CN112592908B - Lipase-supported photocatalyst with magnetic core-shell structure, and preparation method and application thereof - Google Patents
Lipase-supported photocatalyst with magnetic core-shell structure, and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a lipase-supported photocatalyst with a magnetic core-shell structure, a preparation method and application thereof, wherein the catalyst modifies amino core-shell structure Fe with aldehyde group 3 O 4 @Cu 2 S is taken as a substrate, and lipase is taken as an active ingredient; the substrate is made of nano Fe 3 O 4 Is nuclear, aldehyde group modified amino Cu 2 S is a shell. The preparation method comprises the following steps: firstly, synthesizing nano Fe by ferric chloride, ammonium acetate and disodium ethylenediamine tetraacetate 3 O 4 The core is reacted with cuprous acetate and hydrogen sulfide to obtain Fe with a core-shell structure 3 O 4 @Cu 2 S substrate is modified by 3-aminopropyl-triethoxysilane and glutaraldehyde to obtain aldehyde modified amino core-shell structure Fe 3 O 4 @Cu 2 S substrate, adding lipase, and freeze drying to obtain the lipase-carried photocatalyst with magnetic core-shell structure. The catalyst is irradiated by a 5-100W incandescent lamp for 10-30 min, and the yield of the 1, 3-diacylglycerol can be remarkably improved.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a lipase-supported photocatalyst with a magnetic core-shell structure, and a preparation method and application thereof.
Background
Along with the increasing number of people suffering from obesity in the world, the high blood pressure, high blood lipid and high blood sugar brought by obesity threaten the health of human bodies. The world health organization suggests reducing consumption of high fat and fat diets, ingesting healthy fat and balancing various fats. Diacylglycerols (DAG) are an ingredient in natural vegetable oils, also intermediates for lipid metabolism in humans, and have two configurations, 1, 3-diacylglycerols (1, 3-DAG) and 1, 2-diacylglycerols (1, 2-DAG). Experimental study shows that the oil rich in 1,3-DAG can inhibit accumulation of visceral fat, prevent weight increase, and is beneficial to health. In the natural grease, the content of 1,3-DAG is relatively low, and the grease can be modified by adopting a chemical method to improve the content of 1, 3-DAG.
Compared with a chemical method, the process condition for synthesizing the 1,3-DAG by using lipase is mild, the method has higher yield and selectivity, and the obtained product can keep better taste and texture. However, free lipases are generally susceptible to inactivation, poor stability and difficult recovery for reuse, which have prevented their industrial application. To overcome this disadvantage, immobilization techniques are widely used in this field to immobilize lipases. The lipase is immobilized, and the activity, stability and selectivity of the lipase are improved. Chinese patent CN111621494a discloses a preparation method of magnetic immobilized lipase, and the activity of the product can reach more than 80% of the activity of free lipase. That is, the immobilized enzyme causes a decrease in enzyme activity, and the lipase activity decreases significantly with the increase in reaction time.
Takes a magnetic immobilized substrate as a core and Cu as a core 2 S is a shell, a novel photo-enzyme catalyst is synthesized, enzyme can be rapidly separated under the condition of an external magnetic field, and photo-generated electrons improve the catalytic activity and stability of lipase under the condition of illumination, so that the yield and selectivity of diglyceride are improved. Such studies have not been reported.
Disclosure of Invention
In order to overcome the defects and shortcomings in the prior art, the primary aim of the invention is to provide a lipase-supported photocatalyst with a magnetic core-shell structure.
The invention also aims to provide a preparation method of the lipase-supported photocatalyst with the magnetic core-shell structure.
It is still another object of the present invention to provide the use of the lipase-supported photocatalyst of magnetic core-shell structure in the preparation of diacylglycerol.
The aim of the invention is achieved by the following technical scheme.
A lipase-carried photocatalyst with magnetic core-shell structure, which uses aldehyde group to modify amino core-shell structure Fe 3 O 4 @Cu 2 S is taken as a substrate, and lipase is taken as an active ingredient; the substrate is made of nano Fe 3 O 4 Is nuclear, aldehyde group modified amino Cu 2 S is a shell.
