CN104131048A - Biological preparation method of R-3-aminobutanol - Google Patents
Biological preparation method of R-3-aminobutanol Download PDFInfo
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- CN104131048A CN104131048A CN201410375457.0A CN201410375457A CN104131048A CN 104131048 A CN104131048 A CN 104131048A CN 201410375457 A CN201410375457 A CN 201410375457A CN 104131048 A CN104131048 A CN 104131048A
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- 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
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Abstract
The invention provides a biological preparation method of R-3-aminobutanol. The method includes: conducting whole-gene synthesis according to an artificially designed sequence shown as SEQ ID NO:1, and performing cloning into pET24a so as to obtain a recombinant expression vector; transferring the recombinant expression vector into E.coli to obtain a recombinant D-transaminase genetic engineering bacterium; culturing the bacterium to prepare recombinant D-transaminase; adding 3-carbonyl butanol with a final concentration of 10-300mmol/L, dimethyl sulfoxide or acetonitrile with a mass percentage concentration of 2-15%, pyridoxal phosphate with a final concentration of 0.1-1mmol/L, isopropylamine or D-alanine with a final concentration of 0.02-1mol/L, and recombinant D-transaminase with a mass percentage concentration of 0.01-1% are added into the reaction solution to compose a reaction system for reaction; and at the end of the reaction, extracting R-3-aminobutanol from the reaction solution. With a low cost, the method provided by the invention can achieve a conversion rate of more than 97% and a product yield of greater than 87%, and does not generate by-product, thus being more suitable for industrial application.
Description
Technical field
The present invention relates to a kind of R-3-amino butanol, specifically a kind of biological preparation method of R-3-amino butanol.
Background technology
The features such as biocatalysis has reaction conditions gentleness, efficiency is high, stereoselectivity is strong, environmental friendliness, so biocatalysis is widely used in the research and development of new drug.In addition, utilize design and rational and orthogenesis to carry out transformation and optimization to biological catalyst, as improved stability and the substrate selective of catalyzer, can be applied to better production technique.
R-3-amino butanol has purposes widely in organic synthesis and drug manufacture, and structural formula is as follows:
At present by chemical process, at ammonium salt, can synthesising racemation 3-amino butanol (Chem.Abstr., 1947, p. 6199) under the catalysis of nickel, but still not with the synthetic report of biocatalysis.
Summary of the invention
The present invention seeks to the deficiency for solving the problems of the technologies described above, a kind of biological preparation method of R-3-amino butanol is provided, the method is compared with chemical method, there is environmental friendliness, optical selective advantages of higher, raw materials cost is cheap, easy and simple to handle, be more suitable for industrial mass production.
The present invention solves the problems of the technologies described above adopted technical scheme to be:
A biological preparation method for R-3-amino butanol, its preparation method comprises the following steps:
The preparation of step 1, restructuring D-aminotransferase gene engineering bacteria
Selection derives from
arthrobacter.sp.the wild sequence of D-transaminase, manually design, the gene order after design is as shown in SEQ ID NO:1; This sequence is synthesized by full gene, be cloned into Nde I and the Hind III site of pET24a; Transform bacillus coli DH 5 alpha competent cell; Picking positive transformant order-checking obtain recombinant expression vector after identifying; Recombinant expression vector is proceeded in e. coli bl21 (DE3) bacterial strain, acquisition can abduction delivering the restructuring D-aminotransferase gene engineering bacteria of restructuring D-transaminase;
The preparation of step 2, restructuring D-transaminase
Restructuring D-aminotransferase gene engineering bacteria is inoculated in the LB liquid nutrient medium that contains kalamycin resistance, in 37 ℃ of cultivation 16h, obtains seed culture fluid; Seed culture fluid is inoculated in the TB liquid nutrient medium containing kalamycin resistance, and inoculum size is 1% of the TB liquid nutrient medium volume containing kalamycin resistance; Then be placed at 37 ℃ and be cultured to OD600 value for 0.4-1.0, add sec.-propyl-β-D-galactoside (IPTG) or lactose, be placed in 28-30 ℃ and continue to cultivate after 16-24h, centrifugal collection thalline; Adopt phosphate buffered saline buffer resuspended after the thalline after collecting is washed once, suspension is placed in after ice bath ultrasonication centrifugal again, gained supernatant liquor is restructuring D-transaminase, adopts Bradford method to detect protein content wherein, standby;
The concrete grammar of described ultrasonication is: suspension is placed in to ice bath and adopts ultrasonic disruption, parameter is work 5s, 15s intermittently, cycle number 30.
