CN102676607A - Method for enzymatic synthesis of feruloyl oligosaccharide in mixed solvent - Google Patents
Method for enzymatic synthesis of feruloyl oligosaccharide in mixed solvent Download PDFInfo
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- CN102676607A CN102676607A CN2012101769981A CN201210176998A CN102676607A CN 102676607 A CN102676607 A CN 102676607A CN 2012101769981 A CN2012101769981 A CN 2012101769981A CN 201210176998 A CN201210176998 A CN 201210176998A CN 102676607 A CN102676607 A CN 102676607A
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Abstract
The invention relates to a method for enzymatic synthesis of feruloyl oligosaccharide in a mixed solvent. The method includes: (1) adding an activated molecular sieve in an organic solvent, and performing dehydration processing for 24-96 hours to obtain the dehydrated organic solvent; (2) performing enzymatic transesterification reaction, adding ferulic acid vinyl ester and glucose in the dehydrated organic solvent according to molar ratio of 2:1-1:6, then adding enzyme and the activated molecular sieve, performing oscillatory reaction for 10-144 hours at the temperature of 30-70 DEG C, filtering after the reaction is finished, taking clear filter liquid to perform rotary evaporation so as to remove the organic solvent, and purifying through a silicagel column. The method is simple to operate and environment-friendly, catalyst enzyme can be reused, and a structure of an obtained product is good in unicity.
Description
Technical field
The invention belongs to the preparation field of ferulic acid glycolipid, the method for the synthetic ferulic acid glycolipid of enzymatic in particularly a kind of mixed solvent.
Background technology
Ferulic acid glycolipid has good antioxidant effect, and it can chelated metal ions, removes radical ion DPPH, and OH and H
2O
2, and the metering of its antioxidant property and ferulic acid glycolipid presents dose-dependence, and the FLA of specific ionization has higher antioxidation property.Ferulic acid glycolipid has the amphipathic performance, makes its range of application expand to fat-solublely from water-soluble, makes it may be used in the oxidation resistant makeup.
Ferulic acid glycolipid is through enzyme process at first, acid system, and cooking process extracts from finished fibers such as Testa Tritici, and the ferulic acid glycolipid that extracts is the FLA oligose mostly.And, relate to a selective binding of glucose hydroxyl about the chemical preparation method of FLA glucose ester, and bad control need be carried out the protection of hydroxyl and go protection, and complex steps and conditional request are harsh.In the enzymatic synthesis method about ferulic acid glycolipid, the FLA of utilization directly and glucose synthesize.Owing to FLA has suitable anti-two kinds of structures, structural instability is prone to take place the change along antistructure, still goes on foot FLA to the FLA vinyl acetate through one, will avoid the generation of this situation, has improved fat-soluble simultaneously.
Summary of the invention
Technical problem to be solved by this invention provides the method for the synthetic ferulic acid glycolipid of enzymatic in a kind of mixed solvent, and this method steps is simply environmentally friendly, and the catalyzer enzyme can reuse.
It is following that the present invention prepares the reaction of ferulic acid glycolipid:
The method of the synthetic ferulic acid glycolipid of enzymatic in a kind of mixed solvent of the present invention comprises:
(1) molecular sieve after the activation is joined in the organic solvent, carry out processed 24-96h, the organic solvent after obtaining dewatering;
(2) enzymatic transesterification reaction: FLA vinyl acetate and glucose joined the organic solvent after the above-mentioned dehydration in 2: 1 in molar ratio ~ 1: 6, add the molecular sieve after enzyme and the activation then, in 30 ~ 70 ℃ of concussion reaction 10 ~ 144h down; Reaction finishes after-filtration, gets clarifying filtrating rotary evaporation and goes out organic solvent, and purifying through silicagel column obtains pure FLA glucose ester.
Molecular sieve described in the step (1) is
molecular sieve.
The preparation method of the molecular sieve after the activation described in step (1) and (2) be with molecular sieve in 500 ℃ of activation 4 hours, vacuum-drying afterwards also is cooled to room temperature, and is subsequent use.