Preferably, the mass ratio of the substrate to the lipase is 100 (1-10); the mass ratio of the core shell in the substrate is 1 (10-50).
The preparation method of the lipase-supported photocatalyst with the magnetic core-shell structure comprises the following steps:
(1) Nano Fe 3 O 4 Dispersing in ethanol solution, adding cuprous acetate and hydrogen sulfide water solution, centrifuging, separating precipitate to obtain Fe 3 O 4 @Cu 2 S；
(2) Fe is added to 3 O 4 @Cu 2 S is dispersed in ethanol water solution, 3-aminopropyl-triethoxysilane is added for reaction, and after the reaction, the reaction product is filtered to obtain the amino core-shell structure Fe 3 O 4 @Cu 2 S solid; by amination of Fe of core-shell structure 3 O 4 @Cu 2 S solid is dissolved in sodium phosphate buffer solution, glutaraldehyde water solution is added, stirring, centrifugation, separation and precipitation are carried out, and aldehyde group modified amino core-shell structure Fe is obtained by freeze drying 3 O 4 @Cu 2 S substrate;
(3) Modification of amino-core-shell structure Fe with aldehyde group 3 O 4 @Cu 2 Dispersing S substrate in phosphoric acid buffer solution, adding lipase, stirring, centrifuging, separating solid, and freeze drying to obtain magnetic core-shell structure lipaseIs a photocatalyst of (a).
Preferably, the nano Fe of step (1) 3 O 4 The mass volume ratio of the ethanol solution is 1g to 1-5 mL; the volume fraction of the ethanol solution is 60-80%; the quality of the cuprous acetate is nano Fe 3 O 4 10 to 50 times of the mass of the nucleus; the mass of the hydrogen sulfide aqueous solution is nano Fe 3 O 4 10-100 times of the mass of the nucleus; the mass concentration of the hydrogen sulfide aqueous solution is 20-30%; the centrifugation is carried out at 8000-20000 rpm for 15-30 min.
Preferably, the nano Fe of step (1) 3 O 4 The preparation of the composition comprises the following steps:
dissolving ferric chloride in 30-60 times of ethylene glycol, and adding ammonium acetate 1.5-3 times of ferric chloride and disodium ethylenediamine tetraacetate 0.5-1.5 times of ferric chloride; stirring at 1000-3000 rpm for 2-5 hr, reacting at 150-250 deg.c in sealed reactor for 8-16 hr, naturally cooling, centrifuging at 8000-20000 rpm for 15-30 min, separating precipitate to obtain nanometer Fe 3 O 4 。
Preferably, the Fe of step (2) 3 O 4 @Cu 2 The mass volume ratio of S to the ethanol water solution is 1 g:1-10 mL; the volume fraction of the ethanol water solution is 60-80%; the addition amount of the 3-aminopropyl-triethoxysilane is Fe 3 O 4 @Cu 2 1 to 10 times of the mass of S; the reaction temperature is 50-60 ℃, and the reaction time is 5-6 h; the aminated core-shell structure Fe 3 O 4 @Cu 2 The mass volume ratio of S solid to sodium phosphate buffer solution is 1 g:10-100 mL; the pH of the sodium phosphate buffer solution=6.5 to 8; the addition amount of the glutaraldehyde aqueous solution is amino core-shell structure Fe 3 O 4 @Cu 2 0.2 to 1 time of the mass of S; the mass concentration of the glutaraldehyde aqueous solution is 15-25%; the stirring is carried out for 2-4 hours at 50-500 rpm; the centrifugation is carried out at 8000-20000 rpm for 15-30 min.
