CN112280810A - Preparation method of medium-long chain triglyceride rich in polyunsaturated fatty acid - Google Patents
Preparation method of medium-long chain triglyceride rich in polyunsaturated fatty acid Download PDFInfo
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Abstract
The invention discloses a preparation method of medium-long chain triglyceride rich in polyunsaturated fatty acid, belonging to the field of deep processing of grease. The method comprises the steps of taking oil containing polyunsaturated fatty acid, lipase, ethanol and water to react in a reactor, and removing non-target products to obtain glyceride rich in the polyunsaturated fatty acid; and then taking the obtained glyceride rich in polyunsaturated fatty acid, medium-chain fatty acid (C8-12) and lipase to react in a reactor, and removing the medium-chain fatty acid in the mixture to obtain medium-and long-chain triglyceride rich in polyunsaturated fatty acid. Compared with the traditional chemical catalysis, the method has the advantages of mild reaction conditions of the enzyme method, less by-products and environmental protection. Compared with a direct esterification method, the loss of PUFA is lower, and the yield is higher; compared with acidolysis and ester exchange, the method can reduce the proportion of saturated and monounsaturated fatty acids, so that the method has more medicinal and health-care values.
Description
Technical Field
The invention belongs to the field of deep processing of grease, and particularly relates to a preparation method of medium-long chain triglyceride rich in polyunsaturated fatty acid.
Background
Polyunsaturated fatty acid (PUFA) has important biological significance to human organisms, and ARA, DHA and EPA are proved to be capable of remarkably preventing and treating cardiovascular diseases and inhibiting the growth of cancer cells in medicine; in addition, these substances have functions of promoting normal development of sensory organs of infants and preventing and treating senile dementia, and thus are widely used in health foods.
Medium-and long-chain triglycerides (MLCT) refer to structural lipids containing both Medium-chain fatty acids (MCFA, C8-12) and long-chain fatty acids (LCFA) on the glycerol backbone. Medium carbon chain triglycerides (MCT) are hydrolyzed and absorbed by the body more rapidly than long carbon chain triglycerides (LCT). And the MLCT contains MCFA and LCFA at the same time, so that the synchronous transportation of fatty acid can be realized, the MCFA is released into blood at a more controllable hydrolysis rate, the absorption of the LCFA can be improved, and the MCT and the LCT have the advantages. Research shows that the MLCT can effectively reduce the blood fat and cholesterol level and prevent and treat vascular diseases such as thrombus and the like. In addition, the medium-chain fatty acid and the monoglyceride thereof generated by the hydrolysis of the MLCT also have strong bacteriostatic and antiviral activities. In addition, the chain length of saturated fatty acids affects their antimicrobial activity, and too long or too short a carbon chain results in reduced activity.
The PUFA products on the market mainly have three forms of ethyl ester type, glyceride type and free fatty acid type, and according to research reports, the ethyl ester type products are difficult to digest and absorb in human bodies, the free fatty acid type has poor storage stability, and the glyceride type has the advantages of the ethyl ester type, the glyceride type and the free fatty acid type. However, the content of the glyceride-type PUFA directly extracted in nature is relatively low, and many researches have been made to enrich glyceride-type PUFA with high concentration, and the core content is that saturated or monounsaturated fatty acid in oil is removed or replaced by PUFA through an enzyme method, so that the value of the oil is improved. However, most of the LCFAs in the MLCT prepared by the current research are saturated and monounsaturated fatty acids of C14-18, and the research is biased to MLCT in infant formula. Is mainly prepared by the ester exchange of long-chain soybean oil or rapeseed oil and medium-chain coconut oil. There are relatively few reports on the research on the preparation of PUFA-enriched MLCT, and the MLCT obtained from the reported research results has many redundant and unremoved fatty acids, such as saturated fatty acids or monounsaturated fatty acids, which affect the biological value. Therefore, there is a need in the art for a method that meets the green and safe philosophy and can efficiently and precisely prepare PUFA-rich MLCTs.
