CN107586770A - A kind of porcine pancreatic lipase inorganic hybrid nano flower catalyst, preparation method and its application in biodiesel preparation - Google Patents
A kind of porcine pancreatic lipase inorganic hybrid nano flower catalyst, preparation method and its application in biodiesel preparation Download PDFInfo
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- CN107586770A CN107586770A CN201710726842.9A CN201710726842A CN107586770A CN 107586770 A CN107586770 A CN 107586770A CN 201710726842 A CN201710726842 A CN 201710726842A CN 107586770 A CN107586770 A CN 107586770A
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- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
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Abstract
A kind of porcine pancreatic lipase inorganic hybrid nano flower catalyst, preparation method and its application in biodiesel preparation, belong to bioenergy technical field.It is by 1~5mL, 80~300mM CuSO4The aqueous solution is added in 100mL, pH=6 dissolved with 10~250mg porcine pancreatic lipases~8 PBS cushioning liquid, 24~72h is incubated at 0~30 DEG C, it is then centrifuged for separating, and clean centrifugation product 3~5 times with deionized water, porcine pancreatic lipase inorganic hybrid nano flower catalyst is obtained after lyophilized, is stored at 20 DEG C；Wherein, weight percentage of the porcine pancreatic lipase in porcine pancreatic lipase inorganic hybrid nano spends catalyst is the weight % of 12 weight %~31.This method avoids the use of organic reagent preparing the nanocatalyst stage, therefore can not only preserve the activity of enzyme well, while can improve the tolerance of enzyme.In addition, the use of this new catalyst can avoid complicated separation process during biodiesel synthesis well, while there is preferable recycling rate of waterused.
Description
Technical field
The invention belongs to bioenergy technical field, and in particular to porcine pancreatic lipase-nothing prepared by a kind of biomimetic mineralization method
Machine hybridized nanometer flower catalyst, preparation method and its application in biodiesel preparation, the catalyst material can be urged effectively
Change sunflower oil to change into biodiesel and there is good recycling rate of waterused.
Background technology
Biodiesel is a kind of fatty acid alkyl esters based on animal tallow or vegetable oil, because it is good renewable
Property, biological degradability and low emission environmental pollution, it is considered to be a kind of Eco-friendly and regenerative resource, can be by aliphatic acid
Esterification is prepared with the esterification of short-chain alcohol ester oil ＆ fat.The production method of biodiesel, including acid or base catalysis turn
Change etc. is several.Acid catalyzed reaction is adapted to glycerine of the catalysis containing a large amount of free fatties, but reaction rate is generally relatively low；Base is urged
It is higher to change reaction rate in reaction, but it needs higher energy consumption.In addition, the residue of these catalyst can cause environment dirty
Dye, and be very difficult to remove or isolate.Compared with these methods, the biodiesel synthetic technology based on enzyme, which has to simplify, to be produced
Technique, reduce energy consumption, the purity for improving glycerin by-product product, the advantages that being formed in systems without soap.Wherein porcine pancreatic lipase
(PPL) obtained due to its moderate price, easily, higher stability and the bioconversion to non-natural matrix have extensive spy
The opposite sex, therefore cause extensive concern.One of most important application of porcine pancreatic lipase is catalytic esterification/ester exchange gentle
Under the conditions of prepare biodiesel.However, the high cost of lipase is still to realize a huge challenge of its commercial Application.Cause
This, improves the recuperability of lipase, it is considered to be a kind of available strategy increased economic efficiency.In addition, given birth in biodiesel
Low tolerance in production to lipase is also a key issue for needing to solve.
Enzyme immobilization is that solve the effective way that lipase prepares above mentioned problem in biodiesel process, main at present to consolidate
Surely changing zymotechnic includes " trapping " and " single enzyme nano-particle " technology, and these immobilised enzymes are compared to resolvase with higher steady
It is qualitative, but their activity often can all reduce a lot, this mainly has following reason：1) organic solvent in immobilization process
Use；2) chemical reaction occurred between fixed material and protein；3) the obvious mass transfer between enzyme and immobilization material.