Preferably, the aldehyde group modified amino core-shell structure Fe of step (3) 3 O 4 @Cu 2 The mass volume ratio of the S substrate to the phosphoric acid buffer solution is 1 g:1-10 mL; the phosphoric acid is slowPh=6.5-8 of the flushing solution; the addition amount of the lipase is 1-10% of the mass of the substrate; the stirring is carried out for 4 to 8 hours at the temperature of between 0 and 4 ℃ and at the speed of between 50 and 500 rpm; the centrifugation is carried out at 8000-20000 rpm for 15-30 min.
Preferably, the lipase is one or more than one of Novozym435,Lipozyme RMIM,Lipozyme TLIM,Lipase G50,Lipase F-AP.
The application of the lipase-supported photocatalyst with the magnetic core-shell structure in preparing diacylglycerol is provided.
Preferably, the application comprises the steps of:
c is C 12 ～C 18 Mixing fatty acid and glycerin in the molar ratio of 2-10 to 1, adding the lipase-carried photocatalyst with the magnetic core-shell structure, wherein the mass of the mixture is 1-10%, and illuminating a 5-100W incandescent lamp for 10-30 min to obtain the 1, 3-diacylglycerol.
Preferably, the fatty acid is one or more of caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, oleic acid and linoleic acid.
The principle of the invention is as follows: firstly synthesizing Fe with a magnetic core-shell structure by adopting a precipitation-deposition wrapping mode 3 O 4 @Cu 2 S carrier, which is subjected to surface modification through amino and aldehyde groups to obtain Fe with a plurality of fixing sites on the surface 3 O 4 @Cu 2 S carrier, which is advantageous for lipase in modified Fe 3 O 4 @Cu 2 And S is covalently bound on the carrier. Cu (Cu) 2 The S nano-particle is a p-type semiconductor, has a forbidden band width of about 1.2eV, and can absorb visible light to generate electron-hole pairs (h + ，e - ) The band gap and high electron mobility of the photoexcited electron migration have synergistic effect with lipase, thus improving the yield of diacylglycerol.
Compared with the prior art, the invention has the following advantages and effects:
(1) After the lipase is covalently immobilized, the stability of the lipase is improved, and a foundation is laid for subsequent homogeneous reaction.
(2) The invention uses magnetic Fe 3 O 4 The nano particles are cores, and lipase can be conveniently and rapidly separated under the action of an external magnetic field.
(3) The invention uses Cu 2 S is a shell, and photocatalysis and enzyme catalysis are combined, and the synergistic effect of the two can not only not reduce the enzyme activity, but also improve the catalytic activity.
Drawings
FIG. 1a is Fe 3 O 4 TEM image of nanoparticles;
FIG. 1b is Fe 3 O 4 TEM image of the @ CuS nanoparticles;
FIG. 1c is Fe 3 O 4 Particle size distribution map of (2);
FIG. 1d is Fe 3 O 4 Particle size distribution profile of the @ CuS (b) nanoparticles;
FIG. 1e is Fe 3 O 4 CuS and Fe 3 O 4 Infrared spectrogram of @ CuS;
FIG. 1f is Fe 3 O 4 CuS and Fe 3 O 4 XRD pattern of @ CuS;
FIG. 2 is a graph showing the content of 1, 3-diacylglycerol produced by lipase catalysis.
Detailed Description
The present invention will be described in further detail with reference to examples, but embodiments of the present invention are not limited thereto.
