CN108315372A - A kind of method of enzyme process removing trichloro-cane-6-ethyl ester acetyl group - Google Patents
A kind of method of enzyme process removing trichloro-cane-6-ethyl ester acetyl group Download PDFInfo
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- CN108315372A CN108315372A CN201810075298.0A CN201810075298A CN108315372A CN 108315372 A CN108315372 A CN 108315372A CN 201810075298 A CN201810075298 A CN 201810075298A CN 108315372 A CN108315372 A CN 108315372A
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Abstract
The invention discloses a kind of methods that enzyme process removes 6 ethyl ester acetyl group of Sucralose, belong to biocatalysis technology field.For the method for the present invention in organic solvent system, 6 ethyl ester of enzymatic Sucralose removes acetyl group, and Sucralose is made.The advantages that reaction condition of the present invention is mild, and good reaction selectivity, reaction product is single, high conversion rate, product safety prepares Sucralose for biology method and provides new feasible way.
Description
Technical field
The present invention relates to the methods that enzyme process prepares Sucralose, belong to biocatalysis technology field.
Background technology
Sucralose（sucralose）It is a kind of novel functional sweetener, Sucralose is one kind of chloro sucrose.
Its sugariness is 600-800 times of sucrose, and safe, pure taste, energy metabolism, energy value zero do not have well human body
Dissolubility and stability, these advantages make the sweetener of forcing of food, beverage etc., at present by more than 30 countries batch
Standard uses.
The synthetic method of Sucralose is primarily now chemical method comprising two class of full radical protection method and mono-esterification method.
Full radical protection method（Such as United States Patent (USP) US4617269）It is the steric hindrance using triphenyl methane, by 3 primary hydroxyls of sucrose
（6-, 1 '-, 6 '-positions）Tritylation, 5 secondary hydroxyls of acylated residue obtain, and five Deprotection, chloro, removing acyl groups obtain
Sucralose, the notable disadvantage of this kind of reaction is exactly that process is cumbersome.Mono-esterification method（US 4889928、US5023329、
US4950746、CN1453285A）It is with chemical means by the shielding of 6 hydroxyls, production list on saccharose group in sucrose molecule
Ester obtains Sucralose using chlorination, deprotection group acetyl group.
No matter which kind of mode to prepare Sucralose by, deacetylation is obtained as final for sucralose compounds,
All it is wherein necessary processing step.Therefore optimization deacetylating step is the important way for improving Sucralose synthesis technology efficiency
Diameter.
In currently available technology, our generally use strong base solutions realize the deacetylation of trichloro-cane-6-ethyl ester
Purpose.This method is usually to be carried out in sodium methoxide or sodium hydroxide are as the methanol solution of catalyst.Using this technique,
It needs a large amount of sodium ions introducing reaction system, resin adsorption removing is also needed to after obtaining the reaction mixture containing Sucralose
Sodium ion.It thus necessarily will produce many downstream processes and purifying products step.Simultaneously as its harsh pH conditions and temperature
Degree may cause glycosidic bond fracture, chlorine atom to fall off, cause by-product more, yield is low.
Invention content
The present invention is directed in currently available technology prepares deficiency existing for Sucralose using chemical method, discloses a kind of enzyme process system
The method of standby Sucralose.Specifically, be by enzymatic reaction by trichloro-cane-6-ethyl ester 6 acetyl group removing from
And the method for obtaining Sucralose product.
Sucralose is prepared using enzyme process reaction, mild condition, high selectivity, high conversion rate, product are easy to purify.
Preferably, which is happened in the reaction system of organic solvent formation.
The present invention, as reaction system, on the one hand can improve reactant solubility using organic solvent, still further aspect,
Using organic solvent as reaction system, the enzyme in enzymatic reaction is suspended in organic solvent, is easily recycled after the completion of reaction.
