CN112921060B - Method for synthesizing sucrose fatty acid ester by lipase catalysis in organic solvent - Google Patents
Method for synthesizing sucrose fatty acid ester by lipase catalysis in organic solvent Download PDFInfo
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- sucrose
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- amyl alcohol
- fatty acid
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- 239000005720 sucrose Substances 0.000 title claims abstract description 87
- 229930006000 Sucrose Natural products 0.000 title claims abstract description 86
- -1 sucrose fatty acid ester Chemical class 0.000 title claims abstract description 28
- 108090001060 Lipase Proteins 0.000 title claims abstract description 26
- 102000004882 Lipase Human genes 0.000 title claims abstract description 26
- 239000004367 Lipase Substances 0.000 title claims abstract description 26
- 235000019421 lipase Nutrition 0.000 title claims abstract description 26
- 235000014113 dietary fatty acids Nutrition 0.000 title claims abstract description 22
- 239000000194 fatty acid Substances 0.000 title claims abstract description 22
- 229930195729 fatty acid Natural products 0.000 title claims abstract description 22
- 238000000034 method Methods 0.000 title claims abstract description 17
- 239000003960 organic solvent Substances 0.000 title claims abstract description 15
- 230000002194 synthesizing effect Effects 0.000 title abstract description 8
- 238000006555 catalytic reaction Methods 0.000 title abstract description 6
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims abstract description 58
- 239000000126 substance Substances 0.000 claims abstract description 22
- GLVVKKSPKXTQRB-UHFFFAOYSA-N ethenyl dodecanoate Chemical compound CCCCCCCCCCCC(=O)OC=C GLVVKKSPKXTQRB-UHFFFAOYSA-N 0.000 claims abstract description 16
- 231100000053 low toxicity Toxicity 0.000 claims abstract description 9
- MSXVEPNJUHWQHW-UHFFFAOYSA-N 2-methylbutan-2-ol Chemical group CCC(C)(C)O MSXVEPNJUHWQHW-UHFFFAOYSA-N 0.000 claims description 79
- 238000006243 chemical reaction Methods 0.000 claims description 36
- 108010084311 Novozyme 435 Proteins 0.000 claims description 28
- BTURAGWYSMTVOW-UHFFFAOYSA-M sodium dodecanoate Chemical group [Na+].CCCCCCCCCCCC([O-])=O BTURAGWYSMTVOW-UHFFFAOYSA-M 0.000 claims description 20
- 229940082004 sodium laurate Drugs 0.000 claims description 20
- 239000002808 molecular sieve Substances 0.000 claims description 18
- URGAHOPLAPQHLN-UHFFFAOYSA-N sodium aluminosilicate Chemical compound [Na+].[Al+3].[O-][Si]([O-])=O.[O-][Si]([O-])=O URGAHOPLAPQHLN-UHFFFAOYSA-N 0.000 claims description 18
- 238000001035 drying Methods 0.000 claims description 14
- 239000000843 powder Substances 0.000 claims description 14
- AFSIMBWBBOJPJG-UHFFFAOYSA-N ethenyl octadecanoate Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC=C AFSIMBWBBOJPJG-UHFFFAOYSA-N 0.000 claims description 13
- 150000004665 fatty acids Chemical class 0.000 claims description 12
- 229920001567 vinyl ester resin Polymers 0.000 claims description 12
- DKGAVHZHDRPRBM-UHFFFAOYSA-N Tert-Butanol Chemical compound CC(C)(C)O DKGAVHZHDRPRBM-UHFFFAOYSA-N 0.000 claims description 11
- 230000015572 biosynthetic process Effects 0.000 claims description 7
- 238000003786 synthesis reaction Methods 0.000 claims description 7
- 239000003054 catalyst Substances 0.000 claims description 4
- 238000001914 filtration Methods 0.000 claims description 2
- 239000002245 particle Substances 0.000 claims description 2
- 238000005406 washing Methods 0.000 claims description 2
- 239000012429 reaction media Substances 0.000 abstract description 8
- 238000004519 manufacturing process Methods 0.000 abstract description 7
- 230000003197 catalytic effect Effects 0.000 abstract description 6
- GCSPRLPXTPMSTL-IBDNADADSA-N [(2s,3r,4s,5s,6r)-2-[(2s,3s,4s,5r)-3,4-dihydroxy-2,5-bis(hydroxymethyl)oxolan-2-yl]-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl] dodecanoate Chemical compound CCCCCCCCCCCC(=O)O[C@@]1([C@]2(CO)[C@H]([C@H](O)[C@@H](CO)O2)O)O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O GCSPRLPXTPMSTL-IBDNADADSA-N 0.000 abstract description 3