Example 1
(1) Ferric chloride is dissolved in glycol with the mass of 30 times of that of the ferric chloride, and ammonium acetate with the mass of 1.5 times of that of the ferric chloride and disodium ethylenediamine tetraacetate with the mass of 0.5 time of that of the ferric chloride are added; stirring at 1000rpm for 5h, reacting at 150 ℃ for 16h in a sealed reaction kettle, naturally cooling, centrifuging at 8000rpm for 30min, and separating precipitate to obtain nano Fe 3 O 4 A core; dispersing the nano Fe in ethanol solution (volume fraction 60%) according to mass-volume ratio of 1:1 (g/mL), and adding nano Fe 3 O 4 10 times of the nuclear mass of cuprous acetate and 10 times of 20% hydrogen sulfide water solution, centrifuging at 8000rpm for 30min, and separating precipitate to obtain Fe 3 O 4 @Cu 2 S。
(2) Fe is added to 3 O 4 @Cu 2 S is dispersed in the mass-volume ratio of 1:1 (g/m)L) ethanol aqueous solution (volume fraction 60%), fe was added 3 O 4 @Cu 2 S is 3-aminopropyl-triethoxysilane with the mass of 1 time, and is filtered after reaction for 6 hours at 50 ℃ to obtain the amino core-shell structure Fe 3 O 4 @Cu 2 S solid; dissolving the mixture in sodium phosphate buffer solution (pH=6.5) in a mass-volume ratio of 1:10 (g/mL), and adding the amino core-shell structure Fe 3 O 4 @Cu 2 Glutaraldehyde aqueous solution (mass fraction 25%) with the mass of 0.2 times of S is stirred at 50rpm for 4h at room temperature, centrifuged at 8000rpm for 30min, separated and precipitated, and freeze-dried to obtain aldehyde modified amino core-shell structure Fe 3 O 4 @Cu 2 S substrate.
(3) Aldehyde group modified amino core-shell structure Fe 3 O 4 @Cu 2 S substrate is dispersed in phosphoric acid buffer solution (pH=6.5) in a mass-volume ratio of 1:1 (g/mL), lipase Novozym with the mass of 1% of the substrate is added, 50rpm is used for stirring for 8 hours at 435,0 ℃, and 8000rpm is used for centrifugation for 30 minutes, solids are separated, and the magnetic core-shell structure lipase-carrying photocatalyst is obtained through freeze drying.
(4) Oleic acid and glycerin are mixed in a molar ratio of 10:1, a magnetic core-shell structure lipase-carrying photocatalyst with a mass of 1% of the mixture is added, 50W incandescent light irradiates for 10min, and the yield of 1, 3-diacylglycerol is 68%.
Example 2
(1) Ferric chloride is dissolved in glycol with the mass of 60 times of that of the ferric chloride, and ammonium acetate with the mass of 3 times of that of the ferric chloride and disodium ethylenediamine tetraacetate with the mass of 1.5 times of that of the ferric chloride are added; stirring at 3000rpm for 2h, reacting at 250deg.C in a sealed reaction kettle for 8h, naturally cooling, centrifuging at 20000rpm for 15min, and separating precipitate to obtain nanometer Fe 3 O 4 A core; dispersing the nano Fe in ethanol solution (volume fraction 80%) according to mass-volume ratio of 1:5 (g/mL), and adding nano Fe 3 O 4 50 times of the nuclear mass of cuprous acetate and 100 times of 30% hydrogen sulfide aqueous solution by mass concentration, centrifuging at 20000rpm for 15min, separating precipitate to obtain Fe 3 O 4 @Cu 2 S。
(2) Fe is added to 3 O 4 @Cu 2 S was dispersed in an aqueous ethanol solution (volume fraction 80%) having a mass to volume ratio of 1:10 (g/mL), and Fe was added 3 O 4 @Cu 2 S is 3-aminopropyl-triethoxysilane with the mass of 10 times, and is filtered after being reacted for 5 hours at 60 ℃ to obtain the amino core-shell structure Fe 3 O 4 @Cu 2 S solid; dissolving the mixture in sodium phosphate buffer solution (pH=8) in a mass-volume ratio of 1:100 (g/mL), and adding the amino core-shell structure Fe 3 O 4 @Cu 2 S is glutaraldehyde aqueous solution (mass fraction 15%) with 1 time of the mass, stirring is carried out for 2h at 500rpm at room temperature, centrifugation is carried out for 15min at 20000rpm, precipitation is separated, and freeze drying is carried out, thus obtaining aldehyde group modified amino core-shell structure Fe 3 O 4 @Cu 2 S substrate.
