GB2617451A - Method for obtaining 1,3-diglyceride from high oleic sunflower oil - Google Patents
Method for obtaining 1,3-diglyceride from high oleic sunflower oil Download PDFInfo
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- GB2617451A GB2617451A GB2302197.5A GB202302197A GB2617451A GB 2617451 A GB2617451 A GB 2617451A GB 202302197 A GB202302197 A GB 202302197A GB 2617451 A GB2617451 A GB 2617451A
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- sunflower oil
- high oleic
- distillation
- oleic sunflower
- diglyceride
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- 235000019486 Sunflower oil Nutrition 0.000 title claims abstract description 74
- 239000002600 sunflower oil Substances 0.000 title claims abstract description 74
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid group Chemical group C(CCCCCCC\C=C/CCCCCCCC)(=O)O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 title claims abstract description 63
- 238000000034 method Methods 0.000 title claims abstract description 44
- 238000004821 distillation Methods 0.000 claims abstract description 71
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 40
- 238000000199 molecular distillation Methods 0.000 claims abstract description 28
- 101710098556 Lipase A Proteins 0.000 claims abstract description 25
- 101710099648 Lysosomal acid lipase/cholesteryl ester hydrolase Proteins 0.000 claims abstract description 25
- 102100026001 Lysosomal acid lipase/cholesteryl ester hydrolase Human genes 0.000 claims abstract description 25
- 241001661345 Moesziomyces antarcticus Species 0.000 claims abstract description 24
- 239000008367 deionised water Substances 0.000 claims abstract description 17
- 239000003921 oil Substances 0.000 claims abstract description 12
- 235000019198 oils Nutrition 0.000 claims abstract description 12
- 239000000047 product Substances 0.000 claims description 32
- 239000006228 supernatant Substances 0.000 claims description 29
- 238000001514 detection method Methods 0.000 claims description 25
- 229910021641 deionized water Inorganic materials 0.000 claims description 16
- 238000006243 chemical reaction Methods 0.000 claims description 13
- 238000005119 centrifugation Methods 0.000 abstract description 3
- 241000208818 Helianthus Species 0.000 abstract 1
- 238000007654 immersion Methods 0.000 abstract 1
- 238000002474 experimental method Methods 0.000 description 26
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 16
- 102000004190 Enzymes Human genes 0.000 description 11
- 108090000790 Enzymes Proteins 0.000 description 11
- 229940088598 enzyme Drugs 0.000 description 11
- 239000000243 solution Substances 0.000 description 10
- 239000012467 final product Substances 0.000 description 9
- 238000011282 treatment Methods 0.000 description 9
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 7
- 239000012528 membrane Substances 0.000 description 7
- 238000007792 addition Methods 0.000 description 6
- 238000002360 preparation method Methods 0.000 description 6
- 235000020238 sunflower seed Nutrition 0.000 description 5
- 230000007071 enzymatic hydrolysis Effects 0.000 description 4
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- 239000000839 emulsion Substances 0.000 description 3
- 235000011187 glycerol Nutrition 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 230000010355 oscillation Effects 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 108010059892 Cellulase Proteins 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- 102000004882 Lipase Human genes 0.000 description 2
- 108090001060 Lipase Proteins 0.000 description 2
- 239000004367 Lipase Substances 0.000 description 2
- 229940106157 cellulase Drugs 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 235000019421 lipase Nutrition 0.000 description 2
- 238000004811 liquid chromatography Methods 0.000 description 2
- 230000035484 reaction time Effects 0.000 description 2
- IKRZCYCTPYDXML-UHFFFAOYSA-N 2-hydroxypropane-1,2,3-tricarboxylic acid;hydrochloride Chemical compound Cl.OC(=O)CC(O)(C(O)=O)CC(O)=O IKRZCYCTPYDXML-UHFFFAOYSA-N 0.000 description 1
- 108091005508 Acid proteases Proteins 0.000 description 1
- 101710121765 Endo-1,4-beta-xylanase Proteins 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 108010059820 Polygalacturonase Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 235000015140 cultured milk Nutrition 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 229940079919 digestives enzyme preparation Drugs 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- ONKWXUPJLQMGFJ-UHFFFAOYSA-N ethyl acetate;hexane;propan-2-ol Chemical compound CC(C)O.CCCCCC.CCOC(C)=O ONKWXUPJLQMGFJ-UHFFFAOYSA-N 0.000 description 1
- 238000000105 evaporative light scattering detection Methods 0.000 description 1
- 108010093305 exopolygalacturonase Proteins 0.000 description 1
- 108010026195 glycanase Proteins 0.000 description 1
- 125000005456 glyceride group Chemical group 0.000 description 1
- -1 glycerol diols Chemical class 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000010298 pulverizing process Methods 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 239000002002 slurry Substances 0.000 description 1
- 238000003756 stirring 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
