CN106755154A - A kind of method that use micro passage reaction prepares phosphatidylserine - Google Patents
A kind of method that use micro passage reaction prepares phosphatidylserine Download PDFInfo
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Abstract
The present invention relates to a kind of method that use micro passage reaction prepares phosphatidylserine, by the following steps：Step one, prepares enzymic catalytic reaction liquid；Step 2, prepares phospholipase D solution；Step 3, the phospholipase D solution that the reaction solution and step 2 that step one is prepared are prepared pumps into micro passage reaction by high-pressure pump, is reacted upon mixing；Step 4, after reaction terminates, high temperature passivation phospholipase D activity then makes reaction system flow through refrigerated module, and final reaction system outflow micro passage reaction obtains phosphatidylserine product.The present invention has the advantages that reaction speed is fast, yield is high.
Description
Technical field
The invention belongs to technical field of biochemical industry, and in particular to one kind prepares phosphatidyl silk ammonia in micro passage reaction
The method of acid.
Background technology
Phosphatide (Phospholipid) is the lipids of phosphorous acid group, and structure is the fatty acid glyceride of phosphinylidyne two.Phosphorus
Fat is different by its molecular structure, can be divided into glyceric alcohol phosphatides and the major class of nerve conduction studies two.Mainly include phosphatidylserine
(Phosphatidylserine, PS), phosphatid ylcholine (lecithin, Phosphatidylcholine, PC), phosphatidyl ethanol
Amine (cephalin, Phosphatidylethanolamine, PE), phosphatidylinositols (Phosphatidylinositol, PI) and
Phosphatidic acid (Phosphatidic acid, PA).
Phosphatidylserine is the phosphatide of a class generally existing, is usually located at the internal layer of cell membrane, with a series of film work(
Can be relevant.Used as the confactor of many enzymes, phosphatidylserine contacts in close relations with cytoactive and iuntercellular.Phosphatidyl
Serine also shows many activity relevant with neuroendocrine response, including acetylcholine, dopamine, norepinephrine
Release.In various phosphatide of human body, phosphatidylserine content is few but function is unique, can influence intracerebral chemistry message
Transmission, and help brain cell to store and reading data.Increase with the age, phosphatidylserine and other important intracerebral chemicals
Matter can be reduced, so as to cause memory, cognitive power etc. to weaken.Phosphatidylserine can increase brain bur number, brain cell membrane
Mobility and promotion glucose metabolism of brain cell, thus make brain cell more active, nowadays it is increasingly being applied to medicine
And healthcare field.
In the prior art, the preparation method about phosphatidylserine has various, wherein being with extraction method and enzyme transforming process
It is main.Extraction method is that phosphatidylserine is extracted from animal phospholipids, and mainly the brain with animal is as raw material.Patent CN1583766A
It is exactly to extract phosphatidylserine by raw material of animal brain.It is existing but due to reasons such as rabid ox diseases, its Product Safety is suspected
It has been eliminated.Enzyme technology is widely used in the preparation of phosphatidylserine in recent years, for example patent
CN101230365A, is that material, enzyme method prepares phosphatidylserine exactly with soybean lecithin.CN101818179A is with two steps
Enzyme process synthesizes the phosphatidylserine of the fatty acyl group of double bond containing how unsaturated from common soybean lecithin.Patent
It to extract from the phosphatide in fish liver is raw material that CN100402656C is, patent CN101157946A is extracting from the phosphatide in squid
It is raw material, is reacted with serine using the phosphatid ylcholine in phospholipase D enzyme law catalysis fish liver or the phosphatid ylcholine in squid
Prepare the phosphatidylserine rich in how unsaturated double bond fatty acyl group.
But, current enzymatic prepares phosphatidylserine technology and all suffers from product purity problem not high, main former
Because being that current enzymic catalytic reaction is all, using traditional tank reactor, due to the characteristic of interfacial reaction, to be limited by still reaction
Device heat and mass relatively slow, reactant is difficult to be fully contacted and cause catalytic reaction not thorough with enzyme, and product phosphatidyl silk ammonia
Acid easily generates the accessory substances such as phosphatidic acid after reaching finite concentration by phospholipase D hydrolysis, has thus resulted in phosphatidyl in final products
The content of serine is not high, and substrate residual quantity is high and the accessory substance such as phosphatidic acid containing higher amount.Therefore, how to be effectively increased anti-
The mass transfer effect of system is answered, the efficiency of catalytic reaction is improved, accelerates reaction process, so as to shorten the reaction time, reduce phosphatidic acid
The generation of hydrolysising by-product, is always that enzymatic prepares the crucial problem that phosphatidylserine technology needs to solve.
Micro passage reaction is a kind of pipeline reactor of continuous flowing, and the wherein internal diameter of the pipeline size general control exists
It is a kind of device chemistry or enzymic catalytic reaction controlled in microresponse space between 10-1000 microns.Microchannel plate should
The microfluid that the structures shape that utensil has passes through wherein can greatly shorten time and the distance of mass transfer, while increasing
Bigger serface, it is possible to achieve the quick mixing of material, so as to greatly improve mass-transfer efficiency.Narrow micro- logical of micro passage reaction
Road also increases temperature transmission gradient simultaneously, and the specific surface area of increase greatly improves the heat-transfer capability of reactor, exchanged heat with tradition
Device greatly improves heat transfer efficiency compared at least order of magnitude greater.The unique texture of micro passage reaction and efficient biography
Matter heat transfer efficiency so that the precise control to reaction process turns into possible.Work is reacted relative to traditional tank reactor gap
Skill, micro passage reaction can accurately adjust reaction process condition, such as to the precise control of reaction temperature, to the reaction time
The precise control of precise control and material mixed proportion, can greatly improve reaction efficiency, reduce the generation of accessory substance, shorten
Reaction time.Due to the advantage of micro passage reaction, flexile chemistry can be met by with reference to different supplemental functionalities
Technique or enzymatic technological requirement, are particularly suited for strong exothermal reaction, the reaction of reactant or unstable products, to reaction
The inapplicable special reaction of the routine tank reactor such as very strict fast reaction of ratio requirement.
The present inventor has found that micro passage reaction is applied to enzyme catalysis method prepares phosphatidylserine tool through studying for many years
There is the advantage that tank reactor is incomparable, be mainly reflected in：1- can greatly improve the efficiency of catalytic reaction, when shortening reaction
Between, compared with the reaction in tank reactor, it is same under conditions of reach more than 95% catalytic rate, micro passage reaction can save
Save for more than 50% time；2- can be greatly reduced the generation of hydrolysising by-product phosphatidic acid, target product phosphatidylserine
Content can improve 5%-15%；3- to reaction temperature and post-processing temperature can quickly and accurately control, temperature during reaction
Degree can keep accurately constant, after reaction terminates, using high temperature module, rapidly be warming up to reaction system in very short time
Uniform temperature, Rapid deactivation phospholipase D activity so that can effectively reduce target product the later stage process and preserve during because
The enzyme of residual and hydrolyze generation accessory substance phosphatidic acid, it can be ensured that the stability of finished product；In subsequent unit operation, by low temperature
Module, can be cooled to uniform temperature by reaction system rapidly in very short time, it is ensured that target product will not be because of for a long time
High-temperature process and degrade.The phosphatidylserine for preparing by this method, target product purity is high, accessory substance phosphatidic acid
Content is substantially reduced, and shortens the reaction time, substantially increases reaction efficiency, thus can be with actual industrial metaplasia product
Cost is substantially reduced, the phosphatidylserine product of high-quality is obtained.