It is also worth noting that using organic solvent as reaction system, the stability of relevant enzyme can be significantly improved.And it utilizes
Under the reaction system of organic solvent, by process regulations such as concentration, temperature, times, may be implemented in enzymatic reaction
The specificity and selective regulation of enzyme.
In addition to this, solvent can also be facilitated to remove and recycle using organic solvent as reaction system, simplifies follow-up step
Suddenly.
Further, the method that enzyme process disclosed by the invention prepares Sucralose is specifically described as：It is reacted in organic solvent
In system, using the trichloro-cane-6-ethyl ester of a concentration of 10 ~ 50g/L as substrate, the enzyme of 2 ~ 10mg/ml is added thereto, 20 ~
It under conditions of 80 DEG C, reacts 6-24 hours, obtains Sucralose.
Preferably, it at the uniform velocity stirs in the reaction.
Preferably, the enzyme, which is further disclosed, in the present invention is selected from Lipase AS Amano（from
Aspergillus niger）、Pig liver esterase、Lipase sp-435（Candide antarctica）、CAL-B
（Lipase B from Candida antarctica）、Lipase G Amano(from Penicillium
Camemberti any one or a few in).
Meanwhile the present invention further discloses the organic solvent and is selected from acetonitrile, tetrahydrofuran, methanol, ethyl alcohol, uncle
In butanol any one or mutually it is soluble several.
Here a kind of organic solvent can be selected as reaction system, two kinds or two or more organic can also be selected
Solution as reaction system, and it is of note that when select two kinds or two or more organic solutions as reaction system
When, it should ensure that each organic solution is mutually solvable.
In addition, the present invention is further disclosed including step：After reaction, it filtering, reaction clear liquid, which passes through, to be concentrated in vacuo,
Solvent is removed, Sucralose crude product is obtained.
Preferably, the temperature of vacuum concentration is 50 ~ 80 DEG C.
Above-mentioned steps can also be：After reaction, it centrifuges, reaction clear liquid removes solvent, obtain three by being concentrated in vacuo
Chlorine sucrose crude product.
The temperature being wherein concentrated in vacuo is 50 ~ 80 DEG C.
Preferably, enzyme is recycled after filtering or centrifuge.Wherein solids portion is enzyme.
After technical solution disclosed by the invention, compare chemical method, this reaction has reaction condition mild, reaction choosing
The advantages that selecting property is high, and reaction product is single, high conversion rate.
Specific implementation mode
With reference to embodiment, the claim of the present invention is described in further detail.
Not specified, agents useful for same is conventional commercial reagent manufacture in the present invention.
Lipase AS Amano（from Aspergillus niger）、Lipase G Amano(from
Penicillium camemberti) purchased from Japanese amano enzyme company.Pig liver esterase、Lipase sp-435
（Candide antarctica）、CAL-B（Lipase B from Candida antarctica）It is public purchased from Aladdin reagent
Department.
The synthesis of 1 Sucralose of embodiment
Trichloro-cane-6-ethyl ester 5g is dissolved in 100ml acetonitriles, 0.8gLipase AS Amano are added（from
Aspergillus niger）, reacted 24 hours under the conditions of 40 DEG C, 1g activated carbon decolorizings be added, are stirred at room temperature 30 minutes, mistake
Enzyme is recycled in filter, and filter vacuum concentrates removing acetonitrile under the conditions of 70 DEG C.Sucralose crude product 4.3g is obtained, 5ml water is added, heating is stirred
Mix dissolving.It is at the uniform velocity cooled to room temperature in 5 hours, obtains Sucralose crystallization.Filtering, dry finished product 4.0g（Purity is 91%,
Yield 90%）.
