CN103014081A - Biological preparation method of 3-amino-4-(2,4,5-trifluorophenyl)methyl butyrate - Google Patents
Biological preparation method of 3-amino-4-(2,4,5-trifluorophenyl)methyl butyrate Download PDFInfo
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- trifluorophenyl
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- JGFBQFKZKSSODQ-UHFFFAOYSA-N Isothiocyanatocyclopropane Chemical compound S=C=NC1CC1 JGFBQFKZKSSODQ-UHFFFAOYSA-N 0.000 title claims abstract description 29
- PWLNAUNEAKQYLH-UHFFFAOYSA-N butyric acid octyl ester Natural products CCCCCCCCOC(=O)CCC PWLNAUNEAKQYLH-UHFFFAOYSA-N 0.000 title claims abstract description 29
- UUIQMZJEGPQKFD-UHFFFAOYSA-N n-butyric acid methyl ester Natural products CCCC(=O)OC UUIQMZJEGPQKFD-UHFFFAOYSA-N 0.000 title claims abstract description 29
- 238000002360 preparation method Methods 0.000 title claims abstract description 22
- 238000006243 chemical reaction Methods 0.000 claims abstract description 17
- 239000000758 substrate Substances 0.000 claims abstract description 11
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims abstract description 10
- 239000000047 product Substances 0.000 claims abstract description 10
- 238000000034 method Methods 0.000 claims abstract description 8
- 102000003929 Transaminases Human genes 0.000 claims description 26
- 108090000340 Transaminases Proteins 0.000 claims description 26
- NGVDGCNFYWLIFO-UHFFFAOYSA-N pyridoxal 5'-phosphate Chemical group CC1=NC=C(COP(O)(O)=O)C(C=O)=C1O NGVDGCNFYWLIFO-UHFFFAOYSA-N 0.000 claims description 12
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N dimethyl sulfoxide Natural products CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 10
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 8
- 238000013016 damping Methods 0.000 claims description 8
- 239000012530 fluid Substances 0.000 claims description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 8
- 239000007788 liquid Substances 0.000 claims description 7
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 6
- XUWPJKDMEZSVTP-LTYMHZPRSA-N kalafungina Chemical compound O=C1C2=C(O)C=CC=C2C(=O)C2=C1[C@@H](C)O[C@H]1[C@@H]2OC(=O)C1 XUWPJKDMEZSVTP-LTYMHZPRSA-N 0.000 claims description 6
- 235000007682 pyridoxal 5'-phosphate Nutrition 0.000 claims description 6
- 239000011589 pyridoxal 5'-phosphate Substances 0.000 claims description 6
- 229960001327 pyridoxal phosphate Drugs 0.000 claims description 6
- 230000004913 activation Effects 0.000 claims description 5
- 239000000872 buffer Substances 0.000 claims description 5
- 239000003054 catalyst Substances 0.000 claims description 5
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical group CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 claims description 5
- 238000003756 stirring Methods 0.000 claims description 5
- 239000000725 suspension Substances 0.000 claims description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 4
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 claims description 4
- 238000001914 filtration Methods 0.000 claims description 4
- 238000004128 high performance liquid chromatography Methods 0.000 claims description 4
- JJWLVOIRVHMVIS-UHFFFAOYSA-N isopropylamine Chemical group CC(C)N JJWLVOIRVHMVIS-UHFFFAOYSA-N 0.000 claims description 4
- 229910052757 nitrogen Inorganic materials 0.000 claims description 4
- 239000002953 phosphate buffered saline Substances 0.000 claims description 4
- 238000010926 purge Methods 0.000 claims description 4
- 238000002390 rotary evaporation Methods 0.000 claims description 4
- 238000004108 freeze drying Methods 0.000 claims description 3
- 238000011081 inoculation Methods 0.000 claims description 3
- 239000006228 supernatant Substances 0.000 claims description 3
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 claims description 2
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 claims description 2
- GSEJCLTVZPLZKY-UHFFFAOYSA-N Triethanolamine Chemical compound OCCN(CCO)CCO GSEJCLTVZPLZKY-UHFFFAOYSA-N 0.000 claims description 2
- 238000012258 culturing Methods 0.000 claims description 2
- 238000001514 detection method Methods 0.000 claims description 2
- 238000001704 evaporation Methods 0.000 claims description 2
- 230000008020 evaporation Effects 0.000 claims description 2
- 239000000706 filtrate Substances 0.000 claims description 2
- 239000008101 lactose Substances 0.000 claims description 2
- 238000001556 precipitation Methods 0.000 claims description 2
- 239000002904 solvent Substances 0.000 claims description 2
- 229960004418 trolamine Drugs 0.000 claims description 2
- -1 (2,4,5-trifluorophenyl) methyl-butyrate Chemical compound 0.000 claims 6
- 230000003287 optical effect Effects 0.000 abstract description 5
- 102000004190 Enzymes Human genes 0.000 abstract description 4
- 108090000790 Enzymes Proteins 0.000 abstract description 4
- 239000006227 byproduct Substances 0.000 abstract description 2
