CN104711298B - A kind of method that triglycerides enzymolysis prepares 1,3 diglycerides - Google Patents
A kind of method that triglycerides enzymolysis prepares 1,3 diglycerides Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 32
- 150000003626 triacylglycerols Chemical class 0.000 title claims abstract description 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 36
- 241000321538 Candidia Species 0.000 claims abstract description 23
- 101710098556 Lipase A Proteins 0.000 claims abstract description 23
- 101710099648 Lysosomal acid lipase/cholesteryl ester hydrolase Proteins 0.000 claims abstract description 23
- 102100026001 Lysosomal acid lipase/cholesteryl ester hydrolase Human genes 0.000 claims abstract description 23
- 229910021645 metal ion Inorganic materials 0.000 claims abstract description 22
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 17
- 239000004367 Lipase Substances 0.000 claims description 9
- 235000012424 soybean oil Nutrition 0.000 claims description 9
- 239000003549 soybean oil Substances 0.000 claims description 9
- 230000015572 biosynthetic process Effects 0.000 claims description 4
- VTLYFUHAOXGGBS-UHFFFAOYSA-N Fe3+ Chemical compound [Fe+3] VTLYFUHAOXGGBS-UHFFFAOYSA-N 0.000 claims description 3
- WAEMQWOKJMHJLA-UHFFFAOYSA-N Manganese(2+) Chemical compound [Mn+2] WAEMQWOKJMHJLA-UHFFFAOYSA-N 0.000 claims description 3
- 229910001447 ferric ion Inorganic materials 0.000 claims description 3
- 229910001437 manganese ion Inorganic materials 0.000 claims description 3
- 230000007062 hydrolysis Effects 0.000 abstract description 18
- 238000006460 hydrolysis reaction Methods 0.000 abstract description 18
- 230000000694 effects Effects 0.000 abstract description 11
- 230000035484 reaction time Effects 0.000 abstract description 5
- 230000001737 promoting effect Effects 0.000 abstract 1
- 238000006243 chemical reaction Methods 0.000 description 27
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 25
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 18
- 239000000839 emulsion Substances 0.000 description 15
- 239000012071 phase Substances 0.000 description 15
- DRCWOKJLSQUJPZ-DZGCQCFKSA-N (4ar,9as)-n-ethyl-1,4,9,9a-tetrahydrofluoren-4a-amine Chemical compound C1C2=CC=CC=C2[C@]2(NCC)[C@H]1CC=CC2 DRCWOKJLSQUJPZ-DZGCQCFKSA-N 0.000 description 12
- 101001008429 Homo sapiens Nucleobindin-2 Proteins 0.000 description 12
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 12
- 102100027441 Nucleobindin-2 Human genes 0.000 description 12
- 235000011187 glycerol Nutrition 0.000 description 12
- 239000003921 oil Substances 0.000 description 11
- 235000019198 oils Nutrition 0.000 description 11
- 102000004882 Lipase Human genes 0.000 description 8
- 108090001060 Lipase Proteins 0.000 description 8
- 229940040461 lipase Drugs 0.000 description 8
- 235000019421 lipase Nutrition 0.000 description 8
- 238000002360 preparation method Methods 0.000 description 8
- LDVVTQMJQSCDMK-UHFFFAOYSA-N 1,3-dihydroxypropan-2-yl formate Chemical compound OCC(CO)OC=O LDVVTQMJQSCDMK-UHFFFAOYSA-N 0.000 description 7
- 238000000199 molecular distillation Methods 0.000 description 7
- 239000008280 blood Substances 0.000 description 6
- 210000004369 blood Anatomy 0.000 description 6
- 235000009508 confectionery Nutrition 0.000 description 6
- 150000002148 esters Chemical class 0.000 description 6
- 238000013459 approach Methods 0.000 description 5
- 238000000265 homogenisation Methods 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 102000004190 Enzymes Human genes 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- 229940088598 enzyme Drugs 0.000 description 4
- 230000032050 esterification Effects 0.000 description 4
- 238000005886 esterification reaction Methods 0.000 description 4
- 239000003925 fat Substances 0.000 description 4
- 235000013305 food Nutrition 0.000 description 4
- 230000036541 health Effects 0.000 description 4
- NQXWGWZJXJUMQB-UHFFFAOYSA-K iron trichloride hexahydrate Chemical class O.O.O.O.O.O.[Cl-].Cl[Fe+]Cl NQXWGWZJXJUMQB-UHFFFAOYSA-K 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 239000002994 raw material Substances 0.000 description 4
- 101100494530 Brassica oleracea var. botrytis CAL-A gene Proteins 0.000 description 3
- 241000222120 Candida <Saccharomycetales> Species 0.000 description 3
- 108050006759 Pancreatic lipases Proteins 0.000 description 3
- 102000019280 Pancreatic lipases Human genes 0.000 description 3
- 125000002252 acyl group Chemical group 0.000 description 3
- 239000004519 grease Substances 0.000 description 3