- 238000000746 purification Methods 0.000 abstract description 3
- 238000000926 separation method Methods 0.000 abstract description 3
- 235000019626 lipase activity Nutrition 0.000 abstract description 2
- 238000010170 biological method Methods 0.000 abstract 1
- 239000000758 substrate Substances 0.000 abstract 1
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 21
- 239000000243 solution Substances 0.000 description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 13
- 238000000227 grinding Methods 0.000 description 12
- 238000004128 high performance liquid chromatography Methods 0.000 description 12
- 239000000203 mixture Substances 0.000 description 10
- 239000000047 product Substances 0.000 description 9
- 150000003445 sucroses Chemical class 0.000 description 8
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 6
- 235000013305 food Nutrition 0.000 description 5
- 230000000694 effects Effects 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 2
- 239000004280 Sodium formate Substances 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 2
- POULHZVOKOAJMA-UHFFFAOYSA-N dodecanoic acid Chemical compound CCCCCCCCCCCC(O)=O POULHZVOKOAJMA-UHFFFAOYSA-N 0.000 description 2
- 238000006911 enzymatic reaction Methods 0.000 description 2
- 238000009776 industrial production Methods 0.000 description 2
- 239000003495 polar organic solvent Substances 0.000 description 2
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 description 2
- 239000001632 sodium acetate Substances 0.000 description 2
- 235000017281 sodium acetate Nutrition 0.000 description 2
- HLBBKKJFGFRGMU-UHFFFAOYSA-M sodium formate Chemical compound [Na+].[O-]C=O HLBBKKJFGFRGMU-UHFFFAOYSA-M 0.000 description 2
- 235000019254 sodium formate Nutrition 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 241000055915 Heterocoma lanuginosa Species 0.000 description 1
- 239000005639 Lauric acid Substances 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- JXASPPWQHFOWPL-UHFFFAOYSA-N Tamarixin Natural products C1=C(O)C(OC)=CC=C1C1=C(OC2C(C(O)C(O)C(CO)O2)O)C(=O)C2=C(O)C=C(O)C=C2O1 JXASPPWQHFOWPL-UHFFFAOYSA-N 0.000 description 1
- SZYSLWCAWVWFLT-UTGHZIEOSA-N [(2s,3s,4s,5r)-3,4-dihydroxy-5-(hydroxymethyl)-2-[(2r,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyoxolan-2-yl]methyl octadecanoate Chemical compound O([C@@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)[C@]1(COC(=O)CCCCCCCCCCCCCCCCC)O[C@H](CO)[C@@H](O)[C@@H]1O SZYSLWCAWVWFLT-UTGHZIEOSA-N 0.000 description 1
- 125000002252 acyl group Chemical group 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 150000005690 diesters Chemical class 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 235000003084 food emulsifier Nutrition 0.000 description 1
- 231100000086 high toxicity Toxicity 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000007794 irritation Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 231100001231 less toxic Toxicity 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000000693 micelle Substances 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 239000002736 nonionic surfactant Substances 0.000 description 1
- 231100000956 nontoxicity Toxicity 0.000 description 1
- 229910000027 potassium carbonate Inorganic materials 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000004809 thin layer chromatography Methods 0.000 description 1
- 150000005691 triesters Chemical class 0.000 description 1
Abstract
The invention discloses a method for catalyzing and synthesizing sucrose fatty acid ester by lipase in an organic solvent. The method is characterized in that low-toxicity organic solvent is used as a reaction medium, substrate sucrose and vinyl laurate are added, sucrose laurate monoester is synthesized under the catalysis of lipase, and alkaline substances are added to deprotonate the lipase, so that the lipase activity is greatly improved. The invention realizes the aim of obtaining higher sucrose ester yield in low-toxicity organic solvent by using biological method, and the catalytic product is almost all sucrose monoester, thus greatly simplifying the separation and purification of the product, and being a green, safe and low-cost production method.