- 238000005265 energy consumption Methods 0.000 abstract description 2
- 239000000126 substance Substances 0.000 abstract description 2
- 108010065394 aminopherase Proteins 0.000 abstract 2
- 239000011942 biocatalyst Substances 0.000 abstract 2
- 238000005215 recombination Methods 0.000 abstract 2
- 230000006798 recombination Effects 0.000 abstract 2
- 239000007853 buffer solution Substances 0.000 abstract 1
- 239000006184 cosolvent Substances 0.000 abstract 1
- 230000036983 biotransformation Effects 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- MFFMDFFZMYYVKS-SECBINFHSA-N sitagliptin Chemical compound C([C@H](CC(=O)N1CC=2N(C(=NN=2)C(F)(F)F)CC1)N)C1=CC(F)=C(F)C=C1F MFFMDFFZMYYVKS-SECBINFHSA-N 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 150000001576 beta-amino acids Chemical class 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000003810 ethyl acetate extraction Methods 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 229960004034 sitagliptin Drugs 0.000 description 2
- 230000009466 transformation Effects 0.000 description 2
- JKTORXLUQLQJCM-UHFFFAOYSA-N 4-phosphonobutylphosphonic acid Chemical compound OP(O)(=O)CCCCP(O)(O)=O JKTORXLUQLQJCM-UHFFFAOYSA-N 0.000 description 1
- 241000186063 Arthrobacter Species 0.000 description 1
- 108010016626 Dipeptides Proteins 0.000 description 1
- KJTLSVCANCCWHF-UHFFFAOYSA-N Ruthenium Chemical compound [Ru] KJTLSVCANCCWHF-UHFFFAOYSA-N 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 230000002924 anti-infective effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229960000074 biopharmaceutical Drugs 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 125000004435 hydrogen atom Chemical class [H]* 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 229940090473 januvia Drugs 0.000 description 1
- 125000000468 ketone group Chemical group 0.000 description 1
- 238000010907 mechanical stirring Methods 0.000 description 1
- 208000030159 metabolic disease Diseases 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 229910052707 ruthenium Inorganic materials 0.000 description 1
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/52—Improvements relating to the production of bulk chemicals using catalysts, e.g. selective catalysts
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to a biological preparation method of 3-amino-4-(2,4,5-trifluorophenyl)methyl butyrate. According to the biological preparation method, 3-carbonyl-4-(2,4,5-trifluorophenyl)methyl butyrate is used as a substrate; under the condition that a biocatalyst, a cofactor, an amino donor and a cosolvent exist, the substrate performs the reaction to generate a target product 3-amino-4-(2,4,5-trifluorophenyl)methyl butyrate; the biocatalyst is recombination aminopherase; and the reaction is carried out in water-phase buffer solution with the pH of 6.0 to 9.0. The biological preparation method realizes that 3-carbonyl-4-(2,4,5-trifluorophenyl)methyl butyrate is converted into 3-amino-4-(2,4,5-trifluorophenyl)methyl butyrate by adopting the recombination aminopherase for the first time; compared with a conventional chemical method, the biological preparation method has low energy consumption, is simple and convenient to operate, generates a small number of by-products; and moreover, the product has high optical activity.
Description
Technical field
The invention belongs to bio-pharmaceuticals and Green Chemistry field, be specifically related to the biological preparation method of a kind of 3-amino-4-(2,4,5-trifluorophenyl) methyl-butyrate.
Background technology
Biocatalysis has obvious green, efficient and economic dispatch characteristics.Utilizing the transaminase selectivity that ketone group is converted into amino, is one of most important bioconversion reaction.
Transaminase (transaminase, amino transferase) is for the synthesis of chirality ammonia or amino acid whose class of enzymes.Wherein the application of beta-amino acids material in medicine is synthetic is very extensive, can be for the synthesis of a series of dipeptide peptidases-IV inhibitor, the medicine (Pharmacol.2004 of important antitumor, anti-infective and treatment metabolism disorder, 404:239, BMC Endocr.Disord.2008,8:14).Prepare in two kinds of main selections of beta-amino acids material in the method for utilizing bio-transformation, compared to complex steps, enzyme Split Method (the Org.Biomol.Chem.2010 that yield is lower, 8:793-799, product yield 42%, optical purity 96%), the method for utilizing transaminase directly to synthesize has fairly obvious superiority.And in the report that utilizes synthetic such material of chemical synthesis process, its yield lower (68%), and need under the condition that high pressure and hydrogen, metal ruthenium catalyst exist, react security and environment friendly relatively poor (Bioorg.Med.Chem.Lett.2007,17:2622).
