CN109837271A - A kind of load enzyme filler and its preparation method and application - Google Patents
A kind of load enzyme filler and its preparation method and application Download PDFInfo
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- CN109837271A CN109837271A CN201811646651.2A CN201811646651A CN109837271A CN 109837271 A CN109837271 A CN 109837271A CN 201811646651 A CN201811646651 A CN 201811646651A CN 109837271 A CN109837271 A CN 109837271A
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- colloidal sol
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- 108090000790 Enzymes Proteins 0.000 title claims abstract description 164
- 102000004190 Enzymes Human genes 0.000 title claims abstract description 164
- 239000000945 filler Substances 0.000 title claims abstract description 151
- 238000002360 preparation method Methods 0.000 title claims abstract description 30
- 238000005507 spraying Methods 0.000 claims abstract description 66
- 238000000034 method Methods 0.000 claims abstract description 42
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims abstract description 21
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims abstract description 21
- 238000006243 chemical reaction Methods 0.000 claims abstract description 13
- 150000002148 esters Chemical group 0.000 claims abstract description 5
- 239000007921 spray Substances 0.000 claims description 23
- 229910052751 metal Inorganic materials 0.000 claims description 14
- 239000002184 metal Substances 0.000 claims description 14
- 238000001035 drying Methods 0.000 claims description 5
- 230000003197 catalytic effect Effects 0.000 claims description 3
- 238000000066 reactive distillation Methods 0.000 claims description 3
- 238000005809 transesterification reaction Methods 0.000 claims description 3
- 230000002255 enzymatic effect Effects 0.000 abstract description 17
- 230000000694 effects Effects 0.000 abstract description 16
- 238000001802 infusion Methods 0.000 abstract description 6
- 238000004821 distillation Methods 0.000 abstract description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 18
- 239000011259 mixed solution Substances 0.000 description 16
- 230000000052 comparative effect Effects 0.000 description 14
- 239000000243 solution Substances 0.000 description 13
- 229940053652 fluorinse Drugs 0.000 description 10
- PUZPDOWCWNUUKD-UHFFFAOYSA-M sodium fluoride Chemical compound [F-].[Na+] PUZPDOWCWNUUKD-UHFFFAOYSA-M 0.000 description 10
- 239000005457 ice water Substances 0.000 description 9
- LFQCEHFDDXELDD-UHFFFAOYSA-N tetramethyl orthosilicate Chemical compound CO[Si](OC)(OC)OC LFQCEHFDDXELDD-UHFFFAOYSA-N 0.000 description 8
- 239000002202 Polyethylene glycol Substances 0.000 description 6
- 229920001223 polyethylene glycol Polymers 0.000 description 6
- YUYCVXFAYWRXLS-UHFFFAOYSA-N trimethoxysilane Chemical compound CO[SiH](OC)OC YUYCVXFAYWRXLS-UHFFFAOYSA-N 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 108010031797 Candida antarctica lipase B Proteins 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 238000006555 catalytic reaction Methods 0.000 description 4
- 238000000576 coating method Methods 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 238000005054 agglomeration Methods 0.000 description 3
- 230000002776 aggregation Effects 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 239000011248 coating agent Substances 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- XVOWIUNYLLPRPG-UHFFFAOYSA-N C(CCCCCCCCCCCCCCC)(=O)O.[N+](=O)([O-])C1=CC=CC=C1 Chemical compound C(CCCCCCCCCCCCCCC)(=O)O.[N+](=O)([O-])C1=CC=CC=C1 XVOWIUNYLLPRPG-UHFFFAOYSA-N 0.000 description 2
- 239000000919 ceramic Substances 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000012856 packing Methods 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 238000013112 stability test Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 239000002699 waste material Substances 0.000 description 2
- BTJIUGUIPKRLHP-UHFFFAOYSA-N 4-nitrophenol Chemical compound OC1=CC=C([N+]([O-])=O)C=C1 BTJIUGUIPKRLHP-UHFFFAOYSA-N 0.000 description 1
- 206010054949 Metaplasia Diseases 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 239000003462 bioceramic Substances 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000009514 concussion Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000010931 ester hydrolysis Methods 0.000 description 1
- 125000004494 ethyl ester group Chemical group 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000015689 metaplastic ossification Effects 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 238000011056 performance test Methods 0.000 description 1
- 229910052573 porcelain Inorganic materials 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
Landscapes
- Enzymes And Modification Thereof (AREA)
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
Abstract
The present invention relates to a kind of load enzyme filler and its preparation method and application, the preparation method for carrying enzyme filler is that will contain enzyme colloidal sol by spray coating method to be sprayed on filler surface, obtains the load enzyme filler；The load enzyme filler that the enzymatic activity and stability of the load enzyme filler that preparation method of the present invention obtains are obtained compared with infusion process significantly improves, after storing 50 days at room temperature, its loss of enzyme activity < 5%, and the enzymatic activity of its ester exchange reaction distillation process for being catalyzed ethyl acetate and n-butanol is 599-621U/g.