The preparation of step 3, R-3-amino butanol
In reaction soln, adding successively final concentration is the substrate of 10-300mmol/L, the solubility promoter that mass concentration is 2-15%, the cofactor that final concentration is 0.1-1mmol/L, the restructuring D-transaminase anabolic reaction system that the amino donor that final concentration is 0.02-1mol/L and mass concentration are 0.2-0.4%; This reaction system is reacted to 24-26h under the condition of 25-30 ℃; After reaction finishes, the content of R-3-amino butanol in detection reaction liquid; Then with the R-3-amino butanol in ethyl acetate or dichloromethane extraction reaction solution, with rotary evaporation, remove ethyl acetate or methylene dichloride, obtain R-3-amino butanol; The reaction process of reaction system, sampling employing high performance liquid phase detects the growing amount of R-3-amino butanol in sample, to monitor its reaction process.
Described substrate is 3-carbonyl butanols; Described reaction soln is that pH is the 50-100mmol/L phosphate buffered saline buffer of 7.0-10.0, Tirs damping fluid or bicarbonate buffer; Described solubility promoter is dimethyl sulfoxide (DMSO) or acetonitrile; Described cofactor is pyridoxal phosphate; Described amino donor is Isopropylamine or D-alanine.
Beneficial effect is:
The biological preparation method of R-3-amino butanol provided by the invention, initial feed is 3-carbonyl butanols, low price.After reaction finishes, the ammonia of generation, water, oxygen are easy to remove, and product separation purifying is not disturbed, and in aqueous phase system, only remaining R-3-amino butanol, is easy to separation, contributes to improve product purity and yield, reduces separation costs.Required restructuring D-transaminase can, by structure Recombinant organism, then, by a large amount of preparations of fermentation, be easy to get and cheapness relatively.Described reaction is " treating different things alike " formula, and substrate and all enzymes add simultaneously and start reaction, directly obtain end product R-3-amino butanol, middle without separating-purifying.Process costs is low, and transformation efficiency and production concentration are high, and there is no by product impact, and this processing method is applicable to industrial application.
Accompanying drawing explanation
Fig. 1 is the reaction principle figure of the biological preparation method of R-3-amino butanol of the present invention.
Embodiment
A biological preparation method for R-3-amino butanol, its preparation method comprises the following steps:
The preparation of step 1, restructuring D-aminotransferase gene engineering bacteria
Selection derives from
arthrobacter.sp.the wild sequence of D-transaminase, manually design, the gene order after design is as shown in SEQ ID NO:1; This sequence is synthesized by full gene, be cloned into Nde I and the Hind III site of pET24a; Transform bacillus coli DH 5 alpha competent cell; Picking positive transformant order-checking obtain recombinant expression vector after identifying; Recombinant expression vector is proceeded in e. coli bl21 (DE3) bacterial strain, acquisition can abduction delivering the restructuring D-aminotransferase gene engineering bacteria of restructuring D-transaminase;
The preparation of step 2, restructuring D-transaminase
Restructuring D-aminotransferase gene engineering bacteria is inoculated in the LB liquid nutrient medium that contains kalamycin resistance, in 37 ℃ of cultivation 16h, obtains seed culture fluid; Seed culture fluid is inoculated in the TB liquid nutrient medium containing kalamycin resistance, and inoculum size is 1% of the TB liquid nutrient medium volume containing kalamycin resistance; Then be placed at 37 ℃ and be cultured to OD600 value for 0.4-1.0, add sec.-propyl-β-D-galactoside (IPTG) or lactose, be placed in 28-30 ℃ and continue to cultivate after 16-24h, centrifugal collection thalline; Adopt phosphate buffered saline buffer resuspended after the thalline after collecting is washed once, suspension is placed in after ice bath ultrasonication centrifugal again, gained supernatant liquor is restructuring D-transaminase, adopts Bradford method to detect protein content wherein, standby;
The concrete grammar of described ultrasonication is: suspension is placed in to ice bath and adopts ultrasonic disruption, parameter is work 5s, 15s intermittently, cycle number 30.
