CN103320479A - Method for catalysis synthesis of L-octadecanol serinate by using immobilized lipase - Google Patents
Method for catalysis synthesis of L-octadecanol serinate by using immobilized lipase Download PDFInfo
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- CN103320479A CN103320479A CN2013101680328A CN201310168032A CN103320479A CN 103320479 A CN103320479 A CN 103320479A CN 2013101680328 A CN2013101680328 A CN 2013101680328A CN 201310168032 A CN201310168032 A CN 201310168032A CN 103320479 A CN103320479 A CN 103320479A
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- serine
- stearyl alcohol
- lipase
- alcohol ester
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Abstract
The present invention relates to a method for catalysis synthesis of L-octadecanol serinate by using immobilized lipase, and belongs to the technical field of biocatalysis. The method comprises that: in a phosphate buffer solution and n-hexane composite system, L-serine and octadecyl alcohol are subjected to an esterification reaction through lipase catalysis to synthesize the L-octadecanol serinate, wherein a reaction temperature is 30-50 DEG C. The preparation method has characteristics of mild reaction conditions, safety, single product, easy purification process, good selectivity and the like, and provides a new approach for L-octadecanol serinate preparation.
Description
Technical field
The invention belongs to the preparation field of Serine stearyl alcohol ester, particularly relate to the method for the synthetic Serine stearyl alcohol ester of a kind of enzyme-catalyzed change, belong to the biocatalysis technology field.
Background technology
Research finds that ceramide type compound has had since the potent active anticancer component; this series products is from numerous marine organisms such as sea anemone; coral; sponge; separate in the starfish etc. and obtain; also from some plants, extract such as the onion soybean etc.; ceramide type compound is acknowledged as the tool resisting pathogenic microbes at present; the functions such as antitumor and class Nerve Growth Factor Activity; in recent years; ceramide type compound is become again the moisturizer of a new generation by the exploitation of external macro-organism technology company; because its hydrophilic lipid that is tool; relatively lower similar with the Person's skin stratum corneum; can with stratum corneum in the water combination; thereby the moisture in the pinning skin; has the function of moistening and protecting skin; have result of study to show, can the check melanin brown class of this compounds also can prevent anaphylaxis dermatosis.After the nineties in last century, ceramide type compound is added in beginning gradually in the cosmetics of super quality that Japan produces, and with annual 15% rate of growth fast development to Europe and even American market, demand constantly increases.As in Europe, this compounds is basically annual to be developed rapidly with the speed about 25-30%, and up to the present, developed into the widespread use in the functional foodstuff by Cosmetic Market, become a large hot fields of European functional foodstuff development, by einnehmen function food, enter the blood circulation of human body at the small intestine position, can promote recovery and the cell regeneration of skin, delay senility.
Serine stearyl alcohol ester (1-Octadecanol Serinate), synthetic new ceramide compounds such as Ceramide HO3TM(Trihydroxypalmitamido dydrmyristyl ether, France Sederma S. A. exploitation), PMS (Palmitoyl myristyl serinate, CAS:15602-32-9, Korea S Pacific Ocean company develops) and Ceramide PC-104(1,3-bis-(N-(2-hydroxyethyl)-palmitoylamino)-2-hydroxypropane, Korea S Pacific Ocean company exploitation) key intermediate of analogue such as, by L-L-Serine and long chain alkane alcohol (stearyl alcohol, 1-Octadecanol; Stearyl alcohol) esterification and generating, following formula is the structural formula of Serine stearyl alcohol ester.
The present domestic Serine stearyl alcohol ester of also not producing, the production of this compounds mainly is in more American-European developed countries, production method is to utilize concentrated hydrochloric acid as catalyzer, high temperature reflux (85 ~ 95 ℃) 9-24 hour, the main drawback of this method is that productive rate is low, the later stage purification process is loaded down with trivial details, selectivity relatively poor (Serine easily lactonizes), contaminate environment, consume energy high, synthetic product colour is darker, product purity is low etc., lipase has stereospecificity and high-efficiency catalytic activity in present method, subsequent disposal is simple, product purity is high, is fit to suitability for industrialized production.