Description
Field of the art
The invention relates to a method for synthesizing sucrose fatty acid ester by lipase catalysis in an organic solvent.
(II) background art
The sucrose fatty acid ester is a nonionic surfactant and has the advantages of no toxicity, good biocompatibility, no irritation, biodegradability and the like. By adjusting the chain length and substitution degree of the acyl donor, various functional characteristics of the sucrose ester, such as Critical Micelle Concentration (CMC), hydrophilic-lipophilic balance (HLB) and the like, can be adjusted in a larger range, so that the sucrose ester meets specific application conditions. Therefore, sucrose esters are widely used in the industries of foods, daily chemicals, pharmaceuticals and the like. Sucrose esters are, for example, recommended food emulsifiers for use by the national food and agricultural and world health organizations; the lauric acid sucrose monoester has strong inhibition effect on harmful bacteria such as staphylococcus aureus and the like.
The current industrial production method of sucrose ester is mainly chemical method. The chemical method uses strong polar organic solvents such as DMSO, DMF and the like as reaction media and uses potassium carbonate and the like as catalysts to synthesize sucrose ester. Although the yield of sucrose esters synthesized by chemical methods is high, it has the following drawbacks: (1) The chemical catalyst has poor regioselectivity, and the catalytic product has by-products such as diester, triester and the like besides monoester, so that the utilization rate of raw materials is greatly reduced, the separation and purification of main products are not facilitated, and the cost of downstream engineering is greatly increased; (2) The strong polar organic solvents such as DMSO and the like have high toxicity and high boiling point, and are difficult to remove by the existing separation technology, so that the application of the obtained sucrose ester in the fields of food, daily chemicals and the like is limited; (3) The chemical method has harsh reaction conditions, usually needs to react under conditions of high temperature, reduced pressure and the like, has high production cost and high requirements on production equipment, and can easily denature and color sucrose esters at high temperature, thereby reducing the yield.
In recent years, enzymatic synthesis of sucrose esters has attracted attention due to the great limitations of chemical methods, wherein lipase is the enzyme used for the synthesis of sugar esters at the earliest. The lipase has higher regioselectivity on the synthesis of the sucrose monoester, and the catalytic product is almost all 6-O-sucrose monoester. In addition, the reaction condition for synthesizing sucrose ester by an enzyme method is mild, and the production cost can be greatly reduced; moreover, the reaction medium for synthesizing sucrose ester by an enzymatic method does not contain or contains less toxic organic solvents such as DMSO and the like, thereby greatly reducing the difficulty and cost of downstream engineering of the product. However, there is no report on the industrial production of sucrose esters by an enzymatic method at present, one of the reasons for this is that the solubility of sucrose in medium-low polarity organic media is very low, thereby greatly limiting the improvement of sucrose ester yield. Therefore, few reports have been made on the use of a single low-toxicity organic system such as t-amyl alcohol as the reaction medium. For this reason, most researchers have used a mixed system of t-amyl alcohol/DMSO (V: V=4:1) as the reaction medium to promote the dissolution of sucrose, e.g., manuel Ferrer et al reported H.lanuginosa lipase/Celite to catalyze the synthesis of sucrose laurate in a mixed t-amyl alcohol/DMSO system, and sucrose monoester yields can reach 30g/L. However, a small portion of DMSO contained in the mixed system still increases the difficulty in isolation and purification of the product and limits the use of sucrose esters. On the other hand, whether lipase can maintain an active conformation in an organic solvent, perform its normal catalytic function, is still another key factor affecting sucrose ester yield. The ionisation state of the lipase largely determines its active conformation, however, little has been reported to date about the effect of the ionisation state of the lipase on the synthesis of sucrose esters in organic media.
(III) summary of the invention
In order to solve the problems, the invention adopts tertiary amyl alcohol or tertiary butyl alcohol as a reaction medium, synthesizes fatty acid sucrose ester by lipase catalysis, and initially explores the effect of adding alkaline substances on improving lipase activity, thereby realizing a green, safe and low-energy consumption sucrose ester production mode, improving the safety of sucrose ester, being applied to the fields of food and daily chemicals, simplifying downstream engineering of products and reducing the production cost of sucrose ester.