The preparation of step 3, R-3-amino butanol
In reaction soln, adding successively final concentration is the substrate of 10-300mmol/L, the whole mass percentage concentration solubility promoter that is 2-15%, the cofactor that final concentration is 0.1-1mmol/L, and final concentration is the restructuring D-transaminase anabolic reaction system that the amino donor of 0.02-1mol/L and whole mass percentage concentration are 0.2-0.4%; This reaction system is reacted to 24-26h under the condition of 25-30 ℃; After reaction finishes, the content of R-3-amino butanol in detection reaction liquid; Then with the R-3-amino butanol in ethyl acetate or dichloromethane extraction reaction solution, with rotary evaporation, remove ethyl acetate or methylene dichloride, obtain R-3-amino butanol; The reaction process of reaction system, sampling employing high performance liquid phase detects the growing amount of R-3-amino butanol in sample, to monitor its reaction process.
Described substrate is 3-carbonyl butanols; Described reaction soln is that pH is the 50-100mmol/L phosphate buffered saline buffer of 7.0-10.0, Tirs damping fluid or bicarbonate buffer; Described solubility promoter is dimethyl sulfoxide (DMSO) or acetonitrile; Described cofactor is pyridoxal phosphate; Described amino donor is Isopropylamine or D-alanine.
embodiment 1
A biological preparation method for R-3-amino butanol, its preparation method comprises the following steps:
The preparation of step 1, restructuring D-aminotransferase gene engineering bacteria
Selection derives from
arthrobacter.sp.the wild sequence of D-transaminase, manually design, the gene order after design is as shown in SEQ ID NO:1; This sequence is synthesized by full gene, be cloned into Nde I and the Hind III site of pET24a; Transform bacillus coli DH 5 alpha competent cell; Picking positive transformant order-checking obtain recombinant expression vector after identifying; Recombinant expression vector is proceeded in e. coli bl21 (DE3) bacterial strain, acquisition can abduction delivering the restructuring D-aminotransferase gene engineering bacteria of restructuring D-transaminase;
The preparation of step 2, restructuring D-transaminase
Restructuring D-aminotransferase gene engineering bacteria is inoculated in the LB liquid nutrient medium that contains kalamycin resistance, in 37 ℃ of cultivation 16h, obtains seed culture fluid; Seed culture fluid is inoculated in the TB liquid nutrient medium containing kalamycin resistance, and inoculum size is 1% of the TB liquid nutrient medium volume containing kalamycin resistance; Then being placed in and being cultured to OD600 value at 37 ℃ is 0.4, adds sec.-propyl-β-D-galactoside (IPTG) or lactose, is placed in 28 ℃ and continues to cultivate after 16h, centrifugal collection thalline; Adopt phosphate buffered saline buffer resuspended after the thalline after collecting is washed once, suspension is placed in after ice bath ultrasonication centrifugal again, gained supernatant liquor is restructuring D-transaminase, adopts Bradford method to detect protein content wherein, standby;
The concrete grammar of described ultrasonication is: suspension is placed in to ice bath and adopts ultrasonic disruption, parameter is: work 5s, 15s intermittently, cycle number 30.