Summary of the invention
The method that the purpose of this invention is to provide the synthetic Serine octadecyl ester of a kind of water and organic phase compound system enzyme-catalyzed change, synthetic method of the present invention improves the selectivity of esterification, the productive rate of Serine octadecyl ester reaches more than 80%, and reaction conditions is gentle, the method reaction conditions is safe, gentle, purifying is easy, and step is few, is convenient to suitability for industrialized production.Reaction equation:
Technical scheme of the present invention: take 0.05M phosphoric acid salt and normal hexane as reaction medium, carry out esterification with lipase-catalyzed Serine and stearyl alcohol, synthetic Serine stearyl alcohol ester.
A kind of method of fixed lipase catalyzed synthetic Serine stearyl alcohol ester, carry out according to following step:
(1) carries out esterification with lipase-catalyzed Serine and stearyl alcohol in the double solvents system, the condition of enzymatic reaction is that substrate Serine concentration is 30 ~ 160g/L, the mol ratio of Serine and stearyl alcohol is 1:0.9-1:4, the mass ratio of Serine and lipase is, the enzymatic reaction temperature is 30 ~ 50 ℃, 18 ~ 70 hours reaction times;
(2) after reaction finishes, reclaim lipase by the method for filtering, reaction solution revolves steaming with the ethyl acetate extraction twice of equal volume with extraction liquid to 40 ~ 60 a ℃ vacuum, removes normal hexane and ethyl acetate, obtains Serine stearyl alcohol ester crude product;
(3) Serine stearyl alcohol ester crude product by the silica gel G column chromatography, obtains purity and is the Serine stearyl alcohol ester more than 93%.
In the step of the present invention (1) in the double solvents system for pH is 6.98 0.05M phosphate buffered saline buffer and the mixing solutions of hexane, volume ratio is 1:1 ~ 4:3.
The present invention has the reaction conditions gentleness, good reaction selectivity, and reaction product is single, and transformation efficiency is high, and product safety, subsequent purification are processed the plurality of advantages such as simple easy.Therefore, the present invention provides new approach for Biological preparation Serine stearyl alcohol ester.
Description of drawings
Fig. 1 is the HPLC figure of Serine stearyl alcohol ester;
Fig. 2 is the mass spectrum of Serine stearyl alcohol ester;
Fig. 3 is the infared spectrum of Serine stearyl alcohol ester;
The nuclear magnetic resonance map (1H NMR, 400MHz, CH3OH) of Fig. 4 Serine stearyl alcohol ester.
Embodiment
The present invention adopts HPLC to analyze the content of product Serine stearyl alcohol ester in the assaying reaction, and chromatographic condition is the OD-H chiral column, 30 ℃ of column temperatures, and moving phase is V (normal hexane): V (Virahol)=90:10, flow velocity is 1mL/min.Detector is light scattering detector (ELSD).
Lipase of the present invention is that Novozym 435(derives from
Candida antarctica), available from Novozymes Company.The below is described further technical scheme of the present invention with specific embodiment.
Embodiment 1
Serine stearyl alcohol ester synthesis: take by weighing the 3gL-Serine and add dissolving in the 10mL 0.05M phosphate buffered saline buffer (pH=6.98), be placed in the 50mL tool plug ground Erlenmeyer flask, and then adding 8mL normal hexane and 6.9 g stearyl alcohol (mol ratio 1:0.9), lipase 0.1g, place 30 ℃ of constant-temperature tables, 120rpm, reaction result follow the tracks of by thin-layer chromatography TLC and detect and sampling HPLC analysis.React after 24 hours, productive rate reaches more than 35%.Remove by filter lipase, with 10 ~ 15mL ethyl acetate extraction, ethyl acetate and normal hexane are removed in 40-50 ℃ of underpressure distillation, obtain white Serine stearyl alcohol ester crude product.
With 200-300 order silica gel wet method dress post, elutriant is normal hexane: ether: acetic acid=88:10:2, with a small amount of elutriant dissolving Serine stearyl alcohol ester crude product, the wet method loading, TLC follows the tracks of the wash-out process, the elutriant that contains single product that obtains is merged evaporate to dryness, obtain the monoesters crystal of white, obtain Serine stearyl alcohol ester sterling (purity is more than 93%).
Lipase of the present invention comprises that Novozym 435(derives from
Candida antarctica) (derive from
Rhizomucor miehei) all available from Novozymes Company.