Bionic mineralising is that the self assembling process in a kind of system using natural biology is converted into biology work outside protection
The strategy of property molecule.By the various enzymes of the construction of strategy-inorganic mixing nano flower, the activity of enzyme, stability and can be improved
Cyclicity, and it is successfully applied to the fields such as biology sensor, living things catalysis and tumor thermal therapy.Therefore, we pass through biology
The lipase of immobilization prepared by biomimetic mineralization technology can be as effective biocatalyst of production of biodiesel, it is contemplated that with compared with
High catalytic activity, stability and reusability.This enzyme-inorganic hybrid nano flower catalyst is used for biodiesel synthesis not yet
Appear in the newspapers, our result show that this enzyme-inorganic hybrid nano flower catalyst can very well catalytically synthesizing biological diesel oil and
With preferably recycling rate of waterused.
The content of the invention
In order to overcome the enzyme occurred in the complex operation occurred in traditional catalyst use, pollution, waste and enzymatic
Tolerance is low, expensive and the problems such as can not recycle, and is prepared the invention provides one kind using bionic mineralising method
New porcine pancreatic lipase-inorganic hybrid nano flower catalyst and preparation method thereof, this method is preparing the nanocatalyst stage
The use of organic reagent is avoided, therefore the activity of enzyme can not only be preserved well, while the tolerance of enzyme can be improved.Separately
On the one hand, the use of this new catalyst can avoid complicated separation process during biodiesel synthesis well, together
When there is preferable recycling rate of waterused.
To realize object above, one kind of the present invention prepares porcine pancreatic lipase-inorganic hybridization using biomimetic mineralization and received
The method of popped rice catalyst, its step are as follows：
(1) it is a kind of to utilize bionic mineralising method using porcine pancreatic lipase as organic moiety, using cupric phosphate as inorganic part system
The method of standby porcine pancreatic lipase-inorganic hybrid nano flower catalyst
By 1~5mL, 80~300mM CuSO4The aqueous solution is added to the 100mL dissolved with 10~250mg porcine pancreatic lipases
In PBS (pH=6~8) cushioning liquid, 24~72h is incubated at 0~30 DEG C, is then centrifuged for separating, and cleaned with deionized water
Centrifuge product 3~5 times, porcine pancreatic lipase of the present invention-inorganic hybrid nano flower catalyst is obtained after lyophilized, in -20 DEG C
Lower storage.Weight percentage of the porcine pancreatic lipase in porcine pancreatic lipase-inorganic hybrid nano flower catalyst is 12 weight %
~31 weight %, when it is 10mg to add porcine pancreatic lipase amount, porcine pancreatic lipase is urged in porcine pancreatic lipase-inorganic hybrid nano flower
Weight percentage in agent is 12 weight %, and when addition porcine pancreatic lipase amount is 150mg, porcine pancreatic lipase is in pig pancreas fat
Weight percentage in enzyme-inorganic hybrid nano flower catalyst is~31 weight %.
Specifically operating procedure is：
1. the use of cupric sulfate pentahydrate and deionized water compound concentration is 80~300mM copper sulfate solutions；
2. by 1.44g Na2HPO4、0.24g KH2PO4, 0.2g KCl, 8.0g NaCl be dissolved into 100mL deionized waters
In, then with deionized water 1L is settled to, pH=6~8 then are adjusted with 1M HCl and NaOH, obtain PBS cushioning liquid；
3. weighing 10~250mg porcine pancreatic lipases is dissolved in the above-mentioned PBS cushioning liquid of 100mL, 6000~10000rad/s from
The heart, gained supernatant is poured into 100mL wide-mouth bottles；Pipette the above-mentioned CuSO of 1~5mL4The aqueous solution is added in wide-mouth bottle, 0
24~72h is incubated at~30 DEG C；
4. 6000~10000rad/s of reaction solution is centrifuged, centrifugation product is cleaned 3~5 times using deionized water, is sunk
Porcine pancreatic lipase-inorganic hybrid nano flower catalyst is obtained at starch -50~-70 DEG C after vacuum freeze-drying, is stored at -20 DEG C.