Disclosure of Invention
The present invention has been made in view of the above and/or problems occurring in the existing preparation methods of medium-and long-chain triglycerides rich in polyunsaturated fatty acids. The invention provides a preparation method of medium-long chain triglyceride rich in polyunsaturated fatty acid, which comprises the steps of firstly utilizing fatty acid specific lipase to alcoholyze saturated and monounsaturated fatty acids in grease to obtain high-concentration PUFA glyceride; secondly, the glyceride product obtained by the first-step hydrolysis and medium-chain fatty acid are esterified into MLCT under the catalysis of lipase, and the proportion of saturated and monounsaturated fatty acid in the product is very low, so that the product has more medicinal and health-care values.
Specifically, the first object of the present invention is to provide a process for producing medium-and long-chain triglycerides rich in polyunsaturated fatty acids, which comprises,
(1) adding oil and fat containing polyunsaturated fatty acid, lipase, alcohol and water into a reactor, reacting for a certain time at 20-50 ℃ to obtain a mixture, and removing the lipase, the alcohol, fatty acid ester and free fatty acid in the mixture to obtain glyceride rich in the polyunsaturated fatty acid; wherein the lipase in step (1) comprises one or more of Candida antarctica lipase A derived from Candida antarctica and NovozymeET2.0 derived from Aspergillus sp;
(2) taking the glyceride rich in polyunsaturated fatty acid, the medium-chain fatty acid and the lipase obtained in the step (1) to react in a reactor for a certain time, and removing the medium-chain fatty acid in the mixture to obtain medium-long chain triglyceride rich in the polyunsaturated fatty acid; wherein the lipase in step (2) comprises one or more of Lipozyme RM IM derived from Rhizomucormieihei, Novozyme 435 and Lipozyme 435 derived from Candida antarctica B, Lipozyme TL IM derived from Thermomyces lanuginosus, and NS 40086 derived from Aspergillus oryzae.
As a preferable mode of the method for producing medium-and long-chain triglycerides rich in polyunsaturated fatty acids, the polyunsaturated fatty acid-containing fat includes a fat containing one or more of eicosapentaenoic acid (EPA), arachidonic acid (ARA), and docosahexaenoic acid (DHA).
As a preferable mode of the method for producing medium-and long-chain triglycerides rich in polyunsaturated fatty acids, the fat or oil containing polyunsaturated fatty acids is preferably fish meat or algal oil.
As a preferable scheme of the preparation method of the medium-long chain triglyceride rich in polyunsaturated fatty acid, the medium-chain fatty acid is fatty acid with a carbon chain length of 8-12; preferably comprising one or more of caprylic acid, pelargonic acid, capric acid, lauric acid.
As a preferred embodiment of the method for preparing the medium-long chain triglyceride rich in the polyunsaturated fatty acids, the alcohol includes one or more of methanol, ethanol and isobutanol.
As a preferable scheme of the preparation method of the medium-long chain triglyceride rich in polyunsaturated fatty acid, in the step (1), the mass ratio of the ethanol to the fat containing the polyunsaturated fatty acid is 0.1-30: 1; the water-alcohol mass ratio is 1-250%.
As a preferable scheme of the preparation method of the medium-long chain triglyceride rich in polyunsaturated fatty acid, in the step (1), the reaction time is 4-48 h, and the addition amount of the lipase is 100-10000U/g of the grease.
As a preferable scheme of the preparation method of the medium-long chain triglyceride rich in the polyunsaturated fatty acid, in the step (2), the mass ratio of the glyceride rich in the polyunsaturated fatty acid to the medium-chain fatty acid is 1: 0.2-10.
As a preferable scheme of the preparation method of the medium-long chain triglyceride rich in polyunsaturated fatty acid, in the step (2), the reaction time is 6-72 hours, and the reaction temperature is 30-70 ℃; the addition amount of the lipase is 50-8000U/g glyceride.
The second purpose of the invention is to provide the application of the method in the field of grease processing.
The invention has the beneficial effects that:
(1) the invention provides a preparation method of medium-long chain triglyceride rich in polyunsaturated fatty acid, which comprises the following two steps of catalytic methods of lipase: firstly, carrying out alcoholysis on saturated and monounsaturated fatty acids in grease by using fatty acid specific lipase to obtain glyceride of high-concentration PUFA; secondly, the product of the first step is esterified with medium-chain fatty acids under lipase catalysis to MLCT. Compared with the traditional chemical catalysis, the enzymatic reaction has mild conditions, few byproducts and environmental protection.