The product that the present invention synthesizes is white powder, molecular weight 357, and fusing point 101.5-102.5 ℃, purity reaches the HPLC figure that 93%, Fig. 1 is Serine stearyl alcohol ester; Fig. 2 is the mass spectrum of Serine stearyl alcohol ester; Fig. 3 is the infared spectrum of Serine stearyl alcohol ester; The nuclear magnetic resonance map (1H NMR, 400MHz, CH3OH) of Fig. 4 Serine stearyl alcohol ester.LC-MS surveys
M/z358 [M+H]
+, IR vmax (KBr) is 3356,2916,2849,1747,1495,1467,1315,1248,1090,1026,721,602 cm
-1 1H NMR (400MHz, CH
3OH) δ 4.25 (2H, br, CH
2), 4.10 (H, br, CH), 3.92 (2H, br, CH
2), 1.70 (2H, br, CH
2), 1.28(30H), 0.89(3H, br, CH
3), according to above-mentioned data, determine that this compound is Serine stearyl alcohol ester (C
21H
43O
3N).
Embodiment 2
Serine stearyl alcohol ester synthesis: take by weighing respectively 3g left and right sides Serine and 30.8 g stearyl alcohol (mol ratio 1:4), add the different solvent (acetone, normal hexane, cyclohexane and tetrahydrofuran (THF)) of 20mL, place 4 50mL tool plug ground Erlenmeyer flasks, then add respectively lipase 0.2g, place 30 ℃ of constant-temperature tables, 120rpm, reaction result follow the tracks of by thin-layer chromatography TLC and detect and sampling HPLC analysis.React after 24 hours, productive rate reaches respectively 12%, 31%, 24% and 19%.Remove by filter lipase and Serine, 40-50 ℃ of underpressure distillation desolventizing obtains white Serine stearyl alcohol ester and stearyl alcohol mixture.Cross silicagel column by embodiment 1 method, obtain Serine stearyl alcohol ester sterling (purity is more than 93%).
Claims (3)
1. the method for a fixed lipase catalyzed synthetic Serine stearyl alcohol ester is characterized in that carrying out according to following step:
(1) carries out esterification with lipase-catalyzed Serine and stearyl alcohol in the double solvents system, the condition of enzymatic reaction is that substrate Serine concentration is 30 ~ 160g/L, the mol ratio of Serine and stearyl alcohol is 1:0.9-1:4, the mass ratio of Serine and lipase is, the enzymatic reaction temperature is 30 ~ 50 ℃, 18 ~ 70 hours reaction times;
(2) after reaction finishes, reclaim lipase by the method for filtering, reaction solution revolves steaming with the ethyl acetate extraction twice of equal volume with extraction liquid to 40 ~ 60 a ℃ vacuum, removes normal hexane and ethyl acetate, obtains Serine stearyl alcohol ester crude product;
(3) Serine stearyl alcohol ester crude product by the silica gel G column chromatography, obtains purity and is the Serine stearyl alcohol ester more than 93%.
2. the method for a kind of fixed lipase catalyzed synthetic Serine stearyl alcohol ester according to claim 1, it is characterized in that in the step (1) in the double solvents system that volume ratio is 1:1 ~ 4:3 for pH is 6.98 0.05M phosphate buffered saline buffer and the mixing solutions of hexane.
3. the method for a kind of fixed lipase catalyzed synthetic Serine stearyl alcohol ester according to claim 1 is characterized in that lipase of the present invention is Novozym 435.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1680281A (en) * | 2004-02-19 | 2005-10-12 | 戈尔德施米特股份公司 | Process for preparing amino acid esters and their acid addition salts |
| CN101429529A (en) * | 2008-12-11 | 2009-05-13 | 江南大学 | Ester zymoid method production process of azeotropy water elimination coupling simulated moving bed |
| CN102876741A (en) * | 2012-09-03 | 2013-01-16 | 常州大学 | Method for preparing myristyl serinate under lipase catalysis |
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2013
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1680281A (en) * | 2004-02-19 | 2005-10-12 | 戈尔德施米特股份公司 | Process for preparing amino acid esters and their acid addition salts |
| CN101429529A (en) * | 2008-12-11 | 2009-05-13 | 江南大学 | Ester zymoid method production process of azeotropy water elimination coupling simulated moving bed |
| CN102876741A (en) * | 2012-09-03 | 2013-01-16 | 常州大学 | Method for preparing myristyl serinate under lipase catalysis |
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Application publication date: 20130925 |