Description
Technical field
The present invention relates to enzyme catalysis fields, and in particular to a kind of load enzyme filler and its preparation method and application.
Background technique
Immobilised enzymes is the mobility that enzyme is reduced with physics or chemical method.General stability increases after enzyme immobilization
Add, easily separated from reaction system, and is easily controllable, it can repeated multiple times use.It is readily transported and stores, be conducive to automatic metaplasia
It produces, but activity reduces, use scope reduces.
CN105176962A discloses a kind of method of bio-enzyme immobilization, in particular to a kind of biological enzyme is fixed on pottery
Method in porcelain filling.This method comprises: Step 1: the process that biological enzyme is handled；Step 2: ceramic packing is carried out
The process of processing；Step 3: treated biological enzyme and ceramic packing to be mixed to prepare to the process of bioceramic filler；This scheme
Load enzyme filler being unevenly distributed in filler surface of preparation is unable to satisfy reality to cause the activity for carrying enzyme filler insufficient
Production needs.
CN107115889A discloses a kind of reactive distillation enzymatic filler and its coating method and application.The enzyme
Catalytic filler is made by following methods, comprising the following steps: is immersed in the colloidal sol containing biological enzyme filler 20~30 seconds, is taken
It is 10~20 seconds dry after out；Then it is submerged into again in colloidal sol 20~30 seconds, it is 10~20 seconds dry after taking out again；It repeats to soak
Not yet, drying steps 8~15 times；Finally paddle dryer is filled out to weight is constant to get to the enzymatic for being attached with biological enzyme coating
Material；The enzyme for the load enzyme filler that this scheme obtains and the binding force of filler are insufficient, reaction process or when storing at room temperature, carry enzyme filler
Mass loss it is larger.
CN106955736A discloses the coating in situ of biocatalysis coating in a kind of enzymic catalytic reaction rectifying and updates
Method.This method cleans the filler inside tower body by sodium hydroxide solution first, and the biological enzyme after inactivation is washed off；Again from tower
Bottom is coated in new degumming enzyme on filler in a manner of being bubbled；This scheme provides just for the replacing options that optimization carries enzyme filler
Benefit, but the method for not providing the stability deficiency and poor activity that solve to carry enzyme filler.
Although above-mentioned document discloses some preparation methods for carrying enzyme filler, but there are still the load enzyme fillers being prepared
Activity it is insufficient, the problem of stability difference, therefore develop a kind of with preferable enzymatic activity and excellent in stability carries enzyme filler
Preparation method is still of great significance.
Summary of the invention
The purpose of the present invention is to provide a kind of preparation method and purposes for carrying enzyme filler, the method is to pass through spray coating method
Enzyme colloidal sol will be contained and be sprayed on filler surface, obtain the load enzyme filler；The load enzyme filler that preparation method of the present invention obtains
The load enzyme filler that activity and stability are obtained compared with infusion process significantly improves, after storing 50 days at room temperature, loss of enzyme activity <
5% (illustratively including 1%, 2%, 3%, 4% or 4.5% etc.), and it is catalyzed ethyl acetate and the transesterification of n-butanol is anti-
The enzymatic activity for answering distillation process is 599-621U/g.