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11C—FATTY ACIDS FROM FATS, OILS OR WAXES; CANDLES; FATS, OILS OR FATTY ACIDS BY CHEMICAL MODIFICATION OF FATS, OILS, OR FATTY ACIDS OBTAINED THEREFROM
- C11C1/00—Preparation of fatty acids from fats, fatty oils, or waxes; Refining the fatty acids
- C11C1/02—Preparation of fatty acids from fats, fatty oils, or waxes; Refining the fatty acids from fats or fatty oils
- C11C1/04—Preparation of fatty acids from fats, fatty oils, or waxes; Refining the fatty acids from fats or fatty oils by hydrolysis
- C11C1/045—Preparation of fatty acids from fats, fatty oils, or waxes; Refining the fatty acids from fats or fatty oils by hydrolysis using enzymes or microorganisms, living or dead
-
- 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
- C12P7/6454—Glycerides by esterification
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11C—FATTY ACIDS FROM FATS, OILS OR WAXES; CANDLES; FATS, OILS OR FATTY ACIDS BY CHEMICAL MODIFICATION OF FATS, OILS, OR FATTY ACIDS OBTAINED THEREFROM
- C11C1/00—Preparation of fatty acids from fats, fatty oils, or waxes; Refining the fatty acids
- C11C1/08—Refining
- C11C1/10—Refining by distillation
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Zoology (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Biochemistry (AREA)
- Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
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Abstract
Method for obtaining 1,3-diglyceride from high oleic sunflower oil, wherein the high oleic sunflower oil is enzymolysed by Candida antarctica lipase A, and the 1,3-diglyceride product is obtained using molecular distillation; and the enzymolysis is one-step. Preferably the dosage of lipase A is 2500 – 3500 u/g. The method may involve adding C. antarctica lipase A to high oleic oil from Helianthus and subsequent addition of deionised water followed by immersion in a constant temperature water bath shaker for 8.5 hours to nine and a half hours at 65 - 75 °C (e.g. 70 degC) and a rotational speed of 250 r.min-1, wherein the amount of DI water accounts for 25-35 wt.% (e.g. 30 weight per cent) of the high oleic sunflower oil, followed by centrifugation of the product at 3000 r/min for 10 minutes. The heavy phase obtained from a primary distillation may be fed into a feed port for a secondary distillation.
Description
METHOD FOR OBTAINING 1,3-DIGLYCERIDE FROM HIGH
OLEIC SUNFLOWER OIL
IECHNICAL FIELD
[0001] The present application relates to the technical field of sunflower oil processing, and in particular relates to a method for obtaining 1,3-diglyceride from high oleic sunflower oil.
BACKGROUND
[0002] Regarding the method for enzymolysis of vegetable oil, the following patent documents have been disclosed: Leng Yuxian etc. disclose a kind of technique of extracting sunflower seed oil by enzymatic hydrolysis method in" the technology of extracting sunflower seed oil by enzymatic hydrolysis method ", concrete production process is: Screen the shelled sunflower seeds, clean and pulverize them, add citrate hydrochloric acid buffer solution after pulverization, stir to form a slurry and press a certain enzyme, add the enzyme preparation into it according to a certain enzyme: seed ratio, carry out enzymolysis, centrifuge after the enzymolysis, and obtain free oil, emulsion layer, hydrolyzate and sediment after centrifugation; take out the emulsion layer and carry out secondary centrifugation to separate the free oil in the emulsion layer; the selected enzyme preparations include protease, compound cellulase, pectinase, acid protease, glycanase and acid xylanase. Finally, compound cellulase has the best effect, the free rate reaches about 90%, and the extracted sunflower oil is light yellow in color and cool, and has a fragrant smell.
[0003] The unresolved problem of the above-mentioned method is: the solution in the above-mentioned document only discloses the extraction and preparation method of sunflower oil, and does not disclose the further enzymolysis of sunflower oil and its enzymatic products.
[0004] Zhang Xiuxiu disclosed a method for preparing diglyceride (DAG) from sunflower oil in the article "Preparation of Sunflower Oil Diglyceride and Its Characteristics in Fermented Milk Application". The specific preparation steps are as follows: weigh sunflower oil, glycerin, and lipase in a conical flask, shake at a constant temperature at 53°C, take the supernatant and centrifuge, collect the supernatant and heat to inactivate the residual enzymes to obtain sunflower Seed Oil DAG.
[0005] Finally, the optimal process conditions for the preparation of DAG from sunflower oil were determined as follows: the mass ratio of alcohol to oil was 1:10, the amount of enzyme added was 7.13%, and the reaction time was 12.35 h. At this time, the DAG content in the reaction product was 49.52%.
[0006] Though the solution in this document discloses the extraction preparation method of sunflower seed oil, there are still following defects: 1) The reaction time is too long; 2) The addition of glycerol to the raw materials makes it impossible to determine whether all the glycerol diols finally obtained come from sunflower oil.