The technical scheme adopted by the invention is as follows:
A method for lipase-catalyzed synthesis of sucrose fatty acid esters in an organic solvent, the method comprising:
(1) Adding sucrose and fatty acid vinyl ester into a low-toxicity organic solvent, taking lipase as a catalyst, simultaneously adding alkaline substances and molecular sieves, and reacting for 18-24 hours at the temperature of 60-85 ℃; the low-toxicity organic solvent is tertiary amyl alcohol or tertiary butyl alcohol; the alkaline substance is one of the following: sodium laurate, triethylamine, sodium formate, sodium acetate, disodium hydrogen phosphate; the tertiary amyl alcohol or tertiary butyl alcohol is a low-toxicity and low-boiling point organic solvent, and can be used as a reaction medium to greatly improve the safety of the sucrose ester product; the added alkaline substance can regulate the ionization state of the lipase to enable the lipase to be in an active conformation, thereby improving the catalytic activity.
(2) And filtering the reaction solution, recovering lipase, washing the lipase with tertiary amyl alcohol, drying in vacuum, and recovering for later use, and obtaining the fatty acid sucrose ester in the tertiary amyl alcohol.
The sucrose is ground fine powder sucrose with small particle size, and the adding amount is 0.1-0.6 mmol/15mL, and most preferably 0.4mmol/15mL.
The fatty acid vinyl ester is lauric acid vinyl ester or stearic acid vinyl ester, and the adding amount is 1.7-3.4 mmol/15mL, and most preferably 2.25mmol/15mL.
The lipase is Novozym435 added in an amount of 75-125 mg/15mL, most preferably 100mg/15mL.
The addition amount of the alkaline substance is 30-90 mg/15mL (preferably 90mg/15 mL), and sodium laurate is most preferred.
Preferably, the molecular sieve isOr/>Molecular sieves.
The beneficial effects of the invention are mainly as follows: the invention adopts low-toxicity and low-boiling-point tertiary amyl alcohol or tertiary butyl alcohol as a reaction medium for synthesizing sucrose ester by lipase catalysis, so that the safety of the sucrose ester product is greatly improved, the invention can be applied to the fields of food and daily chemicals, and meanwhile, the downstream engineering of the product is simplified, and the production cost is reduced; on the other hand, the alkaline substance added in the invention can adjust the ionization state of the lipase to enable the lipase to be in an active conformation, thereby improving the catalytic activity, leading the yield of the sucrose ester to be close to a DMSO/tert-amyl alcohol mixed system, and being a novel method for synthesizing the sucrose ester with green safety, high efficiency and low cost.
(IV) description of the drawings
FIG. 1 is a graph showing the results of thin layer chromatography analysis of sucrose stearate.
FIG. 2 is a high performance liquid chromatogram of sucrose laurate in example 1 (t-amyl alcohol system).
(Fifth) detailed description of the invention
The present invention will be described in further detail with reference to the following specific examples, which are only for the purpose of illustrating the invention and are not to be construed as limiting the scope of the invention.
Example 1:
grinding sucrose into fine powder, and drying in vacuum oven at 60deg.C for 6 hr. Into two 50mL round bottom flasks were added sucrose 0.1mmol (0.034 g), 2.25mmol (585. Mu.L) vinyl laurate, and 15mL t-amyl alcohol or t-butanol, respectively, and stirred in a 60℃water bath for 1h. Then 0.1g Novozym435, 0.09g sodium laurate and 0.4g were added respectively Molecular sieve, in a water bath kettle at 60 ℃ for 24 hours. After the reaction was completed, the reaction solution was filtered, novozym435 was recovered, and then washed with t-amyl alcohol, and dried in vacuo for use. The concentration of sucrose monoester in the tertiary amyl alcohol system is 2.32g/L and the yield is 66.9 percent by high performance liquid chromatography; sucrose monoester concentration in the tert-butanol system was 2.04g/L with 58.8% yield.
It can be seen that the yield of the tert-amyl alcohol system is higher than that of the tert-butanol system, so that the subsequent experiment uses tert-amyl alcohol as solvent.
