CN1260368C - Method for catalytic synthesis of vitamin A fatty acid ester using immobilized lipase - Google Patents
Method for catalytic synthesis of vitamin A fatty acid ester using immobilized lipase Download PDFInfo
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- CN1260368C CN1260368C CN 200310116834 CN200310116834A CN1260368C CN 1260368 C CN1260368 C CN 1260368C CN 200310116834 CN200310116834 CN 200310116834 CN 200310116834 A CN200310116834 A CN 200310116834A CN 1260368 C CN1260368 C CN 1260368C
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Abstract
The present invention relates to a method for catalytic synthesis of vitamin A fatty acid ester using immobilized lipase. Substrate vitamin A acetate and C10 to C18 fatty acid or fatty ester are mixed in the molar ratio of 1: 1 to 1: 7 as well as organic solvent and immobilized lipase in the amount of 0.2 to 5 times mass of the vitamin A acetate for reaction under 20 to 50 DEG C for 9 to 50 hours. The immobilized lipase is taken out. Reaction liquid is separated and crystallized to obtain vitamin A fatty ester. The organic solvent is C6 to C10 saturated alkane containing 0 to 1 % of water, and the immobilized lipase is yeast type lipase on the carrier of diatomite, resin or a fabric membrane. The immobilized lipase of the present invention has the characteristic of large catalysis area, high catalysis efficiency, little enzyme dosage, high stability and low cost and can be recycled and reused. Reaction solvent is non-toxic. An obtained product has the advantages of light color and luster and high quality.
Description
(1) technical field
The present invention relates to a kind of method, particularly in non-aqueous system, use the method for the lipase-catalyzed reaction synthesise vitamins of immobilized microorganism A fatty acid ester with and longer chain fatty acid or long chain fatty acid ester reaction synthesise vitamins A fatty acid ester.
(2) background technology
Vitamin A fatty acid ester is common chemical materials such as the many industries of daily-use chemical industry such as food, medicine, dietary supplements, makeup, fodder additives, the indispensable a kind of material of human homergy, it can keep and promote people's bulk-growth, growth, reproduction and cell membrane stability, to the tangible effect of being formed with of vision.And vitamin A fatty acid ester also helps the breeding of cell and distinguishes non-viable non-apoptotic cell, and can produce the combined with radical of transgenation with those as a kind of antioxidant.But for a long time, industrial vitamin A fatty acid ester all is to produce by chemical method, carries out a series of catalyzed reactions with catalyzer such as sodium methylate, Grignard reagent etc. under High Temperature High Pressure and finishes.Not only total conversion rate is not high, but also has many shortcomings that are difficult to overcome.As: catalyzer is toxic, high-temperature high-voltage reaction energy consumption height, side reaction are many, and the product color and luster is dark, separation and refining difficulty etc., and acid is serious to equipment corrosion, is unfavorable for producing.Along with development of biology, the particularly research of enzyme engineering is for the biological catalyst Acrawax provides new method.Compare with traditional chemical method, enzyme process is synthetic to have the reaction conditions gentleness, high specificity, and side reaction is few, characteristics such as product quality height.Particularly fatty enzymatic reaction in the nonaqueous phase, the range of application and the field of having widened enzyme, the focus of enzymology over becoming year.
Numerous research reports and general survey have showed that all the Enzyme Catalysis in Non-Aqueous Media effect is better than the enzyme in the water.Organic solvent can keep a lower water-activity, can reduce the thermokinetics and the kinetics obstacle of esterification and hydrolysis reaction.And organic substrates has higher solubleness and can improve speed of response in organic solvent.Therefore, the esterification in non-aqueous system can realize high conversion, simplifies separating technology.
Enzyme reactor is the container of resolvase, immobilized enzyme or immobilized cell catalyzed reaction.There is various ways in the immobilized lipase enzyme reactor, and traditional reactor comprises stirred-tank reactor, fixed bed type reactor (being called the filling bed type reactor again), fluidized bed reactor and membrane reactor.
