CN110358804A

CN110358804A - The enzymatic production process of R-3- amino n-butanol

Info

Publication number: CN110358804A
Application number: CN201810313800.7A
Authority: CN
Inventors: 陶荣盛; 王博; 沈青; 朱傅赟; 孙梁栋; 沈正权; 郑云; 潘震华; 刘萍; 王亚夫
Original assignee: Huzhou Yi Hui Biotechnology Co Ltd
Current assignee: Huzhou Yi Hui Biotechnology Co Ltd
Priority date: 2018-04-10
Filing date: 2018-04-10
Publication date: 2019-10-22
Anticipated expiration: 2038-04-10
Also published as: CN110358804B

Abstract

The invention discloses a kind of enzymatic production process of R-3- amino n-butanol, it is using 3- amino n-butanol racemic modification and pyruvic acid as substrate, S-3- amino n-butanol is catalyzed by microbe-derived ω-transaminase stereospecificity to react with pyruvic acid, then the R-3- amino n-butanol for obtaining having neither part nor lot in reaction is recycled from reaction system, the enantiomer R-3- amino n-butanol of high-optical-purity is obtained, industrialized production is suitable for.

Description

The enzymatic production process of R-3- amino n-butanol

Technical field

The invention belongs to biocatalysis technology fields, are related to a kind of enzymatic production process of R-3- amino n-butanol, specifically Ground says, is related to a kind of preparing R-3- amino n-butanol by the transaminase-catalyzed fractionation 3- amino n-butanol racemic modification of ω- Method.

Background technique

R-3- amino n-butanol is also known as (R) -3- amino butanol, R-3- amino-n-butyl alcohol, is synthesizing anti-AIDS integrase The important intermediate of inhibitor-Du Lutewei, with existing hiv integrase inhibitor Merck, angstrom for compared with lattice Wei, the medicine The safety is improved for object, and compared with the AntiHIV1 RT activity of Mo Shadong/AIDS drug draws for drawing for Wei, Du Lutewei is not only in three phases clinic Reached the curative effect being mutually equal to it in test, and its do not need with drug enhancers drug combination, with very potent resistance to Medicine attribute, and its dosage takes once for day.Rapid development is presented in Du Lutewei sales volume since listing, and R-3- amino is just Optical purity and the price height of butanol have an important influence the quality and production cost of Du Lutewei.

Based on chemical synthesis process, one is obtained using Kinetic Resolution for the production of R-3- amino n-butanol at present The 3- amino butanol (Journal of Organic Chemistry, 42,1650,1977) of chiral purity, but chemical resolution method is deposited It is low in yield, the defects of resolving agent consumption is big and severe reaction conditions.Another kind is the D- using chipal compounds as starting material Alanine or R type phenyl ethylamine obtained by multistep reaction chiral purity 3- amino n-butanol (Tetrahedron, 61,9031, 2005 Hes

CN101417954B), chiral purity cost of material is more expensive in such methods, and reaction step is tediously long, production cost compared with It is high.

In recent years, bioanalysis preparation R-3- amino n-butanol has become a research hotspot.With chemical synthesis phase Than biological synthesis process has many advantages, such as that mild reaction condition, high conversion efficiency and stereoselectivity are strong.Patent CN104131048A, CN104178533A and CN106754806A, which are disclosed, utilizes transaminase and its mutant asymmetry catalysis 4- The method that hydroxy-2-butanone synthesizes R-3- amino butanol, but since reaction density is low, the reasons such as unstable are converted, are not yet realized Industrial applications.

Summary of the invention

For overcome the deficiencies in the prior art, the R-3- amino n-butanol for being suitable for industrialized production high-optical-purity is explored Technique, inventor carried out a large amount of research to transaminase-catalyzed method, it has unexpectedly been found that the ω-for being derived partly from microorganism turns Adnosine deaminase (ω-TA) has 3- amino n-butanol enantiomer the stereospecificity of height, and S-3- amino n-butanol and third The dynamic balance issue generally existing there is no transaminase when ketone acid reacts, S-3- amino n-butanol almost can fully reacting, This discovery constitutes basis of the invention.Specifically, described in technical scheme is as follows.