In order to achieve that object of the invention, the invention adopts the following technical scheme:
In a first aspect, the method is that will be contained by spray coating method the present invention provides a kind of preparation method for carrying enzyme filler
Enzyme colloidal sol is sprayed on filler surface, obtains the load enzyme filler.
The preparation method of the present invention for carrying enzyme filler, will contain enzyme colloidal sol by spray coating method and be sprayed on filler surface, obtain
To the load enzyme filler, the stability for carrying enzyme filler and catalytic activity are obviously mentioned compared to load enzyme filler prepared by infusion process
Height, after the load enzyme filler that preparation method of the present invention obtains stores 50 days at room temperature, the loss < 5% of enzymatic activity, and
The enzymatic activity of its ester exchange reaction distillation process for being catalyzed ethyl acetate and n-butanol is 599-621U/g.
Preferably, the air pressure of the spraying be 0.1-0.3MPa, such as 0.1MPa, 0.15MPa, 0.2MPa, 0.25MPa or
0.3MPa etc., preferably 0.15-0.25MPa.
Above-mentioned air pressure is used during spraying of the present invention, and it is higher can both to have guaranteed that spraying process colloidal sol containing enzyme had
Fogging degree, and can be to avoid the generation of the agglomeration when spraying to filler surface of colloidal sol containing enzyme；It is molten containing enzyme to obtain
The effect that glue is evenly distributed in filler surface.
During spraying of the present invention, as spraying air pressure < 0.1MPa, colloidal sol containing enzyme is atomized during spraying
Degree is low, and the colloidal sol containing enzyme of ejection is watery or drop-wise, causes spraying uneven, the immobilization effect being not achieved；Work as spray
When applying air pressure > 0.3MPa, because the impact force of spraying process is larger, agglomeration can occur for the filler after spraying, to influence
Carry the activity of enzyme filler.
Preferably, the flow of the spraying is 1.5-2.5kg/h, such as 1.5kg/h, 1.6kg/h, 1.7kg/h, 1.8kg/
H, 1.9kg/h, 2kg/h, 2.1kg/h, 2.2kg/h, 2.3kg/h, 2.4kg/h or 2.5kg/h etc., preferably 1.9-2.3kg/h.
The flow control of spraying of the present invention can guarantee that spraying colloidal sol has foot in filler surface in above-mentioned range of flow
Enough adhesive forces, while avoiding the waste of colloidal sol；As the flow < 1.5kg/h of spraying, colloidal sol containing enzyme is in the glutinous of filler surface
Attached power is poor, and the stability for carrying enzyme filler is insufficient, and the performance of reuse is poor, as the flow > 2.5kg/h of spraying, then will lead to
The waste of the colloidal sol containing enzyme.
Preferably, during the spraying distance of the spray gun apart from filler surface be 10-25cm, such as 10cm, 12cm,
15cm, 17cm, 20cm, 22cm or 25cm etc., preferably 15-20cm.
The distance of spraying of the present invention uses above-mentioned distance range, and enzyme can be sprayed unevenly and contained to avoid colloidal sol containing enzyme
The low problem of colloidal sol utilization rate；As spray distance < 10cm, the colloidal sol containing enzyme of spraying can reunite in filler surface, thus
The stability and activity of load enzyme filler are influenced, as spray distance > 25cm, it will cause 30% or more colloidal sols containing enzyme can not
Filler surface is reached, the utilization rate of colloidal sol containing enzyme is caused to decline.
Preferably, direction is sprayed during the spraying perpendicular to filler surface.
Preferably, filler is fixed on mechanical rot with wire mesh during the spraying.
Preferably, the mechanical rot carries out uniform rotation.
Preferably, the filler includes metal Θ ring filler.
Preferably, it is described spraying the following steps are included:
(1) filler is fixed on mechanical rot, adjustment revolving speed makes its uniform rotation；
(2) enzyme colloidal sol will be contained using spray gun and is sprayed on filler surface, and obtain the load enzyme filler.