As a further improvement on the present invention, step 1. in, the CuSO of preparation4The concentration of the aqueous solution is 100~200mM；
As a further improvement on the present invention, step 2. in be with 1M HCl and NaOH regulation pH=6.8~7.4, obtain
PBS cushioning liquid；
As a further improvement on the present invention, step 3. in be to weigh 10~200mg porcine pancreatic lipases to be dissolved in 100mL PBS
In cushioning liquid, 6000~10000rad/s centrifugations, gained supernatant is poured into 100mL wide-mouth bottles, it is above-mentioned to pipette 1~3mL
CuSO4The aqueous solution is added in wide-mouth bottle, and 12~24h is incubated at 4~25 DEG C；
(2) it is substrate biodiesel synthesis by catalyst, sunflower oil of porcine pancreatic lipase-inorganic hybrid nano flower
1. weighing porcine pancreatic lipase-inorganic hybrid nano flower 50~200mg of catalyst, ultrasonic disperse is in 10~50mL sunflowers
In seed oil；
2. adding methanol into above-mentioned dispersion liquid, the volume ratio of sunflower oil and methanol is 1~10：1, and use 1~10M
NaOH adjusts pH=8~12 of reaction solution, and 1~12h of reaction under (600~800 turns/s) is then stirred vigorously at 40~50 DEG C；
3. being filtered to remove porcine pancreatic lipase-inorganic hybrid nano flower catalyst, supernatant liquor is separated simultaneously using separatory funnel
Rotary evaporation removes unreacted methanol in supernatant liquor, obtains product biodiesel；
As a further improvement on the present invention, step (2) 1. in, be to weigh porcine pancreatic lipase-inorganic hybrid nano flower
The amount of catalyst is 50~150mg；Step (2) 2. in, the volume ratio of sunflower oil and methanol is 2~8：1, and use 2~
5M NaOH adjust pH=8~10 of reaction solution, be then stirred vigorously at 40~50 DEG C under (600~800 turns/s) reaction 2~
5h；
As a further improvement on the present invention, step (2) 3. in, with the flower catalysis of porcine pancreatic lipase-inorganic hybrid nano
Agent prepares the conversion ratio of biodiesel 70%~98%, and catalytically synthesizing biological diesel oil converts after recycling number is 4~5 times
Rate is still 40%~70%；When preparing biodiesel with porcine pancreatic lipase-inorganic hybrid nano flower catalyst, sunflower
The volume ratio of seed oil and methanol is 2：Biodiesel conversion rate is 98% when 1, and conversion ratio still reaches after reusing 5 times
70%；The volume ratio of sunflower oil and methanol is 8：When 1 biodiesel conversion rate be 70%, recycling 4 times after conversion ratio according to
So reach more than 40%.
The invention has the advantages that while enzymatic activity can be retained, enzyme tolerance is improved.With porcine pancreatic lipase-nothing
Machine hybridized nanometer can obtain very high conversion ratio and with weight well when spending catalyst sunflower oil biodiesel synthesis
Multiple utilization rate.
Brief description of the drawings
Fig. 1：It is the surface topography ESEM of porcine pancreatic lipase-inorganic hybrid nano flower catalyst prepared by the present invention
Figure；Scheme the low power scanning electron microscope (SEM) photograph of (a) porcine pancreatic lipase-inorganic hybrid nano flower catalyst；Figure (b) porcine pancreatic lipase-inorganic
Hybridized nanometer spends the high power scanning electron microscope (SEM) photograph of catalyst；Electron microscope shows that porcine pancreatic lipase-inorganic hybrid nano flower catalyst is in
Now more intercalation flower-like structures；
Fig. 2：It is that porcine pancreatic lipase-inorganic hybrid nano prepared by the present invention spends catalyst with free porcine pancreatic lipase not
With relative activity column diagram (b) under relative activity column diagram (a) under the conditions of pH and condition of different temperatures；
Fig. 3：It is the nuclear-magnetism hydrogen of porcine pancreatic lipase-nano flower catalyst synthetic product biodiesel prepared by the present invention
Spectrogram；
Fig. 4：It is the biodiesel conversion rate cylindricality under the conditions of different sunflower oils and methanol volume ratio prepared by the present invention
Figure；
Fig. 5：It is that porcine pancreatic lipase-nano flower catalyst prepared by the present invention is catalyzed preparation biology in different cycle-indexes
The conversion rate curve figure of diesel oil；
Embodiment
In order that the object, technical solutions and advantages of the present invention are clearer, below in conjunction with the accompanying drawings with specific embodiment pair
The present invention is described in detail.