The preparation of step 3, R-3-amino butanol
In reaction soln, adding successively final concentration is the substrate of 10-300mmol/L, the solubility promoter that mass concentration is 2-15%, the cofactor that final concentration is 0.1mmol/L, the restructuring D-transaminase anabolic reaction system that the amino donor that final concentration is 0.02-1mol/L and mass concentration are 0.2%; This reaction system is reacted to 24h under the condition of 25 ℃; The reaction process of reaction system, sampling employing high performance liquid phase detects the growing amount of R-3-amino butanol in sample, result optical purity (optical purity (ee value)) 99%, transformation efficiency 97%; Reaction finishes with the R-3-amino butanol in ethyl acetate or dichloromethane extraction reaction solution, with rotary evaporation, to remove ethyl acetate or methylene dichloride afterwards, obtains R-3-amino butanol, efficiency of pcr product 88%.
Described substrate is 3-carbonyl butanols; Described reaction soln is that pH is 7.0 50mmol/L phosphoric acid salt, Tirs damping fluid; Described solubility promoter is dimethyl sulfoxide (DMSO); Described cofactor is pyridoxal phosphate; Described amino donor is D-alanine.
embodiment 2
A biological preparation method for R-3-amino butanol, its preparation method comprises the following steps:
The preparation of step 1, restructuring D-aminotransferase gene engineering bacteria
Selection derives from
arthrobacter.sp.the wild sequence of D-transaminase, manually design, the gene order after design is as shown in SEQ ID NO:1; This sequence is synthesized by full gene, be cloned into Nde I and the Hind III site of pET24a; Transform bacillus coli DH 5 alpha competent cell; Picking positive transformant order-checking obtain recombinant expression vector after identifying; Recombinant expression vector is proceeded in e. coli bl21 (DE3) bacterial strain, acquisition can abduction delivering the restructuring D-aminotransferase gene engineering bacteria of restructuring D-transaminase;
The preparation of step 2, restructuring D-transaminase
Restructuring D-aminotransferase gene engineering bacteria is inoculated in the LB liquid nutrient medium that contains kalamycin resistance, in 37 ℃ of cultivation 16h, obtains seed culture fluid; Seed culture fluid is inoculated in the TB liquid nutrient medium containing kalamycin resistance, and inoculum size is 1% of the TB liquid nutrient medium volume containing kalamycin resistance; Then being placed in and being cultured to OD600 value at 37 ℃ is 1.0, adds sec.-propyl-β-D-galactoside (IPTG) or lactose, is placed in 28-30 ℃ and continues to cultivate after 24h, centrifugal collection thalline; Adopt phosphate buffered saline buffer resuspended after the thalline after collecting is washed once, suspension is placed in after ice bath ultrasonication centrifugal again, gained supernatant liquor is restructuring D-transaminase, adopts Bradford method to detect protein content wherein, standby;
The concrete grammar of described ultrasonication is: suspension is placed in to ice bath and adopts ultrasonic disruption, parameter is: work 5s, 15s intermittently, cycle number 30.
The preparation of step 3, R-3-amino butanol
In reaction soln, adding successively final concentration is the substrate of 300mmol/L, the solubility promoter that mass concentration is 15%, the cofactor that final concentration is 1mmol/L, the restructuring D-transaminase anabolic reaction system that the amino donor that final concentration is 1mol/L and mass concentration are 0.2-0.4%; This reaction system is reacted to 26h under the condition of 30 ℃; The reaction process of reaction system, sampling employing high performance liquid phase detects the growing amount of R-3-amino butanol in sample, result optical purity (optical purity (ee value)) 99%, transformation efficiency 98%; After reaction finishes, with the R-3-amino butanol in ethyl acetate or dichloromethane extraction reaction solution, with rotary evaporation, remove ethyl acetate or methylene dichloride, obtain R-3-amino butanol, efficiency of pcr product is 90%.