Preferably, step (2) further includes that filler surface is dried during spraying.
Preferably, the method for the drying is dried up including the use of hair dryer.
The method of the invention will contain enzyme colloidal sol using spray coating method and be sprayed on filler surface, and the colloidal sol containing enzyme of filler surface exists
It is dried to obtain the filler that load has the gel containing enzyme during spraying.
Preferably, the colloidal sol containing enzyme preparation the following steps are included:
(a) methyl orthosilicate, trimethoxy silane and solvent are mixed, obtains the first mixed solution；
(b) Fluorinse, polyethylene glycol, enzyme solutions and water are mixed, obtains the second mixed solution；
(c) the first mixed solution is added in the second mixed solution, obtains colloidal sol containing enzyme.
Preferably, step (a) further includes being stirred the first mixed solution in ice-water bath.
Preferably, the solvent includes methanol.
Preferably, the mass ratio of the methyl orthosilicate, trimethoxy silane and methanol is (1-3): (6-8): (7-9),
Such as 1:6:9,1.5:7:8.6,2.03:7.2:8.49 or 3:8:7 etc., preferably 2.03:7.2:8.49.
Preferably, step (b) further includes being stirred the second mixed solution in ice-water bath.
Preferably, the concentration of the Fluorinse is 0.5-1.5mol/L, such as 0.5mol/L, 0.7mol/L, 1mol/
L, 1.2mol/L or 1.5mol/L etc., preferably 1mol/L.
Preferably, the enzyme solutions are candida antarctica lipase B (CALB) enzyme solutions.
Preferably, the protein content of the enzyme solutions be 9-12mg/mL, such as 9mg/mL, 10mg/mL, 10.9mg/mL or
12mg/mL etc., preferably 10.9mg/mL.
Preferably, the mass ratio of the Fluorinse, polyethylene glycol, enzyme solutions and water is 1:(2-4): (1-3): (3-
5), such as 1:2:3:3,1:3:2:3.4 or 1:4:1:5 etc., preferably 1:3:2:3.4.
Preferably, the mass ratio of the methyl orthosilicate and Fluorinse is (1-3): 1, such as 1:1,2.03:1 or 3:
1 etc., preferably 2.03:1.
Preferably, step (c) further include by the second mixed solution be added the first mixed solution after, obtained mixing is molten
Liquid is stirred in ice-water bath.
Preferably, the time of step (c) stirring is 2-6min, such as 2min, 3min, 4min, 5min or 6min etc..
As currently preferred technical solution, the preparation method of the colloidal sol containing enzyme the following steps are included:
(a) methyl orthosilicate, trimethoxy silane and methanol are mixed in ice-water bath, stirs, it is molten obtains the first mixing
Liquid, the methyl orthosilicate, trimethoxy silane and methanol quality ratio are (1-3): (6-8): (7-9)；
(b) Fluorinse, polyethylene glycol, CALB enzyme solutions and water are mixed in ice-water bath, is stirred, obtain the
Two mixed solutions, the concentration of the Fluorinse are 0.5-1.5mol/L, and the protein content of the CALB enzyme solutions is 9-
12mg/mL, Fluorinse, polyethylene glycol, enzyme solutions and water mass ratio be 1:(2-4): (1-3): (3-5)；
(c) the first mixed solution is added in the second mixed solution, gained mixed solution is stirred 2- in ice-water bath
6min obtains colloidal sol containing enzyme.
As currently preferred technical solution, it is described carry enzyme filler preparation method the following steps are included:
(1) metal Θ ring filler is fixed on mechanical rot using wire mesh, adjustment revolving speed makes its uniform rotation；
(2) enzyme colloidal sol will be contained using spray gun and is sprayed on metal Θ ring filler surface, using blowing during the spraying
Machine dries up metal Θ ring filler surface, obtains the load enzyme filler, and the air pressure of the spraying is 0.1-0.3MPa, the spraying
Flow be 1.5-2.5kg/h, during the spraying distance of the spray gun apart from filler surface be 10-25cm.