- 239000003960 organic solvent Substances 0.000 description 3
- 229940116369 pancreatic lipase Drugs 0.000 description 3
- PORPENFLTBBHSG-MGBGTMOVSA-N 1,2-dihexadecanoyl-sn-glycerol-3-phosphate Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(O)=O)OC(=O)CCCCCCCCCCCCCCC PORPENFLTBBHSG-MGBGTMOVSA-N 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 102000004157 Hydrolases Human genes 0.000 description 2
- 108090000604 Hydrolases Proteins 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 229910021380 Manganese Chloride Inorganic materials 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000008367 deionised water Substances 0.000 description 2
- 229910021641 deionized water Inorganic materials 0.000 description 2
- 235000021588 free fatty acids Nutrition 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 239000011565 manganese chloride Substances 0.000 description 2
- 229940099607 manganese chloride Drugs 0.000 description 2
- 235000002867 manganese chloride Nutrition 0.000 description 2
- BZDIAFGKSAYYFC-UHFFFAOYSA-N manganese;hydrate Chemical compound O.[Mn] BZDIAFGKSAYYFC-UHFFFAOYSA-N 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- 108010004103 Chylomicrons Proteins 0.000 description 1
- 208000001145 Metabolic Syndrome Diseases 0.000 description 1
- 235000019483 Peanut oil Nutrition 0.000 description 1
- 235000019484 Rapeseed oil Nutrition 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 125000001931 aliphatic group Chemical group 0.000 description 1
- 239000010775 animal oil Substances 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 229940099352 cholate Drugs 0.000 description 1
- BHQCQFFYRZLCQQ-OELDTZBJSA-N cholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 BHQCQFFYRZLCQQ-OELDTZBJSA-N 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 230000000536 complexating effect Effects 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000000378 dietary effect Effects 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 210000001842 enterocyte Anatomy 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000010931 ester hydrolysis Methods 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 125000005456 glyceride group Chemical group 0.000 description 1
- 125000003745 glyceroyl group Chemical group C(C(O)CO)(=O)* 0.000 description 1
- JEGUKCSWCFPDGT-UHFFFAOYSA-N h2o hydrate Chemical compound O.O JEGUKCSWCFPDGT-UHFFFAOYSA-N 0.000 description 1
- 235000006486 human diet Nutrition 0.000 description 1
- 239000000413 hydrolysate Substances 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 210000002490 intestinal epithelial cell Anatomy 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 230000037356 lipid metabolism Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- CNFDGXZLMLFIJV-UHFFFAOYSA-L manganese(II) chloride tetrahydrate Chemical class O.O.O.O.[Cl-].[Cl-].[Mn+2] CNFDGXZLMLFIJV-UHFFFAOYSA-L 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000037353 metabolic pathway Effects 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000011017 operating method Methods 0.000 description 1
- 239000000312 peanut oil Substances 0.000 description 1
- 230000000291 postprandial effect Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000010926 purge Methods 0.000 description 1
- 230000036632 reaction speed Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 210000000813 small intestine Anatomy 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- 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
- C12P7/6436—Fatty acid esters
- C12P7/6445—Glycerides
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- Organic Chemistry (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The method for preparing 1,3 diglycerides is digested the present invention relates to a kind of triglycerides, belongs to the preparing technical field of 1,3 diglycerides.The method of the present invention, including the step of ethanol or/and metal ion form hydrolyzation system is added into TAG, water and antarctic candidia lipase A.The metal ion that the present invention is added can significantly improve antarctic candidia lipase A activity, so as to improve the hydrolysis rate of triglycerides；And ethanol and metal ion are added simultaneously；Antarctic candidia lipase A can not only be improved simultaneously to select tendentiousness to 2 of TAG and shorten the reaction time；Moreover, hydrolysis rate increase rate further increases；As can be seen here, ethanol serves the effect for promoting metal ion to improve hydrolysis rate.