[0007] Therefore, it is necessary to optimize the process of enzymatic hydrolysis of sunflower oil, and to invent a method that only uses single sunflower oil as a raw material, can extract diglycerides, and can improve the efficiency of enzymolysis.
SUMMARY OF THE APPLICATION
[0008] In order to solve the above problems, the present application provides a method for obtaining 1,3-diglyceride from high oleic sunflower oil.
100091 A method for obtaining 1,3-diglyceride from high oleic sunflower oil provided by the present application, using high oleic sunflower oil as raw material, specifically comprises the following steps: 100101 1)A method for obtaining 1,3-diglyceride from high oleic sunflower oil, using Candida antarctica lipase A to enzymolyze high oleic sunflower oil; [0011] 2) After the enzymolysis, the product containing 1,3-diglyceride is obtained by using molecular distillation; the enzyme used in the enzymatic hydrolysis is Candida antarctica lipase A. 100121 The enzymolysis is one-step enzymolysis.
[0013] By weight of the high oleic sunflower oil, the dosage of Candida antarctica lipase A is 2500-3500u/g.
[0014] During enzymolysis, weight high oleic sunflower oil, and add Candida antarctica lipase A and then add deionized water, put it in a constant temperature water bath shaker, and react at a rotational speed of 250r/min at 65-75°C for 8.5-9.5h, after the reaction, centrifuge the product at 3000r/min for 10min.
[0015] By weight of the high oleic sunflower oil, the amount of Candida antarctica lipase A is 2500-3500u/g, the amount of deionized water accounts for 25-35% of the weight of the high oleic sunflower oil.
[0016] By weight of the high oleic sunflower oil, the amount of Candida antarctica lipase A is 3000u/g; the amount of deionized water accounts for 30% of the weight of the high oleic sunflower oil.
[0017] Shaking conditions in a constant temperature water bath are as follows: react at a rotational speed of 250r/min at 70°C for 9h, after the reaction, centrifuge the product at 3000r/min for 10min.
[0018] During molecular distillation, pour a centrifuged enzymolysis product supernatant into a feed port of a molecular distillation instrument for a primary distillation, wherein after the distillation, take out the light and heavy phases and clean the instrument; pour the heavy phase obtained from the primary distillation into the feed port for a secondary distillation to obtain a product containing 1,3-diglyceride.
100191 During the primary distillation, a vacuum degree is 2.0*10-2mbr, and temperature of the primary distillation is 110°C, rotational speed of scraper is 220-320r/min, and flow rate is 2.7-3.2mL/min.
[0020] During the primary distillation, a vacuum degree is 2.0*10-2mbr, and temperature of the primary distillation is 100°C, rotational speed of scraper is 300r/min, and flow rate is 3mL/min.
[0021] The method for obtaining 1,3-diglyceride from high oleic sunflower oil comprises the following steps: [0022] 1) Enzymolysis: Add high oleic sunflower oil to 3000u/g Candida antarctica lipase A by weight of oil, then add 30% deionized water by weight of oil, and place in a constant temperature water bath shaker, reacted at 70°C at a rotational speed of 250r/min for 9h, after the reaction, centrifuge the product at 3000r/min for I Omin, and take the supernatant for detection; [0023] 2) Molecular distillation: Pour 50mL of centrifuged enzymolysis product supernatant into a feed port of the molecular distillation instrument, set vacuum degree to 2.0*10-2mbr, temperature of a primary distillation at 110°C, rotational speed of a scraper at 300r/min, and flow rate 3mL/min, take out the light and heavy phase after the distillation, and clean the instrument; pour the heavy phase obtained from the primary distillation into the feed port, adjust the distillation temperature to 150°C, and keep other conditions unchanged, take out the light and heavy phase after the distillation, and carry out detection. [0024] The beneficial effect of the present application is: [0025] I) This method adopts a one-step enzymolysis method to prepare I,3-DAG, the operation steps are simple, and it is suitable for large-scale industrial production; [0026] 2) The Candida antarctica lipase A used in this application is a lipase with a tendency to hydrolyze the sn-2 bond of triglyceride, which can significantly improve the enzymolysis efficiency and increase the content of I,3-DAG.
BRIEF DESCRIPTION OF THE DRAWINGS
[0027] Figure 1 is a photo of the product; [0028] Figure 2 is the impact of different enzymolysis time on the content of 1,3-DAG; [0029] Figure 3 is the impact of different oil-water ratios on the content of 1,3-DAG: [0030] Figure 4 is the impact of different enzymolysis temperatures on the content of 1,3-DAG; [0031] Figure 5 is the impact of different enzyme additions on the 1,3-DAG content; [0032] Figure 6 is the impact of different distillation temperatures on the 1,3-DAG content; [0033] Figure 7 shows the impact of different scraper speeds on the content of 1,3-DAG; [0034] Figure 8 shows the impact of different feed rates on the content of 1,3-DAG.