The synthesis of 2 Sucralose of embodiment
Trichloro-cane-6-ethyl ester 1g is dissolved in the 100ml tert-butyl alcohols, 0.6gPig liver esterase are added, in 80 DEG C of conditions
Lower reaction 14 hours, is added 1g activated carbon decolorizings, is stirred at room temperature 30 minutes, centrifugation recycling enzyme, and filter vacuum is dense under the conditions of 70 DEG C
Contracting removes the tert-butyl alcohol.Sucralose crude product 0.92g is obtained, 5ml water, heating stirring dissolving is added.It is at the uniform velocity cooled to room in 3 hours
Temperature obtains Sucralose crystallization.Filtering, dry finished product 0.84g（Purity is 96%, yield 92%）.
The synthesis of 3 Sucralose of embodiment
Trichloro-cane-6-ethyl ester 4g is dissolved in 100ml methanol, 0.4gLipase sp-435 are added（Candide
antarctica）, reacted 6 hours at 60 DEG C, 1g activated carbon decolorizings be added, is stirred at room temperature 40 minutes, enzyme, 50 DEG C of items is recovered by filtration
It is concentrated in vacuo under part and removes methanol.Obtain Sucralose crude product 3.8g.5ml water, heating stirring dissolving is added.In 5 hours at the uniform velocity
It is cooled to room temperature, obtains Sucralose crystallization.Filtering, dry finished product 3.6g（Purity is 92%, yield 90%）.
Embodiment 4
Trichloro-cane-6-ethyl ester 3g is dissolved in 100ml ethyl alcohol, adds 1.0gCAL-B（Lipase B from Candida
antarctica）, reacted 10 hours under the conditions of 20 DEG C, 1g activated carbon decolorizings be added, is stirred at room temperature 40 minutes, is recovered by filtration
Enzyme is concentrated in vacuo under the conditions of 60 DEG C and removes the tert-butyl alcohol.Obtain Sucralose crude product 2.81g.5ml water, heating stirring dissolving is added.
It is at the uniform velocity cooled to room temperature in 5 hours, obtains Sucralose crystallization.Filtering, dry finished product 2.73g（Purity is 93%, yield
91%）.
Embodiment 5
Trichloro-cane-6-ethyl ester 2g is dissolved in 100ml tetrahydrofurans, 0.2gLipase G Amano (from are added
Penicillium camemberti), it is reacted 24 hours at 50 DEG C, 1g activated carbon decolorizings is added, are stirred at room temperature 40 minutes, filtered
Enzyme is recycled, is concentrated in vacuo under the conditions of 50 DEG C and removes ethyl alcohol.Obtain Sucralose crude product 1.96g.5ml water is added, heating stirring is molten
Solution.It is at the uniform velocity cooled to room temperature in 5 hours, obtains Sucralose crystallization.Filtering, dry finished product 1.8g（Purity is 94%, yield
90%）.
Claims (10)
1. a kind of method of enzyme process removing trichloro-cane-6-ethyl ester acetyl group, it is characterized in that：This method refers to passing through enzymatic reaction
6 in trichloro-cane-6-ethyl ester acetyl group are removed into the method to obtain Sucralose product.
2. a kind of method of enzyme process removing trichloro-cane-6-ethyl ester acetyl group according to claim 1, it is characterized in that：It should
Enzymatic reaction is happened in the reaction system of organic solvent formation.
3. a kind of method of enzyme process removing trichloro-cane-6-ethyl ester acetyl group according to claim 1 or 2, it is characterized in that：
In organic solvent reaction system, using the trichloro-cane-6-ethyl ester of 10 ~ 50g/L of concentration as substrate, thereto be added concentration 2 ~
The enzyme of 10mg/ml reacts 6-24 hours under conditions of 20 ~ 80 DEG C, obtains Sucralose.
4. a kind of method of enzyme process removing trichloro-cane-6-ethyl ester acetyl group according to claim 3, it is characterized in that：Bottom
A concentration of 10 ~ 50g/L of object trichloro-cane-6-ethyl ester.
5. a kind of method of enzyme process removing trichloro-cane-6-ethyl ester acetyl group according to claim 3, it is characterized in that：Institute
The enzyme stated is selected from Lipase AS Amano, Pig liver esterase, Lipase sp-435, CAL-B, Lipase G
Any one or a few in Amano.