So far, there is not yet the preparation technology that phosphatidylserine is carried out in the way of micro passage reaction continuous stream grinds
Study carefully.
Based on this, spy proposes the present invention.
The content of the invention：
The purpose of the present invention is directed to prior art weak point and provides one kind and phosphatid ylcholine is converted into phosphatidyl
The preparation method of serine, a kind of method that especially use micro passage reaction prepares phosphatidylserine.
The purpose of the present invention is achieved through the following technical solutions：
A kind of method that use micro passage reaction prepares phosphatidylserine, it is characterized in that, prepares under passing through
Row step：
Step one, prepares enzymic catalytic reaction liquid；
Step 2, prepares phospholipase D solution；
Step 3, the phospholipase D solution that the reaction solution and step 2 that step one is prepared are prepared is pumped into micro- by high-pressure pump
Channel reactor, is reacted upon mixing；
Step 4, after reaction terminates, high temperature passivation phospholipase D activity then makes reaction system flow through refrigerated module, finally
Reaction system flows out micro passage reaction, obtains phosphatidylserine product.
Further, the micro passage reaction includes mixing module, reaction module, high temperature module and refrigerated module, often
The fluid path length of individual module is 0.1m~300m, preferably 1m~60m；The width and depth of the fluid passage of each module
It is 0.5 μm~10mm, preferably 0.1mm~1mm；The fluid passageway width and depth of each module section link be 0.5 μm-
100μm。
Further, the proportion relation of each material is as follows in reaction solution in the step one：Phosphatid ylcholine is mixed at every liter
It is 100 to close the weight portion in reaction solution；Weight portion of the serine in every liter of mixed reaction solution be 60~300, preferably 200~
250；Weight portion of the calcium salt in every liter of reaction solution is 0.05~10, preferably 2~4；C5-C9 alkane organic solvent is anti-in total mixing
It is 0~95% to answer the percent by volume occupied in liquid；Short hydrocarbon alcohol or short hydrocarbon the ketone possessive volume hundred in total mixed reaction solution
Divide than being 0~80%；.
Further, in the step 3, the reaction solution and phospholipase D solution before micro passage reaction is pumped into,
500 mesh and 0.5 μm of ultrafiltration apparatus need to be sequentially passed through, solid particle is filtered to remove.
Further, in the step 3, the reaction solution and phospholipase D solution by 0.5~500bar, preferably 2~
The compression pump of 50bar enters micro passage reaction.
Further, in the step 3, after reaction system enters the reaction module of micro passage reaction, reaction temperature is
35~55 degrees Celsius, preferably 45-50 degrees Celsius；Residence time of the reaction system in reaction module is 30~1200 seconds, preferably
60-300 seconds.
Further, in the step 4, after reaction terminates, reaction solution is made to flow through high temperature module, the temperature of high temperature module
It is 75~105 degrees Celsius, preferably 85~90 degrees Celsius；Reaction solution residence time in high temperature module is 10~1200 seconds, preferably
15-120 seconds.
Further, in the step 4, reaction solution when refrigerated module is flowed through, for 0~40 take the photograph by the temperature of refrigerated module
Family name's degree, preferably 25-35 degree；Reaction solution is 30~1200 seconds, preferably 30-300 seconds in the refrigerated module residence time.
Further, every kind of module of micro passage reaction connects to be formed for polylith little module, wherein reaction module 2-200
Block, high temperature module 2-20 blocks, refrigerated module 2-20 blocks.
Compared with prior art, the invention has the advantages that：
1) present invention prepares phosphatidylserine using micro passage reaction, in micro passage reaction, due to passing
Matter efficiency high, reaction speed is fast and reaction is thorough, substantially reduces the reaction time, reduces the incidence of enzyme hydrolysis side reaction,
So as to significantly reduce the generation of accessory substance phosphatidic acid, the content of phosphatidylserine obtains 5~15% raising in product.
2) course of reaction mass-and heat-transfer efficiency is greatly improved, and whole production process can be carried out controllable and accurately temperature is adjusted
Control, after particularly reaction terminates, can be processed by short time high temperature enzyme-deactivating and subsequent fast cooling is processed, anti-compared to autoclave
Should, it is processed as not only can easily inactivating the enzyme activity of residual in product but also will not causes to be produced due to long-time high-temperature process
Phosphatidylserine structural damage, ensures that the stabilization of finished product quality, extends the shelf-life of product.
3) by the present invention in that with micro passage reaction without enlarge-effect, it is easy to upgrading amplify, it is possible to achieve from research and development to
The seamless scale-up of production.
The product for finally giving can be used for senior nutrition and health products, can also be added into food, medicine or cosmetics
Use.
Brief description of the drawings:
Fig. 1 is the similarities and differences of the phosphatidylserine molecular structural formula that phosphatid ylcholine is formed with conversion.
Fig. 2 is to generate phosphatidylserine and phosphatidylserine water by phosphatid ylcholine under the catalysis of phospholipase D
The reaction schematic diagram of solution generation phosphatidic acid.
Fig. 3 produces phosphatidylserine process chart for the present invention using continuous micro passage reaction.
Wherein：The material-compound tanks of 1- first, the material-compound tanks of 2- second, 3- mixing modules, 4- reaction modules, 5- high temperature modules, 6- are low
Warm module, 7- product collecting devices.
Specific embodiment
The present invention is described further below in conjunction with accompanying drawing 1-3.
To more fully understand present disclosure, first Fig. 1 is explained：In figure, Sn-1, Sn-2 and Sn-3 refer to difference respectively
1,2 and 3 of glycerine ester group in the phosphatide in source；R1 and R2 is Long carbon chain fatty acyl group.
- the R1 of Sn-1 common are：- C15H31,-C17H35,-C17H33.If it is-C15H31, then illustrate that R1 comes
From Palmitic acid or palmitic acid ,-C15H31Coupled carbonyl group constitutes a 16 carbocyclic aliphatic acyl groups, represents in Sn-1
Position is the ester that a Palmitic acid is formed with Sn-1 alcohol of glycerine；If it is-C17H35, then illustrate R1 from 18 carbonic acid or
Stearic acid ,-C17H35Coupled carbonyl group constitutes a 18 carbocyclic aliphatic acyl groups, represents at Sn-1 it is one 18
The ester that carbonic acid is formed with Sn-1 alcohol of glycerine；If it is-C17H33, then illustrate that R1 comes from 18 carbonic acid or oleic acid ,-C17H33
Coupled carbonyl group constitutes a 18 carbocyclic aliphatic acyl groups, represents at Sn-1 it is 18 carbonic acid and glycerine
The ester that Sn-1 alcohol is formed.