Description
Technical field
The present invention relates to a kind of method that triglycerides enzymolysis prepares 1,3-DAG, belong to the system of 1,3-DAG
Standby technical field.
Background technology
Grease is the important component of human diet, carries and provides energy for body, and in food processing process
Improve the functions such as flavour of food products.With expanding economy and the raising of living standards of the people, China resident grease intake is increasingly
Improve.And many studies have shown that, grease intake can excessively increase body and suffer from atherosclerosis, diabetes B, high fat of blood, height
The risk of the metabolic syndromes such as blood pressure, serious threat body health.
1,3-DAG（DAG）It is the main constituents in natural oil, its color, energy density, smell, stably
Property and the bioavilability in body and traditional triglycerides（TAG）Between without significant difference.Many Dietary frequencies are real
To test and show, 1,3-DAG intake can significantly reduce the indexs such as blood fat and the blood glucose of body, to improving blood fat and metabolism of blood glucose,
Promote health and play an important roll.
Architectural difference between 1,3-DAG and TAG, cause its metabolic pathway to change, be that it plays health efficacy
Basis.Pancreatic lipase in human body is sn-1 positions and sn-3 positions specific lipase, can only be hydrolyzed on TAG molecules 1 and 3
Ester bond.TAG molecules enter small enteral, and the ester bond of its 1 and 3 produces 2- monoacyls first by pancreatic lipase selective hydrolysis
Glyceride（2-MAG）With the free fatty acids of 2 molecules（NEFA）.These products combine to form chylomicron with cholate, are absorbed
TAG is recombined rapidly into intestinal epithelial cell, and by 2- acylglycerols approach, into lower blood circulation system.By
In the speed that 2-MAG approach synthesizes TAG quickly, therefore after body intake TAG, postprandial lipid metabolism concentration is caused to raise, and at 4 hours
Left and right reaches peak value.After 1,3-DAG enters small intestine, glycerine and NEFA are decomposed into the presence of pancreatic lipase, both enter small
After enterocyte, TAG is recombined by phosphatidic acid approach.Due to phosphatidic acid approach synthesis TAG speed it is very slow, it is all not
TAG is easily recombined in vivo, therefore can just improve blood lipids metabolism.
1,2-DAG and 2,3-DAG, although being all diglyceride, after both single hydrolysis in vivo, it can all generate 2-MAG, institute
TAG can also be recombined rapidly in body by 2-MAG approach by having, therefore not possess 1, health efficacy as 3-DAG.
Because contents of 1, the 3-DAG in natural oil is few, it is therefore desirable to by changing to existing oil resource
Make, to obtain the 1,3-DAG of high-purity.Existing 1,3-DAG preparation methods mainly have following several：
1st, direct esterification
Direct esterification is with aliphatic acid or fatty acid donors, is raw material with glycerine or glyceroyl donor, necessarily to compare
After example mixing, under conditions of micro- aqueous phase or organic solvent, 1,3- specific fats Enzyme catalyzed synthesis 1,3-DAG is utilized.Its is anti-
Answer formula as follows：
There are many patents and paper to report at present and the 1,3-DAG of high-purity is can obtain using this method, such as country's hair
A kind of bright patent " preparation method of diglyceride（200410015348.4）, a kind of national inventing patent " preparation of diglyceride
Method（20131025590.9）Deng.Direct esterification has that reaction is simple, can a step complete, product purity it is high, be easy to purifying point
It is short from, reaction time, the advantages that utilization rate of relevant enzyme and instrument is high.
But direct esterification is difficult to mass produce, some following problem is primarily present：First, required raw material is
Free fatty acids or glycerine price are very high；Second, the reaction is carried out typically in organic solvent environment, organic solvent residual be present
The problem of.
, glycerine solution
From the point of view of existing data, glycerol rhizolomy seemingly produces the most economical methods of 1,3-DAG, and current industrial production
Most important method.Glycerine solution reaction equation is as follows：
Also there are Patents to report glycerine solution at present, such as the national inventing patent " preparation method of diglyceride
（201310199809.7）" and national inventing patent " intracellular lipase producing strains and its application machine screening technique and application method
（201310183968.8）" etc..