Utilize transaminase to carry out in the report of bio-transformation, the people such as Savile of Codexis company transform and are used to transaminase ATA-117 from Arthrobacter, catalysis 1-(3-(trifluoromethyl)-5,6-dihydro-[1,2,4] triazole [4,3-a] pyrazine-7 (8H)-yl)-4-(2,4,5-trifluorophenyl) butane-1, the 3-diketone is converted into 3-amino-1-(3-(trifluoromethyl)-5,6-dihydro-[1,2,4] triazole [4,3-a] pyrazine-7 (8H)-yl)-4-(2,4,5-trifluorophenyl) butane-1-ketone, i.e. the treatment diabetes medicament sitagliptin (Sitagliptin of Merck company, trade(brand)name JANUVIA) be to utilize transaminase to carry out the outstanding example (Science2010,329:305) of bio-transformation.
3-carbonyl-4-(2,4,5-trifluorophenyl) methyl-butyrate is one of model substrates of above-mentioned reaction, also is important potential chiral precursor.But, up to the present, also utilize transaminase 3-carbonyl-4-(2,4,5-trifluorophenyl) methyl-butyrate to be converted into the report of 3-amino-4-(2,4,5-trifluorophenyl) methyl-butyrate.
Summary of the invention
Technical problem to be solved by this invention provides the biological preparation method that a kind of utilization restructuring transaminase is converted into 3-carbonyl-4-(2,4,5-trifluorophenyl) methyl-butyrate 3-amino-4-(2,4,5-trifluorophenyl) methyl-butyrate.
For solving above technical problem, the present invention adopts following technical scheme:
A kind of 3-amino-4-(2,4, the 5-trifluorophenyl) biological preparation method of methyl-butyrate, it is with 3-carbonyl-4-(2,4, the 5-trifluorophenyl) methyl-butyrate is substrate, this substrate is at biological catalyst, cofactor, react under the existence of amino donor and solubility promoter and generate target product 3-amino-4-(2,4, the 5-trifluorophenyl) methyl-butyrate, described biological catalyst is the restructuring transaminase, the preparation method of described restructuring transaminase is: will contain single colony inoculation of transaminase in the liquid LB substratum that contains kalamycin resistance, in 35-40 ℃ of lower activation 8-12 hour, the culture that obtains after the activation is transferred in the liquid LB substratum that contains kalamycin resistance, reach 0.6-0.8 in 35-40 ℃ of lower enlarged culturing to OD600 value, add inductor, continue to cultivate 8-12 hour in 25-35 ℃, centrifugal, the collecting precipitation thing adds phosphate buffered saline buffer and obtains suspension, suspension was placed the ice bath ultrasonic disruption 8-12 minute, centrifugal again, be-10 ~-25 ℃ with supernatant liquor pre-freeze to temperature, and then freeze-drying 24-48 hour, namely get the restructuring transaminase, described reaction is carried out in pH is the water damping fluid of 6.0-9.0.
Further, in the reaction system when initial, described restructuring transaminase: described cofactor: described amino donor: the mass ratio of described 3-carbonyl-4-(2,4,5-trifluorophenyl) methyl-butyrate is 0.4-0.6:0.01-0.1:5-7:1.
Preferably, described inductor is isopropyl-β-D-thiogalactoside(IPTG) or lactose.
Preferably, described cofactor is pyridoxal phosphate.
Preferably, described amino donor is Isopropylamine.
Preferably, described solubility promoter is dimethyl sulfoxide (DMSO).
Preferably, described water damping fluid is phosphate buffered saline buffer, Tris-HCl buffered soln or trolamine hydrochloric acid buffered soln.
Preferably, described reaction is carried out in pH is 7.0 water damping fluid.
Further, described preparation method's implementation process is as follows: add successively described water damping fluid in reaction vessel, substrate 3-carbonyl-4-(2,4, the 5-trifluorophenyl) methyl-butyrate, described amino donor, described solubility promoter, described cofactor and described restructuring transaminase, stir, under the condition of 25-45 ℃ and nitrogen purging, react, utilize HPLC detection reaction process, when rate to be transformed reaches 90-99%, regulate pH to 2-3, diatomite filtration, add ethyl acetate and repeatedly extract in filtrate, solvent is sloughed in the rotary evaporation evaporation, namely gets 3-amino-4-(2,4,5-trifluorophenyl) methyl-butyrate product.