(2) The two lipases selected in the first step of alcoholysis have PUFA discrimination in the aspect of alcoholysis speed, and especially have better and obvious effects on ARA, EPA and DHA; the alcoholysis rate of the selected lipase on triglyceride containing saturated fatty acid and low unsaturated fatty acid is much higher than that of triglyceride containing polyunsaturated fatty acid, so that the saturated fatty acid and the monounsaturated fatty acid are removed by moderate alcoholysis by utilizing the characteristic, while the protein of the general lipase in an alcohol solution is denatured to make the lipase lose reactivity,
(3) compared with a one-step direct esterification method, the method has the advantages that the loss of PUFA in the prepared product is lower, the yield is higher, and the content of saturated fatty acid and monounsaturated fatty acid is greatly reduced; in addition, compared with one-step acidolysis and ester exchange, the acidolysis possibly occurring in the second step of the invention further reduces the saturated and monounsaturated fatty acids in the product, and finally the content of the saturated and monounsaturated fatty acids in the product is lower than 9 percent and can be reduced to 4 percent at least, so that the product has more medicinal and health-care values.
Drawings
FIG. 1 is a gas chromatographic chart of the glyceride fatty acid component before reaction of algal oil in example 13 of the present invention.
FIG. 2 is a gas chromatography chromatogram of the glyceride fatty acid component after the algal oil reaction in example 13 of the present invention.
FIG. 3 is a gas chromatography chromatogram of the fatty acid composition of a glyceride after esterification of a glyceride and a medium-chain fatty acid in example 25 of the present invention.
Detailed Description
In order to make the aforementioned objects, features and advantages of the present invention more comprehensible, specific embodiments thereof are described in detail below with reference to examples of the specification.
In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention, but the present invention may be practiced in other ways than those specifically described and will be readily apparent to those of ordinary skill in the art without departing from the spirit of the present invention, and therefore the present invention is not limited to the specific embodiments disclosed below.
The detection method comprises the following steps:
(1) method for measuring fatty acid content
Putting 50mg of sample into a 10mL graduated tube, adding 2mL of 0.5mol/L potassium hydroxide-methanol solution, saponifying at 65 ℃ for 30min, cooling, adding 2mL of 25% volume fraction boron trifluoride-methanol solution, and carrying out water bath at 70 ℃ for 5 min; adding 2mL of n-hexane, oscillating for 3-4min to extract fatty acid methyl ester, adding 4mL of saturated NaCl solution, taking the upper layer solution, adding anhydrous sodium sulfate, oscillating (centrifuging at 10000rpm for 5min), sucking by a syringe, passing through a membrane, and detecting by using a gas chromatography, wherein the gas chromatography has the following operation parameters: selecting a 7890 gas chromatograph and a Flame Ionization Detector (FID); the gas chromatographic column is 60m × 0.32mm × 2.5 μm; the nitrogen flow rate was set to 1.0mL/min and the temperature of the injector and detector were set to 250 ℃. The initial column temperature was maintained at 80 ℃ for 0.5min and then increased from 80 ℃ to 165 ℃ at a rate of 40 ℃/min. The column temperature was raised to 230 ℃ at a rate of 4 ℃/min and held at 230 ℃ for 4 min. And calculating the content of the fatty acid by using a peak area normalization method.
The lipases Candida antarctica lipase A (6000LUN/g), Novozyme 2.0(1000LUN/g), Novozym435 (10000PLUN/g), Lipozyme 435(10000UN/g), NS 40086(275IUN/g), Lipozyme RM IM (275IUN/g), Lipozyme TL IM (250IUN/g) of the present invention were purchased from Novoxin Biotechnology, Inc. AY "Amano"30SD, 30000U/g, available from Japan Nature Bio-enzyme preparations Ltd.
The oil used in the invention is commercially available, wherein the content of n-3PUFA in the fish oil is 34.3%; the content of n-3PUFA in algae oil is 46.0%. Other reagents are not specifically indicated and are all commercially available.