A method of R-3- amino n-butanol is prepared, is included the following steps: with 3- amino n-butanol racemic modification and third Ketone acid is substrate, is catalyst with ω-transaminase, is catalyzed to stereospecificity S-3- amino n-butanol and reacts with pyruvic acid, so The enantiomer R-3- amino n-butanol for having neither part nor lot in reaction is recycled from reaction system afterwards.Reaction equation is as described below:

Above-mentioned ω-transaminase preferably derives from microorganism selected from the group below: (people is grey by Ochrobactrum anthropic White bacillus), Bradyrhizobium sp.Ec3.3 (slow raw rhizobium sp.Ec3.3), Brucella neotomae (sarin mouse Kind of brucella), Novosphingobium acidiphilum (the new sphingolipid bacterium of acidophilus), Ochrobactrum Intermedium (intermediate anthropi), Pseudaminobacter salicylatoxidans (false amino acidfast bacilli), Brucella abortus (B. abortus), Paracoccus denitrificans PD1222 (Paracoccus denitrificans PD1222).More preferably derive from Ochrobactrum anthropic, Brucella neotomae, Novosphingobium acidiphilum、Brucella abortus。

Preferably, above-mentioned ω-transaminase is selected from the group: SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:7。

In a preferred embodiment, also phosphopyridoxal pyridoxal phosphate (PLP) is added in reaction system.Phosphopyridoxal pyridoxal phosphate is made Transaminase-catalyzed transamination can be promoted to react for coenzyme.The additive amount of phosphopyridoxal pyridoxal phosphate in the reaction system is 0.1- 1mmol/L (i.e. 0.1-1mM).

Preferably, reaction solution used in above-mentioned reaction system is aqueous solution, phosphate buffer, Tirs buffer or carbon Sour hydrogen salt buffer.For example it can be 10-100mM phosphate buffer, 10-100mM Tirs buffer or 10-100mM carbon Sour hydrogen salt buffer.

Above-mentioned ω-transaminase can be in enzyme form, can also express microorganism form in ω-transaminase, for example be expression ω-transaminase recombination bacillus coli.When using recombination bacillus coli, ω-transaminase expressive host can be large intestine bar Bacterium BL21 (DE3).

In one embodiment, reaction system pH6.5-pH9.0, preferably pH6.8-pH8.5, more preferable pH7.0- PH8.0, more preferable pH7.0-pH7.8, more preferable pH7.0-pH7.5.

In one embodiment, reaction temperature be 25-45 DEG C, preferably 30-40 DEG C, it is 32-40 DEG C more preferable, more preferably 35-38 DEG C, such as 37 DEG C.

Preferably, from the method that R-3- amino n-butanol is recycled in reaction system can be conversion fluid concentration after with methanol or Ethyl alcohol dissolves out R-3- amino n-butanol, the method then crystallized at salt, can also be chromatographed and be recycled by column, can also adopted It is recycled under alkaline condition by extraction with organic solvent such as ethyl acetate or methylene chloride.

Since the present invention is to prepare R- configuration enantiomer by the method for Its Enzymatic Resolution 3- amino butanol racemic modification, The conversion ratio of substrate is higher, and the optical purity of R-3- amino n-butanol is also higher, therefore can pass through the conversion of control substrate Rate controls the optical purity of final product R-3- amino n-butanol, reaches the optical purity to meet the expected requirements.Enzymatic Not only process is controllable for method for splitting, but also has the advantages that reaction speed is fast, easy to operate and safe.

Meanwhile ω-transaminase reaction substrate 3- amino n-butanol and pyruvic acid allow with high concentration, it can be according to change The feed molar ratio that equilibrium principle adjusts the two reaction substrates is learned, advantageously reduces cost, it is easy to accomplish industrialized production.

Specific embodiment

The present invention is described in further details below in conjunction with specific embodiment.It should be understood that following embodiment is only used for The bright present invention is not for limiting the scope of the invention.

Additive amount, content and the concentration of many kinds of substance is referred to herein, wherein the percentage composition, except special instruction Outside, all refer to mass percentage.

For the sake of for convenience of description, ω-transaminase of Ochrobactrum anthropic will be derived from sometimes herein Referred to as OATA, amino acid sequence are SEQ ID NO:1, and encoding gene is NCBI accession number: NC_009667；It will derive from ω-transaminase of Bradyrhizobium sp.Ec3.3 is referred to as BSTA, and amino acid sequence is SEQ ID NO:2, encodes base Because of NCBI accession number: WP027523883；It will be referred to as BNTA from ω-transaminase of Brucella neotomae, Amino acid sequence is SEQ ID NO:3, and encoding gene is NCBI accession number: WP004687895；It will derive from ω-transaminase of Novosphingobium acidiphilum is referred to as NATA, and amino acid sequence is SEQ ID NO:4, compiles Code gene is NCBI accession number: WP028641684；It will be from the ω of Ochrobactrum intermedium-transaminase letter Referred to as OITA, amino acid sequence are SEQ ID NO:5, and encoding gene is NCBI accession number: WP006465915；It will derive from ω-transaminase of Pseudaminobacter salicylatoxidans is referred to as PSTA, and amino acid sequence is SEQ ID NO:6, encoding gene are NCBI accession number: WP019171585；It will be from the ω of Brucella abortus-transaminase abbreviation For BATA, amino acid sequence is SEQ ID NO:7, and encoding gene is NCBI accession number: WP006212579；It will derive from ω-transaminase of Paracoccus denitrificans PD1222 is referred to as PDTA, and amino acid sequence is SEQ ID NO: 8, encoding gene is NCBI accession number: ABL72050.