Embodiment 1：
Porcine pancreatic lipase-inorganic hybrid nano flower catalyst preparation：Using bionic mineralising method, comprise the following steps that：
1) it is 200mM copper sulfate solutions first by cupric sulfate pentahydrate and deionized water compound concentration；2) by 1.44g Na2HPO4、
0.24g KH2PO4, 0.2g KCl, 8.0g NaCl be dissolved into 100mL deionized waters, then 1L be settled to deionized water, then
PH=7.4 is adjusted with 1M HCl and NaOH, obtains PBS cushioning liquid；3) weighing 200mg porcine pancreatic lipases, to be dissolved in 100mL above-mentioned
PBS cushioning liquid, 6000rad/s centrifugations, gained supernatant is poured into 100mL wide-mouth bottles；Pipette the above-mentioned CuSO of 3mL4Water
Solution is added in wide-mouth bottle, and 24h is incubated at 4 DEG C；4) reaction solution 8000rad/s is centrifuged, centrifugation product use go from
Sub- water cleans 4 times, obtains 260mg porcine pancreatic lipases-inorganic hybrid nano flower catalyst at -50 DEG C of sediment after vacuum freeze-drying,
Stored at -20 DEG C.Weight percentage of the porcine pancreatic lipase in porcine pancreatic lipase-inorganic hybrid nano flower catalyst is 31
Weight %；During the addition increase of enzyme, weight hundred of the porcine pancreatic lipase in porcine pancreatic lipase-inorganic hybrid nano flower catalyst
Point content can also increase, but at maximum up to 31 weight %.
Porcine pancreatic lipase-inorganic hybrid nano flower catalyst Biodiesel Produced by Sunflower Seeds Oil method specific steps are such as
Under：1) porcine pancreatic lipase-inorganic hybrid nano flower catalyst 100mg is weighed, ultrasonic disperse is in 20mL sunflower oils；2) upwards
State and methanol is added in dispersion liquid, the volume ratio of sunflower oil and methanol is 2：1, and use the pH=of 5M NaOH regulation reaction solutions
10, (600 turns/s) reaction 4h are then stirred vigorously down at 45 DEG C；3) it is filtered to remove porcine pancreatic lipase-inorganic hybrid nano flower
Catalyst, separate supernatant liquor using separatory funnel and rotary evaporation removes unreacted methanol in supernatant liquor, obtain product
Biodiesel, the conversion ratio of biodiesel is spent as 98% using porcine pancreatic lipase-inorganic hybrid nano, recycling
Catalytically synthesizing biological diesel oil conversion ratio is still 70% after number is 5 times；
Biodiesel is prepared as a comparison case using free porcine pancreatic lipase, tested.
Test 1：Using the pattern of ESEM (SEM) observation sample.Scanning electron microscopic picture is obtained by HITACHI SU4800
.As shown in figure 1, porcine pancreatic lipase-inorganic hybrid nano spends catalyst particle size that multi-level intercalation is presented at 0.8~1 μm or so
Flower-like structure.
Test 2：With 10mM p- p-nitrophenyls caprylates (p-NPC) for substrate, tested and swum using alternating temperature ultraviolet (UV2550)
It is faster from porcine pancreatic lipase and porcine pancreatic lipase-inorganic hybrid nano flower catalyst hydrolysing activity and stability, absorbance change
Represent that porcine pancreatic lipase-inorganic hybrid nano flower catalyst hydrolysing activity is higher.Test condition is：Hydrolyzed under different temperatures, pH
Absorbance change of the product at 405nm.As shown in Fig. 2 (a), it can be seen that free esterase and porcine pancreatic lipase-inorganic hybridization
The optimal pH of nano flower catalyst is 7.5, high pH (>8) under the conditions of, porcine pancreatic lipase-inorganic hybrid nano flower catalyst
Specific ionization esterase is more stable and vigor is higher, Fig. 2 (b) displays, is optimal activity, pig pancreas at 40 DEG C compared to free-fat enzyme
Lipase-inorganic hybrid nano flower catalyst is optimal activity at 50 DEG C, illustrates that porcine pancreatic lipase-inorganic hybrid nano flower is urged
Agent has higher temperature stability；
Test 3：Using nucleus magnetic hydrogen spectrum (1H NMR) test sign biodiesel structure.Nuclear-magnetism spectrum by Bruker400M with
CDCl3Obtained for solvent.As shown in figure 3, singlet δ=3.65 and triplet δ=2.3 in nuclear-magnetism, can confirm that methyl esters knot
The generation of structure and methyl ester conversion rate, calculated using this and understand biodiesel conversion rate as 98%.As shown in Figure 4.