According to the present invention, except the restructuring transaminase, all the other raw materials all can be commercially available by general channel.
Beneficial effect of the present invention is:
The present invention has realized adopting the restructuring transaminase with 3-carbonyl-4-(2 first, 4, the 5-trifluorophenyl) methyl-butyrate is converted into 3-amino-4-(2,4,5-trifluorophenyl) methyl-butyrate is compared with traditional chemical method, energy consumption of the present invention is low, simple to operation, by product is few, and the optical activity of product is higher.
Embodiment
Reaction formula of the present invention is as follows:
The following examples can make the present invention of professional and technical personnel's comprehend, but do not limit the present invention in any way.
Embodiment one
From the glycerine pipe or transform dull and stereotyped single colony inoculation and spend the night (37 ℃ to the liquid LB substratum activation that 4mL contains kalamycin resistance, 200rpm), from overnight culture with 1/100(v/v) inoculum size is transferred to the liquid LB substratum that 100mL contains kalamycin resistance, 37 ℃, 200rpm shaking culture to OD600 value reaches 0.6-0.8, add IPTG and continue overnight incubation in 30 ℃, centrifugal collecting cell, with 10mL phosphoric acid buffer (2mM, pH7.0) suspension cell, cell suspending liquid placed the ice bath ultrasonic disruption 10 minutes, centrifugal again, supernatant liquor pre-freeze spend the night to temperature be-10 ~-25 ℃, freeze-drying 24h-48h again.
Embodiment two
In the 20mL there-necked flask, add successively 0.1M pH 7.0 phosphate buffer soln 3.85mL, substrate 100mg, Isopropylamine 900 μ L, DMSO 0.25mL, pyridoxal phosphate (PLP) 1mg, transaminase 50mg is in 30 ℃, the 200rpm stirring rake stirs, 0.01MPa under the condition of nitrogen purging, reaction 24h, HPLC detects transformation efficiency〉95%.Add the salt acid for adjusting pH to 2-3, diatomite filtration adds equal-volume ethyl acetate extraction twice, and rotary evaporation obtains product 90mg, purity〉95%, optical purity〉99%.
Embodiment three
In reactor, add successively 0.1M pH 7.0 phosphate buffer soln 3.85L, substrate 100g, Isopropylamine 900mL, DMSO 0.25L, pyridoxal phosphate (PLP) 1g, transaminase 50g is in 30 ° of C, the mechanical stirring oar stirs, 0.01MPa under the condition of nitrogen purging, reaction 24h, HPLC detects transformation efficiency〉95%.Add the salt acid for adjusting pH to 2-3, diatomite filtration adds equal-volume ethyl acetate extraction twice, and rotary evaporation obtains about product 90g, purity〉95%, optical purity〉99%.
Above-described embodiment only is explanation technical conceive of the present invention and characteristics, and its purpose is to allow the personage who is familiar with technique can understand content of the present invention and according to this enforcement, can not limit protection scope of the present invention with this.All equivalences that spirit is done according to the present invention change or modify, and all should be encompassed within protection scope of the present invention.
Claims (9)
1. 3-amino-4-(2,4, the 5-trifluorophenyl) biological preparation method of methyl-butyrate, it is characterized in that: it is with 3-carbonyl-4-(2,4, the 5-trifluorophenyl) methyl-butyrate is substrate, this substrate is at biological catalyst, cofactor, react under the existence of amino donor and solubility promoter and generate target product 3-amino-4-(2,4, the 5-trifluorophenyl) methyl-butyrate, described biological catalyst is the restructuring transaminase, the preparation method of described restructuring transaminase is: will contain single colony inoculation of transaminase in the liquid LB substratum that contains kalamycin resistance, in 35-40 ℃ of lower activation 8-12 hour, the culture that obtains after the activation is transferred in the liquid LB substratum that contains kalamycin resistance, reach 0.6-0.8 in 35-40 ℃ of lower enlarged culturing to OD600 value, add inductor, continue to cultivate 8-12 hour in 25-35 ℃, centrifugal, the collecting precipitation thing, add phosphate buffered saline buffer and obtain suspension, suspension was placed the ice bath ultrasonic disruption 8-12 minute, centrifugal again, it is-10 ~-25 ℃ with supernatant liquor pre-freeze to temperature, and then freeze-drying 24-48 hour, namely get the restructuring transaminase, described reaction is carried out in pH is the water damping fluid of 6.0-9.0.