DETAILED DESCRIPTION
[0035] In order to enable those skilled in the art to better understand the present application, the present application will now be further described in conjunction with specific embodiments.
[0036] Embodiment 1A1-A5 [0037] Embodiment 1A1-A5 focuses on the impact of deion zed water with different oil-water ratios on the content of the final product 1,3-diglycer de. The specific solution is as follows: [0038] The method for obtaining 1,3-diglyceride from high oleic sunflower oil comprises the following steps: [0039] 1) Enzymolysis: Accurately weigh 5g of high oleic sunflower oil, add 3000u/g Candida antarctica lipase A by weight of high oleic sunflower oil, and then add deionized water with different oil-to-water ratios (see Table 1), place in a constant temperature water bath shaker, reacted at 70°C at a rotational speed of 250r/min for 9h, centrifuge the product at 3000r/min for I Omin after the reaction, and take the supernatant to detect the content of 1,3-DAG (1,3-diglyceride).
[0040] Table I Impact of different oil-water ratio (%) treatments on I,3-DAG content Embodiment Oil- I,3-D A G CONTENT Error Water ratio/% Experiment Experiment Experiment Average 1 2 3 value Al 10 24.65 23.89 24.15 24.23 0.18 A2 20 27.74 27.26 28.1 27.7 0.2 A3 30 30.59 31.78 31.89 31.42 0.34 A4 40 29.1 28.38 28.26 28.58 0.21 A5 50 24.01 23.77 24.25 24.01 0.11 100411 2) Molecular distillation: Pour 50mL of centrifuged enzymolysis product supernatant into a feed port of the molecular distillation instrument, set vacuum degree to 2.0*10'mbr, temperature of a primary distillation at 110°C, rotational speed of a scraper at 300r/min, and flow rate 3mL/min, take out the light and heavy phase after the distillation, and clean the instrument; pour the heavy phase obtained from the primary distillation into the feed port, adjust the distillation temperature to 150°C, and keep other conditions unchanged, take out the light and heavy phase after the distillation, and carry out detection. [0042] 3) Detection: Sample treatment: Centrifuge the sample at 3000r/min for I Omin, take 100pL supernatant and add 24004_, n-hexane, vortex for 1min and pass through a 0.45u filter membrane.
[0043] Detection conditions: [0044] Liquid Chromatography Evaporative Light Detector [0045] Chromatographic column: ChromSpHer 5 Lipids column, 250*4.6mm (part number 28313) [0046] Mobile phase A: n-hexane-isopropanol-ethyl acetate (820:40:140) [0047] Mobile phase B: n-hexane: isopropanol: acetonitrile (956:40:4) [0048] Flow rate: 0.8mL/min, column temperature: 30°C, injection volume I CML. Before the next injection, the HPLC system was kept in the initial mobile phase for 2 minutes to ensure the stability of the injection.
[0049] Atomizer pressure: 35psi; atomizer flow rate 6L/min, atomizer temperature 36°C; ELSD drift tube temperature: 40°C; pressure: 2.5105Pa. The elution procedure is as follows: [0050] Table 2 Liquid chromatography mobile phase and gradient elution conditions Time ( min) Mobile Phase (%) Mobile Phase B (%) Flow rate/ (mL/min) 0 100 0 0.8 8 100 0 0.8 0 100 0.8 18 100 0 0.8 100 0 0.8 [0051] A product containing 1,3-d glyceride is finally obtained.
[0052] Embodiment 1B1-B5 [0053] Embodiment 1111 -B5focuses on the impact of water bath oscillation at different temperatures on the content of the final product 1,3-diglyceride, the specific solution is as follows: [0054] The method for obtaining 1,3-diglyceride from high oleic sunflower oil comprises the following steps: 100551 1) Enzymolysis: Accurately weigh 7g of high oleic sunflower oil, add 3000u/g Candida antarctica lipase A by weight of high oleic sunflower oil, and then add deionized water accounting for 30% by weight of high oleic sunflower oil, place in a constant temperature water bath shaker, reacted at different temperatures (see Table 3) at a rotational speed of 250r/min for 9h, centrifuge the product at 3000r/min for 10min after the reaction, and take the supernatant to detect the content of 1,3-DAG (1,3-diglyceride). [0056] Table 3 Impacts of water bath shaking treatment at different temperatures (°C) on the content of 1,3-DAG Embodiment Temperature I,3-D A G CONTENT Error °C Experiment Experiment Experiment Average 1 2 3 value B1 60 26.25 25.57 26.21 26.01 0.18 B2 65 27.78 28.11 28.5 28.13 0.17 B3 70 30.58 32.12 31.62 31.44 0.37 B4 75 29.15 29.24 28.52 28.97 0.18 B5 80 24.66 23.21 23.07 23.65 0.42 [0057] 2) Molecular distillation: Pour 50mL of centrifuged enzymolysis product supernatant into a feed port of the molecular distillation instrument, set vacuum degree to 2.0*10-2mbr, temperature of a primary distillation at 110°C, rotational speed of a scraper at 300r/min, and flow rate 3mL/min, take out the light and heavy phase after the distillation, and clean the instrument; pour the heavy phase obtained from the primary distillation into the feed port, adjust the distillation temperature to 150°C, and keep other conditions unchanged, take out the light and heavy phase after the distillation, and carry out detection. [0058] 3) Detection: Sample treatment: Centrifuge the sample at 3000r/min for 10min, take 100uL supernatant and add 24001,11 n-hexane, vortex for lmin and pass through a 0.45u filter membrane.