6. a kind of method of enzyme process removing trichloro-cane-6-ethyl ester acetyl group according to claim 3, it is characterized in that：Have
Solvent in acetonitrile, tetrahydrofuran, methanol, ethyl alcohol, the tert-butyl alcohol any one or mutually it is soluble several.
7. a kind of method of enzyme process removing trichloro-cane-6-ethyl ester acetyl group according to claim 3, it is characterized in that：Also
Including step, after reaction, filtering, reaction clear liquid removes solvent, obtains Sucralose crude product by being concentrated in vacuo.
8. a kind of method of enzyme process removing trichloro-cane-6-ethyl ester acetyl group according to claim 3, it is characterized in that：Instead
After answering, centrifugation, reaction clear liquid removes solvent, obtains Sucralose crude product by being concentrated in vacuo.
9. a kind of method of enzyme process removing trichloro-cane-6-ethyl ester acetyl group according to claim 7 or 8, it is characterized in that：
The temperature of vacuum concentration is 50 ~ 80 DEG C.
10. a kind of method of enzyme process removing trichloro-cane-6-ethyl ester acetyl group according to claim 7 or 8, feature
It is：Enzyme is recycled after filtering or centrifugation.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0043649A1 (en) * | 1980-07-08 | 1982-01-13 | TATE & LYLE PUBLIC LIMITED COMPANY | Process for the preparation of 4, 1',6'-trichloro-4,1',6'-trideoxygalactosucrose (TGS) |
| CN101260127A (en) * | 2007-03-06 | 2008-09-10 | 盐城捷康三氯蔗糖制造有限公司 | Method for synthesizing trichlorosucrose by deacetylation of trichlorosucrose-6-ethyl ester |
| CN101709069A (en) * | 2009-08-06 | 2010-05-19 | 浙江工业大学 | Novel method for separating and purifying sucralose-6-ethyl ester |
| CN101886100A (en) * | 2010-07-12 | 2010-11-17 | 江南大学 | Method for preparing sucrose-6-acetic ester by enzymatic method |
| CN101928738A (en) * | 2010-08-23 | 2010-12-29 | 浙江工业大学 | Method for synthesizing cane sugar-6-acetic ester by using lipase for catalyzing |
-
2018
- 2018-01-26 CN CN201810075298.0A patent/CN108315372A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0043649A1 (en) * | 1980-07-08 | 1982-01-13 | TATE & LYLE PUBLIC LIMITED COMPANY | Process for the preparation of 4, 1',6'-trichloro-4,1',6'-trideoxygalactosucrose (TGS) |
| CN101260127A (en) * | 2007-03-06 | 2008-09-10 | 盐城捷康三氯蔗糖制造有限公司 | Method for synthesizing trichlorosucrose by deacetylation of trichlorosucrose-6-ethyl ester |
| CN101709069A (en) * | 2009-08-06 | 2010-05-19 | 浙江工业大学 | Novel method for separating and purifying sucralose-6-ethyl ester |
| CN101886100A (en) * | 2010-07-12 | 2010-11-17 | 江南大学 | Method for preparing sucrose-6-acetic ester by enzymatic method |
| CN101928738A (en) * | 2010-08-23 | 2010-12-29 | 浙江工业大学 | Method for synthesizing cane sugar-6-acetic ester by using lipase for catalyzing |
Non-Patent Citations (3)
| Title |
|---|
| CHAUBEY A等: "Bioconversion of sucralose-6-acetate to sucralose using immobilized microbial cells", 《JOURNAL OF MOLECULAR CATALYSIS B ENZYMATIC》 * |
| PALMER D C等: "Regioselective enzymatic deacetylation of sucrose octaacetate in organic solvents", 《TETRAHEDRON LETTERS》 * |
| 张德华: "《蛋白质与酶工程》", 30 September 2015 * |
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Application publication date: 20180724 |