CN103468754A - Method for enzymatic synthesis of propyl caffeate by utilizing microreactor - Google Patents
Method for enzymatic synthesis of propyl caffeate by utilizing microreactor Download PDFInfo
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- CN103468754A CN103468754A CN2013103585826A CN201310358582A CN103468754A CN 103468754 A CN103468754 A CN 103468754A CN 2013103585826 A CN2013103585826 A CN 2013103585826A CN 201310358582 A CN201310358582 A CN 201310358582A CN 103468754 A CN103468754 A CN 103468754A
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- propyl ester
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Abstract
A method for enzymatic synthesis of propyl caffeate by utilizing a microreactor is provided. A lipase Novozym435 is filled into an immobilization area, an ionic liquid containing substrates of methyl caffeate and propyl alcohol is injected into the immobilization area by an injection pump, wherein the concentration of methyl caffeate is in the range of 0.1 mg/mL to 20 mg/mL, the molar ratio of methyl caffeate to propyl alcohol is in the range of 1:1 to 1:200, the flow velocity is in the range of 0.1 to 100 [mu]L/min, and the reaction temperature is in the range of 30 to 80 DEG C; and an enzymatic transesterification reaction is carried out to prepare propyl caffeate. Compared with a traditional batch-type reactor, the method greatly accelerates the reaction efficiency, shortens the reaction time, and enables the propyl caffeate product to have high purity.
Description
Technical field
The present invention relates to the biochemical pharmacy field, be specifically related to a kind of method of utilizing the synthetic coffic acid propyl ester of microreactor enzymatic.
Background technology
The coffic acid propyl ester, chemical name (E)-propyl group-3-(3,4-dihydroxyl) acrylate, there is good oxidation-resistance and anti-tumor activity (Bioorganic& Medicinal Chemistry, 2004,12,3581-3589), can effectively extend the oxidation induction period of edible oil, can be used as a kind of novel natural origin food antioxidant.
The method for preparing the coffic acid propyl ester mainly contains two kinds of chemical method and biological enzymes.
(1) chemical method: the people such as Uwai have proposed with disulfide chloride catalysis coffic acid and propyl alcohol, and after backflow 17h, the yield of coffic acid propyl ester is 86% (Bioorganic& Medicinal Chemistry, 2008,16,7795-7803); Fiuza etc. utilize the Fischer esterification process, with H
2sO
4for catalyzer, direct esterification coffic acid and propyl alcohol, do not mark concrete yield (Bioorganic& Medicinal Chemistry, 2004,12,3581-3589); Xia Chunnian adopts one kettle way to synthesize the coffic acid propyl ester, after isopropylidene malonate is dissolved in to toluene, adds propyl alcohol, after reaction for some time, adds 0412, pyridine and piperidines, and final yield is 67.8%; CN103086880A discloses a kind of method with Catalyzed by p-Toluenesulfonic Acid coffic acid and propyl alcohol, and yield reaches 67.8%.Yet, utilize that chemical method is synthetic exists byproduct of reaction many, equipment is corrosive, the problem such as environmental pollution is large, be not suitable with the development in epoch.
(2) enzyme process: CN103086880A has related to a kind of with lipase direct esterification coffic acid and propyl alcohol, and yield only has 41.0%; The people such as Pang have proposed to synthesize the coffic acid propyl ester with the lipase transesterification, take Methyl caffeoate and propyl alcohol as substrate, after reaction 24h, yield can reach 98.5% (Bioresource Technology, 2013,139,337-342).But the reaction times is longer, efficiency is lower, consumes energy larger.Therefore, be badly in need of finding a kind of new technology for preparing efficiently the coffic acid propyl ester.