Herein, for the sake of for convenience of description, R-3- amino n-butanol is referred to as R- enantiomer sometimes, by S-3- amino N-butanol is referred to as S- enantiomer, and (R, S) -3- amino n-butanol or 3- amino n-butanol are referred to as racemic modification.It is aobvious and easy See, ω-transaminase catalysis substrate that the present invention selects is S- enantiomer.

ω used in the present invention-transaminase structure is clear, therefore those skilled in the art can be readily available its volume Code gene, the expression cassette comprising these genes and plasmid and the transformant comprising the plasmid.These genes, expression cassette, matter Grain, transformant can be obtained by genetic engineering building mode well-known to those skilled in the art.

When as being that biocatalyst is used to produce R- enantiomer, the form of enzyme can be presented in ω-transaminase of the invention Or the form of thallus.The form of the enzyme includes resolvase, immobilised enzymes, including purifying enzyme, thick enzyme, fermentation liquid, carrier are solid Fixed enzyme etc..And these enzymes isolate and purify including immobilised enzymes technology of preparing is also well-known to those skilled in the art. The form of the thallus includes survival thallus and dead thallus.

In a preferred embodiment, reaction system can be added with 0.1-1mmol/L phosphopyridoxal pyridoxal phosphate, as ω-transaminase coenzyme promotes resolution reaction.Specific additional amount depends on enzyme amount and substrate additional amount, can be by simple real It tests and is determined.

Embodiment

Material and method

Full genome synthesis, primer synthesis and sequencing herein is all completed by Nanjing Genscript Biotechnology Co., Ltd..

Molecular biology experiment herein include plasmid construction, digestion, competent cell preparation, conversion etc. referring especially to " Molecular Cloning:A Laboratory guide " (third edition), J. Pehanorm Brooker, D.W. Russell (beauty) write, and Huang Peitang etc. is translated, and science goes out Version society, Beijing, 2002) it carries out.It can be operated according to related kit specification.It can be determined when necessary by simple experiment specific Experiment condition such as PCR condition.

TB culture medium: 24g/L yeast extract, 12g/L tryptone, 16.43g/L K₂HPO₄.3H₂O、2.31g/L KH₂PO₄, 5g/L glycerol, pH 7.0-7.5,121 DEG C of autoclave sterilization 20min.

The HPLC determination condition of reaction substrate and product:

Mobile phase composition A:0.05M sodium acetate aqueous solution

Mobile phase composition B: methanol

Mobile phase: A:B=70:30 (v/v)

Sample volume: 5.0 μ L

Flow velocity: 1.0mL/min

Acquisition time: 30min

Chromatographic column: Agilent C8 (180 × 4.6)

Column temperature: 30 DEG C

Detection wavelength: 334nm

Retention time: alanine 3.8min

R-3- amino n-butanol 26.7min.

The building of 1 ω of embodiment-transaminase expression bacterial strain

From NCBI accession number be respectively NC_009667, WP027523883, WP004687895, WP028641684, WP006465915, WP019171585, WP006212579, ABL72050 obtain coding ω-transaminase OATA, BSTA, BNTA, The polynucleotide sequence of NATA, OITA, PSTA, BATA and PDTA.Full genome synthesizes these ω-aminotransferase gene sequence, respectively In both ends design limitation restriction enzyme site NdeI and BamHI, it is subcloned into corresponding positions on carrier pET24a (purchased from Novagen) Point obtains recombinant plasmid pET24a.The recombinant plasmid built is respectively converted into Bacillus coli expression host with Calcium Chloride Method BL21 (DE3) is expressed the recombination bacillus coli of each ω-transaminase respectively.

