and 1Pa, respectively.
5. The enzymatic process for the production of n-3PUFA in the 2-monoglyceride type according to claim 1, wherein the immobilized lipase used in step 2 and step 3 is Novozym 435 or Lipozyme 435.
6. The enzymatic method for producing 2-monoglyceride type n-3PUFA according to claim 1, wherein in step 2, the molar ratio of n-3 PUFA-containing fatty acid ethyl ester to glycerol is 3:1, the amount of enzyme added is 2 to 5% of the total mass of the substrate, the reaction temperature is 55 to 65 ℃, and the reaction time is 12 to 24 hours.
7. The enzymatic method for preparing 2-monoglyceride type n-3PUFA according to claim 1, wherein in step 3, the alcohol used in the alcoholysis reaction is ethanol, the mass ratio of the upper oil phase to the ethanol is 1:2.5 to 3, the amount of enzyme added is 6 to 10 wt% of the total mass of the substrates, the reaction temperature is 25 to 30 ℃, and the reaction time is 1.5 to 3 hours.
8. The enzymatic process for producing 2-monoglyceride type n-3PUFA according to claim 1, characterized in that the specific steps of extraction in step 3 are: adding n-hexane into the alcoholysis reaction product, adding ethanol, and washing with n-hexane for several times.
9. The enzymatic process for producing 2-monoglyceride type n-3PUFA according to claim 8, wherein the volume fraction of ethanol is 85%.
10. The enzymatic process for producing 2-monoglyceride type n-3PUFA according to claim 1, wherein in step 3, the separation is performed by rotary evaporation or distillation under reduced pressure.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101161819A (en) * | 2007-10-12 | 2008-04-16 | 广东海洋大学 | Method for preparing n-3PUFA ocean glycerin ester by enzymatical process |
| CN103880672A (en) * | 2014-03-20 | 2014-06-25 | 江苏中邦制药有限公司 | High-purity DHA algae oil ethyl ester and preparation method for transferring high-purity DHA algae oil ethyl ester into glyceride |
| CN105483170A (en) * | 2016-01-08 | 2016-04-13 | 江南大学 | Method for synthesizing Sn-2-monoglyceride through enzymic method |
| CN105821088A (en) * | 2016-05-26 | 2016-08-03 | 暨南大学 | Method for preparing glyceride rich in EPA and DHA through enzyme catalysis |
| CN106496026A (en) * | 2016-10-21 | 2017-03-15 | 武汉藤欣生物工程有限公司 | In the method that acer truncatum buge oil prepares neural acetoacetic ester as raw material |
-
2020
- 2020-03-11 CN CN202010167155.XA patent/CN111172208A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101161819A (en) * | 2007-10-12 | 2008-04-16 | 广东海洋大学 | Method for preparing n-3PUFA ocean glycerin ester by enzymatical process |
| CN103880672A (en) * | 2014-03-20 | 2014-06-25 | 江苏中邦制药有限公司 | High-purity DHA algae oil ethyl ester and preparation method for transferring high-purity DHA algae oil ethyl ester into glyceride |
| CN105483170A (en) * | 2016-01-08 | 2016-04-13 | 江南大学 | Method for synthesizing Sn-2-monoglyceride through enzymic method |
| CN105821088A (en) * | 2016-05-26 | 2016-08-03 | 暨南大学 | Method for preparing glyceride rich in EPA and DHA through enzyme catalysis |
| CN106496026A (en) * | 2016-10-21 | 2017-03-15 | 武汉藤欣生物工程有限公司 | In the method that acer truncatum buge oil prepares neural acetoacetic ester as raw material |
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