Example 1
Accurately weighing 3.0g of fish oil, 1.5g of absolute ethyl alcohol, 0.30g of water and 1400U of Candida antarctica lipase A, adding into a reaction kettle, placing into a magnetic rotor, and sealing. The reaction kettle is placed on a magnetic stirrer, circulating water is introduced into the reaction kettle, the temperature of the water is constant at 35 ℃, and the reaction kettle is kept for 10 hours. And after the reaction is finished, removing fatty acid ethyl ester and free fatty acid in the mixture by molecular distillation to obtain the fish oil glyceride rich in n-3 PUFAs. The fatty acid composition of the glyceride product was increased from 34.3% to 56.6% after alcoholysis by n-3PUFAs, as shown in Table 1.
The experimental conditions and results for examples 2-24 are shown in Table 1 below, with the operating parameters being the same as in example 1 except for the conditions noted.
Comparative example 1
Accurately weighing 3.0g of fish oil, 1.5g of absolute ethyl alcohol (alcohol-oil molar ratio is 48:1), 0.0.30g of water and AY 'Amano' 30SD lipase 1400U, adding into a reaction kettle, placing into a magnetic rotor, and sealing. The reaction kettle is placed on a magnetic stirrer, circulating water is introduced into the reaction kettle, the temperature of the water is constant at 35 ℃, and the reaction kettle is kept for 10 hours. And after the reaction is finished, removing fatty acid ethyl ester and free fatty acid in the mixture by molecular distillation to obtain the fish oil glyceride rich in n-3 PUFAs. In the fatty acid composition of the glyceride product, the n-3PUFAs accounts for 34.8 percent of the improvement from 34.3 percent of the crude oil after alcoholysis.
Comparative example 2
The same procedure as in comparative example 1 was repeated except that the lipase type in the reaction system was changed to Novozym435, and the n-3PUFAs content in the obtained glycerides was as shown in Table 1.
Comparative example 3
The same as in comparative example 1, except that the reaction raw material in the reaction system was changed to algal oil, the other operations were the same, and the n-3PUFAs content in the obtained glyceride was as shown in Table 1.
Comparative example 4
The same procedure as in comparative example 3 was repeated except that the lipase type in the reaction system was changed to Novozym435, and the n-3PUFAs content in the obtained glycerides was as shown in Table 1.
The reaction conditions and n-3PUFAs content in the glyceride products of examples 1 to 24 and comparative examples 1 to 4 are shown in Table 1.
TABLE 1 reaction conditions and n-3PUFAs content in glycerides for examples 1-24 and comparative examples 1-4
As can be seen from Table 1, the lipase Candida antarctica lipase A and NovozymeT2.0 can carry out alcoholysis of sunflower oil or fish oil in ethanol, and the glyceride product n-3PUFA is obviously enriched, and the enrichment effect is more than 1.5 times that before alcoholysis.
Example 25
2g of the product obtained in example 13 was taken, and 1.2g of capric acid (purity. gtoreq.99%) was added thereto. And adding 1000U of Lipozyme RM IM lipase into the reaction kettle, adding a magnetic rotor, and sealing. The reaction kettle is placed on a magnetic stirrer, circulating water is introduced into the reaction kettle, the water temperature is constant at 40 ℃, and the reaction kettle is kept for 12 hours. And after the reaction is finished, removing free fatty acid in the mixture by molecular distillation to obtain an esterified product, namely the medium-long chain triglyceride rich in n-3 PUFAs. In the fatty acid composition of the product, the content of n-3PUFAs is 60.3%, the content of medium-chain fatty acid is 23.4%, and the content of saturated and monounsaturated fatty acid is 4.7%.
Example 26
2g of the product obtained in example 13 was taken, and 1.2g of octanoic acid (purity. gtoreq.99%) was added. Adding 1000U of Novozym435 lipase into the reaction kettle, adding a magnetic rotor, and sealing the reaction kettle. The reaction kettle is placed on a magnetic stirrer, circulating water is introduced into the reaction kettle, the water temperature is constant at 40 ℃, and the reaction kettle is kept for 12 hours. And after the reaction is finished, removing free fatty acid in the mixture by molecular distillation to obtain an esterified product, namely the medium-long chain triglyceride rich in n-3 PUFAs. The n-3PUFAs and medium chain fatty acid content of the obtained product are shown in Table 2.
Example 27
The same procedure as in example 25 was repeated except for changing the medium-chain fatty acid in the reaction system to pelargonic acid (purity. gtoreq.99%) to give a product having n-3PUFAs and medium-chain fatty acid contents as shown in Table 2.