Second aspect, the present invention also provides the load enzyme fillers that method as described in relation to the first aspect is prepared.
The third aspect, the present invention also provides the purposes for carrying enzyme filler as described in second aspect, the load enzyme filler is used for
Catalyzed transesterification rectifying.
Preferably, the ester exchange reaction rectifying for carrying enzyme filler and being used to be catalyzed ethyl acetate and n-butanol.
Compared with the existing technology, the invention has the following advantages:
(1) the load enzyme that the activity and stability for the load enzyme filler that preparation method of the present invention obtains are obtained compared with infusion process is filled out
Material significantly improves, and after storing 50 days at room temperature, loss of enzyme activity < 5% (illustratively includes 1%, 2%, 3%, 4%
Or 4.5% etc.), and the enzymatic activity of its ester exchange reaction distillation process for being catalyzed ethyl acetate and n-butanol is 599-621U/g；
(2) preparation process of preparation method of the present invention is simple, is easy to industrial applications.
Detailed description of the invention
Fig. 1 is the schematic diagram that the preparation method of enzyme filler is carried described in the specific embodiment of the invention；
Fig. 2 is the load enzyme filler that the embodiment of the present invention 1 is prepared.
Specific embodiment
The technical scheme of the invention is further explained by means of specific implementation.Those skilled in the art should be bright
, the described embodiments are merely helpful in understanding the present invention, should not be regarded as a specific limitation of the invention.
The schematic diagram of the preparation method of the present invention for carrying enzyme filler is as shown in Figure 1, as seen from the figure, the load enzyme
The preparation method of filler the following steps are included:
(1) filler is fixed on mechanical rot using wire netting, adjustment revolving speed makes its uniform rotation；
(2) enzyme colloidal sol will be contained using pneumatic spray gun in the side of mechanical rot and is sprayed on filler surface, in other side benefit
It is dried up with hair dryer, obtains the load enzyme filler.
Following example 1-18 is all made of the preparation method of above-mentioned load enzyme filler.
Embodiment 1
The present embodiment load enzyme filler preparation method the following steps are included:
(1) metal Θ ring filler is fixed on mechanical rot using wire mesh, adjustment revolving speed makes its uniform rotation；
(2) enzyme colloidal sol will be contained using spray gun and is sprayed on filler surface, spraying process is sprayed using " it " word, the spraying
Metal Θ ring filler surface is dried up using hair dryer in the process, obtains the load enzyme filler, the air pressure of the spraying is
The flow of 0.15MPa, the spraying are 1.82kg/h, and distance of the spray gun apart from filler surface is during the spraying
15cm。
The specification of metal Θ ring filler used by the present embodiment is 3cm × 3cm, and following embodiment and comparative example are all made of
The metal Θ ring filler of this specification.
The preparation method of the colloidal sol containing enzyme of the present embodiment the following steps are included:
(a) methyl orthosilicate, trimethoxy silane and methanol are mixed in ice-water bath, stirs, it is molten obtains the first mixing
Liquid, the methyl orthosilicate, trimethoxy silane and methanol quality ratio are 2.03:7.2:8.49；
(b) Fluorinse of 1mol/L, polyethylene glycol, CALB enzyme solutions and water are mixed in ice-water bath, is stirred
It mixes, obtains the second mixed solution, the protein content of the enzyme solutions is 10.9mg/mL, and Fluorinse, polyethylene glycol, enzyme are molten
The mass ratio of liquid and water is 1:3:2:3.4, and the mass ratio of the Fluorinse of step (a) methyl orthosilicate and 1mol/L is
2.03:1；
(c) the first mixed solution is added in the second mixed solution, gained mixed solution is stirred into 5min in ice-water bath, is obtained
To the colloidal sol containing enzyme.
Load enzyme filler that the present embodiment is prepared is as shown in Fig. 2, the load enzyme filler that is prepared as seen from the figure
The colloidal sol containing enzyme of filler surface is evenly distributed, and apparent agglomeration does not occur.