This novel concept of " Miniaturizing Biocatalysis " (" miniature organism catalysis ") is proposed in 2010 in the world first, and rapidly by internationally recognized for be bioengineering field first develop one of direction (Chemical Engineering Journal, 2013,223,497-506).With traditional reactor, compare, microreactor has heat transfer, advantage that mass-transfer efficiency is high, improves greatly productive rate and Reaction time shorten.
deng the oxidation of having studied in microreactor with alcoholdehydrogenase catalysis hexanol, it is high 33 times that speed of reaction is compared than traditional reactor, after 7.8s productive rate almost reach 100% (Engineering in Life Sciences, 2012,12,49-56).Microreactor more easily amplifies, and can, by the simple quantity that increases microreactor, and realization is large-scale, produce.Continuous reaction replaces intermittent reaction, easily-controlled reaction conditions, contribute to overcome simultaneously and solve tradition synthetic in reactor volume large, catalytic selectivity low and problem (Chemical Engineering Journal, 2012 such as product component complexity, 189 – 190,376-382).So far, there is not yet the report of microreactor for the synthetic coffic acid propyl ester of enzymatic.
Therefore, the present invention, by the novel method of using the synthetic coffic acid propyl ester of a kind of microreactor enzymatic is provided, is conducive to improve combined coefficient, the shortening time, reduce power consumption, and for promoting its widespread use tool in food antioxidant, be of great significance.
Summary of the invention
The technical problem solved:
The purpose of this invention is to provide a kind of method of utilizing the synthetic coffic acid propyl ester of microreactor enzymatic, with low, the consuming time length of speed of reaction that solves the synthetic coffic acid propyl ester existed in prior art and can not serialization production etc. problem.
Technical scheme:
A kind of method of utilizing the synthetic coffic acid propyl ester of microreactor enzymatic, lipase Novozym435 is filled into to immobilized zone, the ionic liquid that will contain substrate Methyl caffeoate and propyl alcohol with syringe pump is injected in immobilized zone, Methyl caffeoate's concentration range is 0.1mg/mL～20mg/mL, the molar ratio range of Methyl caffeoate and propyl alcohol is 1:1～1:200, flow rates is 0.1～100 μ L/min, and range of reaction temperature is 30～80 ℃, carries out the enzymatic transesterification and prepares the coffic acid propyl alcohol; Described sandwich type microreactor comprises syringe pump (1) and reactor body, described reactor body material is polydimethylsiloxane (polydimethylsiloxane), and formed by Sptting plate (2) and cover plate (3), the Sptting plate center is provided with immobilized zone (4), the wide 1cm in immobilized zone, dark 500 μ m, long 50mm-200mm.
Be provided with retaining clip between described Sptting plate and cover plate.
Described lipase Novozym435 adopts the mode of tiling to be filled in immobilized zone.
The filling enzyme amount of described lipase Novozym435 is 50mg-200mg.
The positive and negative ionic type combination of described ionic liquid comprises that cation type is [Bmim], [Hmim], [Emim], [TOMA] or [Omim], and anionic type is [CF
3sO
3], [PF
6], [Tf
2n], [BF
4] or [Cl].
The reaction formula of the synthetic coffic acid propyl ester of enzymatic transesterification:
Beneficial effect:
The present invention selects to utilize the microreactor enzymatic to synthesize the coffic acid propyl ester, and the reaction times is shortened to 2.5h by traditional 24h, and reaction efficiency improves greatly, and power consumption reduces.Simultaneously, the purity of product is higher, and productive rate reaches 99.5%.In addition, the apparatus structure of the present invention's employing is simple, only needs simply number to be amplified and gets final product expanding production capacity of equal proportion, has good industrial applications prospect, can meet the industrial requirement in food antioxidant market.
The accompanying drawing explanation
Fig. 1 is apparatus structure schematic diagram of the present invention.
Embodiment
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only are not used in and limit the scope of the invention for the present invention is described.Should be understood that in addition those skilled in the art can make various changes or modifications the present invention after the content of having read the present invention's instruction, these equivalent form of values fall within the application's appended claims limited range equally.
The sandwich type microreactor, comprise syringe pump 1 and reactor body, and described reactor body material is polydimethylsiloxane (polydimethylsiloxane), and be comprised of Sptting plate 2 and cover plate 3, and the Sptting plate center is provided with immobilized regional 4.Be provided with retaining clip between Sptting plate and cover plate.The 0.1mg Methyl caffeoate is joined in [Omim] [Cl] ionic liquid of 1mL, be made into reaction system according to Methyl caffeoate and propyl alcohol mol ratio 1:1, this reaction system is continuously pumped in microreactor and carries out transesterification by micro-injection pump with the flow velocity of 20 μ L/min.The wide 1cm of microchannel in the middle of microreactor, high 500 μ m, long 75mm, one deck Novozym435(90mg) be filled in this microchannel.Control 30 ℃ of temperature of reaction, reaction raw materials residence time in reactor is about 15min.Collect outlet material, the yield of being combined to the coffic acid propyl ester with the transesterification of HPLC detection enzymatic is 0.7%.