Example 28
The same procedure as in example 26 was repeated except for changing the medium-chain fatty acid in the reaction system to lauric acid (purity. gtoreq.99%) to give a product having n-3PUFAs and medium-chain fatty acid contents as shown in Table 2.
Example 29
The same procedure as in example 25 was repeated except for changing the lipase in the reaction system to Lipozyme 435, and the n-3PUFAs and medium-chain fatty acid contents in the obtained product were as shown in Table 2.
Example 30
The same procedure as in example 25 was repeated except for changing the lipase in the reaction system to Lipozyme TL IM, and the n-3PUFAs and medium-chain fatty acid contents in the obtained product were as shown in Table 2.
Example 31
The same procedure as in example 25 was repeated except for changing the lipase in the reaction system to NS 40086, and the n-3PUFAs and medium-chain fatty acid contents in the obtained product were as shown in Table 2.
Example 32
The same procedure as in example 25 was repeated except that the mass of the medium-chain fatty acid in the reaction system was changed to 10g, and the n-3PUFAs and the medium-chain fatty acid content in the obtained product were as shown in Table 2.
Example 33
The same procedure as in example 25 was repeated except that the mass of the medium-chain fatty acid in the reaction system was changed to 0.6g, and the contents of n-3PUFAs and medium-chain fatty acids in the obtained glyceride were as shown in Table 2.
Example 34
2g of the product obtained in example 13 was taken, and 1.2g of lauric acid (purity: 99%) was added thereto. Adding 1000U of Novozym435 lipase into the reaction kettle, adding a magnetic rotor, and sealing the reaction kettle. The reaction kettle is placed on a magnetic stirrer, circulating water is introduced into the reaction kettle, the water temperature is constant at 40 ℃, and the reaction kettle is kept for 6 hours. And after the reaction is finished, removing free fatty acid in the mixture by molecular distillation to obtain the medium-long chain triglyceride rich in n-3 PUFAs. The n-3PUFAs and medium chain fatty acid content of the obtained product are shown in Table 2.
Example 35
The same procedure as in example 34, except that the reaction time in the reaction system was changed to 48 hours, was repeated, and the n-3PUFAs and the medium-chain fatty acid content in the obtained glyceride were as shown in Table 2.
Example 36
2g of the product obtained in example 13 was taken, and 1.2g of octanoic acid (purity. gtoreq.99%) was added. Then adding 100U of Lipozyme 435 lipase, adding into the reaction kettle, adding a magnetic rotor, and sealing. The reaction kettle is placed on a magnetic stirrer, circulating water is introduced into the reaction kettle, the water temperature is constant at 40 ℃, and the reaction kettle is kept for 12 hours. And after the reaction is finished, removing free fatty acid in the mixture by molecular distillation to obtain the medium-long chain triglyceride rich in n-3 PUFAs. The n-3PUFAs and medium chain fatty acid content of the obtained product are shown in Table 2.
Example 37
The same procedure as in example 36 was repeated except that the amount of the enzyme added to the reaction system was changed to 6000U, and the contents of n-3PUFAs and medium-chain fatty acids in the obtained glyceride were as shown in Table 2.
Example 38
The same procedure as in example 25 was repeated except that the reaction temperature in the reaction system was changed to 25 ℃ and the n-3PUFAs and the medium-chain fatty acid content in the obtained glyceride were as shown in Table 2.
Example 39
2g of the product obtained in example 13 was taken, and 1.2g of octanoic acid (purity. gtoreq.99%) was added. Adding 1000U of Novozym435 lipase into the reaction kettle, adding a magnetic rotor, and sealing the reaction kettle. The reaction kettle is placed on a magnetic stirrer, circulating water is introduced into the reaction kettle, the temperature of the water is constant at 60 ℃, and the reaction kettle is kept for 12 hours. And after the reaction is finished, removing free fatty acid in the mixture by molecular distillation to obtain the medium-long chain triglyceride rich in n-3 PUFAs. The n-3PUFAs and medium chain fatty acid content of the obtained product are shown in Table 2.