Organic solvent described in the step (1) is that volume ratio is the trimethyl carbinol of 1: 1 (v/v) and the mixed solvent of pyridine.
The consumption of molecular sieve is 200mg/mL in the step (1).
Enzyme described in the step (2) is the Sumizyme MP (200,000 of producing bacillus subtilis; Wuxi Xue Mei Science and Technology Ltd.), Lipase from Candida Cylindacea (5.18u/mg; Sigma-Aldrich), Lipase from Candida rugosa (1104u/mg; Sigma), Lipase from Rhizopus oryzae (55.7u/mg, sigma-Aldrich), lypase Novozym435 (7u/mg, the bright industry and trade far away in Guangzhou ltd), Lipase from Pseudomonas fluorescens (2.2u/mg; Sigma-Aldrich), Lipase from Aspergillus niger (187u/g; Fluka) a kind of in, wherein the ratio of the add-on of every kind of enzyme and organic solvent is 5mg ~ 40mg: 1mL, the mass ratio of the add-on of every kind of enzyme and FLA vinyl acetate is 1: 2-2: 1.
The add-on of the molecular sieve after the activation described in the step (2) and the ratio of organic solvent are 200mg: 1mL.
Concussion speed during reaction described in the step (2) is 220r/min.
Substrate FLA vinyl acetate is the laboratory chemosynthesis among the present invention; Compound method is: with FLA 5g, mercuric acetate 0.22g, vinyl acetate 68ml join in the 150ml reaction flask successively; Drip the 0.2ml vitriol oil, refluxed 3h in 80 ℃ of oil bath pans after stirring 30min.Reaction finishes to be cooled to room temperature and adds the 2g sodium-acetate and fully stir the back hold over night, crosses the leaching supernatant and revolves steaming, obtains yellow oil, and through the pure FLA vinyl acetate that obtains of column chromatography for separation, purity reaches 98%.
Reaction solvent is the pyridine and the trimethyl carbinol of AG, and solvent parameter is respectively: 0.65 and 1.31.Two kinds of substrates are to be dissolved in respectively in these two kinds of organic solvents, and having avoided direct misciblely has the substrate can not consoluet embarrassment, make substrate under the state of a CL through the synthetic ferulic acid glycolipid of enzyme catalysis.
Reaction substrate FLA vinyl acetate of the present invention is to utilize commercially available FLA synthetic, and purity is up to 98%; Glucose also is commercially available glucose; Various enzyme preparation enzyme activity units have nothing in common with each other; Reaction solvent is a pyridine, the trimethyl carbinol, or solvent-free system is directly done dissolvant of reaction system with liquid enzyme.
The method of the synthetic FLA glucose ester of enzymatic of the present invention mainly is to utilize the incompatible synthetic FLA glucose ester verivate with parents' characteristic of different enzymes and different group of solvents.
The present invention has used a kind of comparatively gentle enzymatic means; The antioxidant that has synthesized a kind of parents' of having characteristic; Protection deprotection process with respect to sugared hydroxyl numerous and diverse in the chemosynthesis; Experimental procedure is simply environmentally friendly, and the catalyzer enzyme can reuse, and synthetic product structure unicity is good.
Beneficial effect
(1) adopt enzymatic synthetic method, reaction conditions is gentle, with respect to chemical reaction, has avoided the reaction conditions of harsh danger;
(2) selectivity of enzymatic reaction is stronger, has avoided hydroxy group of glucose protection and deprotection steps, and reactions step is simple;
(3) the reaction solvent boiling point of reaction process application is all lower, is easy to separate;
(4) reaction is under a state that does not have a free-water, to carry out, and has prevented because the existence of water causes the generation of the hydrolytic action of enzyme;
(5) synthetic FLA glucose ester has improved the oxidation-resistance of FLA simultaneously, has obtained parents' the sugar compounds that contains, and has widened range of application.
Description of drawings
Fig. 1: the infrared spectrogram of ferulic acid glycolipid;
Fig. 2: the chemical structural formula of ferulic acid glycolipid.