[0059] Detection conditions are the same as in Embodiment A. [0060] Embodiment 1C1-05 [0061] Embodiment I CI -05 focuses on the impact of different enzyme addition amounts on the content of the final product 1,3-diglyceride. The specific solution is as follows: [0062] The method for obtaining I,3-diglyceride from high oleic sunflower oil comprises the following steps: [0063] 1) Enzymolysis experiment: Accurately weigh llg of high oleic sunflower oil, add Candida antarctica lipase A with different contents (see Table 4), and then add deionized water accounting for 30% by weight of high oleic sunflower oil, place in a constant temperature water bath shaker, reacted at 70°C at a rotational speed of 250r/min for 9h, centrifuge the product at 3000r/min for 10min after the reaction, and take the supernatant to detect the content of 1,3-DAG (1,3-diglyceride).
[0064] Table 4 Impacts of different enzyme addition amounts (u/g) on the content of 1,3-DAG Embodiment Enzyme I,3-D A G CONTENT Error addition amount (u/g) Experiment Experiment Experiment Average 1 2 3 ' value Cl 1000 21.76 22.61 21.89 22.09 0.22 C2 2000 25.76 26.36 26.13 26.08 0.14 C3 3000 31.03 31.94 30.76 31.24 0.29 C4 4000 32.17 32.36 32.88 32.47 0.17 C5 5000 33.26 34.88 33.97 34.04 0.38 [0065] 2) Molecular distillalon: Pour 50mL of centrifuged enzymolysis product supernatant into a feed port of the molecular distillation instrument, set vacuum degree to 2.0*10-2mbr, temperature of a primary distillation at 110°C, rotational speed of a scraper at 300r/min, and flow rate 3mL/min, take out the light and heavy phase after the distillation, and clean the instrument; pour the heavy phase obtained from the primary distillation into the feed port, adjust the distillation temperature to 150°C, and keep other conditions unchanged, take out the light and heavy phase after the distillation, and carry out detection. [0066] 3) Detection: Sample treatment: Centrifuge the sample at 3000r/min for 10min, take I 0Opt supernatant and add 2400pL n-hexane, vortex for I min and pass through a 0.45u filter membrane.
[0067] Detection conditions are the same as in Embodiment A. [0068] Embodiment ID I -D5 [0069] Embodiment 1D1-D5 focuses on the impact of different constant temperature oscillation times on the content of the final product 1,3-diglyceride. The specific solution is as follows: [0070] The method for obtaining 1,3-diglyceride from high oleic sunflower oil comprises the following steps: [0071] I) Enzymolysis experiment: Accurately weigh 9g of high oleic sunflower oil, add 3000u/g Candida antarctica lipase A by weight of high oleic sunflower oil, and then then add deionized water accounting for 30% by weight of high oleic sunflower oil, place in a constant temperature water bath shaker, reacted at 70°C at a rotational speed of 250r/min for different times (see Table 5 for specific times), centrifuge the product at 3000r/min for I Omin after the reaction, and take the supernatant to detect the content of I,3-DAG (1,3-diglyceride).
[00721 Table 5 Impacts of different constant temperature shaking times (h) on the content of 1,3-DAG Embodiment Time/h 1,3-DAG CONTENT Error Experiment Experiment Experiment Average 1 2 3 value DI 1 16.91 17.34 18.96 17.74 0.51 D2 2 22.59 23.51 21.65 22.58 0.44 D3 3 25.97 24.33 25.01 25.1 0.39 D4 4 27.28 25.36 26.93 26.52 0.48 D5 5 29.87 27.24 28.08 28.4 0.63 D6 6 29.35 29.25 28.48 29.03 0.22 D7 7 30.15 30.66 30.81 30.54 0.16 D8 8 30.37 31.95 31.13 31.15 0.37 D9 9 31.32 32.06 31.67 31.68 0.17 DIO 10 31.79 32.16 32.00 31.98 0.09 Dll 11 31.33 31.90 32.22 31.82 0.21 D12 12 32.67 31.99 32.43 32.36 0.16 [0073] 2) Molecular distillation: Pour 50mL of centrifuged enzymolysis product supernatant into a feed port of the molecular distillation instrument, set vacuum degree to 2.0*10-2mbr, temperature of a primary distillation at I I 0°C, rotational speed of a scraper at 300r/min, and flow rate 3mL/min, take out the light and heavy phase after the distillation, and clean the instrument; pour the heavy phase obtained from the primary distillation into the feed port, adjust the distillation temperature to 150°C, and keep other conditions unchanged, take out the light and heavy phase after the distillation, and carry out detection. 100741 3) Detection: Sample treatment: Centrifuge the sample at 3000r/min for 10min, take 1004 supernatant and add 2400pL n-hexane, vortex for lmin and pass through a 0.45u filter membrane.