Example 2:
Grinding sucrose into fine powder, and drying in vacuum oven at 60deg.C for 6 hr. Into five 50mL round bottom flasks were added sucrose 0.08/0.1/0.4/0.6/0.8mmol, and 2.25mmol (585. Mu.L) vinyl laurate and 15mL t-amyl alcohol, respectively, and stirred in a 60℃water bath for 1h. Then, 0.1g of Novozym435, 0.09g of sodium laurate and 0.4g of the above-mentioned materials were added to the above-mentioned mixed solution Molecular sieve, in a water bath kettle at 60 ℃ for 24 hours. After the reaction was completed, the reaction solution was filtered, novozym435 was recovered, and then washed with t-amyl alcohol, and dried in vacuo for use. The concentration of sucrose monoester in the tertiary amyl alcohol system is 2.3/3.61/4.26/4.28/4.13g/L respectively by high performance liquid chromatography.
As can be seen, the sucrose monoester yield increases with increasing sucrose concentration, but when the sucrose concentration in the system is greater than 0.6mmol/15mL, the sucrose monoester yield decreases, so that the sucrose concentration in the system is preferably 0.1 to 0.6mmol/15 mL.
Example 3:
Grinding sucrose into fine powder, and drying in vacuum oven at 60deg.C for 6 hr. Into five 50mL round bottom flasks were added 1.51/1.70/2.25/3.40/3.61mmol of vinyl laurate, and 0.4mmol (0.137 g) of sucrose and 15mL of t-amyl alcohol, respectively, and stirred in a 60℃water bath for 1h. To the mixture were added 0.1g Novozym435, 0.09g sodium laurate and 0.4g Molecular sieve, in a 75 ℃ water bath kettle for reaction for 24 hours. After the reaction was completed, the reaction solution was filtered, novozym435 was recovered, and then washed with t-amyl alcohol, and dried in vacuo for use. The concentration of sucrose monoester in the tertiary amyl alcohol system is measured by high performance liquid chromatography to be 4.86/5.40/6.42/6.40/6.37g/L respectively.
As can be seen, the yield of sucrose monoesters increases with increasing vinyl laurate concentration, but when the vinyl laurate concentration in the system is greater than 3.4mmol/15mL, the yield of sucrose monoesters decreases, so that the vinyl laurate concentration in the system is preferably 1.7 to 3.4mmol/15 mL.
Example 4:
Grinding sucrose into fine powder, and drying in vacuum oven at 60deg.C for 6 hr. Into five 50mL round bottom flasks were added sucrose 0.08/0.1/0.4/0.6/0.8mmol, and 2.25mmol (585. Mu.L) of vinyl stearate and 15mL of t-amyl alcohol, respectively, and stirred in a 60℃water bath for 1h. To the mixture were added 0.1g Novozym435, 0.09g sodium laurate and 0.4g Molecular sieve, in a 75 ℃ water bath kettle for reaction for 24 hours. After the reaction was completed, the reaction solution was filtered, novozym435 was recovered, and then washed with t-amyl alcohol, and dried in vacuo for use. The concentration of sucrose monoester in the tertiary amyl alcohol system is 15.8/18.7/28.8/27.9/27.2g/L respectively by high performance liquid chromatography.
As can be seen, when the fatty acid vinyl ester is vinyl stearate, the concentration of sucrose in the system is preferably 0.1-0.6 mmol/15 mL.
Example 5:
grinding sucrose into fine powder, and drying in vacuum oven at 60deg.C for 6 hr. Into five 50mL round bottom flasks were added 1.51/1.70/2.25/3.40/3.61mmol of vinyl stearate, 0.4mmol of sucrose (0.137 g) and 15mL of t-amyl alcohol, respectively, and stirred in a 75℃water bath for 1h. To the mixture were added 0.1g Novozym435, 0.09g sodium laurate and 0.4g Molecular sieve, in a 75 ℃ water bath kettle for reaction for 24 hours. After the reaction was completed, the reaction solution was filtered, novozym435 was recovered, and then washed with t-amyl alcohol, and dried in vacuo for use. The concentration of sucrose monoester in the tertiary amyl alcohol system is 17.2/19.3/28.8/28.4/28.0g/L respectively by high performance liquid chromatography.