Example 40
2g of the product obtained in example 1 was taken, and 1.2g of capric acid (purity: 99% or more) was added thereto. And adding 1000U of Lipozyme RM IM lipase into the reaction kettle, adding a magnetic rotor, and sealing. The reaction kettle is placed on a magnetic stirrer, circulating water is introduced into the reaction kettle, the water temperature is constant at 40 ℃, and the reaction kettle is kept for 12 hours. And after the reaction is finished, removing free fatty acid in the mixture by molecular distillation to obtain the medium-long chain triglyceride rich in n-3 PUFAs. The n-3PUFAs and medium chain fatty acid content of the obtained product are shown in Table 2.
EXAMPLE 41
2g of the product obtained in example 1 was taken, and 1.2g of octanoic acid (purity. gtoreq.99%) was added. Adding 1000U of Novozym435 lipase into the reaction kettle, adding a magnetic rotor, and sealing the reaction kettle. The reaction kettle is placed on a magnetic stirrer, circulating water is introduced into the reaction kettle, the water temperature is constant at 40 ℃, and the reaction kettle is kept for 12 hours. And after the reaction is finished, removing free fatty acid in the mixture by molecular distillation to obtain the medium-long chain triglyceride rich in n-3 PUFAs. The n-3PUFAs and medium chain fatty acid content of the obtained product are shown in Table 2.
Comparative example 5
The same procedure as in example 25, but changing the lipase in the reaction system to Candida antarctica lipase A, was repeated in the same manner, and the n-3PUFAs and medium-chain fatty acid contents in the obtained product were as shown in Table 2.
Comparative example 6
The same procedure as in example 25 was repeated except for changing the lipase in the reaction system to AY "Amano"30SD, and the resulting products had n-3PUFAs and medium-chain fatty acid contents as shown in Table 2.
Comparative example 7
2g of algae oil is taken, and 1.2g of capric acid (the purity is more than or equal to 99%) is added. And adding 1000U of Lipozyme RM IM lipase into the reaction kettle, adding a magnetic rotor, and sealing. The reaction kettle is placed on a magnetic stirrer, circulating water is introduced into the reaction kettle, the water temperature is constant at 40 ℃, and the reaction kettle is kept for 12 hours. And after the reaction is finished, removing free fatty acid in the mixture by molecular distillation to obtain the medium-long chain triglyceride. The n-3PUFAs and medium chain fatty acid content of the obtained product are shown in Table 2.
Comparative example 8
The same as in comparative example 7, except that the lipase in the reaction system was changed to Novozym435, the same was conducted, and the n-3PUFAs and medium-chain fatty acid contents in the obtained product were as shown in Table 2.
Comparative example 9
The same as in comparative example 7, except that the algal oil in the reaction system was changed to fish oil, the same operation was carried out, and the contents of n-3PUFAs and medium-chain fatty acids in the obtained product were as shown in Table 2.
Comparative example 10
The same as in comparative example 9, except that the lipase in the reaction system was changed to Novozym435, the same was conducted, and the n-3PUFAs and medium-chain fatty acid contents in the obtained product were as shown in Table 2.
The reaction conditions, the contents of n-3PUFAs and medium-chain fatty acids in the products of examples 25-41 and comparative examples 5-10 are shown in Table 2, wherein the reaction raw materials of examples 25-39 and comparative examples 5-6 are the products of example 13, and the reaction raw materials of examples 40-41 are the products of example 1.
Table 2 reaction conditions, n-3PUFAs and medium chain fatty acid contents in the products, in examples 25 to 41 and comparative examples 5 to 10
Wherein a represents the content of saturated fatty acids and monounsaturated fatty acids in the product
As can be seen from examples 25-41 in Table 2, the medium-long chain triglyceride prepared by the two-step method disclosed by the invention is rich in n-3PUFAs, the content of a target product is further improved in the enzymolysis process, and compared with the one-step acidolysis (comparative examples 7-10), the method disclosed by the invention is capable of greatly reducing the content of saturated fatty acids and monounsaturated fatty acids in the product and improving the content of PUFAs in the triglyceride, so that the content of non-target products of algae oil is reduced to 4.0% and is far less than 15.7% of that of the one-step method; the content of the fish oil non-target products is reduced to 22.9 percent, which is far less than 34.8 percent of the content of the fish oil non-target products in the one-step method.