100751 Detection conditions are the same as in Embodiment A. [0076] Embodiment 1E1-E5 100771 Embodiment 1E1-E5 focuses on the impact of different secondary distillation temperatures on the content of the final product 1,3-diglyceride. The specific solution is as follows: [0078] The method for obtaining I,3-diglyceride from high oleic sunflower oil comprises the following steps: [0079] I) Enzymolysis experiment: Accurately weigh I Og of high oleic sunflower oil, add 3000u/g Candida antarctica lipase A by weight of high oleic sunflower oil, and then add deionized water accounting for 30% by weight of high oleic sunflower oil, place in a constant temperature water bath shaker, reacted at 70°C at a rotational speed of 250r/min for 9h, centrifuge the product at 3000r/min for I Omin after the reaction, and take the supernatant to detect the content of I,3-DAG (1,3-diglyceride).
[0080] 2) Molecular distillation: Pour 50mL of centrifuged enzymolysis product supernatant into a feed port of the molecular distillation instrument, set vacuum degree to 2.0*10-2mbr, temperature of a primary distillation at 110°C, rotational speed of a scraper at 300r/min, and flow rate 3mL/min, take out the light and heavy phase after the distillation, and clean the instrument; pour the heavy phase obtained from the primary distillation into the feed port, adjust different distillation temperatures (see Table 6), and keep other conditions unchangedtake out the light and heavy phase after the distillation, and carry out detection. [0081] Table 6 Impacts of different secondary distillation temperatures (°C) on the content of 1,3-DAG Embodiment Secondary distillation temperature °C 1,3-DAG CON1ENT Error Experiment Experiment Experiment Average 1 2 3 value El 130 34.72 35.54 35.68 35.31 0.24 E2 140 37.5 36.29 37.51 37.1 0.33 E3 150 39.81 40.35 40.18 40.11 0.13 E4 160 39.03 38.77 38.64 38.81 0.09 E5 170 38.42 37.68 38.12 38.08 0.18 100821 3) Detection: Sample treatment: Centrifuge the sample at 3000r/min for 10min, take 1004 supernatant and add 24004 n-hexane, vortex for lmin and pass through a 0.45u filter membrane.
[0083] Detection conditions are the same as in Embodiment A. [0084] Embodiment 1F1-F5 [0085] Embodiment 1F1-F5 focuses on the impacts of different flow rates on the content of the final product 1,3-diglyceride in the molecular distillation process. The specific solution is as follows: [0086] The method for obtaining 1,3-diglyceride from high oleic sunflower oil comprises the following steps: [0087] 1) Enzymolysis experiment: Accurately weigh lOg of high oleic sunflower oil, add 3000u/g Candida antarctica lipase A by weight of high oleic sunflower oil, and then add deionized water accounting for 30% by weight of high oleic sunflower oil, place in a constant temperature water bath shaker, reacted at 70°C at a rotational speed of 250r/min for 9h, centrifuge the product at 3000r/min for 10min after the reaction, and take the supernatant to detect the content of I,3-DAG (1,3-diglyceride).
100881 2) Molecular distillation: Pour 50mL of centrifuged enzymolys s product supernatant into a feed port of the molecular distillation instrument, set vacuum degree to 2.0*10-2mbr, temperature of a primary distillation at 110°C, rotational speed of a scraper at 300r/min, and set different flow rates (see Table 7), take out the light and heavy phase after the distillation, and clean the instrument; pour the heavy phase obtained from the primary distillation into the feed port, adjust the distillation temperature to 150°C, and keep other conditions unchanged, take out the light and heavy phase after the distillation, and carry out detection.
[0089] Table 7 impacts of different flow rates (mL/min) on the content of 1,3-DAG Embodiment Flow rates I,3-D A G CONTENT Error (mL/min) Experiment Experiment Experiment Average 1 2 3 value Fl 2 37.07 36.89 36.55 36.84 0.12 F2 2.5 38.05 38.44 38.33 38.27 0.09 F3 3 39.77 40.31 39.72 39.93 0.15 F4 3.5 39.04 38.59 38.63 38.75 0.12 F5 4 37.21 38.26 37.59 37.69 0.25 [0090] 3) Detection: Sample treatment: Centrifuge the sample at 3000r/min for 10min, take 1004 supernatant and add 2400pL n-hexane, vortex for lmin and pass through a 0.45u filter membrane.