Embodiment 2：
Porcine pancreatic lipase-inorganic hybrid nano flower catalyst preparation：Porcine pancreatic lipase-inorganic hybrid nano flower catalyst system
It is standby：Using bionic mineralising method, comprise the following steps that：1) it is first by cupric sulfate pentahydrate and deionized water compound concentration
100mM copper sulfate solutions；2) by 1.44g Na2HPO4、0.24g KH2PO4, 0.2g KCl, 8.0g NaCl be dissolved into 100mL
In deionized water, then with deionized water 1L is settled to, then adjusts pH=7.4 with 1M HCl and NaOH, it is molten to obtain PBS bufferings
Liquid；3) 10mg porcine pancreatic lipases are weighed and are dissolved in the above-mentioned PBS cushioning liquid of 100mL, 6000rad/s centrifugations, gained supernatant are fallen
Enter in 100mL wide-mouth bottles；Pipette the above-mentioned CuSO of 3mL4The aqueous solution is added in wide-mouth bottle, and 24h is incubated at 4 DEG C；4) will reaction
Liquid 8000rad/s is centrifuged, and centrifugation product is cleaned 4 times using deionized water, is obtained at -50 DEG C of sediment after vacuum freeze-drying
83mg porcine pancreatic lipases-inorganic hybrid nano flower catalyst, are stored at -20 DEG C.Porcine pancreatic lipase is in porcine pancreatic lipase-inorganic
Weight percentage in hybridized nanometer flower catalyst is 12 weight %；
Porcine pancreatic lipase-inorganic hybrid nano flower catalyst Biodiesel Produced by Sunflower Seeds Oil method specific steps are such as
Under：1) porcine pancreatic lipase-inorganic hybrid nano flower catalyst 100mg is weighed, ultrasonic disperse is in 20mL sunflower oils；2) upwards
State and methanol is added in dispersion liquid, the volume ratio of sunflower oil and methanol is 2：1, and use the pH=of 5M NaOH regulation reaction solutions
10, then it is stirred vigorously at 45 DEG C under (600 turns/s) and reacts 4h；3) it is filtered to remove porcine pancreatic lipase-inorganic hybrid nano flower
Catalyst, separate supernatant liquor using separatory funnel and rotary evaporation removes unreacted methanol in supernatant liquor, obtain product
Biodiesel, the conversion ratio of biodiesel is spent as 90% using porcine pancreatic lipase-inorganic hybrid nano, recycling
Catalytically synthesizing biological diesel oil conversion ratio is still 61% after number is 4 times；
Embodiment 3：
Porcine pancreatic lipase-inorganic hybrid nano flower catalyst preparation：Porcine pancreatic lipase-inorganic hybrid nano flower catalyst system
It is standby：Using bionic mineralising method, comprise the following steps that：1) it is first by cupric sulfate pentahydrate and deionized water compound concentration
200mM copper sulfate solutions；2) by 1.44g Na2HPO4、0.24g KH2PO4, 0.2g KCl, 8.0g NaCl be dissolved into 100mL
In deionized water, then with deionized water 1L is settled to, then adjusts pH=7.4 with 1M HCl and NaOH, it is molten to obtain PBS bufferings
Liquid；3) 200mg porcine pancreatic lipases are weighed and are dissolved in the above-mentioned PBS cushioning liquid of 100mL, 6000rad/s centrifugations, gained supernatant are fallen
Enter in 100mL wide-mouth bottles；Pipette the above-mentioned CuSO4 aqueous solution of 3mL to add in wide-mouth bottle, 24h is incubated at 4 DEG C；4) will reaction
Liquid 8000rad/s is centrifuged, and centrifugation product is cleaned 4 times using deionized water, is obtained at -50 DEG C of sediment after vacuum freeze-drying
260mg porcine pancreatic lipases-inorganic hybrid nano flower catalyst, are stored at -20 DEG C.Porcine pancreatic lipase is in porcine pancreatic lipase-nothing