(3) Aldehyde group modified amino core-shell structure Fe 3 O 4 @Cu 2 S substrate is dispersed in phosphoric acid buffer solution (pH=8) in a mass-volume ratio of 1:10 (g/mL), lipase RMIM with 5% of substrate mass and lipase TLIM with 5% of substrate mass are added, stirring is carried out at 500rpm for 4h at 4 ℃, centrifugation is carried out at 20000rpm for 15min, solid is separated, and the magnetic core-shell structure lipase-carried photocatalyst is obtained by freeze drying.
(4) Palmitic acid and glycerin are mixed in a molar ratio of 5:1, a magnetic core-shell structure lipase-carrying photocatalyst with the mass of 10% of the mixture is added, 5W incandescent light is irradiated for 30min, and the yield of 1, 3-diacylglycerol is 87%.
Example 3
(1) Ferric chloride is dissolved in glycol with the mass being 45 times of that of the ferric chloride, and ammonium acetate with the mass being 2 times of that of the ferric chloride and disodium ethylenediamine tetraacetate with the mass being 1 time of that of the ferric chloride are added; stirring at 2000rpm for 3h, reacting at 200deg.C in a sealed reaction kettle for 12h, naturally cooling, centrifuging at 10000rpm for 20min, separating precipitate to obtain nanometer Fe 3 O 4 A core; dispersing the nano Fe in ethanol solution (volume fraction 70%) according to a mass-to-volume ratio of 1:3 (g/mL), and adding nano Fe 3 O 4 The method comprises the steps of (1) centrifuging copper acetate with a nuclear mass of 30 times and hydrogen sulfide aqueous solution with a mass concentration of 25% with 10000rpm for 20min, separating precipitate to obtain Fe 3 O 4 @Cu 2 S。
(2) Fe is added to 3 O 4 @Cu 2 S is dispersed in ethanol water solution (volume fraction 70%) with the mass-volume ratio of 1:5g/mL, and Fe is added 3 O 4 @Cu 2 S mass 5 times of 3-aminopropylReacting the base-triethoxysilane for 5.5 hours at 55 ℃, and filtering to obtain the amino core-shell structure Fe 3 O 4 @Cu 2 S solid; dissolving the mixture in sodium phosphate buffer solution (pH=7) in a mass-volume ratio of 1:50 (g/mL), and adding the amino core-shell structure Fe 3 O 4 @Cu 2 Glutaraldehyde aqueous solution (mass fraction 20%) with the mass of 0.5 times of S is stirred for 3h at 300rpm at room temperature, centrifuged for 20min at 10000rpm, separated and precipitated, and freeze-dried to obtain aldehyde modified amino core-shell structure Fe 3 O 4 @Cu 2 S substrate.
(3) Aldehyde group modified amino core-shell structure Fe 3 O 4 @Cu 2 S substrate is dispersed in phosphoric acid buffer solution (pH=7) in a mass-volume ratio of 1:5 (G/mL), lipase G50 with 3% of substrate mass and Lipase F-AP with 4% of substrate mass are added, stirring is carried out at 200rpm for 6h at 2 ℃, centrifugation is carried out at 10000rpm for 20min, solid is separated, and the magnetic core-shell structure Lipase-carried photocatalyst is obtained through freeze drying.
(4) Octanoic acid and glycerin are mixed in a molar ratio of 5:1, a magnetic core-shell structure lipase-carrying photocatalyst with the mass of 5% of the mixture is added, 100W incandescent light irradiates for 10min, and the yield of 1, 3-diacylglycerol is 73%.
Test 1
Core-shell structural characterization of the products prepared in examples 1-3
The method comprises the following steps: mixing a proper amount of the product prepared in the experimental examples 1-3 with potassium bromide, tabletting, and carrying out infrared spectrum scanning; the sample was dispersed in water and then subjected to particle size analysis, and another sample was taken for X-ray diffraction (XRD) analysis and Transmission Electron Microscope (TEM) observation.