Glycerine solution equally has necessarily restricted, is mainly reflected in the following aspects：First, reaction substrate glycerine
Price it is relatively higher；It could react and finish second, the reaction speed is slow, generally requires 10h or longer time；Third, product
In 1,3-DAG purity it is not high, purge process is complicated；Fourth, 1,3-DAG preparation rate is not high, the triglycerides of 1 molecule and 1 molecule
Glycerine, it can be only generated the 1,3-DAG of 1 molecule.
, direct hydrolysis method
Refer to using TAG as raw material, in the presence of with 2 acyl hydrolase tendentiousness lipase, direct part hydrolysis life
Into 1,3-DAG and NEFA.Antarctic candidia lipase A（CAL-A,Candida Antarcticalipases-A）It is all
To 2 acyl hydrolase tendentiousness highest enzymes of triglycerides in lipase；It can be used for preparing direct hydrolysis TAG preparations 1,3-
DAG.The purity of 1 prepared, 3-DAG is about 42%；Preparation time is about 72h；Purity still needs to improve, and the time still needs to shorten.
The content of the invention
It is an object of the invention to provide a kind of TAG in antarctic candidia lipase A（CAL-A）The lower direct hydrolysis of effect
Prepare high-purity 1,3-DAG method.
Technical scheme
A kind of method that triglycerides enzymolysis prepares 1,3-DAG, including it is fatty to TAG, water and antarctic candida
The step of ethanol or/and metal ion formation hydrolyzation system are added in enzyme A；
The mol ratio of TAG and water is 1:2-10；
Antarctic candidia lipase A dosage is the 0.5-5% of TAG mass；
The addition of ethanol is the 4-20% of TAG mass；
The content of metal ion is 5-50mmol/L in hydrolyzation system；
The metal ion is ferric ion or/and manganese ion.
The content of 1,3-DAG in reaction system is detected according to instant sampling, determines optimum reacting time, by reaction system from
The heart, then oil phase is used into molecular distillation, separate monoglyceride and NEFA, diglyceride and triglycerides（The separation, distillation side
Method is conventional method, and technological parameter used, those skilled in the art can determine according to the performance of material）.Wherein, diglyceride
1,3-DAG ratio is up to more than 80% in phase.
The present invention by experimental studies have found that, add ethanol in TAG hydrolyzation systems, improve the hydrophobic of TAG hydrolyzation systems
Property, it can be fully extended antarctic candidia lipase A spaces, improve and select tendentiousness to 2 of TAG, so as to effectively carry
1,3-DAG content in high hydrolysate（That is, 1,3-DAG purity is improved）.
Metal ion can influence the activity that antarctic candidia lipase A hydrolyzes to TAG；Different metal ions can be to the South Pole
Lipase from candida sp A activity produces different influence results；Some metal ions can improve antarctic candidia lipase A
Activity, and some metal ions can suppress antarctic candidia lipase A activity；Wherein, the metal ion that the present invention is added
Antarctic candidia lipase A activity can be significantly improved, so as to improve the hydrolysis rate of triglycerides.
And ethanol and metal ion are added simultaneously；Antarctic candidia lipase A can not only be improved simultaneously to 2 of TAG
Select tendentiousness and shorten the reaction time；Moreover, hydrolysis rate increase rate further increases；As can be seen here, ethanol serves
Metal ion is promoted to improve the effect of hydrolysis rate.
The present invention by experimental studies have found that, to hydrolyzation system, with 5000-10000rpm rotating speed high speed homogenate 10-
After 30min, relative to the hydrolyzation system of non-homogenate, hydrolysis rate improves 30%, and hydrolysis time shortens 50h, it is only necessary to 22h.
So the above method, preferably also includes：To hydrolyzation system with 5000-10000rpm rotating speed high-speed homogenization 10-30min's
Step.
Antarctic candidia lipase A optimum active temperature is 50-70 DEG C, optimum activity pH is 5.5-7；Institute
With in order to give full play to the catalytic activity that antarctic candidia lipase A hydrolyzes to TAG, to improve hydrolysis rate, TAG hydrolysis
The temperature of system is 50-70 DEG C, pH 5.5-7.0；For preferred temperature with 70 DEG C, adjustment pH is 7.0.
The above method, it is preferred that the mol ratio of TAG and water is 1:5.
The above method, it is preferred that antarctic candidia lipase A dosage is the 2% of TAG mass.