Weight percentage in machine hybridized nanometer flower catalyst is 31 weight %；
Porcine pancreatic lipase-inorganic hybrid nano flower catalyst Biodiesel Produced by Sunflower Seeds Oil method specific steps are such as
Under：1) porcine pancreatic lipase-inorganic hybrid nano flower catalyst 100mg is weighed, ultrasonic disperse is in 20mL sunflower oils；2) upwards
State and methanol is added in dispersion liquid, the volume ratio of sunflower oil and methanol is 8：1, and use the pH=of 5M NaOH regulation reaction solutions
10, then it is stirred vigorously at 45 DEG C under (600 turns/s) and reacts 4h；3) it is filtered to remove porcine pancreatic lipase-inorganic hybrid nano flower
Catalyst, separate supernatant liquor using separatory funnel and rotary evaporation removes unreacted methanol in supernatant liquor, obtain product
Biodiesel, the conversion ratio of biodiesel is spent as 70% using porcine pancreatic lipase-inorganic hybrid nano, recycling
Catalytically synthesizing biological diesel oil conversion ratio is still 40% after number is 4 times；
The above embodiments are merely illustrative of the technical solutions of the present invention and it is unrestricted, although with reference to preferred embodiment to this hair
It is bright to be described in detail, it will be understood by those within the art that, technical scheme can be modified
Or equivalent substitution, without departing from the spirit and scope of technical solution of the present invention.
Claims (10)
- A kind of 1. preparation method of porcine pancreatic lipase-inorganic hybrid nano flower catalyst, it is characterised in that：It is by 1~5mL, 80 ~300mM CuSO4The PBS bufferings that the aqueous solution is added to 100mL, pH=6 dissolved with 10~250mg porcine pancreatic lipases~8 are molten In liquid, 24~72h is incubated at 0~30 DEG C, is then centrifuged for separating, and centrifugation product is cleaned 3~5 times with deionized water, freezed After obtain porcine pancreatic lipase-inorganic hybrid nano flower catalyst, stored at -20 DEG C；Wherein, porcine pancreatic lipase is in pig pancreas fat Weight percentage in enzyme-inorganic hybrid nano flower catalyst is the weight % of 12 weight %~31.
- 2. a kind of preparation method of porcine pancreatic lipase as claimed in claim 1-inorganic hybrid nano flower catalyst, its feature exist In：1. the use of cupric sulfate pentahydrate and deionized water compound concentration is 80~300mM copper sulfate solutions；2. by 1.44g Na2HPO4、0.24g KH2PO4, 0.2g KCl, 8.0g NaCl be dissolved into 100mL deionized waters, then use Deionized water is settled to 1L, then adjusts pH=6~8 with 1M HCl and NaOH, obtains PBS cushioning liquid；3. weighing 10~250mg porcine pancreatic lipases is dissolved in the above-mentioned PBS cushioning liquid of 100mL, 6000~10000rad/s centrifugations will Gained supernatant is poured into 100mL wide-mouth bottles；Pipette the above-mentioned CuSO of 1~5mL4The aqueous solution is added in wide-mouth bottle, at 0~30 DEG C 24~72h of lower incubation；4. 6000~10000rad/s of reaction solution is centrifuged, centrifugation product is cleaned 3~5 times using deionized water, sediment- Porcine pancreatic lipase-inorganic hybrid nano flower catalyst is obtained at 50~-70 DEG C after vacuum freeze-drying, is stored at -20 DEG C.
- 3. a kind of preparation method of porcine pancreatic lipase as claimed in claim 2-inorganic hybrid nano flower catalyst, its feature exist In：Step 1. in, the CuSO of preparation4The concentration of the aqueous solution is 100~200mM.