" Inada, Yuji " at " modification lipase ", JP 60156395 (1984), disclose the partial amino-acid molecule that the derivative that is connected to the polyalkylene glycol of hydrophobic group with molecular end replaces lipase, and lipase is modified in preparation.Lipase after the modification not only can improve the solubility in organic solvent, and can keep the activity of enzyme.This method is to use 2 of the Pseudomonas fluorescens quasi-lipase of 10mg, 400mg, 4-two (methoxy poly (ethylene glycol))-6-S-triazine adds the borate buffer solution of 2ml 0.4M pH=10, stirred one hour at 37 ℃, then the lipase molecule that can have been modified with conventional method of purification.Wherein the lipase molecule has 52% amino acid to be modified by the derivative of triazine, and the hydrophobicity of the modification lipase that obtains after the lyophilize strengthens greatly.What this method was used is the lipase of modifying, modifying method complexity, cost height, and after reaction finished, catalyzer can't separate with product, and catalyzer is reused difficulty.
" Inada, Yuji " in " preparation of Davitin A ", JP 62248495 (1986), disclose the Pseudomonas fluorescens quasi-lipase of modifying with the O-methoxy poly (ethylene glycol), catalysis Davitin A synthetic method.The characteristics of this method be lipase after modifying with the O-methoxy poly (ethylene glycol) as catalyzer, in water saturated benzene solvent, temperature of reaction is 25～30 ℃, utilizes and C
5～C
20Lipid acid comprises valeric acid, sad, capric acid, lauric acid, tetradecanoic acid, palmitinic acid, oleic acid etc., and mol ratio is a synthesise vitamins A ester under 1: 10 the condition, and the Davitin A yield reaches 80～85%.This method Preparation of catalysts complexity, the cost height; And with an organic solvent benzene is made reaction medium, and benzene is toxic, and environment is had pollution.And, use lipid acid too much can increase the difficulty that later separation is purified, must cause the product cost height.
(3) summary of the invention
The purpose of this invention is to provide a kind of method of utilizing immobilized lipase synthesise vitamins A fatty acid ester.The present invention has adopted novel immobilized lipase, uses avirulent reaction solvent, and it is few to have an enzyme dosage, and immobilized lipase and reaction solvent all reclaim easily and reuse, and can reduce production costs, and help serialization production.
The method of the fixed lipase catalyzed synthesise vitamins A of a kind of usefulness of the present invention fatty acid ester is with substrate and C
10～C
18Lipid acid or fatty acid ester are in molar ratio 1: 1～1: 7 scope, mix with organic solvent and immobilized lipase, the consumption of immobilized lipase is 0.2～5 times of quality, under 20～50 ℃ condition, reacted 9～50 hours, take out immobilized lipase, reaction solution obtains vitamin A fatty acid ester through separation, crystallization; Organic solvent is that water content is 0～1% C
6～C
10Saturated alkane; Immobilized lipase is the yeast quasi-lipase that is fixed on the carrier, and carrier is diatomite, resin or textiles film; The yeast quasi-lipase, for example: separate ester lipase from candida sp, South Pole first silk yeast fat enzyme etc.
The preparation method of immobilized lipase of the present invention has following two kinds of methods: (A) with carrier and altogether fixing agent press mass volume ratio 1: 1～1: 3 scope mixing, room temperature is dried and is obtained activated carrier; Fixing agent is gelatin, Yelkin TTS, polyoxyethylene glycol, polysorbate altogether, the lipid acid sorb is smooth and the mixture of magnesium salts, its mass ratio is: gelatin: lecithin: polyoxyethylene glycol: polysorbate: the lipid acid sorb is smooth: magnesium salts=5: 1: 1: 2: 2: 1, magnesium salts was magnesium chloride or sal epsom; With the aqueous solution of yeast quasi-lipase by 1000～30000 units/gram activated carriers with the activated carrier immersion, mix, room temperature dries, make enzyme and live and be the immobilized lipase of 8000IU/g; (B) fermented liquid with the yeast quasi-lipase soaks carrier, mix, and room temperature is dried, and making enzyme alive is the immobilized lipase of 8000IU/g.
The C that the present invention is used
10～C
18Lipid acid or fatty acid ester are: capric acid, lauric acid, tetradecanoic acid, palmitinic acid, stearic acid, oleic acid, methyl caprate, Laurate methyl, Myristicin acid methylester, Uniphat A60, methyl stearate, Witconol 2301, ethyl decylate, Laurate ethyl, ethyl myristate, ethyl palmitate, Stearic ethyl stearate or ethyl oleate.
The organic solvent C that the present invention adopts
6～C
10Saturated alkane, for example: normal hexane, hexanaphthene, sherwood oil, normal heptane, octane, positive nonane, n-decane etc.