Described substrate is 3-carbonyl butanols; Described reaction soln is that pH is 10.0 100mmol/L bicarbonate buffer; Described solubility promoter is acetonitrile; Described cofactor is pyridoxal phosphate; Described amino donor is D-alanine.
embodiment 3
A biological preparation method for R-3-amino butanol, its preparation method comprises the following steps:
The preparation of step 1, restructuring D-aminotransferase gene engineering bacteria
Selection derives from
arthrobacter.sp.the wild sequence of D-transaminase, manually design, the gene order after design is as shown in SEQ ID NO:1; This sequence is synthesized by full gene, be cloned into Nde I and the Hind III site of pET24a; Transform bacillus coli DH 5 alpha competent cell; Picking positive transformant order-checking obtain recombinant expression vector after identifying; Recombinant expression vector is proceeded in e. coli bl21 (DE3) bacterial strain, acquisition can abduction delivering the restructuring D-aminotransferase gene engineering bacteria of restructuring D-transaminase;
The preparation of step 2, restructuring D-transaminase
Restructuring D-aminotransferase gene engineering bacteria is inoculated in the LB liquid nutrient medium that contains kalamycin resistance, in 37 ℃ of cultivation 16h, obtains seed culture fluid; Seed culture fluid is inoculated in the TB liquid nutrient medium containing kalamycin resistance, and inoculum size is 1% of the TB liquid nutrient medium volume containing kalamycin resistance; Then being placed in and being cultured to OD600 value at 37 ℃ is 0.6, adds sec.-propyl-β-D-galactoside (IPTG) or lactose, is placed in 29 ℃ and continues to cultivate after 20h, centrifugal collection thalline; Adopt phosphate buffered saline buffer resuspended after the thalline after collecting is washed once, suspension is placed in after ice bath ultrasonication centrifugal again, gained supernatant liquor is restructuring D-transaminase, adopts Bradford method to detect protein content wherein, standby;
The concrete grammar of described ultrasonication is: suspension is placed in to ice bath and adopts ultrasonic disruption, parameter is work 5s, 15s intermittently, cycle number 30.
The preparation of step 3, R-3-amino butanol
In reaction soln, adding successively final concentration is that the substrate of 200mmol/L, solubility promoter, the final concentration that mass concentration is 10% are 0.15mmol/L cofactor, the restructuring D-transaminase anabolic reaction system that the amino donor that final concentration is 0.5mol/L and mass concentration are 0.3%; This reaction system is reacted to 25h under the condition of 28 ℃; The reaction process of reaction system, sampling employing high performance liquid phase detects the growing amount of R-3-amino butanol in sample, result optical purity (optical purity (ee value)) 99%, transformation efficiency 98%; Reaction finishes with the R-3-amino butanol in ethyl acetate or dichloromethane extraction reaction solution, with rotary evaporation, to remove ethyl acetate or methylene dichloride afterwards, obtains R-3-amino butanol, efficiency of pcr product 90%.
Described substrate is 3-carbonyl butanols; Described reaction soln is that pH is 8.0 90mmol/L phosphate buffered saline buffer; Described solubility promoter is dimethyl sulfoxide (DMSO); Described cofactor is pyridoxal phosphate; Described amino donor is Isopropylamine.
Reagent described in the present invention is common agents, can buy and obtain from market; Described e. coli bl21 (DE3), for conventional bacterial classification, can be bought and be obtained by market.
The composition of described LB liquid nutrient medium: contain by weight percentage 1% Tryptones, 0.5% yeast extract, 3% sodium-chlor in LB liquid nutrient medium, surplus is water.
The composition of described TB substratum: peptone 12g/L, yeast powder 24g/L, glycerine 4g/L, potassium primary phosphate 17mmol/L, dipotassium hydrogen phosphate 72mmol/L, surplus is water.