Embodiment 2
The present embodiment the difference from embodiment 1 is that, the air pressure of spraying is replaced with into 0.2MPa, the flow of spraying replaces with
2.05kg/h, distance of the spray gun apart from filler surface replace with 20cm, and other conditions are identical compared with Example 1.
The colloidal sol containing enzyme that the present embodiment uses is same as Example 1.
Embodiment 3
The present embodiment the difference from embodiment 1 is that, the air pressure of spraying is replaced with into 0.3MPa, the flow of spraying replaces with
2.32kg/h, distance of the spray gun apart from filler surface replace with 25cm, and other conditions are identical compared with Example 1.
The colloidal sol containing enzyme that the present embodiment uses is same as Example 1.
Embodiment 4
The air pressure of spraying in embodiment 1 is replaced with 0.1MPa by the present embodiment, and other conditions are complete compared with Example 1
It is identical.
The colloidal sol containing enzyme that the present embodiment uses is same as Example 1.
Embodiment 5
The air pressure of spraying in embodiment 1 is replaced with 0.25MPa by the present embodiment, and other conditions are complete compared with Example 1
It is exactly the same.
The colloidal sol containing enzyme that the present embodiment uses is same as Example 1.
Embodiment 6
The air pressure of spraying in embodiment 1 is replaced with 0.3MPa by the present embodiment, and other conditions are complete compared with Example 1
It is identical.
The colloidal sol containing enzyme that the present embodiment uses is same as Example 1.
Embodiment 7
The air pressure of spraying in embodiment 1 is replaced with 0.5MPa by the present embodiment, and other conditions are complete compared with Example 1
It is identical.
The colloidal sol containing enzyme that the present embodiment uses is same as Example 1.
Embodiment 8
The air pressure of spraying in embodiment 1 is replaced with 0.05MPa by the present embodiment, and other conditions are complete compared with Example 1
It is exactly the same.
The colloidal sol containing enzyme that the present embodiment uses is same as Example 1.
Embodiment 9
The flow of spraying in embodiment 1 is replaced with 1.5kg/h by the present embodiment, and other conditions are complete compared with Example 1
It is exactly the same.
The colloidal sol containing enzyme that the present embodiment uses is same as Example 1.
Embodiment 10
The flow of spraying in embodiment 1 is replaced with 2.5kg/h by the present embodiment, and other conditions are complete compared with Example 1
It is exactly the same.
The colloidal sol containing enzyme that the present embodiment uses is same as Example 1.
Embodiment 11
The flow of spraying in embodiment 1 is replaced with 2.3kg/h by the present embodiment, and other conditions are complete compared with Example 1
It is exactly the same.
The colloidal sol containing enzyme that the present embodiment uses is same as Example 1.
Embodiment 12
The flow of spraying in embodiment 1 is replaced with 2.7kg/h by the present embodiment, and other conditions are complete compared with Example 1
It is exactly the same.
The colloidal sol containing enzyme that the present embodiment uses is same as Example 1.
Embodiment 13
The flow of spraying in embodiment 1 is replaced with 1.2kg/h by the present embodiment, and other conditions are complete compared with Example 1
It is exactly the same.
The colloidal sol containing enzyme that the present embodiment uses is same as Example 1.
Embodiment 14
Distance of the present embodiment by the spray gun in embodiment 1 apart from filler surface replaces with 20cm, other conditions and implementation
Example 1 is compared to identical.
The colloidal sol containing enzyme that the present embodiment uses is same as Example 1.
Embodiment 15
Distance of the present embodiment by the spray gun in embodiment 1 apart from filler surface replaces with 10cm, other conditions and implementation
Example 1 is compared to identical.
The colloidal sol containing enzyme that the present embodiment uses is same as Example 1.
Embodiment 16
Distance of the present embodiment by the spray gun in embodiment 1 apart from filler surface replaces with 25cm, other conditions and implementation
Example 1 is compared to identical.
The colloidal sol containing enzyme that the present embodiment uses is same as Example 1.