The used textiles film of the present invention is silk floss, nylon, silk, polyester or cellulosic woven.
The immobilized lipase of diatomite of the present invention or resin is used for stirred-tank reactor or fixed bed type reactor.
The immobilized lipase of textiles film of the present invention is used for stirred-tank reactor, and immobilized lipase is fixed on the cylindrical rack, and rack is fixed in the rotating shaft and does axially rotation as stirring rake in the lump.
The immobilized lipase of textiles film of the present invention is used for fixing hearth reactor, and immobilized lipase is fixed on the cylindrical rack, and cylindrical rack is fixed in the reactor.
Shallow by the product color that the present invention obtains, the quality height, cost is low.For the suitability for industrialized production of fatty acid ester provides a kind of novel method.
The used yeast quasi-lipase of the present invention is that commercially available or preparation certainly, commonly used, known yeast quasi-lipase all can be used, for example: separate ester candiyeast, South Pole first silk yeast etc.Used carrier diatomite and resin also are known common carrier, and the textiles film is the woven of selling on the market.The used fermented liquid of the present invention is a yeast quasi-lipase fermented liquid, with commercially available or all can of preparation certainly.For example: adopt and separate the preparation of ester gum silk yeast fermentation, fermentation condition is that 26 ℃ of temperature, pH are natural, mixing speed is that 300rpm, air flow are 1VVM, and substratum consists of: compositions such as organic nitrogen source, organic carbon source, inorganic salt.
The shaking table that reactor is used with this area stirred-tank reactor, fixed bed type reactor or laboratory commonly used.When the immobilized lipase of textiles film is used for stirred-tank reactor, do not need the former agitating vane that is provided with, need immobilized lipase is fixed on the rack, rack is fixed in the rotating shaft and does axially rotation as stirring rake in the lump.Rotating speed is identical with the rotating speed of original agitator of common stirred-tank reactor.The immobilized lipase of algae soil or resin can be directly used in existing stirred-tank reactor or fixed bed type reactor.When the immobilized lipase of textiles film is used for fixing hearth reactor, do not need to change the reactor primary structure, only need be immobilized lipase is fixed on the rack, rack is fixed in the reactor.
Enzyme is lived and is defined as: under 40 ℃ of conditions, the enzyme amount that hydrolysis sweet oil in the phosphate buffer solution of pH8.0, per minute discharge 1 μ mol lipid acid is an enzyme unit alive.The present invention for simplicity, sets the enzyme 8000IU/g of being alive when the preparation immobilized lipase, the enzyme that also can be set at other numerical value is lived.This is the known knowledge of those skilled in the art.
The present invention compares with traditional chemical synthesis, and method reaction conditions gentleness of the present invention, energy consumption are low, greatly reduce production cost.The present invention simultaneously uses biological catalyst, and atopic is strong, by product is few.Product has lighter color, characteristics that quality is high.
The present invention compares with similar biosynthetic means, and advantage is:
1, selected new immobilized lipase for use, because immobilized lipase can stop the congregation of enzyme molecule in organic solvent, thereby the active area that increases enzyme has reduced enzyme dosage.
2, by fixing agent and enzyme are fixing altogether altogether, can well improve the hydrophilic and hydrophobic property of carrier, strengthened contacting and diffusion effect of substrate and enzyme,, thereby improved catalytic activity and the thermodynamic stability of enzyme greatly for enzyme provides good and microenvironment substrate-function.Above factor degree has reduced the use cost of enzyme.
3, adopt textiles film immobilization lipase, and it is fixed in the rack of rotation, the textiles film can not only increase the catalysis area, the catalytic efficiency height, reduce enzyme dosage, and have that adsorptivity is strong, good stability, low price, recovery easily and the characteristics that can reuse.Adopt the lipase fermented liquid to prepare immobilized lipase, can reduce production costs greatly.The present invention compares with resolvase or granular immobilized enzyme, be easier to separate with reaction solution, and it is multiplexing to help the recovery of serialization production and enzyme.Be used for stirred-tank reactor or fixed bed type reactor and compare with other reactors (as tubular type, membrane reactor), structure and simple to operate, equipment interoperability is strong.
4, the reaction solvent nontoxicity of the present invention's use is easy to reclaim, and particularly the sherwood oil cheapness can reduce production costs widely.