The above method, it is preferred that the addition of ethanol is the 10% of TAG weight.
The above method, the content of metal ion is 20mmol/L in hydrolyzation system.
In order to further reduce production cost, the TAG preferably uses vegetable oil or animal oil, it is furthermore preferred that using low
The soybean oil of value.
The water, can be running water or deionized water；Make to improve promotion of the metal ion to enzymatic activity
With raising hydrolysis rate, preferentially using deionized water.
If 1,3-DAG prepared by the method for the present invention as food or for food, with chelating agent remove metal from
Son.
Beneficial effect
First, reaction raw materials are cheap；Only with the peanut oil such as soybean oil, rapeseed oil, and lard etc., with other preparation sides
Glycerine, NEFA etc. are compared used by method, and cost substantially reduces；
Second, course of reaction is simple, cost is low；This reaction with the higher CAL-A of 2 tendentiousness, as lipase, to pass through
2 acyl groups of single step reaction hydrolyzing triglyceride, prepare 1,3-DAG, and process is simple, reduces reaction cost, improves effect
Rate；
Third, adding the ethanol of certain volume in course of reaction, change the hydrophobicity be clear and answer system, improve hydrolysis
2 selection tendentiousness of reaction, improve the purity of 1,3-DAG；
Fourth, adding a complexing metal ion in reaction system, lipase active is improved, shortens glycerine three
The ester hydrolysis time；
Fifth, being fully homogenized, water and oil phase are sufficiently mixed, and increase response area, improve reaction rate；
Sixth, being prepared for NEFA, price is high, and the added value of production technology greatly improved.
Embodiment
Embodiment 1
1. 1000g soybean oils are mixed with 100g water, 100g ethanol is added；With 5000rpm rotating speed high-speed homogenization
20min, prepare emulsion；
2. emulsion is heated into 70 DEG C and keeps constant temperature, the pH of emulsion is adjusted to 7.0 with hydrochloric acid or sodium hydroxide；
3. add antarctic candidia lipase A 20g；
4. after reacting 22h, reaction terminates；
5. reaction system is centrifuged, then oil phase used into molecular distillation, separate monoglyceride and NEFA, diglyceride and sweet
Oily three esters；1,3-DAG ratio is 83% wherein in diglyceride phase.
Embodiment 2
1. 1000g soybean oils are mixed with 100g water, 2.7g Iron(III) chloride hexahydrates and 1.98g tetrahydrate manganese chlorides are added；
With 5000rpm rotating speed high-speed homogenization 20min, emulsion is prepared；
2. emulsion is heated into 70 DEG C and keeps constant temperature, the pH of emulsion is adjusted to 7.0 with sodium hydroxide or hydrochloric acid；
3. add antarctic candidia lipase A 20g；
4. after reacting 18h, reaction terminates；
5. reaction system is centrifuged, then oil phase used into molecular distillation, separate monoglyceride and NEFA, diglyceride and sweet
Oily three esters；1,3-DAG ratio is 78% wherein in diglyceride phase.
Embodiment 3
1. 1000g soybean oils are mixed with 100g water, 100g ethanol, 2.7g Iron(III) chloride hexahydrates and 1.98g tetra- are added
Water manganese chloride；With 5000rpm rotating speed high-speed homogenization 20min, emulsion is prepared；
2. emulsion is heated into 70 DEG C and keeps constant temperature, the pH of emulsion is adjusted to 7.0 with sodium hydroxide or hydrochloric acid；
3. add antarctic candidia lipase A 20g；
4. after reacting 16h, reaction terminates；
5. reaction system is centrifuged, then oil phase used into molecular distillation, separate monoglyceride and NEFA, diglyceride and sweet
Oily three esters；1,3-DAG ratio is 85% wherein in diglyceride phase.