As can be seen, when the fatty acid vinyl ester is vinyl stearate, the vinyl stearate in the system is preferably 1.7-3.4 mmol/15 mL.
Example 6:
Grinding sucrose into fine powder, and drying in vacuum oven at 60deg.C for 6 hr. Into six 50mL round bottom flasks were added sodium carbonate/sodium formate/sodium acetate/sodium laurate/disodium hydrogen phosphate 0.09g and 50. Mu.L triethylamine, 0.4mmol sucrose (0.136 g), 2.25mmol (585. Mu.L) vinyl laurate and 15mL t-amyl alcohol, respectively, and stirred in a water bath at 60℃for 1h. To the mixture was added 0.1g Novozym435 and 0.4g Molecular sieve, in a 75 ℃ water bath kettle for reaction for 24 hours. After the reaction was completed, the reaction solution was filtered, novozym435 was recovered, and then washed with t-amyl alcohol, and dried in vacuo for use. The concentration of sucrose monoester in the tertiary amyl alcohol system is 2.04/5.83/6.08/6.37/5.44/5.96g/L respectively by high performance liquid chromatography.
It can be seen that when the fatty acid vinyl ester is vinyl laurate, the alkaline substance in the system is most preferably sodium laurate.
Example 7:
grinding sucrose into fine powder, and drying in vacuum oven at 60deg.C for 6 hr. To five 50mL round bottom flasks were added 0/15/30/90/100/120mg sodium laurate, 0.4mmol sucrose (0.136 g), 2.25mmol vinyl laurate (585. Mu.L) and 15mL t-amyl alcohol, respectively, and stirred in a 60℃water bath for 1h. To the mixture was added 0.1g Novozym435 and 0.4g Molecular sieve, in a water bath kettle at 60 ℃ for 24 hours. After the reaction was completed, the reaction solution was filtered, novozym435 was recovered, and then washed with t-amyl alcohol, and dried in vacuo for use. The concentration of sucrose monoester in the tertiary amyl alcohol system is 1.04/1.54/1.88/2.18/2.15/2.09g/L respectively by high performance liquid chromatography.
As can be seen, the amount of the alkaline substance sodium laurate added is preferably 30 to 90mg/15mL, and most preferably 90mg/15mL.
Example 8:
Grinding sucrose into fine powder, and drying in vacuum oven at 60deg.C for 6 hr. To five 50mL round bottom flasks were added 0/15/30/90/100/120mg sodium laurate, 0.4mmol sucrose (0.136 g), 2.25mmol vinyl stearate (585. Mu.L) and 15mL t-amyl alcohol, respectively, and stirred in a 60℃water bath for 1h. To the mixture was added 0.1g Novozym435 and 0.4g Molecular sieve, in a water bath kettle at 60 ℃ for 24 hours. After the reaction was completed, the reaction solution was filtered, novozym435 was recovered, and then washed with t-amyl alcohol, and dried in vacuo for use. The concentration of sucrose monoester in the tertiary amyl alcohol system is 2.03/3.88/4.26/4.65/4.53/4.50g/L respectively measured by high performance liquid chromatography.
Similarly, when the fatty acid vinyl ester is vinyl stearate, the amount of sodium laurate as an alkaline substance added in the system is preferably 30 to 90mg/15mL, and most preferably 90mg/15mL.
Example 9:
Grinding sucrose into fine powder, and drying in vacuum oven at 60deg.C for 6 hr. Into five 50mL round bottom flasks were added 0.4mmol sucrose (0.136 g), 2.25mmol vinyl laurate (585. Mu.L) and 15mL t-amyl alcohol, respectively, and stirred in a 60℃water bath for 1h. 50/75/100/125/150mg Novozym435, 90mg sodium laurate and 0.4g were added to the mixture Molecular sieve, in a 75 ℃ water bath kettle for reaction for 24 hours. After the reaction was completed, the reaction solution was filtered, novozym435 was recovered, and then washed with t-amyl alcohol, and dried in vacuo for use. The concentration of sucrose monoester in the tertiary amyl alcohol system is 2.06/4.55/6.42/6.45/6.50g/L respectively measured by high performance liquid chromatography.
As can be seen, when the fatty acid vinyl ester is lauric acid vinyl ester, the amount of Novozym435 added in the system is preferably 75-150 mg/15 mL.