Embodiment
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used to the present invention is described and be not used in the restriction scope of the present invention.Should be understood that in addition those skilled in the art can do various changes or modification to the present invention after the content of having read the present invention's instruction, these equivalent form of values fall within the application's appended claims institute restricted portion equally.
Synthesizing of different enzymatic FLA glucose esters, concrete steps are following:
(1) with after
molecule activation drying; Add in the reaction solvent for use; Carry out the drying of solvent; The organic solvent that obtains dewatering after four days, the consumption of molecular sieve are 200mg/ml.
(2) enzymatic transesterification reaction: get concussion dissolving back mixing in equal-volume organic solvent pyridine that 3.8g glucose and 1g FLA vinyl acetate be added to 100ml dehydration respectively and the trimethyl carbinol; The various enzymes that add 2g again; The Sumizyme MP of producing bacillus subtilis (200,000, Wuxi Xue Mei Science and Technology Ltd.); Lipase from Candida Cylindacea (5.18u/mg, sigma-Aldrich); Lipase from Candida rugosa (1104u/mg, sigma); Lipase from Rhizopus oryzae (55.7u/mg, sigma-Aldrich); Lypase Novozym 435 (7u/mg, the bright industry and trade far away in Guangzhou ltd); Lipasefrom Pseudomonas fluorescens (2.2u/mg, sigma-Aldrich); Lipase from Aspergillus niger (187u/g, fluka).Add molecular sieve 20g, under 50 ℃, 220r/min shakes 72h.Reaction finishes the back and removes molecular sieve and enzyme with filter paper filtering, and the filtrating that obtains is evaporated away organic solvent through revolving the steaming appearance, and purifying through silicagel column afterwards obtains FLA glucose ester, and purity is 90% ~ 95%.
Embodiment 2
The enzymatic building-up reactions of different organic solvents catalysis ferulic acid glycolipid, concrete steps are following:
(1) with after
molecular sieve activation 4 drying; Add in the reaction solvent for use; Carry out the drying of solvent; The organic solvent that obtains dewatering after four days, the consumption of molecular sieve are 200mg/ml.
(2) enzymatic transesterification reaction: get 3.8g ~ 58g glucose and be added to the different dehydrated organic solvent pyridine of 100ml respectively with the 1g FLA; The trimethyl carbinol; And in the isopyknic trimethyl carbinol of 100ml and the pyridine mixed solvent, add the lypase Novozyon 435 (7u/mg) of 2g again; Add molecular sieve 20g, under 50 ℃, 220r/min shakes 72h.Reaction finishes the back and removes molecular sieve and enzyme with filter paper filtering, and the filtrating that obtains is evaporated away organic solvent through revolving the steaming appearance, and purifying through silicagel column afterwards obtains FLA glucose ester, and purity is 90% ~ 95%.
Claims (8)
1. the method for the synthetic ferulic acid glycolipid of enzymatic in the mixed solvent comprises:
(1) molecular sieve after the activation is joined in the organic solvent, carry out processed 24-96h, the organic solvent after obtaining dewatering;
(2) FLA vinyl acetate and glucose joined the organic solvent after the above-mentioned dehydration in 2: 1 in molar ratio ~ 1: 6, add the molecular sieve after enzyme and the activation then, in 30 ~ 70 ℃ of concussion reaction 10 ~ 144h down; Reaction finishes after-filtration, gets clarifying filtrating rotary evaporation and goes out organic solvent, purifies through silicagel column, gets final product.
2. the method for the synthetic ferulic acid glycolipid of enzymatic in a kind of mixed solvent according to claim 1, it is characterized in that: the molecular sieve described in the step (1) is
molecular sieve.
3. the method for the synthetic ferulic acid glycolipid of enzymatic in a kind of mixed solvent according to claim 1; It is characterized in that: the preparation method of the molecular sieve after the activation described in step (1) and (2) is in 500 ℃ of activation 4 hours with molecular sieve; Vacuum-drying afterwards also is cooled to room temperature, and is subsequent use.
4. the method for the synthetic ferulic acid glycolipid of enzymatic in a kind of mixed solvent according to claim 1, it is characterized in that: the organic solvent described in the step (1) is the mixed solvent of the trimethyl carbinol and pyridine, wherein the volume ratio of the trimethyl carbinol and pyridine is 1: 1.