Embodiment 17
Distance of the present embodiment by the spray gun in embodiment 1 apart from filler surface replaces with 30cm, other conditions and implementation
Example 1 is compared to identical.
The colloidal sol containing enzyme that the present embodiment uses is same as Example 1.
Embodiment 18
Distance of the present embodiment by the spray gun in embodiment 1 apart from filler surface replaces with 8cm, other conditions and embodiment
1 compared to identical.
The colloidal sol containing enzyme that the present embodiment uses is same as Example 1.
Comparative example 1
This comparative example load enzyme filler preparation method the following steps are included:
(1) metal Θ ring filler is immersed in 0.5min in colloidal sol containing enzyme；
(2) the metal Θ ring filler of step (1) is taken out into dry 20s；
(3) above-mentioned submergence and drying steps 15 times are repeated in；
(4) the metal Θ ring filler that step (3) obtains is dried it is constant to weight, obtain carry enzyme filler.
The colloidal sol containing enzyme that this comparative example uses is same as Example 1.
Performance test methods:
The activity for the load enzyme filler that embodiment 1-18 and comparative example 1 obtain and the test method of stability are as follows:
The step of carrying the enzymatic activity test of enzyme filler: it is tested with the ester hydrolysis reaction of the 4- nitrobenzene palmitate of 5mg/mL
20mg load enzyme filler is put into the buffer solution of 3mL pH=7.5,200 microlitres of 4- nitrobenzene palmitates is added by enzymatic activity,
Concussion 30 seconds is reacted 2.5 minutes in the water-bath that temperature is 65 DEG C, then carries out gas chromatographic analysis, defines hydrolysis per minute
Enzyme amount needed for generating 1 μm of ol 4- nitrophenol is 1 unit of activity (U).
It carries the stability test step of enzyme filler: enzyme filler will be carried and be filled into the glass that tower internal diameter is 50mm, tower height is 960mm
In glass rectifying column, the ethyl acetate and n-butanol that molar ratio is 2:1, tower pressure interior force 10kPa is added, bottom temperature is 65 DEG C,
The quality for carrying the gel containing enzyme on enzyme filler is 49.9g, reaction time 3.5h；Above-mentioned steps are (primary in triplicate altogether
Experiment, secondary experiment and experiment three times), and during repeating to test, it is changed without and carries enzyme filler；Record experiment every time respectively
After load enzyme filler mass loss and n-butanol conversion ratio.
Carry the calculation method of the mass loss of enzyme filler:
The enzymatic activity for the load enzyme filler that embodiment 1-18 and comparative example 1 obtain is as shown in table 1:
Table 1
As can be seen from Table 1, the enzymatic activity of the load enzyme filler of embodiment 1-18 >=599U/g, and the load enzyme of comparative example 1
The enzymatic activity of filler is only 575U/g, it can be seen that the load enzyme that the preparation method of the present invention for carrying enzyme filler is prepared
The enzymatic activity of filler is substantially better than the enzymatic activity prepared by infusion process for carrying enzyme filler, and comparative example 1,4-8 can be seen that and work as
When spraying air pressure is 0.1-0.3MPa, the equal > 610U/g of enzymatic activity of the load enzyme filler sprayed, and optimal spraying air pressure
For 0.15-0.25MPa；It is 1.9-2.3kg/h that comparative example 1,9-13, which can be seen that optimal spraying flow,；Comparison is implemented
It is 15-20cm that example 1,14-18, which can be seen that optimal spray distance,.
The results are shown in Table 2 for the stability test for the load enzyme filler that embodiment 1-18 and comparative example 1 obtain:
Table 2
"-" refers to mass loss less than 0.05%.
As can be seen from the above table, the load enzyme filler catalysis acetic acid that the preparation method of the present invention for carrying enzyme filler is prepared
The activity and stability of ethyl ester and n-butanol ester exchange reaction are substantially better than described in comparative example 1 through load enzyme prepared by infusion process
Filler；In reactive distillation processes, using the load enzyme filler of phase homogenous quantities, the load enzyme filler that the method for the invention is prepared is urged
Change the conversion ratio of n-butanol conversion up to 99% or more, and the conversion ratio for carrying enzyme filler catalysis n-butanol of comparative example 1 then≤
98.2%, and the stability of load enzyme filler that the method for the invention obtains is also significantly better than comparative example 1, in one experiment
Load enzyme filler mass loss≤12.1%, and comparative example 1 load enzyme filler loss be 19.8%.