- the R2 of Sn-2 common are：- C17H31,-C19H29,-C19H31,-C21H31,-C17H33, if it is-
C17H31, then illustrate that R2 comes from the diluted acid of 18 carbon two or linoleic acid ,-C17H31Coupled carbonyl group constitutes 18 carbon
Two dilute fatty acyl groups, represent at Sn-2 it is ester of the diluted acid of 18 carbon two with the Sn-2 alcohol formation of glycerine；If it is-
C19H29, then illustrate that R2 comes from eicosapentaenoic acid or EPA ,-C19H29Coupled carbonyl group constitutes 20 light dydrocarbons
Dilute fatty acyl group, represents at Sn-2 it is ester of the eicosapentaenoic acid with the Sn-2 alcohol formation of glycerine；If it is-
C19H31, then illustrate that R2 comes from the diluted acid of 20 carbon four or arachidonic acid ,-C19H31Coupled carbonyl group constitutes one two
The dilute fatty acyl group of ten carbon four, represents at Sn-2 it is ester of the diluted acid of 20 carbon four with the Sn-2 alcohol formation of glycerine；If its
For-C21H31, then illustrate that R2 comes from the diluted acid of 22 carbon six ,-C21H31Coupled carbonyl group constitutes 22 carbon
Six dilute fatty acyl groups, represent at Sn-2 it is ester of the diluted acid of 22 carbon six with the Sn-2 alcohol formation of glycerine；If its
For-C17H33, then illustrate that R2 comes from 18 carbon diluted acids or oleic acid ,-C17H33Coupled carbonyl group constitutes 18 carbon
Dilute fatty acyl group, represents at Sn-2 it is ester of the 18 carbon diluted acids with the Sn-2 alcohol formation of glycerine.
- the X of Sn-3 is different linking groups, if it is-CH2CH2N(CH3)3, then it represents that the phosphatide cpd is
Phosphatid ylcholine；If it is-CH2CH(NH2) COOH, then the phosphatide cpd is phosphatidylserine.
Fig. 2 gives the reaction and phosphatide under the catalysis of phospholipase D by phosphatid ylcholine generation phosphatidylserine
The reaction schematic diagram of acyl serine hydrolysis generation phosphatidic acid.Under reaction condition disclosed by the invention, phospholipase D can be catalyzed phosphorus
Phosphatidylcholine generates phosphatidylserine.However, the phosphatidylserine of generation can also enter one in the case where dealing with improperly
Step hydrolyzes generation phosphatidic acid under the catalysis of phospholipase D.The meaning of Fig. 2 reaction equations refers to that the phosphatid ylcholine of a molecule can
Reacted with the serine with a molecule, one phosphatidylserine of molecule of generation generates a courage for molecule simultaneously
Alkali；One phosphatidylserine of molecule hydrolyzes generation one phosphatidic acid and a molecule of molecule under the catalysis of phospholipase D
Serine.
Fig. 3 is to use continuous micro passage reaction to produce phosphatidylserine process chart, here continuous microchannel plate
Answer the series connection that the module in device can be multiple submodule.Under the method for preparing phosphatidylserine using micro passage reaction includes
Row step：
Step one, prepares enzymic catalytic reaction liquid, and the reaction solution is the reaction system of water phase and organic solvent phase composition, the
Phosphatid ylcholine is first dissolved in C5-C9 alkane organic solvents in one material-compound tank 1, is then added to molten containing serine, calcium salt, buffering
In the aqueous solution that liquid and short hydrocarbon alcohol or short hydrocarbon ketone are configured to, so as to form the reaction of water phase and organic solvent two-phase mixtures
Liquid.
The proportion relation of each material is as follows in two-phase reaction system in the step one：Phosphatid ylcholine is in every liter of mixing
The weight portion added in reaction solution is 100；The weight portion that serine is added in every liter of reaction solution can for 60,80,100,
130th, 150,180,220,260 or 300, i.e. weight portion scope are 60-300, preferably 200~250；Calcium salt is in every liter of reaction solution
The weight portion of addition can be 0.05,0.10,0.20,0.80,1.2,2.5,3.6,4.5,5.0,6.0,7.0,8.0 or 10, i.e.,
Weight portion scope is 0.05-10, preferably 2~4；The percent by volume that C5-C9 alkane organic solvents occupy in mixed reaction solution
Can be 0~95%, short hydrocarbon alcohol or short for 0,5%, 10%, 20%, 40%, 70% or 95%, i.e. volume percent range
Chain hydrocarbon ketone in total mixed reaction solution possessive volume percentage can for 0,5%, 10%, 20%, 40%, 70% or 80%, i.e.,
Volume percent range is 0~80%, preferably 10-15%, and the pH value of reaction system can be 5.0,6.0,7.0,8.0 or 8.5,
I.e. pH value range is 5.0~8.5, preferably 7~7.5.
C5-C9 alkane organic solvent can be from following article optionally：Pentane, hexane, hexamethylene, heptane, octane, nonane,
The mixed solvent that any one of decane or their isomer solvent or these solvents are mixed to form in any proportion.
Short hydrocarbon alcohol or short hydrocarbon ketone can be from following articles optionally：Ethanol, propyl alcohol, butanol, amylalcohol, acetone, butanone or
The mixed solvent that any one of their isomer solvent or these solvents are mixed to form in any proportion.
Phosphatid ylcholine source in the aqueous solution and organic solvent mixed reaction solution can be plant origin, for example, source
In oilseed plants such as soybean, sunflower, rapeseed, kardiseeds；Can also be animal origin, for example, deriving from milk, goat milk, depth
Ocean fish, deep-sea shrimp, deep-sea scallop etc.；Or microbe-derived etc..Wherein, the phosphatid ylcholine of animal origin is rich in more not
The phosphatid ylcholine of saturation double bond fatty acyl group, can prepare the phosphatide rich in how unsaturated double bond fatty acyl group as raw material
Acyl serine.
Serine source in the aqueous solution and organic solvent reaction solution：The biological product of commercialization；Calcium salt：It is solubility calcium
Salt, for example, calcium chloride, the concentration in water can be 0.1 gram per liter to 20 gram per liters；Buffer solution：It is conventional water buffer solution, example
Such as Phosphate buffered saline or acetic acid brine buffer solution, generally 0.01 to 0.2 mole every liter of salinity system.
The aqueous solution and organic solvent reaction solution, before micro passage reaction is pumped into, need to fully dissolve, and remove insoluble solid
Body, and microporous filter is sequentially passed through, filter impurity elimination.Microporous filter in this step, by coarse filtration, micro-filtration and can surpass
Filter, wherein three-level or cascade filtration composition.
Step 2, prepares phospholipase D solution.A certain amount of phospholipase D is fully dissolved in room temperature in the second material-compound tank 2
In appropriate pure water.In this step, phospholipase D is a kind of enzyme, is derived from the commercialization enzyme system of streptomycete and/or plant
The phospholipase D (phospholipase D) that agent, such as Sigma-Aldrich companies (St.Louis, MO, U.S.A.) are sold, the enzyme
Also the zymotic fluid of streptomycete can be purified from.