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (10)
1. A method for preparing medium-long chain triglyceride rich in polyunsaturated fatty acid, which comprises the following steps,
(1) adding oil and fat containing polyunsaturated fatty acid, lipase, alcohol and water into a reactor, reacting for a certain time at 20-60 ℃ to obtain a mixture, and removing the lipase, the alcohol, fatty acid ester and free fatty acid in the mixture to obtain glyceride rich in the polyunsaturated fatty acid;
wherein the lipase in step (1) comprises one or more of Candida antarctica lipase A derived from Candida antarctica and NovozymeET2.0 derived from Aspergillus sp;
(2) taking the glyceride rich in polyunsaturated fatty acid, the medium-chain fatty acid and the lipase obtained in the step (1) to react in a reactor for a certain time, and removing the medium-chain fatty acid in the mixture to obtain medium-long chain triglyceride rich in the polyunsaturated fatty acid;
wherein the lipase in step (2) comprises one or more of Lipozyme RM IM derived from Rhizomucormieihei, Novozyme 435 and Lipozyme 435 derived from Candida antarctica B, Lipozyme TL IM derived from Thermomyces lanuginosus, and NS 40086 derived from Aspergillus oryzae.
2. The method of claim 1, wherein the polyunsaturated fatty acid-containing fat comprises one or more of eicosapentaenoic acid, arachidonic acid, and docosahexaenoic acid.
3. The method for preparing middle-and-long-chain triglycerides rich in polyunsaturated fatty acids according to claim 1 or 2, wherein the middle-and-long-chain fatty acids are fatty acids with a carbon chain length of 8-12.
4. The method of claim 3, wherein the alcohol comprises one or more of methanol, ethanol and isobutanol.
5. The method for preparing medium-and long-chain triglycerides rich in polyunsaturated fatty acids according to any of claims 1 to 4, wherein in the step (1), the mass ratio of the ethanol to the polyunsaturated fatty acid-containing oil is 0.1-30: 1; the water-alcohol mass ratio is 1-250%.
6. The method for preparing middle-long chain triglyceride rich in polyunsaturated fatty acids according to any one of claims 1 to 5, wherein in the step (1), the reaction time is 4 to 48 hours, and the reaction temperature is 20 to 50 ℃; the addition amount of the lipase is 100-10000U/g of grease.
7. The method for preparing medium-and long-chain triglycerides rich in polyunsaturated fatty acids according to any of claims 1-6, wherein in the step (2), the mass ratio of the glycerides rich in polyunsaturated fatty acids to the medium-and long-chain fatty acids is 1: 0.2-10.
8. The method for preparing middle-long chain triglyceride rich in polyunsaturated fatty acids according to any one of claims 1 to 7, wherein in the step (2), the reaction time is 6 to 72 hours, and the reaction temperature is 30 to 70 ℃.
9. The method for preparing middle-and long-chain triglycerides rich in polyunsaturated fatty acids according to any of claims 1 to 8, wherein the lipase is added in an amount of 50 to 8000U/g of glycerides in step (2).
10. The application of the preparation method of the polyunsaturated fatty acid-rich medium-long chain triglyceride in any one of claims 1 to 9 in the field of oil processing.
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| CN111378701A (en) * | 2020-03-20 | 2020-07-07 | 江南大学 | Preparation method of middle-long carbon chain triglyceride containing polyunsaturated fatty acid and product thereof |
| CN112980898A (en) * | 2021-04-30 | 2021-06-18 | 浙江师范大学 | Method for concentrating DHA in schizochytrium limacinum grease |
| CN113481247A (en) * | 2021-07-20 | 2021-10-08 | 广东浠芮生物科技有限责任公司 | Preparation method of medium-carbon-chain triglyceride |
| CN113881718A (en) * | 2021-10-25 | 2022-01-04 | 江南大学 | Method for preparing free fatty acid-rich grease composition by using two-step enzyme catalysis method and application |
| CN114521655A (en) * | 2022-01-27 | 2022-05-24 | 华南理工大学 | Intestinal tract probiotic promoter containing medium-long chain triglyceride in almond oil and preparation method and application thereof |
| CN115369132A (en) * | 2021-09-23 | 2022-11-22 | 江南大学 | Enzymatic synthesis method of medium-long chain triglyceride |
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| CN114521655A (en) * | 2022-01-27 | 2022-05-24 | 华南理工大学 | Intestinal tract probiotic promoter containing medium-long chain triglyceride in almond oil and preparation method and application thereof |
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