Embodiment 2
Apparatus structure, as embodiment 1, joins the 20mg Methyl caffeoate [Hmim] [PF of 1mL
6] in ionic liquid, according to Methyl caffeoate and propyl alcohol mol ratio 1:200, being made into reaction system, this reaction system is continuously pumped in microreactor and carries out transesterification by micro-injection pump with the flow velocity of 2 μ L/min.The wide 1cm of microchannel in the middle of microreactor, high 500 μ m, long 75mm, one deck Novozym435(90mg) be filled in this microchannel.Control 80 ℃ of temperature of reaction, reaction raw materials residence time in reactor is about 2.5h.Collect outlet material, the yield of being combined to the coffic acid propyl ester with the transesterification of HPLC detection enzymatic is 4.3%.
Apparatus structure, as embodiment 1, joins the 5mg Methyl caffeoate [Bmim] [Tf of 1mL
2n] in ionic liquid, according to Methyl caffeoate and propyl alcohol mol ratio 1:100, be made into reaction system, this reaction system is continuously pumped in microreactor and carries out transesterification by micro-injection pump with the flow velocity of 5 μ L/min.The wide 1cm of microchannel in the middle of microreactor, high 500 μ m, long 75mm, one deck Novozym435(90mg) be filled in this microchannel.Control 60 ℃ of temperature of reaction, reaction raw materials residence time in reactor is about 1h.Collect outlet material, the yield of being combined to the coffic acid propyl ester with the transesterification of HPLC detection enzymatic is 53.2%.
Embodiment 4
Apparatus structure, as embodiment 1, joins the 0.5mg Methyl caffeoate [Emim] [CF of 1mL
3sO
3] in ionic liquid, according to Methyl caffeoate and propyl alcohol mol ratio 1:20, being made into reaction system, this reaction system is continuously pumped in microreactor and carries out transesterification by micro-injection pump with the flow velocity of 5 μ L/min.The wide 1cm of microchannel in the middle of microreactor, high 500 μ m, long 75mm, one deck Novozym435(90mg) be filled in this microchannel.Control 60 ℃ of temperature of reaction, reaction raw materials residence time in reactor is about 1h.Collect outlet material, the yield of being combined to the coffic acid propyl ester with the transesterification of HPLC detection enzymatic is 78.9%.
Embodiment 5
Apparatus structure, as embodiment 1, joins the 1mg Methyl caffeoate [Bmim] [CF of 1mL
3sO
3] in ionic liquid, according to Methyl caffeoate and propyl alcohol mol ratio 1:40, being made into reaction system, this reaction system is continuously pumped in microreactor and carries out transesterification by micro-injection pump with the flow velocity of 2 μ L/min.The wide 1cm of microchannel in the middle of microreactor, high 500 μ m, long 75mm, one deck Novozym435(90mg) be filled in this microchannel.Control 70 ℃ of temperature of reaction, reaction raw materials residence time in reactor is about 2.5h.Collect outlet material, the yield of being combined to the coffic acid propyl ester with the transesterification of HPLC detection enzymatic is 97.6%.
Embodiment 6
Apparatus structure, as embodiment 1, joins the 1mg Methyl caffeoate [Bmim] [CF of 1mL
3sO
3] in ionic liquid, according to Methyl caffeoate and propyl alcohol mol ratio 1:40, being made into reaction system, this reaction system is continuously pumped in microreactor and carries out transesterification by micro-injection pump with the flow velocity of 2 μ L/min.The wide 1cm of microchannel in the middle of microreactor, high 500 μ m, long 75mm, one deck Novozym435(90mg) be filled in this microchannel.Control 60 ℃ of temperature of reaction, reaction raw materials residence time in reactor is about 2.5h.Collect outlet material, the yield of being combined to the coffic acid propyl ester with the transesterification of HPLC detection enzymatic is 99.5%.