The enzyme phospholipase D solution need to fully dissolve before micro passage reaction is pumped into, removal insoluble solids, and according to
It is secondary by microporous filter, filter impurity elimination.Microporous filter in this step, can comprising micro-filtration and ultrafiltration or one of them
Function.
Step 3, the phospholipase D solution that the reaction solution and step 2 that step one is prepared are prepared is existed by high-pressure pump
0.5bar, 1bar, 5bar, 20bar, 50bar, 100bar or 500bar, i.e. 0.5~500bar pressure, it is preferred to use 2-50bar
Pressure, carries out high-pressure delivery, pumps into the mixing module 3 of micro passage reaction, is sufficiently mixed, and mixing temperature can be taken the photograph for 25
Family name's degree, 30 degrees Celsius, 35 degrees Celsius, 40 degrees Celsius or 45 degrees Celsius, i.e. blend temperature range are 25~45 degrees Celsius, in mixing
The residence time of module 3 is 30 seconds, 40 seconds, 50 seconds, 60 seconds, 100 seconds, 150 seconds, 250 seconds or 300 seconds, i.e., residence time ranges are
30~300 seconds, to increase mixed effect.Multiple mixing module 3 can be used herein, or time use is carried out to mixing module 3；So
Afterwards, reaction system is reacted into the reaction module 4 of micro passage reaction, reaction temperature be 35 degrees Celsius, 40 degrees Celsius,
4550 degrees Celsius or 55 degrees Celsius, i.e. range of reaction temperature are 35~55 degrees Celsius, and reaction system is in the residence time of reaction module 4
It is that 30 seconds, 50 seconds, 100 seconds, 200 seconds, 400 seconds, 800 seconds or 1200 seconds, i.e. residence time ranges are 30~1200 seconds, is to ensure
Reaction fully thoroughly, herein can be used multiple reaction module 4, or carry out time use to reaction module 4.Whole course of reaction
In, catalyst phospholipase D consumption is the 0.05%~10% of reaction system weight.
Step 4, reaction terminate after, reaction system is flowed through high temperature module 5, at 30 seconds, 50 seconds, 100 seconds, 200 seconds or
In 300 seconds, i.e. in 30-300 seconds it is rapid by reaction system be warming up to 75 degrees Celsius, 80 degrees Celsius, 85 degrees Celsius, 90 degrees Celsius,
95 degrees Celsius, 100 degrees Celsius or 105 degrees Celsius, that is, it is warming up in the range of 75~105 degrees Celsius, preferably 85~90 degrees Celsius
In the range of, to be passivated phospholipase D activity, reaction system is 30 seconds, 50 seconds, 100 seconds, 200 seconds in the residence time of high temperature module 5
Or 300 seconds, i.e. residence time ranges are 30~300 seconds；Following reaction system flows through refrigerated module 6,30 seconds, 50 seconds, 100
In second, 200 seconds or 300 seconds, i.e., residence time ranges are that reaction system is cooled into 25~35 degrees Celsius rapidly in 30-300 seconds；
Final reaction system flows out micro passage reaction, loads product collecting device 7, obtains phosphatidylserine product.
Step 3 and step 4 are carried out in micro passage reaction, and the whole time is 0.1~8 hour.Gained after reaction
In reaction system, the conversion of raw material phosphatid ylcholine obtains the conversion ratio of phosphatidylserine more than 95%, and phosphatidylserine turns
The conversion ratio that change obtains phosphatidic acid is less than 3%.
Finally, gained reaction solution refined, be dried to obtain phosphatidylserine, complete to prepare.
The most preferred embodiment to the present invention in a particular application is described in detail, it is necessary to illustrate, these realities below
Apply example and row constraint not is entered to claims of the invention.
The fluid passage of each module of micro passage reaction used in the present invention has no unified regulation, length be 0.1m~
300m, preferably 1m~60m, the width and depth of fluid passage are 0.5 μm~10mm, preferably 0.1mm~0.5mm.Manufacture
The material of these modules can be stainless steel, extra hard steel, copper, ruby, diamond, ceramics, carborundum, cermet composite wood
Material or other macromolecular materials.Micro passage reaction in the present invention can be the micro passage reaction of commercialization, for example, the U.S.
The heart-shaped structure G1 glass high flux micro passage reactions of Corning Incorporated's production, or prepare according to same principle other
Commercialization or non-commercial micro passage reaction.
Embodiment 1：The phosphatidylserine of soybean-source is prepared using micro passage reaction.
(1) device：Micro-channel device is stopped according to flow velocity using self-control high pressure micro passage reaction, micro-tunnel length with reaction
The time is stayed to determine, each module channels length used herein is 10m, and caliber is 0.1mm, and heat transferring medium is conduction oil.
(2) preparation of the phosphatidylserine of soybean-source：
The preparation of A- micro passage reactions：Reactant is controlled by the passage length of the flow and microchannel that adjust delivery pump
The residence time of modules is tied up to, wherein, the residence time is 30 seconds in mixing module 3, and the residence time is in reaction module 4
120 seconds, the residence time was 15 seconds in high temperature module 5, and the residence time is 100 seconds in refrigerated module 6.By outer circulation heat conduction oil body
It is to control the temperature of reactor modules, wherein, the temperature of mixing module 3 is 30 degrees Celsius, the temperature of reaction module 4 is
45 degrees Celsius, the temperature of high temperature module 5 is 80 degrees Celsius, and the temperature of refrigerated module 6 is 30 degrees Celsius.
The preparation of B- reaction systems：In the first material-compound tank 1, by 100g soybean lecithins raw material, (wherein phosphatid ylcholine contains
It is 53.7% to measure, and phosphatide acid content is 6.6%, and phosphatidylserine content is 0.4%) 300ml n-hexanes, 50ml isopropanols,
200g serines, 8g calcium chloride is put into 700ml Phosphate buffered saline systems, and the pH value of the aqueous solution is 6.5.Reactant
System passes through 50 Mm filter film process by 500 mesh membrane filtrations through 2bar pressure, it is ensured that insoluble solid in removal raw material
Body impurity.
The preparation of C- catalyst phospholipase Ds：In the second material-compound tank 2, the phospholipase D of 10ml is put into 100ml phosphoric acid
In buffered saline liquid system, the pH value of the aqueous solution is 7.5.And the removal of impurity is gone by 0.5 micron of ultrafiltration membrane treatment through 5bar pressure
And miscellaneous bacteria.