(4) embodiment
Embodiment 1～embodiment 24 lists in the table 1, in the table: A-immobilized lipase B-immobilized lipase/mass ratio C-substrate title (lipid acid or fatty acid ester) D-substrate mol ratio (/ lipid acid or fatty acid ester) E-solvent comprises water amount.
Embodiment 1:
Continuous cloth and common fixing agent are pressed 1: 1 mixed of mass volume ratio, and (W: V), room temperature is dried, and obtains activated carrier; Fixing agent is gelatin, Yelkin TTS, polyoxyethylene glycol, polysorbate altogether, the lipid acid sorb is smooth and the mixture of magnesium salts, and its mass ratio is: gelatin: lecithin: polyoxyethylene glycol: polysorbate: the lipid acid sorb is smooth: magnesium chloride=5: 1: 1: 2: 2: 1; With the aqueous solution of separating ester first silk yeast fat enzyme in the ratio of 8000 units/gram activated carriers with the activated carrier immersion, mix, room temperature dries, make enzyme and live and be the immobilized lipase of 8000IU/g; Be that 0.5% normal hexane solvent and 0.3g immobilized lipase mix again with substrate 0.100g (0.3mmol) and 0.234g (0.9mmol) palmitinic acid and 10ml water content, under the condition of 30 ℃ of temperature, in the shaking table (190r/min), reacted 24 hours, transformation efficiency is 79%; Take out immobilized lipase, reaction solution obtains light yellow Vitamin A Palmitate 1.7 M.I.U/Gram crystal through separation, crystallization.
Embodiment 2:
Operation steps is with embodiment 1, and difference is: continuous cloth is used resin instead; Separate ester first silk yeast fat enzyme and use South Pole first silk yeast instead; Transformation efficiency is 82%.
Embodiment 3:
Operation steps is with embodiment 1, and difference is: continuous cloth is used diatomite instead; Transformation efficiency is 73%.
Embodiment 4:
The fermented liquid of separating ester first silk yeast fat enzyme is soaked continuous cloth, and room temperature is dried, and makes enzyme work and is the immobilized lipase of 8000IU/g; Be that 0.5% normal hexane solvent and 0.3g immobilized lipase mix again with substrate 0.100g (0.3mmol) and 0.234g (0.9mmol) palmitinic acid and 10ml water content, under the condition of 35 ℃ of temperature, in the shaking table (190r/min), reacted 24 hours, transformation efficiency is 83%; Take out immobilized lipase, reaction solution obtains light yellow Vitamin A Palmitate 1.7 M.I.U/Gram crystal through separation, crystallization.
Embodiment 5:
Operation steps is with embodiment 4, and difference is: 0.234g (0.9mmol) palmitinic acid is used the capric acid into 0.155g (0.9mmol) instead; Water content is that 0.5% to use water content instead be 0%; Temperature is used 30 ℃ of temperature instead for 35 ℃; Transformation efficiency is 79%.
Embodiment 6:
Operation steps is with embodiment 4, and difference is: 0.234g (0.9mmol) palmitinic acid is used 0.243g (0.9mmol) Uniphat A60 instead; Water content is that 0.5% to use water content instead be 0%; Temperature is used 30 ℃ of temperature instead for 35 ℃; Transformation efficiency is 84%.
Embodiment 7:
Operation steps is with embodiment 4, and difference is: 0.234g (0.9mmol) palmitinic acid is used 0.256g (0.9mmol) ethyl palmitate instead; Temperature is used 30 ℃ of temperature instead for 35 ℃; Transformation efficiency is 82%.
Embodiment 8:
Operation steps is with embodiment 4, and difference is: 0.234g (0.9mmol) palmitinic acid is used 0.254g (0.9mmol) oleic acid instead; Transformation efficiency is 82%.
Embodiment 9:
Operation steps is with embodiment 4, and difference is: the carrier silk, and 0.234g (0.9mmol) palmitinic acid is used 0.078g (0.3mmol) palmitinic acid instead; Transformation efficiency is 57%.
Embodiment 10:
Operation steps is with embodiment 4, and difference is: 0.234g (0.9mmol) palmitinic acid is used 0.546g (2.1mmol) palmitinic acid instead; Transformation efficiency is 83%.