The comparison of the transaminase-catalyzed 3- amino n-butanol of embodiment 2 fractionation ability

The shake flask fermentation of 2.1 ω-transaminase expression strain: it prepares fluid nutrient medium TB (pH7.0), is sub-packed in 500mL tri- Angle shaking flask, liquid amount 100mL, then in 121 DEG C of heat sterilization 20min in high-pressure sterilizing pot.Strain is expressed from ω-transaminase Plate on choose ring of numbers thallus with oese and be inoculated in TB shaking flask, 100 μ g/mL cards are added in TB culture medium before inoculation, and that is mould Element, in 37 DEG C, 220rpm shaking table culture to OD₆₀₀0.2mM IPTG (isopropylthiogalactoside) is added in 28 DEG C in=5-6 Induce left and right for 24 hours.

The preparation of 2.2 crude enzyme liquids: fermentation liquid 50ml is taken to be fitted into centrifuge tube；8000rpm centrifugation goes supernatant to obtain thallus, Purified water is added to be resuspended by thallus 200g/L, suspension thalline is cooled with an ice bath, progress ultrasonication (voltage 400W, ultrasonic time 3s, Interval time 5s, work times 80 times) obtain crude enzyme liquid.

The reaction of 2.3 Its Enzymatic Resolutions: it weighs 0.88g (0.4M) Sodium Pyruvate and is placed in shaking flask, 0.893g is then added (0.5M) 3- amino n-butanol, 0.1mm/L PLP, water 10ml, with acetic acid tune PH to 7.5, add that 20g/L has been crushed turns ammonia Enzyme crude enzyme liquid adds purified water to be settled to 20ml, and 37 DEG C are converted.It is sampled after converting 1h and 16h respectively, is 1:1 with volume ratio 0.1M hydrochloric acid terminate reaction, HPLC detects alanine, R-3- amino n-butanol and S-3- amino levels of n-butanol, and calculates production The ee value of object R-3- amino n-butanol.

The result that each transaminase-catalyzed 3- amino n-butanol of ω-is split is listed in Tables 1 and 2.

Result after 1 multiple-microorganism source ω of table-transaminase 1h compares

Result after 2 multiple-microorganism source ω of table-transaminase 16h compares

By Tables 1 and 2 as it can be seen that ω-transaminase OATA, BATA, BNTA, NATA catalysis activity is relatively higher.

The fractionation ability of the transaminase-catalyzed high concentration 3- amino n-butanol of embodiment 3 compares

The shake flask fermentation of 3.1 ω-transaminase expression strain: it prepares fluid nutrient medium TB (pH7.0-7.5), is sub-packed in 500mL triangle shake bottle, liquid amount 100mL, then in 121 DEG C of heat sterilization 20min in high-pressure sterilizing pot.From ω-transaminase Ring of numbers thallus is chosen with oese respectively on the plate of OATA, BATA, BNTA and NATA expression strain to be inoculated in each TB shaking flask, 100 μ g/mL kanamycins are added in TB culture medium before inoculation, in 37 DEG C, 220rpm shaking table culture to OD₆₀₀=5-6 is added 0.2mM IPTG induces left and right for 24 hours in 28 DEG C.

The preparation of 3.2 crude enzyme liquids: fermentation liquid 50ml is taken to be fitted into centrifuge tube；8000rpm centrifugation goes supernatant to obtain thallus, Purified water is added to be resuspended by thallus 200g/L, suspension thalline is cooled with an ice bath, progress ultrasonication (voltage 400W, ultrasonic time 3s, Interval time 5s, work times 80 times) obtain crude enzyme liquid.

The reaction of 3.3 Its Enzymatic Resolutions: weighing 1.76g (0.8M) Sodium Pyruvate and be placed in shaking flask, and 2g (100g/ is then added L) 3- amino n-butanol, 0.1mm/L PLP, water 10ml add the transaminase enzyme that 20g/L has been crushed with acetic acid tune PH to 7.5 Liquid adds water to be settled to 20ml, and 37 DEG C are converted.It samples after converting 1h and after 16h respectively, the 0.1M for being 1:1 with volume ratio Hydrochloric acid terminates reaction, and HPLC detects alanine, R-3- amino n-butanol and S-3- amino levels of n-butanol, and calculates product R-3- The ee value of amino n-butanol.

The results are shown in Table 3 for each transaminase-catalyzed 3- amino n-butanol fractionation of ω-.

The comparison that the transaminase-catalyzed high concentration 3- amino n-butanol of 3 difference ω-of table is split

The experimental results showed that allowing to react using the transaminase-catalyzed fractionation 3- amino butanol racemic modification of ω-of the invention Substrate 3- amino n-butanol has high concentration, such as can achieve 100g/L, and reaction speed is fast, advantageously reduces R-3- ammonia Base n-butanol production cost, thus there is industrial prospect.