D- reacts：The high-pressure metering pump that the first material-compound tank 1 and the second material-compound tank 2 will be connected is opened, under 35bar pressure,
High-pressure delivery so that reaction system and catalyst phospholipase D system are fully mixed into the mixing module 3 of micro passage reaction
Close, the temperature of mixing module 3 is 30 degrees Celsius, and mixed liquor subsequently enters 50 groups of cascade reaction modules, and the temperature of reaction module 4 is
45 degrees Celsius, reaction solution subsequently enters high temperature module 5, and module 5 is 5 modules, and the temperature of high temperature module 5 is 85 degrees Celsius, instead
Liquid is answered to subsequently enter refrigerated module 6, module 6 is 2 block coupled in series, and the temperature of refrigerated module 6 is 30 degrees Celsius, finally, reaction
Product enters product reception device 7.
E- is refined：The product that will be obtained is extracted 2 times with normal heptane, is concentrated with rotavapor under vacuum, removes positive heptan
Alkane, is then placed on vacuum drying oven by product, and under 50 degrees celsius, vacuum drying finally obtains 86g faint yellow solids.Through
HPLC liquid phases detect that in product, phosphatidylserine content is 55.7%, and phosphatidylcholine content is 0.5%, phosphatide acid content
It is 6.7%.
Embodiment 2：The phosphatidylserine of soybean-source is prepared using micro passage reaction.
(1) device：Micro-channel device is stopped according to flow velocity using self-control high pressure micro passage reaction, micro-tunnel length with reaction
The time is stayed to determine, each module channels length used herein is 50m, caliber is that 0.5mm heat transferring mediums are conduction oil.
(2) preparation of the phosphatidylserine of soybean-source：
The preparation of A- micro passage reactions：Reactant is controlled by the passage length of the flow and microchannel that adjust delivery pump
The residence time of modules is tied up to, wherein, the residence time is 60 seconds in mixing module 3, and the residence time is in reaction module 4
300 seconds, the residence time was 60 seconds in high temperature module 5, and the residence time is 300 seconds in refrigerated module 6.By outer circulation heat conduction oil body
It is to control the temperature of reactor modules, wherein, the temperature of mixing module 3 is 30 degrees Celsius, the temperature of reaction module 4 is
48 degrees Celsius, the temperature of high temperature module 5 is 90 degrees Celsius, and the temperature of refrigerated module 6 is 25 degrees Celsius.
The preparation of B- reaction systems：In the first material-compound tank 1, by 100g soybean lecithins raw material, (wherein phosphatid ylcholine contains
It is 55.0% to measure, and phosphatide acid content is 6.4%, and phosphatidylserine content is 0.1%) 500ml n-hexanes, 75ml isopropanols,
250g serines, 8g calcium chloride, the pH value of the aqueous solution is 6.0.Reaction system passes through 500 mesh membrane filtrations, and through 5bar pressure
By 50 Mm filter film process, it is ensured that the insoluble solid impurity in removal raw material.
The preparation of C- catalyst phospholipase Ds：In the second material-compound tank 2, the phospholipase D of 10ml is put into 200ml phosphoric acid
In buffered saline liquid system, the pH value of the aqueous solution is 7.0.And the removal of impurity is gone by 0.5 micron of ultrafiltration membrane treatment through 5bar pressure
And miscellaneous bacteria.
D- reacts：The high-pressure metering pump that the first material-compound tank 1 and the second material-compound tank 2 will be connected is opened, under 50bar pressure,
High-pressure delivery so that reaction system and catalyst phospholipase D system are fully mixed into the mixing module 3 of micro passage reaction
Close, the temperature of mixing module 3 is 30 degrees Celsius, and mixed liquor subsequently enters 20 groups of cascade reaction modules, and the temperature of reaction module 4 is
48 degrees Celsius, reaction solution subsequently enters high temperature module 5, and module 5 is 1 module, and the temperature of high temperature module 5 is 90 degrees Celsius, instead
Liquid is answered to subsequently enter refrigerated module 6, module 6 is 10 block coupled in series, and the temperature of refrigerated module 6 is 25 degrees Celsius, finally, reaction
Product enters product reception device 7.
E- is refined：The product that will be obtained is extracted 3 times with normal heptane, is concentrated with rotavapor under vacuum, removes positive heptan
Alkane, is then placed on vacuum drying oven by product, and under 60 degrees celsius, vacuum drying finally obtains 81g faint yellow solids.Through
HPLC liquid phases detect that in product, phosphatidylserine content is 60.7%, and phosphatidylcholine content is 0.3%, phosphatide acid content
It is 7.0%.
Embodiment 3：The phosphatidylserine of soybean-source is prepared using micro passage reaction.
(1) device：Micro-channel device is stopped according to flow velocity using self-control high pressure micro passage reaction, micro-tunnel length with reaction
The time is stayed to determine, each module channels length used herein is 30m, caliber is that 0.25mm heat transferring mediums are conduction oil.
(2) preparation of the phosphatidylserine of soybean-source：
The preparation of A- micro passage reactions：Reactant is controlled by the passage length of the flow and microchannel that adjust delivery pump
The residence time of modules is tied up to, wherein, the residence time is 100 seconds in mixing module 3, and the residence time is in reaction module 4
600 seconds, the residence time was 120 seconds in high temperature module 5, and the residence time is 60 seconds in refrigerated module 6.By outer circulation heat conduction oil body
It is to control the temperature of reactor modules, wherein, the temperature of mixing module 3 is 40 degrees Celsius, the temperature of reaction module 4 is
50 degrees Celsius, the temperature of high temperature module 5 is 95 degrees Celsius, and the temperature of refrigerated module 6 is 35 degrees Celsius.
The preparation of B- reaction systems：In the first material-compound tank 1, by 100g soybean lecithins raw material, (wherein phosphatid ylcholine contains
It is 60.0% to measure, and phosphatide acid content is 5.6%, and phosphatidylserine content is 0.01%) 75ml isopropanols, 250g serines,
8g calcium chloride, the pH value of the aqueous solution is 5.5.Reaction system passes through 50 microns by 500 mesh membrane filtrations through 10bar pressure
Filtering film process, it is ensured that the insoluble solid impurity in removal raw material.
The preparation of C- catalyst phospholipase Ds：In the second material-compound tank 2, the phospholipase D of 10ml is put into 100ml water-soluble
In liquid, the pH value of the aqueous solution is 7.0.And the removal of impurity and miscellaneous bacteria are gone by 0.5 micron of ultrafiltration membrane treatment through 1bar pressure.
D- reacts：The high-pressure metering pump that the first material-compound tank 1 and the second material-compound tank 2 will be connected is opened, under 10bar pressure,
High-pressure delivery so that reaction system and catalyst phospholipase D system are fully mixed into the mixing module 3 of micro passage reaction
Close, the temperature of mixing module 3 is 40 degrees Celsius, and mixed liquor subsequently enters 100 groups of cascade reaction modules, the temperature of reaction module 4
It is 50 degrees Celsius, reaction solution subsequently enters high temperature module 5, module 5 is 1 module, and the temperature of high temperature module 5 is 95 degrees Celsius,
Reaction solution subsequently enters refrigerated module 6, and module 6 is 3 block coupled in series, and the temperature of refrigerated module 6 is 35 degrees Celsius, finally, instead
Product is answered to enter product reception device 7.