Embodiment 11:
Operation steps is with embodiment 4, and difference is: the 0.3g immobilized lipase is used the 0.02g immobilized lipase instead; Water content is that 0.5% to use water content instead be 0%; Transformation efficiency is 54%.
Embodiment 12:
Operation steps is with embodiment 4, and difference is: the 0.3g immobilized lipase is used the 0.5g immobilized lipase instead; Water content is that 0.5% to use water content instead be 0%; Transformation efficiency is 84%.
Embodiment 13:
Operation steps is with embodiment 4, and difference is: the normal hexane solvent is used petroleum ether solvent instead; Transformation efficiency is 79%.
Embodiment 14:
Operation steps is with embodiment 4, and difference is: the normal hexane solvent is used cyclohexane solvent instead; Transformation efficiency is 82%.
Embodiment 15:
Operation steps is with embodiment 4, and difference is: water content is that 0.5% to use water content instead be 0%; The normal hexane solvent is used normal heptane solvent instead; Transformation efficiency is 74%.
Embodiment 16:
Operation steps is with embodiment 4, and difference is: water content is that 0.5% to use water content instead be 1%; The normal hexane solvent is used the n-decane solvent instead; Transformation efficiency is 62%.
Embodiment 17:
Operation steps is with embodiment 4, and difference is: react to change in 24 hours and reacted 9 hours; Transformation efficiency is 76%.
Embodiment 18:
Operation steps is with embodiment 4, and difference is: react to change in 24 hours and reacted 50 hours; Transformation efficiency is 83%.
Embodiment 19:
Operation steps is with embodiment 4, and difference is: temperature is used 20 ℃ of temperature of reaction instead for 35 ℃; Transformation efficiency is 51%.
Embodiment 20:
Operation steps is with embodiment 4, and difference is: temperature is used 25 ℃ of temperature of reaction instead for 35 ℃; Transformation efficiency is 85%.
Embodiment 21:
Operation steps is with embodiment 4, and difference is: temperature is used 50 ℃ of temperature of reaction instead for 35 ℃; Transformation efficiency is 69%.
Embodiment 22:
Continuous cloth and common fixing agent are pressed 1: 1 mixed of mass volume ratio, and (W: V), room temperature is dried, and obtains activated carrier; Fixing agent is gelatin, Yelkin TTS, polyoxyethylene glycol, polysorbate altogether, the lipid acid sorb is smooth and the mixture of magnesium salts, and its mass ratio is: gelatin: lecithin: polyoxyethylene glycol: polysorbate: the lipid acid sorb is smooth: magnesium chloride=5: 1: 1: 2: 2: 1; Dry in ratio and activated carrier immersion, the room temperature of 8000 units/gram activated carriers with the aqueous solution of separating ester first silk yeast fat enzyme, make enzyme work and be the immobilized lipase of 8000IU/g; Be that 0.5% normal hexane solvent and 30g immobilized lipase mix again with substrate 10.00g (30.00mmol) and 23.40g (90.00mmol) palmitinic acid and 1L water content, under the condition of 25 ℃ of temperature, stir in the enzyme reactor (500r/min), reacted 24 hours, transformation efficiency is 71%; Take out immobilized lipase, reaction solution obtains light yellow Vitamin A Palmitate 1.7 M.I.U/Gram crystal through separation, crystallization.
Embodiment 23:
The fermented liquid of separating ester first silk yeast fat enzyme is soaked continuous cloth, and room temperature is dried, and makes enzyme work and is the immobilized lipase of 8000IU/g; With the diameter of the stirring enzyme reactor of 30g immobilized lipase stuck-at-L is on the cylindrical rack of 6.5cm; Add substrate 10.00g (30.00mmol) and 23.40g (90.00mmol) palmitinic acid and 1L water content again and be 0.5% normal hexane solvent, under the condition of 25 ℃ of temperature, stir (mixing speed 500r/min), reacted 24 hours, transformation efficiency is 79%; Take out immobilized lipase, reaction solution obtains light yellow Vitamin A Palmitate 1.7 M.I.U/Gram crystal through separation, crystallization.
Embodiment 24:
Operation steps is with embodiment 23, and difference is: enzyme reactor is used the fixed bed enzyme reactor instead; Transformation efficiency is 82%.