Results: FIGS. 1a and 1b are Fe, respectively 3 O 4 And Fe (Fe) 3 O 4 TEM image of @ CuS nanoparticles, fe 3 O 4 The nanoparticles are burr-like spheres with an average particle size of 105nm (see fig. 1c and 1 d); in the coating of Cu 2 After the S shell, it was found that the particle size of the particles became large; FIGS. 1e and 1f are Fe, respectively 3 O 4 、Cu 2 S and Fe 3 O 4 @Cu 2 S infrared spectrum and XRD pattern, fe 3 O 4 Infrared spectrogram of @ CuS 1200cm -1 One is obviously appeared atThe sharp peak, XRD, also shows characteristic peaks of two substances, which indicates Fe 3 O 4 The @ CuS has a heterostructure, combined with a TEM image. These characterizations illustrate synthesized Fe 3 O 4 The @ CuS has a core-shell structure.
Test 2
Catalytic Activity measurement of the lipase-carried photocatalyst having the magnetic core-Shell Structure prepared in example 1
The method comprises the following steps: mixing oleic acid and glycerin at a molar ratio of 10:1, adding the lipase-carrying photocatalyst with a magnetic core-shell structure, which is prepared in example 1, in an amount of 1% by mass of the mixture, illuminating 50W incandescent light for 10min, and measuring the content of 1, 3-diglyceride in the reaction product by using high performance liquid chromatography equipped with a low-temperature evaporative light scattering detector.
Results: FIG. 2 shows lipase/Fe in columns 1-6 from left to right 3 O 4 @Cu 2 S (no light), lipase/Fe 3 O 4 @Cu 2 S (with illumination), cu 2 S-carried lipase, fe 3 O 4 Lipase-carrying free lipase and pure carrier Fe 3 O 4 @Cu 2 Comparison of the content of 1,3-DAG generated by S catalysis shows that the lipase is subjected to Fe 3 O 4 @Cu 2 After S is fixed, the catalytic activity is obviously improved and is obviously better than Cu 2 S-carried lipase, fe 3 O 4 Lipase-carrying free lipase and pure carrier Fe 3 O 4 @Cu 2 S, S; lipase/Fe under illumination 3 O 4 @Cu 2 The S catalyst has synergistic effect of photocatalysis and enzyme catalysis, and the activity of the catalyst is further improved, which indicates that the photocatalysis and the enzyme catalysis have synergistic effect.
Comparative example 1 (increasing enzyme dosage)
This comparative example differs from example 1 in that: the amount of lipase Novozym435 in step (3) was 20% of the substrate. The catalytic activity was determined according to test 2.
As a result, the content of 1, 3-diglyceride which is the catalytic product was 45.2%, and the yield was lowered because the amount of the lipase Novozym435 was too large to be completely immobilized, resulting in the formation of excessive free lipase.
Comparative example 2 (reduction of the Shell in core-Shell Structure)
This comparative example differs from example 1 in that: the quality of the cuprous acetate in the step (1) is nano Fe 3 O 4 The nuclear mass is 5 times. The catalytic activity was determined according to test 2.
As a result, the content of 1, 3-diglyceride which is a catalytic product was 39.6%, and the yield was lowered because the proportion of the shell in the core-shell structure was too low, resulting in a low photocatalytic activity, and thus the effect of lipase could not be activated effectively.
The above examples of the present invention are merely illustrative of the present invention and are not intended to limit the embodiments of the present invention. Other variations or modifications of the above teachings will be apparent to those of ordinary skill in the art. It is not necessary here nor is it exhaustive of all embodiments. Any modification, equivalent replacement, improvement, etc. which come within the spirit and principles of the invention are desired to be protected by the following claims.