E- is refined：The product that will be obtained is extracted 3 times with normal heptane, is concentrated with rotavapor under vacuum, removes positive heptan
Alkane, is then placed on vacuum drying oven by product, and under 60 degrees celsius, vacuum drying finally obtains 89g faint yellow solids.Through
HPLC liquid phases detect that in product, phosphatidylserine content is 65.6%, and phosphatidylcholine content is 0.2%, phosphatide acid content
It is 6.9%.
Embodiment 4：The phosphatidylserine of soybean-source is prepared using micro passage reaction.
(1) device：Micro-channel device is stopped according to flow velocity using self-control high pressure micro passage reaction, micro-tunnel length with reaction
The time is stayed to determine that heat transferring medium is conduction oil.
(2) preparation of the phosphatidylserine of soybean-source：
The preparation of A- micro passage reactions：Reactant is controlled by the passage length of the flow and microchannel that adjust delivery pump
The residence time of modules is tied up to, wherein, the residence time is 90 seconds in mixing module 3, and the residence time is in reaction module 4
600 seconds, the residence time was 30 seconds in high temperature module 5, and the residence time is 60 seconds in refrigerated module 6.By outer circulation heat conduction oil body
It is to control the temperature of reactor modules, wherein, the temperature of mixing module 3 is 45 degrees Celsius, the temperature of reaction module 4 is
50 degrees Celsius, the temperature of high temperature module 5 is 88 degrees Celsius, and the temperature of refrigerated module 6 is 25 degrees Celsius.
The preparation of B- reaction systems：In the first material-compound tank 1, by 100g soybean lecithins raw material, (wherein phosphatid ylcholine contains
It is 40.0% to measure, and phosphatide acid content is 8.4%, and phosphatidylserine content is 0.1%) 250g serines, 8g calcium chloride, water
The pH value of solution is 5.5.Reaction system passes through 50 Mm filter film process by 500 mesh membrane filtrations through 10bar pressure,
Ensure to remove the insoluble solid impurity in raw material.
The preparation of C- catalyst phospholipase Ds：In the second material-compound tank 2, the phospholipase D of 10ml is put into 100ml water-soluble
In liquid, the pH value of the aqueous solution is 7.0.And the removal of impurity and miscellaneous bacteria are gone by 0.5 micron of ultrafiltration membrane treatment through 1bar pressure.
D- reacts：The high-pressure metering pump that the first material-compound tank 1 and the second material-compound tank 2 will be connected is opened, high under 3bar pressure
Pressure conveying so that reaction system and catalyst phospholipase D system are fully mixed into the mixing module 3 of micro passage reaction
Close, the temperature of mixing module 3 is 40 degrees Celsius, and mixed liquor subsequently enters 20 groups of cascade reaction modules, and the temperature of reaction module 4 is
50 degrees Celsius, reaction solution subsequently enters high temperature module 5, and module 5 is 6 modules, and the temperature of high temperature module 5 is 88 degrees Celsius, instead
Liquid is answered to subsequently enter refrigerated module 6, module 6 is 5 block coupled in series, and the temperature of refrigerated module 6 is 25 degrees Celsius, finally, reaction
Product enters product reception device 7.
E- is refined：The product that will be obtained is extracted 3 times with normal heptane, is concentrated with rotavapor under vacuum, then will be produced
Thing is placed on vacuum drying oven, and under 60 degrees celsius, vacuum drying finally obtains 78g faint yellow solids.Examined through HPLC liquid phases
Survey, in product, phosphatidylserine content is 46.7%, and phosphatidylcholine content is 0.5%, phosphatide acid content is 11.0%.
Embodiment 5：The phosphatidylserine in sunflower seeds source is prepared using micro passage reaction.
(1) device：Micro-channel device using self-control high-pressure glass micro passage reaction, micro-tunnel length according to flow velocity with it is anti-
Should determine the residence time.The length of each reaction module passage is 60m herein, and channel diameter is 0.2mm；Each refrigerated module leads to
Road length is 30m, and channel diameter is 0.2mm；Each high temperature module channels length is 0.5mm, and channel diameter is 1mm.Heat exchange is situated between
Matter is conduction oil.
(2) preparation of the phosphatidylserine in sunflower seeds source：
The preparation of A- micro passage reactions：Reactant is controlled by the passage length of the flow and microchannel that adjust delivery pump
The residence time of modules is tied up to, wherein, residence time of mixing module 3 is 75 seconds, and the residence time of reaction module 4 is
300 seconds, the residence time of high temperature module 5 was 15 seconds, and the residence time of refrigerated module 6 is 120 seconds.By outer circulation heat conduction oil body
It is to control the temperature of reactor modules, wherein, the temperature of mixing module 3 is 45 degrees Celsius, the temperature of reaction module 4 is
47 degrees Celsius, the temperature of high temperature module 5 is 86 degrees Celsius, and the temperature of refrigerated module 6 is 24 degrees Celsius.
The preparation of B- reaction systems：In the first material-compound tank 1, by 100g sunflower seeds lecithin materials (wherein phosphatid ylcholine
Content is 47.6%, and phosphatide acid content is 7.2%, and phosphatidylserine content is 0.3%) 300ml n-hexanes, 50ml isopropyls
Alcohol, 200g serines, 8g calcium chloride is put into 700ml Phosphate buffered saline systems, and the pH value of the aqueous solution is 6.5.Reaction
System passes through 50 Mm filter film process by 500 mesh membrane filtrations through 3bar pressure, it is ensured that insoluble in removal raw material
Solid impurity.
The preparation of C- catalyst phospholipase Ds：In the second material-compound tank 2, the phospholipase D of 10ml is put into 100ml phosphoric acid
In buffered saline liquid system, the pH value of the aqueous solution is 6.5.And the removal of impurity is gone by 0.5 micron of ultrafiltration membrane treatment through 2bar pressure
And miscellaneous bacteria.
D- reacts：The high-pressure metering pump that the first material-compound tank 1 and the second material-compound tank 2 will be connected is opened, under 70bar pressure,
High-pressure delivery so that reaction system and catalyst phospholipase D system are fully mixed into the mixing module 3 of micro passage reaction
Close, the temperature of mixing module 3 is 45 degrees Celsius, and mixed liquor subsequently enters 100 groups of cascade reaction modules 4, the temperature of reaction module 4
It is 47 degrees Celsius, reaction solution subsequently enters high temperature module 5, is 3 groups of series connection, the temperature of high temperature module 5 is 86 degrees Celsius, reaction solution
Refrigerated module 6 is subsequently entered, is 7 groups of series connection, the temperature of refrigerated module 6 is 24 degrees Celsius, and finally, product connects into product
Receiving apparatus 7.
E- is refined：The product that will be obtained is extracted 2 times with normal heptane, is concentrated with rotavapor under vacuum, removes positive heptan
Alkane, is then placed on vacuum drying oven by product, and under 50 degrees celsius, vacuum drying finally obtains 76g faint yellow solids.Through
HPLC liquid phases detect that in product, phosphatidylserine content is 51.3%, and phosphatidylcholine content is 0.7%, phosphatide acid content
It is 7.8%.