Table 1
| Embodiment | A | B | C | D | E | Reaction solvent | Reaction times (hr) | Temperature (℃) | Transformation efficiency (%) |
| 1 | The cotton immobilized enzyme | 3∶1 | Palmitinic acid | 1∶3 | 0.5 | Normal hexane | 24 | 35 | 79 |
| 2 | Resin immobilized enzyme | 3∶1 | Palmitinic acid | 1∶3 | 0.5 | Normal hexane | 24 | 30 | 82 |
| 3 | The diatomite immobilized enzyme | 3∶1 | Palmitinic acid | 1∶3 | 0.5 | Normal hexane | 24 | 30 | 73 |
| 4 | Cotton immobilization fermentation liquid | 3∶1 | Palmitinic acid | 1∶3 | 0.5 | Normal hexane | 24 | 35 | 83 |
| 5 | Cotton immobilization fermentation liquid | 3∶1 | Capric acid | 1∶3 | 0 | Normal hexane | 24 | 30 | 79 |
| 6 | Cotton immobilization fermentation liquid | 3∶1 | Uniphat A60 | 1∶3 | 0 | Normal hexane | 24 | 30 | 84 |
| 7 | Cotton immobilization fermentation liquid | 3∶1 | Ethyl palmitate | 1∶3 | 0.5 | Normal hexane | 24 | 30 | 82 |
| 8 | Cotton immobilization fermentation liquid | 3∶1 | Oleic acid | 1∶3 | 0.5 | Normal hexane | 24 | 30 | 82 |
| 9 | Silk immobilization fermentation liquid | 3∶1 | Palmitinic acid | 1∶1 | 0.5 | Normal hexane | 24 | 35 | 57 |
| 10 | Cotton immobilization fermentation liquid | 3∶1 | Palmitinic acid | 1∶7 | 0.5 | Normal hexane | 24 | 35 | 83 |
| 11 | Cotton immobilization fermentation liquid | 0.2∶1 | Palmitinic acid | 1∶3 | 0 | Normal hexane | 24 | 35 | 54 |
| 12 | Cotton immobilization fermentation liquid | 5∶1 | Palmitinic acid | 1∶3 | 0 | Normal hexane | 24 | 35 | 84 |
| 13 | Cotton immobilization fermentation liquid | 3∶1 | Palmitinic acid | 1∶3 | 0.5 | Sherwood oil | 24 | 35 | 79 |
| 14 | Cotton immobilization fermentation liquid | 3∶1 | Palmitinic acid | 1∶3 | 0.5 | Hexanaphthene | 24 | 35 | 82 |
| 15 | Cotton immobilization fermentation liquid | 3∶1 | Palmitinic acid | 1∶3 | 0 | Normal heptane | 24 | 35 | 74 |
| 16 | Cotton immobilization fermentation liquid | 3∶1 | Palmitinic acid | 1∶3 | 1 | N-decane | 24 | 35 | 62 |
| 17 | Cotton immobilization fermentation liquid | 3∶1 | Palmitinic acid | 1∶3 | 0.5 | Normal hexane | 9 | 35 | 76 |
| 18 | Cotton immobilization fermentation liquid | 3∶1 | Palmitinic acid | 1∶3 | 1 | Normal hexane | 50 | 35 | 83 |
| 19 | Cotton immobilization fermentation liquid | 3∶1 | Palmitinic acid | 1∶3 | 0.5 | Normal hexane | 24 | 20 | 51 |
| 20 | Cotton immobilization fermentation liquid | 3∶1 | Palmitinic acid | 1∶3 | 0.5 | Normal hexane | 24 | 25 | 85 |
| 21 | Cotton immobilization fermentation liquid | 3∶1 | Palmitinic acid | 1∶3 | 0.5 | Normal hexane | 24 | 50 | 69 |
| 22 | The diatomite immobilized enzyme | 3∶1 | Palmitinic acid | 1∶3 | 0.5 | Normal hexane | 24 | 25 | 71 |
| 23 | Cotton immobilization fermentation liquid | 3∶1 | Palmitinic acid | 1∶3 | 0.5 | Normal hexane | 24 | 25 | 79 |
| 24 | Cotton immobilization fermentation liquid | 3∶1 | Palmitinic acid | 1∶3 | 0.5 | Normal hexane | 24 | 25 | 82 |
Claims (8)
1, the method for the fixed lipase catalyzed synthesise vitamins A of a kind of usefulness fatty acid ester is with substrate and C
10～C
18Lipid acid or fatty acid ester are in molar ratio 1: 1～1: 7 scope, mix with organic solvent and immobilized lipase, the consumption of immobilized lipase is 0.2～5 times of quality, under 20～50 ℃ condition, reacted 9～50 hours, take out immobilized lipase, reaction solution obtains vitamin A fatty acid ester through separation, crystallization; Organic solvent is that water content is 0～1% C
6～C
10Saturated alkane; Immobilized lipase is the yeast quasi-lipase that is fixed on the carrier, and carrier is diatomite, resin or textiles film, and the yeast quasi-lipase is for separating ester lipase from candida sp or South Pole first silk yeast fat enzyme.