SEQUENCE LISTING
<110> Luoyang Huarong Bioisystech Co., Ltd
The biological preparation method of a <120> R-3-amino butanol
<130> 1
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 993
<212> DNA
<213> artificial sequence
<400> 1
gtggcattca gcgccgatac ctccgagatc gtctacacgc acgacaccgg cctcgactac 60
atcacttata gcgactacga actcgatcct gctaacccgc tcgcgggagg tgcggcatgg 120
atcgagggtg cattcgtgcc gccgtcggag gcgcggatct cgatcttcga tcagggttac 180
ctccactcgg acgtcaccta cacggtcttc cacgtctgga acggaaatgc attccgcctc 240
gacgaccaca tcgaacgcct cttctccaac gcggagtcga tgcgcatcat ccctccgctc 300
acacaggacg aagtgaagga gattgcgctc gaactcgtcg cgaagaccga attgcgtgag 360
gccttcgtgt ccgtgtcgat tacccgcggt tacagctcga ctccgggcga gcgcgacatc 420
acgaagcacc gcccgcaggt gtacatgtat gccgtcccat atcagtggat cgtgccgttt 480
gaccgaattc gcgacggcgt gcacgccatg gtcgcacaga gcgtgcgccg aaccccgcgc 540
agctcgatcg accctcaggt caagaacttc cagtgggggg atctgatccg tgcggttcaa 600
gagacgcacg accgcgggtt cgaggctccc cttctgctcg acggcgatgg actgcttgcc 660
gagggctcgg ggttcaacgt cgtcgtgatc aaggacggcg tcgtgcgcag cccgggtcga 720
gcggcgctcc ccggcattac gcggaagacc gtgctcgaga tcgccgaatc gctcggacac 780
gaggcgattc tcgccgacat cacgctcgct gaactgctcg acgccgacga agtgctcggc 840
tgcacgactg cgggcggagt gtggccattc gtcagcgtgg acggcaaccc catctcggac 900
ggggttcccg gccccatcac ccagtcgatc atccgtcgtt actgggagct gaatgtcgag 960
agctcgtcgt tgcttacgcc tgtgcagtac tga 993
Claims (4)
1. a biological preparation method for R-3-amino butanol, is characterized in that: its preparation method comprises the following steps:
The preparation of step 1, restructuring D-aminotransferase gene engineering bacteria
It is synthetic that sequence according to artificial design as shown in SEQ ID NO:1 is carried out full gene, is cloned into Nde I and the Hind III site of pET24a; Transform bacillus coli DH 5 alpha competent cell; Picking positive transformant order-checking obtain recombinant expression vector after identifying; Recombinant expression vector is proceeded in e. coli bl21 (DE3) bacterial strain, acquisition can abduction delivering the restructuring D-aminotransferase gene engineering bacteria of restructuring D-transaminase;
The preparation of step 2, restructuring D-transaminase
Restructuring D-aminotransferase gene engineering bacteria is inoculated in the LB liquid nutrient medium that contains kalamycin resistance, in 37 ℃ of cultivation 16h, obtains seed culture fluid; Seed culture fluid is inoculated in the TB liquid nutrient medium containing kalamycin resistance, and inoculum size is 1% of the TB liquid nutrient medium volume containing kalamycin resistance; Then be placed at 37 ℃ and be cultured to OD600 value for 0.4-1.0, add sec.-propyl-β-D-galactoside or lactose, be placed in 28-30 ℃ and continue to cultivate after 16-24h, centrifugal collection thalline; Adopt phosphate buffered saline buffer resuspended after the thalline after collecting is washed once, suspension is placed in after ice bath ultrasonication centrifugal again, gained supernatant liquor is restructuring D-transaminase, detects protein content wherein, standby;
The preparation of step 3, R-3-amino butanol
In reaction soln, add successively the restructuring D-transaminase anabolic reaction system that final concentration is the substrate of 10-300mmol/L, solubility promoter that mass concentration is 2-15%, final concentration is 0.1-1mmol/L cofactor, amino donor that final concentration is 0.02-1mol/L and mass concentration are 0.2-0.4%; This reaction system is reacted to 24-26h under the condition of 25-30 ℃; After reaction finishes, the content of R-3-amino butanol in detection reaction liquid; Then with the R-3-amino butanol in ethyl acetate or dichloromethane extraction reaction solution, with rotary evaporation, remove ethyl acetate or methylene dichloride, obtain R-3-amino butanol;
Described substrate is 3-carbonyl butanols; Described reaction soln is that pH is 50-100mmol/L phosphate buffered saline buffer, Tirs damping fluid or the bicarbonate buffer of 7.0-10.0; Described solubility promoter is dimethyl sulfoxide (DMSO) or acetonitrile; Described cofactor is pyridoxal phosphate; Described amino donor is Isopropylamine or D-alanine.