100911 Detection conditions are the same as in Embodiment A. [0092] Embodiment 1G1-G5 [0093] Embodiment 1G1-G5 focuses on the impacts of different rotational speeds of the scraper on the content of the final product 1,3-diglyceride in the molecular distillation process. The specific solution is as follows: [0094] The method for obtaining 1,3-diglyceride from high oleic sunflower oil comprises the following steps: [0095] 1) Enzymolysis experiment: Accurately weigh lOg of high oleic sunflower oil, add 3000u/g Candida antarctica lipase A by weight of high oleic sunflower oil, and then add deionized water accounting for 30% by weight of high oleic sunflower oil, place in a constant temperature water bath shaker, reacted at 70°C at a rotational speed of 250r/min for 9h, centrifuge the product at 3000r/min for 10min after the reaction, and take the supernatant to detect the content of 1,3-DAG (1,3-diglyceride).
[0096] 2) Molecular distillation: Pour 50mL of centrifuged enzymolysis product supernatant into a feed port of the molecular distillation instrument, set vacuum degree to 2.0*10-2mbr, temperature of a primary distillation at I I 0°C, set different rotational speed of a scraper (see Table 8), and flow rate 3mL/min, take out the light and heavy phase after the distillation, and clean the instrument; pour the heavy phase obtained from the primary distillation into the feed port, adjust the distillation temperature to I50°C, and keep other conditions unchanged, take out the light and heavy phase after the distillation, and carry out detection.
[0097] Table 8 Impacts of different rotational speeds of a scraper (r/min) on the content of 1,3-DAG Embodiment Rotational 1,3-DAG CONTENT Error speed (r/min) Experiment Experiment Experiment Average 1 2 3 value G1 100 37.3 36.64 36.35 36.76 0.23 G2 200 38.14 38.34 37.97 38.15 0.09 G3 300 39.83 40.59 40.02 40.15 0.19 G4 350 38.41 38.77 38.44 38.54 0.09 GS 400 37.66 37.27 36.83 37.25 0.20 [0098] 3) Detection: Sample treatment: Centrifuge the sample at 3000r/min for 10min, take 100kiL supernatant and add 24001,11 n-hexane, vortex for lmin and pass through a 0.45u filter membrane.
[0099] Detection conditions are the same as in Embodiment A. [00100] Embodiments 2-6 [00101] Embodiments 2-6 comprehensively investigated the impact of different parameter ranges on the content of the final product 1,3-diglyceride in the process of enzymolysis and molecular distillation. The specific solution is as follows: [00102] Table 9 1,3-diglyceride content under the range of different parameters Water temperature (SC) Enzyme Oil- Oscillating distillation Secondary Flow rate Rotational 1,3-DAG speed of CONEENT ratio/% addition amount Oscillation time (h) temperature (mL/min) Scraper (u/g) (°C) (r/min) 2 30 70 3000 9 150 3 300 40.03 3 25 65 2500 8.5 145 2.7 220 32.59 4 27 67 2700 8.7 147 2.9 250 36.78 32 72 3200 9.2 152 3.1 280 38.97 6 35 75 3500 9.5 155 3.2 320 37.42 [00103 The method of Embodiments 2-6 is the same as that of Embodiments IA, except that some parameters are adjusted.
[00104] From the above Embodiments, it can be known that deionized water with different oil-water ratios, different Enzymolysis temperatures, different dosages of Candida antarctica lipase A, different temperatures of secondary distillation, different flow rates in the molecular distillation process, different rotational speed of the scraper will have a significant impact on the 1,3-diglyceride content of the final product. Within the parameters range of the method adopted in the present application, the 1,3-diglyceride content of the product under the parameters of the final embodiment 2 is the highest.