Control experiment
In traditional batch reactor, take the Methyl caffeoate as reaction substrate, according to the Methyl caffeoate: the mol ratio of phenylethyl alcohol is 1:5, the Methyl caffeoate: the mass ratio of lipase Novozym435 is 1:20, phenylethyl alcohol: ionic liquid [Bmim] [CF
3sO
3] volume ratio be that 1:10 is made into reaction system, under 60 ℃ of temperature of reaction, under rotating speed 120rpm, react 38h.After reaction finishes, sample is after HPLC detects, and calculating the CAPE yield is 98.5%.
Claims (5)
1. a method of utilizing the synthetic coffic acid propyl ester of microreactor enzymatic, it is characterized in that: lipase Novozym435 is filled into to immobilized zone, the ionic liquid that will contain substrate Methyl caffeoate and propyl alcohol with syringe pump is injected in immobilized zone, Methyl caffeoate's concentration range is 0.1mg/mL～20mg/mL, the molar ratio range of Methyl caffeoate and propyl alcohol is 1:1～1:200, flow rates is 0.1～100 μ L/min, range of reaction temperature is 30～80 ℃, carries out the enzymatic transesterification and prepares the coffic acid propyl ester; Described sandwich type microreactor comprises syringe pump (1) and reactor body, described reactor body material is polydimethylsiloxane (polydimethylsiloxane), and formed by Sptting plate (2) and cover plate (3), the Sptting plate center is provided with immobilized zone (4), the wide 1cm in immobilized zone, dark 500 μ m, long 50mm-200mm.
2. a kind of method of utilizing the synthetic coffic acid propyl ester of microreactor enzymatic according to claim 1, is characterized in that being provided with retaining clip between Sptting plate and cover plate.
3. a kind of method of utilizing the synthetic coffic acid propyl ester of microreactor enzymatic according to claim 1, is characterized in that described lipase Novozym435 adopts the mode of tiling to be filled in immobilized zone.
4. a kind of method of utilizing the synthetic coffic acid propyl ester of microreactor enzymatic according to claim 1, is characterized in that the filling enzyme amount of lipase Novozym435 is 50mg-200mg.
5. a kind of method of utilizing the synthetic coffic acid propyl ester of microreactor enzymatic according to claim 1, the positive and negative ionic type combination that it is characterized in that described ionic liquid comprises that cation type is [Bmim], [Hmim], [Emim], [TOMA] or [Omim], and anionic type is [CF
3sO
3], [PF
6], [Tf
2n], [BF
4] or [Cl].
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104404091A (en) * | 2014-11-19 | 2015-03-11 | 江苏大学 | Enzymatic synthesis method for methyl caffeate by utilizing microreactor |
| CN112375801A (en) * | 2020-10-22 | 2021-02-19 | 复旦大学 | Micro-reaction system and method for continuously preparing (R) -3-hydroxy-5-hexenoic acid ester by using same |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103088078A (en) * | 2013-02-04 | 2013-05-08 | 江苏科技大学 | Lipase synthesis-promoting method of propyl caffeate |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103088078A (en) * | 2013-02-04 | 2013-05-08 | 江苏科技大学 | Lipase synthesis-promoting method of propyl caffeate |
Non-Patent Citations (2)
| Title |
|---|
| ROHAN KARANDE ET AL.: "Enzyme Catalysis in an Aqueous/Organic Segment Flow Microreactor:Ways to Stabilize Enzyme Activity", 《LANGMUIR》 * |
| 李晶等: "离子液体中脂肪酶催化酯类合成的新进展", 《有机化学》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104404091A (en) * | 2014-11-19 | 2015-03-11 | 江苏大学 | Enzymatic synthesis method for methyl caffeate by utilizing microreactor |
| CN112375801A (en) * | 2020-10-22 | 2021-02-19 | 复旦大学 | Micro-reaction system and method for continuously preparing (R) -3-hydroxy-5-hexenoic acid ester by using same |
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