2. the biological preparation method of R-3-amino butanol as claimed in claim 1, is characterized in that: the concrete grammar of ultrasonication described in step 2 is: suspension is placed in to ice bath and adopts ultrasonic disruption, parameter is work 5s, 15s intermittently, cycle number 30.
3. the biological preparation method of R-3-amino butanol as claimed in claim 1, is characterized in that: the method that detects protein content in supernatant liquor described in step 2 is Bradford method.
4. the biological preparation method of R-3-amino butanol as claimed in claim 1, is characterized in that: the reaction process of reaction system in step 3, sampling employing high performance liquid phase detects the growing amount of R-3-amino butanol in sample, to monitor its reaction process.
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| CN106754806A (en) * | 2016-12-20 | 2017-05-31 | 尚科生物医药（上海）有限公司 | A kind of improved transaminase and its application in the preparation of (R) 3 amino butanol |
| CN107586796A (en) * | 2017-07-20 | 2018-01-16 | 暨明医药科技（苏州）有限公司 | (R) synthetic method of 2 (1 amino-ethyl) 4 fluoroanilines |
| CN108823179A (en) * | 2018-06-30 | 2018-11-16 | 浙江工业大学 | A kind of transaminase from actinomyces, mutant, recombinant bacterium and application |
| CN110358804A (en) * | 2018-04-10 | 2019-10-22 | 湖州颐辉生物科技有限公司 | The enzymatic production process of R-3- amino n-butanol |
| CN112852895A (en) * | 2020-12-01 | 2021-05-28 | 中国科学院天津工业生物技术研究所 | Method for synthesizing (R) -3-amino-1-butanol by double-enzyme cascade catalysis |
| CN113105342A (en) * | 2021-03-31 | 2021-07-13 | 湖州柏特生物科技有限公司 | Preparation method of (R) -3-aminobutanol |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106754806A (en) * | 2016-12-20 | 2017-05-31 | 尚科生物医药（上海）有限公司 | A kind of improved transaminase and its application in the preparation of (R) 3 amino butanol |
| CN106754806B (en) * | 2016-12-20 | 2020-04-14 | 尚科生物医药（上海）有限公司 | Improved transaminase and application thereof in preparation of (R) -3-aminobutanol |
| CN107586796A (en) * | 2017-07-20 | 2018-01-16 | 暨明医药科技（苏州）有限公司 | (R) synthetic method of 2 (1 amino-ethyl) 4 fluoroanilines |
| CN107586796B (en) * | 2017-07-20 | 2021-08-06 | 暨明医药科技（苏州）有限公司 | Synthesis method of (R) -2- (1-aminoethyl) -4-fluorophenol |
| CN110358804A (en) * | 2018-04-10 | 2019-10-22 | 湖州颐辉生物科技有限公司 | The enzymatic production process of R-3- amino n-butanol |
| CN108823179A (en) * | 2018-06-30 | 2018-11-16 | 浙江工业大学 | A kind of transaminase from actinomyces, mutant, recombinant bacterium and application |
| CN108823179B (en) * | 2018-06-30 | 2020-10-09 | 浙江工业大学 | Transaminase derived from actinomycetes, mutant, recombinant bacterium and application |
| CN112852895A (en) * | 2020-12-01 | 2021-05-28 | 中国科学院天津工业生物技术研究所 | Method for synthesizing (R) -3-amino-1-butanol by double-enzyme cascade catalysis |
| CN112852895B (en) * | 2020-12-01 | 2021-11-30 | 中国科学院天津工业生物技术研究所 | Method for synthesizing (R) -3-amino-1-butanol by double-enzyme cascade catalysis |
| CN113105342A (en) * | 2021-03-31 | 2021-07-13 | 湖州柏特生物科技有限公司 | Preparation method of (R) -3-aminobutanol |
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