Embodiment 6：The phosphatidylserine of antarctic krill is prepared using micro passage reaction.
(1) device：Micro-channel device is micro- using the self-control heart-shaped structure high-pressure glass high flux of Corning Incorporated's production
Channel reactor, micro-tunnel length determines that heat transferring medium is conduction oil according to flow velocity and reaction time.
(2) preparation of the phosphatidylserine of antarctic krill：
The preparation of A- micro passage reactions：Reactant is controlled by the passage length of the flow and microchannel that adjust delivery pump
The residence time of modules is tied up to, wherein, residence time of mixing module 3 is 30 seconds, and the residence time of reaction module 4 is
300 seconds, the residence time of high temperature module 5 was 20 seconds, and the residence time of refrigerated module 6 is 120 seconds.By outer circulation heat conduction oil body
It is to control the temperature of reactor modules, wherein, the temperature of mixing module 3 is 35 degrees Celsius, the temperature of reaction module 4 is
46 degrees Celsius, the temperature of high temperature module 5 is 80 degrees Celsius, and the temperature of refrigerated module 6 is 35 degrees Celsius.
The preparation of B- reaction systems：In the first material-compound tank 1, by 100g antarctic krill phospholipids raw material (wherein phosphatidyl courage
Alkali content is 41.2%, and phosphatide acid content is 3.5%, and phosphatidylserine content is that 0.4%), 300ml normal heptanes, 50ml is different
Propyl alcohol, 200g serines, 9g calcium chloride is put into 700ml Phosphate buffered saline systems, and the pH value of the aqueous solution is 6.8.Instead
System is answered by 500 mesh membrane filtrations, and passes through 50 Mm filter film process through 5bar pressure, it is ensured that be in removal raw material non-molten
Property solid impurity.
The preparation of C- catalyst phospholipase Ds：In the second material-compound tank 2, the phospholipase D of 10ml is put into 100ml acetic acid
In buffered saline liquid system, the pH value of the aqueous solution is 6.8.And removal of impurities is gone by 0.5 micron of ultrafiltration membrane treatment through 2.5bar pressure
Matter and miscellaneous bacteria.
D- reacts：The high-pressure metering pump that the first material-compound tank 1 and the second material-compound tank 2 will be connected is opened, high under 6bar pressure
Pressure conveying so that reaction system and catalyst phospholipase D system are fully mixed into the mixing module 3 of micro passage reaction
Close, the temperature of mixing module 3 is 30 degrees Celsius, and mixed liquor subsequently enters 10 groups of cascade reaction modules 4, the temperature of reaction module 4
It is 46 degrees Celsius, reaction solution subsequently enters high temperature module 5, is 2 groups of series connection, the temperature of high temperature module 5 is 80 degrees Celsius, reaction solution
3 groups of series connection refrigerated modules 6 are subsequently entered, is 4 groups of series connection, the temperature of refrigerated module 6 is 30 degrees Celsius, and finally, product is entered
Enter product reception device 7.
E- is refined：The product that will be obtained is extracted 2 times with normal heptane, is concentrated with rotavapor under vacuum, removes positive heptan
Alkane, is then placed on vacuum drying oven by product, and under 50 degrees celsius, vacuum drying finally obtains 81g brown-red solids.Through
HPLC liquid phases detect that in product, phosphatidylserine content is 45.8%, and phosphatidylcholine content is 0.8%, phosphatide acid content
It is 4.6%.
Embodiment 7：The phosphatidylserine of milk origin is prepared using micro passage reaction.
(1) device：Micro-channel device using self-control glass high flux micro passage reaction, micro-tunnel length according to flow velocity with
Reaction time determines that heat transferring medium is conduction oil.
(2) preparation of the phosphatidylserine of milk origin：
The preparation of A- micro passage reactions：Reactant is controlled by the passage length of the flow and microchannel that adjust delivery pump
The residence time of modules is tied up to, wherein, the residence time of mixing module 3 is 30 seconds, and the residence time of reaction module 4 is 60
Second, the residence time of high temperature module 5 is 60 seconds, and the residence time of refrigerated module 6 is 100 seconds.By outer circulation heat conduction oil systems
To control the temperature of reactor modules, wherein, the temperature of mixing module 3 is 30 degrees Celsius, and the temperature of reaction module 4 is 45
Degree Celsius, the temperature of high temperature module 5 is 80 degrees Celsius, and the temperature of refrigerated module 6 is 30 degrees Celsius.
The preparation of B- reaction systems：In the first material-compound tank 1, by 100g milk lecithin materials, (wherein phosphatid ylcholine contains
It is 18.7% to measure, and phosphatide acid content is 9.2%, and phosphatidylserine content is 0.6%) 300ml n-hexanes, 50ml isopropanols,
300g serines, 9g calcium chloride is put into 700ml Phosphate buffered saline systems, and the pH value of the aqueous solution is 6.8.Reactant
System passes through 50 Mm filter film process by 500 mesh membrane filtrations through 2bar pressure, it is ensured that insoluble solid in removal raw material
Body impurity.
The preparation of C- catalyst phospholipase Ds：In material-compound tank 2, the phospholipase D of 10ml is put into 100ml acetic acid salt solution
In buffer solution system, the pH value of the aqueous solution is 6.8.And through 2bar pressure by 0.5 micron of ultrafiltration membrane treatment, go the removal of impurity and miscellaneous
Bacterium.
D- reacts：The high-pressure metering pump that material-compound tank 1 and material-compound tank 2 will be connected is opened, under 5bar pressure, high-pressure delivery,
So that reaction system and catalyst phospholipase D system are sufficiently mixed into the mixing module 3 of micro passage reaction, hybrid guided mode
The temperature of block 3 is 30 degrees Celsius, and mixed liquor subsequently enters 7 groups of cascade reaction modules 4, and the temperature of reaction module 4 is 45 degrees Celsius,
Reaction solution subsequently enters high temperature module 5, and the temperature of high temperature module 5 is 80 degrees Celsius, and reaction solution subsequently enters refrigerated module 6, low
The temperature of warm module 6 is 30 degrees Celsius, and finally, product enters product reception device 7.
E- is refined：The product that will be obtained is extracted 2 times with normal heptane, is concentrated with rotavapor under vacuum, removes positive heptan
Alkane, is then placed on vacuum drying oven by product, and under 50 degrees celsius, vacuum drying finally obtains 89g faint yellow solids.Through
HPLC liquid phases detect that in product, phosphatidylserine content is 24.3%, and phosphatidylcholine content is less than 0.3%, and phosphatidic acid is pure
Spend is 10.4%.
Embodiment 8：With the corresponding reference examples of embodiment 1.
(1) device：Using traditional tank reactor, anchor formula agitating paddle is equipped with chuck, and heat transferring medium is conduction oil.