Claims (8)
1. A lipase-supported photocatalyst with a magnetic core-shell structure is characterized in that the photocatalyst is prepared by modifying amino core-shell structure Fe with aldehyde groups 3 O 4 @Cu 2 S is taken as a substrate, and lipase is taken as an active ingredient; the substrate is made of nano Fe 3 O 4 As a nucleus, aldehyde group is used for modifying amino Cu 2 S is a shell;
the lipase-supported photocatalyst with the magnetic core-shell structure is prepared by the following steps:
(1) Nano Fe 3 O 4 Dispersing in ethanol solution, adding cuprous acetate and hydrogen sulfide water solution, centrifuging, separating precipitate to obtain Fe 3 O 4 @Cu 2 S；
(2) Fe is added to 3 O 4 @Cu 2 S is dispersed in ethanol water solution, 3-aminopropyl-triethoxysilane is added for reaction, and after the reaction, the reaction product is filtered to obtain the amino core-shell structure Fe 3 O 4 @Cu 2 S solid; amination ofCore-shell structure Fe 3 O 4 @Cu 2 S solid is dissolved in sodium phosphate buffer solution, glutaraldehyde water solution is added, stirring, centrifugation, separation and precipitation are carried out, and aldehyde group modified amino core-shell structure Fe is obtained by freeze drying 3 O 4 @Cu 2 S substrate;
(3) Modification of amino-core-shell structure Fe with aldehyde group 3 O 4 @Cu 2 S substrate is dispersed in phosphoric acid buffer solution, then lipase is added, stirring and centrifugation are carried out, solid is separated, and the photo-enzyme catalyst of magnetic core-shell structure carrying lipase is obtained through freeze drying;
the nano Fe in the step (1) 3 O 4 The mass volume ratio of the ethanol solution is 1g to 1-5 mL; the volume fraction of the ethanol solution is 60-80%; the quality of the cuprous acetate is nano Fe 3 O 4 10 to 50 times of the mass of the nucleus; the mass of the hydrogen sulfide aqueous solution is nano Fe 3 O 4 10-100 times of the mass of the nucleus; the mass concentration of the hydrogen sulfide aqueous solution is 20-30%; the centrifugation is carried out at 8000-20000 rpm for 15-30 min;
the Fe of step (2) 3 O 4 @Cu 2 The mass volume ratio of S to the ethanol water solution is 1 g:1-10 mL; the volume fraction of the ethanol water solution is 60-80%; the addition amount of the 3-aminopropyl-triethoxysilane is Fe 3 O 4 @Cu 2 1 to 10 times of the mass of S; the reaction temperature is 50-60 ℃, and the reaction time is 5-6 h; the aminated core-shell structure Fe 3 O 4 @Cu 2 The mass volume ratio of S solid to sodium phosphate buffer solution is 1 g:10-100 mL; the pH of the sodium phosphate buffer solution=6.5 to 8; the addition amount of the glutaraldehyde aqueous solution is amino core-shell structure Fe 3 O 4 @Cu 2 0.2 to 1 time of the mass of S; the mass concentration of the glutaraldehyde aqueous solution is 15-25%; the stirring is carried out for 2-4 hours at 50-500 rpm; the centrifugation is carried out at 8000-20000 rpm for 15-30 min;
the addition amount of the lipase is 1-10% of the mass of the substrate.
2. The lipase-supported photocatalyst with a magnetic core-shell structure according to claim 1, wherein the mass ratio of the substrate to the lipase is 100 (1-10); the mass ratio of the core shell in the substrate is 1 (10-50).