Example 10:
Grinding sucrose into fine powder, and drying in vacuum oven at 60deg.C for 6 hr. Into five 50mL round bottom flasks were added 0.4mmol sucrose (0.136 g), 2.25mmol vinyl stearate (585. Mu.L) and 15mL t-amyl alcohol, respectively, and stirred in a 60℃water bath for 1h. 50/75/100/125/150mg Novozym435, 90mg sodium laurate and 0.4g were added to the mixture Molecular sieve, in a 75 ℃ water bath kettle for reaction for 24 hours. After the reaction was completed, the reaction solution was filtered, novozym435 was recovered, and then washed with t-amyl alcohol, and dried in vacuo for use. The concentration of sucrose monoester in the tertiary amyl alcohol system is 15.4/20.2/28.8/29.1/29.6g/L respectively by high performance liquid chromatography.
As can be seen, when the fatty acid vinyl ester is vinyl stearate, the amount of Novozym435 added in the system is preferably 75-150 mg/15 mL.
Example 11:
Grinding sucrose into fine powder, and drying in vacuum oven at 60deg.C for 6 hr. Into five 50mL round bottom flasks were added 0.4mmol sucrose (0.136 g), 2.25mmol vinyl laurate (585. Mu.L) and 15mL t-amyl alcohol, respectively, and stirred in a 60℃water bath for 1h. 100mg of Novozym435, 90mg of sodium laurate and 0.4g of sodium laurate were added to the mixture Molecular sieves were reacted in 70/75/80/85/90 ℃ water baths for 24h, respectively. After the reaction was completed, the reaction solution was filtered, novozym435 was recovered, and then washed with t-amyl alcohol, and dried in vacuo for use. The concentration of sucrose monoester in the tertiary amyl alcohol system is 5.68/6.42/7.44/7.46/7.32g/L respectively as measured by high performance liquid chromatography.
As can be seen, when the fatty acid vinyl ester is vinyl laurate, the reaction temperature of the system is preferably 85 ℃.
Example 12:
Grinding sucrose into fine powder, and drying in vacuum oven at 60deg.C for 6 hr. Into five 50mL round bottom flasks were added 0.4mmol sucrose (0.136 g), 2.25mmol vinyl stearate (585. Mu.L) and 15mL t-amyl alcohol, respectively, and stirred in a 60℃water bath for 1h. 100mg of Novozym435, 90mg of sodium laurate and 0.4g of sodium laurate were added to the mixture Molecular sieves were reacted in 70/75/80/85/90 ℃ water baths for 24h, respectively. After the reaction was completed, the reaction solution was filtered, novozym435 was recovered, and then washed with t-amyl alcohol, and dried in vacuo for use. The concentration of sucrose monoester in the tertiary amyl alcohol system is 24.5/28.8/30.1/30.5/25.4g/L respectively by high performance liquid chromatography.
As can be seen, when the fatty acid vinyl ester is vinyl stearate, the reaction temperature of the system is preferably 85 ℃.
Claims (1)
1. A method for lipase-catalyzed synthesis of sucrose fatty acid esters in an organic solvent, the method comprising:
(1) Adding sucrose and fatty acid vinyl ester into a low-toxicity organic solvent, taking lipase as a catalyst, simultaneously adding an alkaline substance and a molecular sieve, and reacting for 18-24 hours at the temperature of 60-85 ℃; the low-toxicity organic solvent is tertiary amyl alcohol or tertiary butyl alcohol; the alkaline substance is sodium laurate, and the addition amount is 30-90 mg/15 mL; the lipase is Novozym435, and the addition amount is 75-150 mg/15 mL; the sucrose is ground fine powder sucrose with small particle size, and the adding amount is 0.1-0.6 mmol/15 mL; the fatty acid vinyl ester is lauric acid vinyl ester or stearic acid vinyl ester, and the addition amount is 1.7-3.4 mmol/15 mL; the molecular sieve is a 3A or 4A molecular sieve;
(2) And filtering the reaction solution, recovering lipase, washing the lipase with tertiary amyl alcohol, drying in vacuum, and recovering for later use, and obtaining the fatty acid sucrose ester in the tertiary amyl alcohol.
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