- 4. a kind of preparation method of porcine pancreatic lipase as claimed in claim 2-inorganic hybrid nano flower catalyst, its feature exist In：Step 2. in be with 1M HCl and NaOH regulation pH=6.8~7.4, obtain PBS cushioning liquid.
- 5. a kind of preparation method of porcine pancreatic lipase as claimed in claim 2-inorganic hybrid nano flower catalyst, its feature exist In：Step 3. in be to weigh 10~200mg porcine pancreatic lipases to be dissolved in 100mL PBS cushioning liquid, 6000~10000rad/s Centrifugation, gained supernatant is poured into 100mL wide-mouth bottles, pipettes the above-mentioned CuSO of 1~3mL4The aqueous solution is added in wide-mouth bottle, 12~24h is incubated at 4~25 DEG C.
- A kind of 6. porcine pancreatic lipase-inorganic hybrid nano flower catalyst, it is characterised in that：It is by Claims 1 to 5 any one Described method is prepared.
- 7. porcine pancreatic lipase-application of the inorganic hybrid nano flower catalyst in biodiesel preparation described in claim 6.
- 8. porcine pancreatic lipase as claimed in claim 7-application of the inorganic hybrid nano flower catalyst in biodiesel preparation, It is characterized in that：It is the application in sunflower oil conversion biodiesel is catalyzed.
- 9. porcine pancreatic lipase as claimed in claim 8-application of the inorganic hybrid nano flower catalyst in biodiesel preparation, It is characterized in that：Concrete operation step is,1. weighing porcine pancreatic lipase-inorganic hybrid nano flower 50~200mg of catalyst, ultrasonic disperse is in 10~50mL sunflower oils In；2. adding methanol into above-mentioned dispersion liquid, the volume ratio of sunflower oil and methanol is 1~10：1, and use 1~10M NaOH adjusts pH=8~12 of reaction solution, and 1~12h of lower reaction is then stirred vigorously at 40~50 DEG C；3. being filtered to remove porcine pancreatic lipase-inorganic hybrid nano flower catalyst, separate supernatant liquor using separatory funnel and rotate Evaporation removes unreacted methanol in supernatant liquor, obtains product biodiesel.
- 10. porcine pancreatic lipase as claimed in claim 9-inorganic hybrid nano flower catalyst answering in biodiesel preparation With, it is characterised in that：Step 1. in, the amount for weighing porcine pancreatic lipase-inorganic hybrid nano flower catalyst is 50~150mg；Step Suddenly 2. in, the volume ratio of sunflower oil and methanol is 2~8：1, and pH=8~10 of 2~5M NaOH regulation reaction solutions are used, Then 2~5h of lower reaction is stirred vigorously at 40~50 DEG C.
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| CN109988756A (en) * | 2019-03-15 | 2019-07-09 | 浙江工业大学 | N- acyl-homoserine lactonase-inorganic hybrid nano catalyst and its preparation |
| CN110564717A (en) * | 2019-09-05 | 2019-12-13 | 湖北大学 | Alkaline pectinase-inorganic hybrid nanoflower with improved thermal stability and application thereof |
| CN113243392A (en) * | 2021-05-24 | 2021-08-13 | 吉林大学 | Beauveria bassiana spore-hybrid nano flower biological pesticide, preparation method and application |
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| CN109988756A (en) * | 2019-03-15 | 2019-07-09 | 浙江工业大学 | N- acyl-homoserine lactonase-inorganic hybrid nano catalyst and its preparation |
| CN109988756B (en) * | 2019-03-15 | 2021-07-06 | 浙江工业大学 | N-acyl homoserine lactonase-inorganic hybrid nano catalyst and preparation thereof |
| CN110564717A (en) * | 2019-09-05 | 2019-12-13 | 湖北大学 | Alkaline pectinase-inorganic hybrid nanoflower with improved thermal stability and application thereof |
| CN113243392A (en) * | 2021-05-24 | 2021-08-13 | 吉林大学 | Beauveria bassiana spore-hybrid nano flower biological pesticide, preparation method and application |
| CN113243392B (en) * | 2021-05-24 | 2021-11-23 | 吉林大学 | Beauveria bassiana spore-hybrid nano flower biological pesticide, preparation method and application |
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