Claims (1)
- What is claimed s: A method for obtaining 1,3-diglyceride from high oleic sunflower oil, wherein the high oleic sunflower oil is enzymolyzed by Candida antarctica lipase A, and then a 1,3-diglyceride product is obtained by using molecular distillation; and the enzymolysis is one-step enzymolysis.A method for obtaining 1,3-diglyceride from high oleic sunflower oil according to claim 1, wherein by weight of the high oleic sunflower oil, the dosage of Candida antarctica lipase A is 2500-3500u/g A method for obtaining 1,3-diglyceride from high oleic sunflower oil according to claim 1, wherein during enzymolysis, weight high oleic sunflower oil, and add Candida antarctica lipase A and then add deionized water, put it in a constant temperature water bath shaker, and react at a rotational speed of 250r/min at 6575°C for 8.5-9.5h, after the reaction, centrifuge the product at 3000r/min for 10min; by weight of the high oleic sunflower oil, the amount of Candida antarctica lipase A is 2500-3500u/g; the amount of deionized water accounts for 25-35% of the weight of the high oleic sunflower oil.A method for obtaining 1,3-diglyceride from high oleic sunflower oil according to claim 3, wherein by weight of the high oleic sunflower oil, the amount of Candida antarctica lipase A is 3000u/g; the amount of deionized water accounts for 30% of the weight of the high oleic sunflower oil.A method for obtaining 1,3-diglyceride from high oleic sunflower oil according to claim 3, wherein react at a rotational speed of 250r/min at 70°C for 9h, after the reaction, centrifuge the product at 3000r/min for 10min.A method for obtaining 1,3-diglyceride from high oleic sunflower oil according to claim 1, wherein, during molecular distillation, pour a centrifuged enzymolysis product supernatant into a feed port of a molecular distillation instrument for a primary distillation, wherein after the distillation, take out the light and heavy phases and clean the instrument; pour the heavy phase obtained from the primary distillation into the feed port for a secondary distillation to obtain a product containing 1,3-diglyceride.A method for obtaining I,3-diglyceride from high oleic sunflower oil according to claim 6, wherein during the primary distillation, a vacuum degree is 2.0*10-2mbr, and temperature of the primary distillation is 110°C, rotational speed of scraper is 220-320r/min, and flow rate is 2.7-3.2mL/min.A method for obtaining 1,3-diglyceride from high oleic sunflower oil according to claim 6, wherein during the primary distillation, a vacuum degree is 2.0*10-2mbr, and temperature of the primary distillation is 100°C, rotational speed of scraper is 300r/min, and flow rate is 3mL/min.A method for obtaining 1,3-diglyceride from high oleic sunflower oil according to claim 6, wherein during the secondary distillation, a vacuum degree is 2.0*10-2mbr, and temperature of the secondary distillation is 145-155°C, rotational speed of scraper is 300r/min, and flow rate is 3mL/min.10. A method for obtaining 1,3-diglyceride from high oleic sunflower oil according to claim 1, comprises the following steps: 1) Enzymolysis: Add high oleic sunflower oil to 3000u/g Candida antarctica lipase A by weight of oil, then add 30% deionized water by weight of oil, and place in a constant temperature water bath shaker, reacted at 70°C at a rotational speed of 250r/min for 9h, after the reaction, centrifuge the product at 3000r/min for 10min, and take the supernatant for detection; 2) Molecular distillation: Pour 50mL of centrifuged enzymolysis product supernatant into a feed port of the molecular distillation instrument, set vacuum degree to 2.0*10-2mbr, temperature of a primary distillation at 110°C, rotational speed of a scraper at 300r/min, and flow rate 3mL/min, take out the light and heavy phase after the distillation, and clean the instrument; pour the heavy phase obtained from the primary distillation into the feed port, adjust the distillation temperature to 150°C, and keep other conditions unchanged, take out the light and heavy phase after the distillation, and carry out detection.
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|---|---|---|---|---|
| WO1995001450A1 (en) * | 1993-07-02 | 1995-01-12 | Unichema Chemie B.V. | Process for the esterification of carboxylic acids with tertiary alcohols |
| CN102517348A (en) * | 2011-12-14 | 2012-06-27 | 浙江大学 | Method for preparing 1,3-diglyceride from surface active magnetic nanoparticle immobilized lipase |
| CN102634547A (en) * | 2012-03-28 | 2012-08-15 | 江南大学 | Preparation method of symmetric triglyceride |
| CN104630296A (en) * | 2015-03-09 | 2015-05-20 | 杭州铎海科技有限公司 | Method for preparing 1,3-diglyceride through enzymolysis |
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| CN100362107C (en) * | 2006-01-24 | 2008-01-16 | 浙江大学 | Diglyceride edible oil production method |
| CN101407819B (en) * | 2008-11-17 | 2011-05-04 | 浙江大学 | Method for constructing ester bond enzyme engineering bacteria for tendency hydrolysis of TAG sn-2 and corresponding recombinant lipase |
| CN103352059B (en) * | 2013-06-25 | 2015-11-18 | 江南大学 | A kind of preparation method of triglyceride |
| CN104711298B (en) * | 2015-03-09 | 2017-12-01 | 杭州铎海科技有限公司 | A kind of method that triglycerides enzymolysis prepares 1,3 diglycerides |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO1995001450A1 (en) * | 1993-07-02 | 1995-01-12 | Unichema Chemie B.V. | Process for the esterification of carboxylic acids with tertiary alcohols |
| CN102517348A (en) * | 2011-12-14 | 2012-06-27 | 浙江大学 | Method for preparing 1,3-diglyceride from surface active magnetic nanoparticle immobilized lipase |
| CN102634547A (en) * | 2012-03-28 | 2012-08-15 | 江南大学 | Preparation method of symmetric triglyceride |
| CN104630296A (en) * | 2015-03-09 | 2015-05-20 | 杭州铎海科技有限公司 | Method for preparing 1,3-diglyceride through enzymolysis |
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