2, method according to claim 1 is characterized in that: the preparation method of immobilized lipase has following two kinds of methods: (A) with carrier and altogether fixing agent press mass volume ratio 1: 1～1: 3 scope mixing, room temperature is dried and is obtained activated carrier; Fixing agent is gelatin, Yelkin TTS, polyoxyethylene glycol, polysorbate altogether, the lipid acid sorb is smooth and the mixture of magnesium salts, its mass ratio is: gelatin: lecithin: polyoxyethylene glycol: polysorbate: the lipid acid sorb is smooth: magnesium salts=5: 1: 1: 2: 2: 1, magnesium salts was magnesium chloride or sal epsom; With the aqueous solution of yeast quasi-lipase by 1000～30000 units/gram activated carriers with the activated carrier immersion, mix, room temperature dries, make enzyme and live and be the immobilized lipase of 8000IU/g; (B) fermented liquid with the yeast quasi-lipase soaks carrier, mix, and room temperature is dried, and making enzyme alive is the immobilized lipase of 8000IU/g.
3, method according to claim 1 is characterized in that: C
10～C
18Lipid acid or fatty acid ester are: capric acid, lauric acid, tetradecanoic acid, palmitinic acid, stearic acid, oleic acid, methyl caprate, Laurate methyl, Myristicin acid methylester, Uniphat A60, methyl stearate, Witconol 2301, ethyl decylate, Laurate ethyl, ethyl myristate, ethyl palmitate, Stearic ethyl stearate or ethyl oleate.
4, method according to claim 1 is characterized in that: organic solvent is normal hexane, hexanaphthene, sherwood oil, normal heptane, octane, positive nonane or n-decane.
5, method according to claim 1 and 2 is characterized in that: the textiles film is silk floss, nylon, silk, polyester or cellulosic woven.
6, method according to claim 1 is characterized in that: the immobilized lipase of diatomite or resin is used for stirred-tank reactor or fixed bed type reactor.
7, method according to claim 1 is characterized in that: the immobilized lipase of textiles film is used for stirred-tank reactor, and immobilized lipase is fixed on the cylindrical rack, and rack is fixed in the rotating shaft and does axially rotation as stirring rake in the lump.
8, method according to claim 1 is characterized in that: the immobilized lipase of textiles film is used for fixing hearth reactor, and immobilized lipase is fixed on the cylindrical rack, and cylindrical rack is fixed in the reactor.
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| CN100424170C (en) * | 2005-10-11 | 2008-10-08 | 北京化工大学 | Lipase, its gene, yalulipolytic geast for producing said enzyme and its application |
| ES2373340T3 (en) * | 2006-08-30 | 2012-02-02 | Basf Se | METHOD FOR THE PREPARATION OF POLYESTEROLS. |
| US7566795B2 (en) * | 2006-10-06 | 2009-07-28 | Eastman Chemical Company | Preparation of retinyl esters |
| CN101240303B (en) * | 2007-02-05 | 2010-12-15 | 北京化工大学 | Method for synthesizing natural tocopherol ester catalyzed by lipase |
| CN102559785B (en) * | 2011-12-09 | 2013-12-18 | 浙江工业大学 | Method for catalytic synthesis of conjugated linoleic acid (CLA) ethyl ester by immobilized lipase in solvent phase |
| CN104140985B (en) * | 2014-07-10 | 2017-04-05 | 华南理工大学 | A kind of enzyme catalysiss non-oxidizability oils and fatss for preparing in situ and method |
| CN105441521A (en) * | 2014-09-01 | 2016-03-30 | 浙江工业大学 | Synthetic method of vitamin A palmitate |
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