(2) preparation of the phosphatidylserine of soybean-source：
The preparation of A- reaction systems：In the first material-compound tank 1, by 100g soybean lecithins raw material, (wherein phosphatid ylcholine contains
It is 53.7% to measure, and phosphatide acid content is 6.6%, and phosphatidylserine content is 0.4%) to be completely dissolved in 300ml n-hexanes, then
50ml isopropanols are added, 1000ml Phosphate buffered salines (serine containing 300g and 10g calcium chloride), aqueous solution pH is subsequently added
Be worth is 7.0.
The preparation of C- catalyst phospholipase Ds：In the second material-compound tank 2, the phospholipase D of 10ml is put into 100ml phosphoric acid
In buffered saline liquid system, the pH value of the aqueous solution is 7.5.
D- reacts：The measuring pump that the first material-compound tank 1 and the second material-compound tank 2 will be connected is opened so that reaction system and catalysis
Agent phospholipase D system enters tank reactor, and agitator speed is 70rpm, and reaction temperature is 45 degrees Celsius, and reaction stirring is maintained
12h, finally, product enters product reception device 7.
E- is refined：The product that will be obtained is extracted 2 times with 500ml normal heptanes, is concentrated with rotavapor under vacuum, plus the third
Ketone is precipitated, and after the de- organic solvent of filtering decompression, product is placed on into vacuum drying oven, under 50 degrees celsius, vacuum drying, finally
Obtain 89g faint yellow solids.Detect that in product, phosphatidylserine content is 38.2%, and phosphatid ylcholine contains through HPLC liquid phases
It is 0.9% to measure, and phosphatide acid content is 18.4%.
Relative to embodiment 1, phosphatidylserine content is low in the product of embodiment 8, is 38.2%, and phosphatid ylcholine contains
It is 0.9% to measure, and phosphatide acid content is high, is 18.4%.And phosphatidylserine content is 55.7% in the product of embodiment 1, phosphorus
Phosphatidylcholine content is 0.5%, and phosphatide acid content is 6.7%.Illustrate that, using microchannel enzyme technology, same materials can be obtained
To more target product phosphatidylserines, and the content of accessory substance phosphatidic acid is lower.
In this description, the present invention is described with reference to its specific embodiment.But it is clear that can still make
Various modifications and alterations still may be used without departing from the spirit and scope of the present invention, a those of ordinary skill for having this professional knowledge
With the technology taught according to this patent, other embodiments are produced within the scope of the invention, but it is every without departing from this hair
The content of bright technical scheme, any simple modification, the equivalent variations made to above example according to technical spirit of the invention
With modification, still fall within the range of technical solution of the present invention.
Claims (9)
1. a kind of method that use micro passage reaction prepares phosphatidylserine, it is characterised in that prepare by the following steps：
Step one, prepares enzymic catalytic reaction liquid；
Step 2, prepares phospholipase D solution；
Step 3, the phospholipase D solution that the reaction solution and step 2 that step one is prepared are prepared pumps into microchannel by high-pressure pump
Reactor, is reacted upon mixing；
Step 4, after reaction terminates, high temperature passivation phospholipase D activity then makes reaction system flow through refrigerated module, final reaction
System flows out micro passage reaction, obtains phosphatidylserine product.
2. the method for claim 1, it is characterised in that the micro passage reaction include mixing module, reaction module,
High temperature module and refrigerated module, the fluid path length of each module is 0.1m~300m, preferably 1m~60m；Each module
Fluid passage width and depth be 0.5 μm~10mm, preferably 0.1mm~1mm.
3. the method for claim 1, it is characterised in that the proportion relation of each material is such as in reaction solution in the step one
Under：Weight portion of the phosphatid ylcholine in every liter of mixed reaction solution is 100；Weight portion of the serine in every liter of mixed reaction solution
It is 60~300, preferably 200~250；Weight portion of the calcium salt in every liter of reaction solution is 0.05~10, preferably 2~4；C5-C9 alkane
The percent by volume that hydrocarbon organic solvent occupies in total mixed reaction solution is 0~95%；Short hydrocarbon alcohol or short hydrocarbon ketone are mixed always
Possessive volume percentage is 0~80% in closing reaction solution.
4. the method for claim 1, it is characterised in that in the step 3, the reaction solution exists with phospholipase D solution
Before pumping into micro passage reaction, 500 mesh and 0.5 μm of ultrafiltration apparatus need to be sequentially passed through, be filtered to remove solid particle.
5. the method for claim 1, it is characterised in that in the step 3, the reaction solution and phospholipase D solution are logical
The compression pump for crossing 0.5~500bar, preferably 2~50bar enters micro passage reaction.
6. the method as described in claim 1-5 is any, it is characterised in that in the step 3, reaction system enters microchannel
After the reaction module of reactor, reaction temperature is 35~55 degrees Celsius, preferably 45-50 degrees Celsius；Reaction system is in reaction module
In residence time be 30~1200 seconds, preferably 60-300 seconds.
7. the method as described in claim 1-6 is any, it is characterised in that in the step 4, after reaction terminates, makes reaction solution
High temperature module is flowed through, the temperature of high temperature module is 75~105 degrees Celsius, preferably 85~90 degrees Celsius；Reaction solution is in high temperature module
The middle residence time is 10~1200 seconds, preferably 15-120 seconds.
8. the method as described in claim 1-7 is any, it is characterised in that in the step 4, reaction solution is flowing through low temperature mould
During block, the temperature of refrigerated module is 0~40 degree Celsius, preferably 25-35 degree；Reaction solution the refrigerated module residence time be 30~
1200 seconds, preferably 30-300 seconds.
9. the method as described in claim 1-8 is any, it is characterised in that every kind of module of micro passage reaction is the small mould of polylith
Block connects to be formed, wherein reaction module 2-200 blocks, high temperature module 2-20 blocks, refrigerated module 2-20 blocks.
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| CN107034248A (en) * | 2017-06-19 | 2017-08-11 | 成都新柯力化工科技有限公司 | A kind of method of continuous high-efficient enzymatic conversion fabricated food phosphatidylserine |
| CN111073924A (en) * | 2020-01-10 | 2020-04-28 | 福州三合元生物科技有限公司 | Micro-channel continuous synthesis method of vitamin C glucoside |
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| CN206730896U (en) * | 2017-01-13 | 2017-12-12 | 理星（天津）生物科技有限公司 | A kind of high pressure homogenizer |
| CN108271885A (en) * | 2018-02-28 | 2018-07-13 | 理星（天津）生物科技有限公司 | A kind of fruits and vegetables compound powder and its preparation method and application |
| CN115720973A (en) * | 2022-11-21 | 2023-03-03 | 宁夏红果生物科技有限公司 | Whole-fruit Chinese wolfberry fresh fruit puree and preparation method and application thereof |
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- 2017-02-16 CN CN201710083734.4A patent/CN106755154A/en active Pending
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| CN111073924A (en) * | 2020-01-10 | 2020-04-28 | 福州三合元生物科技有限公司 | Micro-channel continuous synthesis method of vitamin C glucoside |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106582340B (en) | 2022-07-12 |
| CN106582340A (en) | 2017-04-26 |
| CN206730896U (en) | 2017-12-12 |
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