2. 3-amino-4-(2 according to claim 1,4, the 5-trifluorophenyl) biological preparation method of methyl-butyrate, it is characterized in that: in the reaction system when initial, described restructuring transaminase: described cofactor: described amino donor: described 3-carbonyl-4-(2,4,5-trifluorophenyl) mass ratio of methyl-butyrate is 0.4-0.6:0.01-0.1:5-7:1.
3. the biological preparation method of 3-amino-4-according to claim 1 (2,4,5-trifluorophenyl) methyl-butyrate, it is characterized in that: described inductor is isopropyl-β-D-thiogalactoside(IPTG) or lactose.
4. the biological preparation method of 3-amino-4-according to claim 1 (2,4,5-trifluorophenyl) methyl-butyrate, it is characterized in that: described cofactor is pyridoxal phosphate.
5. the biological preparation method of 3-amino-4-according to claim 1 (2,4,5-trifluorophenyl) methyl-butyrate, it is characterized in that: described amino donor is Isopropylamine.
6. the biological preparation method of 3-amino-4-according to claim 1 (2,4,5-trifluorophenyl) methyl-butyrate, it is characterized in that: described solubility promoter is dimethyl sulfoxide (DMSO).
7. the biological preparation method of 3-amino-4-according to claim 1 (2,4,5-trifluorophenyl) methyl-butyrate, it is characterized in that: described water damping fluid is phosphate buffered saline buffer, Tris-HCl buffered soln or trolamine hydrochloric acid buffered soln.
8. the biological preparation method of 3-amino-4-according to claim 1 (2,4,5-trifluorophenyl) methyl-butyrate is characterized in that: described reaction is carried out in pH is the water damping fluid of 6.0-9.0.
9. 3-amino-4-(2 according to claim 1 and 2,4, the 5-trifluorophenyl) biological preparation method of methyl-butyrate, it is characterized in that: described preparation method's implementation process is as follows: add successively described water damping fluid in reaction vessel, substrate 3-carbonyl-4-(2,4, the 5-trifluorophenyl) methyl-butyrate, described amino donor, described solubility promoter, described cofactor and described restructuring transaminase, stir, under the condition of 25 ~ 45 ℃ and nitrogen purging, react, utilize HPLC detection reaction process, when rate to be transformed reaches 90 ~ 99%, regulate pH to 2-3, diatomite filtration, add ethyl acetate and repeatedly extract in filtrate, solvent is sloughed in the rotary evaporation evaporation, namely gets 3-amino-4-(2,4,5-trifluorophenyl) methyl-butyrate product.
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Cited By (10)
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| CN104131048A (en) * | 2014-08-01 | 2014-11-05 | 洛阳华荣生物技术有限公司 | Biological preparation method of R-3-aminobutanol |
| CN104803954A (en) * | 2015-04-30 | 2015-07-29 | 上海应用技术学院 | Preparation method for fosamprenir intermediate |
| CN105039450A (en) * | 2015-06-25 | 2015-11-11 | 南京迪缘医药科技有限公司 | Synthetic method of efinaconazole intermediate |
| CN106191148A (en) * | 2016-07-27 | 2016-12-07 | 苏州汉酶生物技术有限公司 | (R) biological preparation method of 3 amino 4 (2,4,5 trifluorophenyl) tert-butyl acetate |
| CN106399418A (en) * | 2015-07-29 | 2017-02-15 | 苏州汉酶生物技术有限公司 | Method for preparing moxifloxacin side chain through biological method |
| WO2019011236A1 (en) * | 2017-07-11 | 2019-01-17 | 上海弈柯莱生物医药科技有限公司 | Use of stereoselective transaminase in asymmetric synthesis of chiral amine |
| CN113846131A (en) * | 2020-06-28 | 2021-12-28 | 尚科生物医药（上海）有限公司 | Method for preparing (R) -3-amino-4- (2,4, 5-trifluorophenyl) -butyric acid |
| CN114702425A (en) * | 2022-03-28 | 2022-07-05 | 苏州汉酶生物技术有限公司 | Preparation method of (S) -2-amino- (S) -3- [ pyrrolidone-2' ] alanine derivative and intermediate |
| US11459549B2 (en) | 2018-05-10 | 2022-10-04 | China Fortune Way Company | Method for biocatalytic synthesis of Sitagliptin and intermediate thereof |
| WO2024125533A1 (en) | 2022-12-16 | 2024-06-20 | 浙江永太科技股份有限公司 | Omega-transaminase mutant and use thereof |
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