The Applicant declares that the foregoing is merely a specific embodiment of the invention, but protection scope of the present invention not office
It is limited to this, it should be clear to those skilled in the art, any to belong to those skilled in the art and take off in the present invention
In the technical scope of dew, any changes or substitutions that can be easily thought of, and all of which fall within the scope of protection and disclosure of the present invention.
Claims (10)
1. a kind of preparation method for carrying enzyme filler, which is characterized in that the method is that will contain enzyme colloidal sol by spray coating method to be sprayed on
Filler surface obtains the load enzyme filler.
2. the method as described in claim 1, which is characterized in that the air pressure of the spraying is 0.1-0.3MPa, preferably 0.15-
0.25MPa；
Preferably, the flow of the spraying is 1.5-2.5kg/h, preferably 1.9-2.3kg/h；
Preferably, distance of the spray gun apart from filler surface is 10-25cm, preferably 15-20cm during the spraying.
3. method according to claim 1 or 2, which is characterized in that spray direction during the spraying perpendicular to filler
Surface.
4. the method according to claim 1, which is characterized in that by filler wire during the spraying
Net is fixed on mechanical rot；
Preferably, the mechanical rot carries out uniform rotation.
5. method according to any of claims 1-4, which is characterized in that the filler includes metal Θ ring filler.
6. the method according to claim 1 to 5, which is characterized in that it is described spraying the following steps are included:
(1) filler is fixed on mechanical rot, adjustment revolving speed makes its uniform rotation；
(2) enzyme colloidal sol will be contained using spray gun and is sprayed on filler surface, and obtain the load enzyme filler.
7. method as claimed in claim 6, which is characterized in that step (2) further includes during spraying by filler surface
It is dried；
Preferably, the method for the drying is dried up including the use of hair dryer.
8. the method according to claim 1 to 7, which is characterized in that the described method comprises the following steps:
(1) metal Θ ring filler is fixed on mechanical rot using wire mesh, adjustment revolving speed makes its uniform rotation；
(2) enzyme colloidal sol will be contained using spray gun and is sprayed on metal Θ ring filler surface, and use hair dryer will during the spraying
The drying of metal Θ ring filler surface, obtains the load enzyme filler, and the air pressure of the spraying is 0.1-0.3Mpa, the stream of the spraying
Amount is 1.5-2.5kg/h, and distance of the spray gun apart from filler surface is 10-25cm during the spraying.
9. the load enzyme filler being prepared such as any one of claim 1-8 the method.
10. carrying the purposes of enzyme filler as claimed in claim 9, which is characterized in that the load enzyme filler is used for catalytic transesterification
Reactive distillation；
Preferably, the ester exchange reaction rectifying for carrying enzyme filler and being used to be catalyzed ethyl acetate and n-butanol.
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| CN201811646651.2A CN109837271A (en) | 2018-12-29 | 2018-12-29 | A kind of load enzyme filler and its preparation method and application |
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| CN201811646651.2A CN109837271A (en) | 2018-12-29 | 2018-12-29 | A kind of load enzyme filler and its preparation method and application |
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| CN109837271A true CN109837271A (en) | 2019-06-04 |
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| CN112760314A (en) * | 2021-01-15 | 2021-05-07 | 河北工业大学 | Preparation method of enzyme-loaded xerogel monolithic column catalytic filler |
| CN113584011A (en) * | 2021-07-28 | 2021-11-02 | 河北工业大学 | Method for immobilizing enzyme on polydopamine-coated plastic filler |
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| CN113584011A (en) * | 2021-07-28 | 2021-11-02 | 河北工业大学 | Method for immobilizing enzyme on polydopamine-coated plastic filler |
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