5. the method for the synthetic ferulic acid glycolipid of enzymatic in a kind of mixed solvent according to claim 1, it is characterized in that: the consumption of molecular sieve is 200mg/mL in the step (1).
6. the method for the synthetic ferulic acid glycolipid of enzymatic in a kind of mixed solvent according to claim 1; It is characterized in that: the enzyme described in the step (2) is a kind of in the Sumizyme MP, Lipase from Candida Cylindacea, Lipase from Candida rugosa, Lipase from Rhizopus oryzae, lypase Novozym 435, Lipase from Pseudomonas fluorescens, Lipase from Aspergillus niger of producing bacillus subtilis; Wherein the ratio of the add-on of every kind of enzyme and organic solvent is 5mg ~ 40mg: 1mL, and the mass ratio of the add-on of every kind of enzyme and FLA vinyl acetate is 1: 2-2: 1.
7. the method for the synthetic ferulic acid glycolipid of enzymatic in a kind of mixed solvent according to claim 1, it is characterized in that: the add-on of the molecular sieve after the activation described in the step (2) and the ratio of organic solvent are 200mg: 1mL.
8. the method for the synthetic ferulic acid glycolipid of enzymatic in a kind of mixed solvent according to claim 1 is characterized in that: the concussion speed during reaction described in the step (2) is 220r/min.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105038450A (en) * | 2015-07-06 | 2015-11-11 | 安徽三联泵业股份有限公司 | Inner wall coating for high-temperature and high-pressure hot water circulating pump |
| CN110938665A (en) * | 2018-09-21 | 2020-03-31 | 南京农业大学 | Production technology of arabinoxylan for improving quality of frozen dough |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101560527A (en) * | 2009-05-22 | 2009-10-21 | 东华大学 | Method for lipase-catalyzed synthesis of feruloylated acylglycerol in solvent-free system |
| CN101608193A (en) * | 2009-07-10 | 2009-12-23 | 东华大学 | The method of enzymatically synthesizing suntan oil ferulic acid tricaprylin by solvent-free system |
| CN101914595A (en) * | 2010-09-07 | 2010-12-15 | 河南工业大学 | Method for synthesizing ferulic acid glycolipid derivatives by enzymatic method |
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2012
- 2012-05-31 CN CN2012101769981A patent/CN102676607A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101560527A (en) * | 2009-05-22 | 2009-10-21 | 东华大学 | Method for lipase-catalyzed synthesis of feruloylated acylglycerol in solvent-free system |
| CN101608193A (en) * | 2009-07-10 | 2009-12-23 | 东华大学 | The method of enzymatically synthesizing suntan oil ferulic acid tricaprylin by solvent-free system |
| CN101914595A (en) * | 2010-09-07 | 2010-12-15 | 河南工业大学 | Method for synthesizing ferulic acid glycolipid derivatives by enzymatic method |
Non-Patent Citations (3)
| Title |
|---|
| YANG YU等: "Enzymatic Synthesis of Feruloylated Lipids: Comparison of the Efficiency of Vinyl Ferulate and Ethyl Ferulate as Substrates", 《JOURNAL OF THE AMERICAN OIL CHEMISTS SOCIETY》, vol. 87, no. 12, 31 December 2010 (2010-12-31) * |
| 周健 等: "非水相脂肪酶催化体系分子筛脱水机制的研究", 《食品与机械》, vol. 21, no. 2, 31 March 2005 (2005-03-31) * |
| 於洋 等: "非水相酶促不可逆转酯反应合成阿魏酸三油酸甘油酯", 《精细化工》, vol. 27, no. 5, 31 May 2010 (2010-05-31) * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105038450A (en) * | 2015-07-06 | 2015-11-11 | 安徽三联泵业股份有限公司 | Inner wall coating for high-temperature and high-pressure hot water circulating pump |
| CN110938665A (en) * | 2018-09-21 | 2020-03-31 | 南京农业大学 | Production technology of arabinoxylan for improving quality of frozen dough |
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Application publication date: 20120919 |