3. The method for preparing the lipase-supported photocatalyst with the magnetic core-shell structure as claimed in claim 1 or 2, which is characterized by comprising the following steps:
(1) Nano Fe 3 O 4 Dispersing in ethanol solution, adding cuprous acetate and hydrogen sulfide water solution, centrifuging, separating precipitate to obtain Fe 3 O 4 @Cu 2 S；
(2) Fe is added to 3 O 4 @Cu 2 S is dispersed in ethanol water solution, 3-aminopropyl-triethoxysilane is added for reaction, and after the reaction, the reaction product is filtered to obtain the amino core-shell structure Fe 3 O 4 @Cu 2 S solid; by amination of Fe of core-shell structure 3 O 4 @Cu 2 S solid is dissolved in sodium phosphate buffer solution, glutaraldehyde water solution is added, stirring, centrifugation, separation and precipitation are carried out, and aldehyde group modified amino core-shell structure Fe is obtained by freeze drying 3 O 4 @Cu 2 S substrate;
(3) Modification of amino-core-shell structure Fe with aldehyde group 3 O 4 @Cu 2 S substrate is dispersed in phosphoric acid buffer solution, then lipase is added, stirring and centrifugation are carried out, solid is separated, and the photo-enzyme catalyst of magnetic core-shell structure carrying lipase is obtained through freeze drying;
the nano Fe in the step (1) 3 O 4 The mass volume ratio of the ethanol solution is 1g to 1-5 mL; the volume fraction of the ethanol solution is 60-80%; the quality of the cuprous acetate is nano Fe 3 O 4 10 to 50 times of the mass of the nucleus; the mass of the hydrogen sulfide aqueous solution is nano Fe 3 O 4 10-100 times of the mass of the nucleus; the mass concentration of the hydrogen sulfide aqueous solution is 20-30%; the centrifugation is carried out at 8000-20000 rpm for 15-30 min;
the Fe of step (2) 3 O 4 @Cu 2 The mass volume ratio of S to the ethanol water solution is 1 g:1-10 mL; the ethanol aqueous solutionThe volume fraction of (2) is 60-80%; the addition amount of the 3-aminopropyl-triethoxysilane is Fe 3 O 4 @Cu 2 1 to 10 times of the mass of S; the reaction temperature is 50-60 ℃, and the reaction time is 5-6 h; the aminated core-shell structure Fe 3 O 4 @Cu 2 The mass volume ratio of S solid to sodium phosphate buffer solution is 1 g:10-100 mL; the pH of the sodium phosphate buffer solution=6.5 to 8; the addition amount of the glutaraldehyde aqueous solution is amino core-shell structure Fe 3 O 4 @Cu 2 0.2 to 1 time of the mass of S; the mass concentration of the glutaraldehyde aqueous solution is 15-25%; the stirring is carried out for 2-4 hours at 50-500 rpm; the centrifugation is carried out at 8000-20000 rpm for 15-30 min;
the addition amount of the lipase is 1-10% of the mass of the substrate.
4. The method according to claim 3, wherein the nano Fe in the step (1) 3 O 4 The preparation of the composition comprises the following steps:
dissolving ferric chloride in 30-60 times of ethylene glycol, and adding ammonium acetate 1.5-3 times of ferric chloride and disodium ethylenediamine tetraacetate 0.5-1.5 times of ferric chloride; stirring at 1000-3000 rpm for 2-5 hr, reacting at 150-250 deg.c in sealed reactor for 8-16 hr, naturally cooling, centrifuging at 8000-20000 rpm for 15-30 min, separating precipitate to obtain nanometer Fe 3 O 4 。
5. The method according to claim 3, wherein the aldehyde-modified aminated core-shell structure Fe in the step (3) 3 O 4 @Cu 2 The mass volume ratio of the S substrate to the phosphoric acid buffer solution is 1 g:1-10 mL; the pH of the phosphate buffer solution=6.5 to 8; the stirring is carried out for 4 to 8 hours at the temperature of between 0 and 4 ℃ and at the speed of between 50 and 500 rpm; the centrifugation is carried out at 8000-20000 rpm for 15-30 min.
6. The method according to claim 3, wherein the Lipase is one or more of Novozym435, lipozyme RMIM, lipozyme TLIM, lipase G50, lipase F-AP.
7. Use of a magnetic core-shell structured lipase-supported photocatalyst according to claim 1 or 2 for the preparation of diacylglycerols.
8. The use according to claim 7, characterized by the steps of:
mixing C12-C18 fatty acid and glycerin in a molar ratio of 2-10:1, adding a lipase-supported photocatalyst with a magnetic core-shell structure, wherein the mass of the mixture is 1-10%, and illuminating a 5-100W incandescent lamp for 10-30 min to obtain the 1, 3-diacylglycerol.
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