Embodiment 4
1. 1000g soybean oils are mixed with 100g water, 100g ethanol, 2.7g Iron(III) chloride hexahydrates and 1.98g tetra- are added
Water manganese chloride, mix, obtain mixed liquor；
2. mixed liquor is heated into 70 DEG C and keeps constant temperature, the pH of mixed liquor is adjusted to 7.0 with sodium hydroxide or hydrochloric acid；
3. add antarctic candidia lipase A 20g；
4. after reacting 28 h, reaction terminates；
5. reaction system is centrifuged, then oil phase used into molecular distillation, separate monoglyceride and NEFA, diglyceride and sweet
Oily three esters；1,3-DAG ratio is 83% wherein in diglyceride phase
Embodiment 5
1. 1000g soybean oils are mixed with 100g water, 100g ethanol and 13.5g Iron(III) chloride hexahydrates are added（System is about
For 1.2L, now iron concentration is 50mmol/L）；With 5000rpm rotating speed high-speed homogenization 20min, emulsion is prepared；
2. emulsion is heated into 70 DEG C and keeps constant temperature, the pH of emulsion is adjusted to 7.0 with sodium hydroxide or hydrochloric acid；
3. add antarctic candidia lipase A 20g；
4. after reacting 18h, reaction terminates；
5. reaction system is centrifuged, then oil phase used into molecular distillation, separate monoglyceride and NEFA, diglyceride and sweet
Oily three esters；1,3-DAG ratio is 83% wherein in diglyceride phase.
Comparative example 1
1. 1000g soybean oils are mixed with 100g water, 20min is mixed with 500rpm rotating speed, prepares emulsion；
2. emulsion is heated into 70 DEG C and keeps constant temperature, the pH of emulsion is adjusted to 7.0 with sodium hydroxide or hydrochloric acid；
3. add antarctic candidia lipase A 20g；
4. after reacting 57 h, reaction terminates；
5. reaction system is centrifuged, then oil phase used into molecular distillation, separate monoglyceride and NEFA, diglyceride and sweet
Oily three esters；1,3-DAG ratio is 45% wherein in diglyceride phase.
Embodiment 6-10
Operating procedure is as shown in the table with embodiment 3, specific process parameter, reaction time, product purity：
| Profit mol ratio | Lipase addition | Ethanol addition | Metal ion addition | Homogenate measure | pH | Temperature | Reaction time | 1,3-DAG purity | |
| Embodiment 5 | 1：5 | 2% | 10% | 50mmol/L | 5000rpm/20min | 7.0 | 70℃ | 18 | 83% |
| Embodiment 6 | 1：5 | 0.5% | 10% | 20mmol/L | 5000rpm/20min | 7.0 | 70℃ | 48 | 79% |
| Embodiment 7 | 1：5 | 0.5% | 10% | 20mmol/L | 10000rpm/20min | 7.0 | 70℃ | 15 | 82% |
| Embodiment 8 | 1：5 | 2% | 4% | 20mmol/L | 5000rpm/20min | 7.0 | 70℃ | 25 | 74% |
| Embodiment 9 | 1：5 | 2% | 20% | 20mmol/L | 5000rpm/20min | 7.0 | 70℃ | 20 | 78% |
| Embodiment 10 | 1：5 | 2% | 10% | 5mmol/L | 5000rpm/20min | 7.0 | 70℃ | 39 | 79% |
| Embodiment 11 | 1：5 | 2% | 10% | 50mmol/L | 5000rpm/20min | 7.0 | 70℃ | 19 | 81% |
In addition, do not tested using manganese ion or ferric ion individually（Other conditions are the same as embodiment 3）, it is between seasonable
16-17h, product 1,3-DAG purity are 84-85%.
Claims (6)
1. a kind of method that triglycerides enzymolysis prepares 1,3-DAG, it is characterised in that including false to TAG, water and the South Pole
The step of ethanol or/and metal ion formation hydrolyzation system are added in silk Yeast-lipase A；
The mol ratio of TAG and water is 1:5；
Antarctic candidia lipase A dosage is the 0.5-5% of TAG mass；
The addition of ethanol is the 4-20% of TAG mass；
The content of metal ion is 5-50mmol/L in hydrolyzation system；
The metal ion is ferric ion or/and manganese ion；
The TAG uses soybean oil.
2. method according to claim 1, it is characterised in that also include：It is high with 5000-10000rpm rotating speed to hydrolyzation system
The step of speed homogenate 10-30min.
3. according to the method for claim 1 or 2, it is characterised in that the temperature of TAG hydrolyzation systems is 50-70 DEG C, pH 5.5-
7.0。
4. method according to claim 3, it is characterised in that antarctic candidia lipase A dosage is the 2% of TAG mass.
5. method according to claim 4, it is characterised in that the addition of ethanol is the 10% of TAG weight.
6. method according to claim 5, it is characterised in that the content of metal ion is 20mmol/L in hydrolyzation system.
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