Sequence table

<110>Huzhou Yi Hui Biotechnology Co., Ltd

<120>enzymatic production process of R-3- amino n-butanol

<130> SHPI810302

<160> 8

<170> SIPOSequenceListing 1.0

<210> 1

<211> 456

<212> PRT

<213> Ochrobactrum anthropic

<400> 1

Met Thr Ala Gln Pro Asn Ser Leu Glu Ala Arg Asp Ile Arg Tyr His

1 5 10 15

Leu His Ser Tyr Thr Asp Ala Val Arg Leu Glu Ala Glu Gly Pro Leu

20 25 30

Val Ile Glu Arg Gly Asp Gly Ile Tyr Val Glu Asp Val Ser Gly Lys

35 40 45

Arg Tyr Ile Glu Ala Met Ser Gly Leu Trp Ser Val Gly Val Gly Phe

50 55 60

Ser Glu Pro Arg Leu Ala Glu Ala Ala Ala Arg Gln Met Lys Lys Leu

65 70 75 80

Pro Phe Tyr His Thr Phe Ser Tyr Arg Ser His Gly Pro Val Ile Asp

85 90 95

Leu Ala Glu Lys Leu Val Ser Met Ala Pro Val Pro Met Ser Lys Ala

100 105 110

Tyr Phe Thr Asn Ser Gly Ser Glu Ala Asn Asp Thr Val Val Lys Leu

115 120 125

Ile Trp Tyr Arg Ser Asn Ala Leu Gly Glu Pro Glu Arg Lys Lys Ile

130 135 140

Ile Ser Arg Lys Arg Gly Tyr His Gly Val Thr Ile Ala Ser Ala Ser

145 150 155 160

Leu Thr Gly Leu Pro Asn Asn His Arg Ser Phe Asp Leu Pro Ile Asp

165 170 175

Arg Ile Leu His Thr Gly Cys Pro His Phe Tyr Arg Glu Gly Gln Ala

180 185 190

Gly Glu Ser Glu Glu Gln Phe Ala Thr Arg Leu Ala Asp Glu Leu Glu

195 200 205

Gln Leu Ile Ile Ala Glu Gly Pro His Thr Ile Ala Ala Phe Ile Gly

210 215 220

Glu Pro Val Met Gly Ala Gly Gly Val Val Val Pro Pro Lys Thr Tyr

225 230 235 240

Trp Glu Lys Val Gln Ala Val Leu Lys Arg Tyr Asp Ile Leu Leu Ile

245 250 255

Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Asn Leu Phe Gly

260 265 270

Ser Gln Thr Phe Asp Met Lys Pro Asp Ile Leu Val Met Ser Lys Gln

275 280 285

Leu Ser Ser Ser Tyr Leu Pro Ile Ser Ala Phe Leu Ile Asn Glu Arg

290 295 300

Val Tyr Ala Pro Ile Ala Glu Glu Ser His Lys Ile Gly Thr Leu Gly

305 310 315 320

Thr Gly Phe Thr Ala Ser Gly His Pro Val Ala Ala Ala Val Ala Leu

325 330 335

Glu Asn Leu Ala Ile Ile Glu Glu Arg Asp Leu Val Ala Asn Ala Arg

340 345 350

Asp Arg Gly Thr Tyr Met Gln Lys Arg Leu Arg Glu Leu Gln Asp His

355 360 365

Pro Leu Val Gly Glu Val Arg Gly Val Gly Leu Ile Ala Gly Val Glu

370 375 380

Leu Val Thr Asp Lys Gln Ala Lys Thr Gly Leu Glu Pro Thr Gly Ala

385 390 395 400

Leu Gly Ala Lys Ala Asn Ala Val Leu Gln Glu Arg Gly Val Ile Ser

405 410 415

Arg Ala Met Gly Asp Thr Leu Ala Phe Cys Pro Pro Leu Ile Ile Asn

420 425 430

Asp Gln Gln Val Asp Thr Met Val Ser Ala Leu Glu Ala Thr Leu Asn

435 440 445

Asp Val Gln Ala Ser Leu Thr Arg

450 455

<210> 2

<211> 460

<212> PRT

<213> Bradyrhizobium sp. Ec3.3

<400> 2

Met Thr Met Leu Pro Asn Ser Gln Glu Ala Arg Asp Val Ala Tyr Gln

1 5 10 15

Leu His Pro Tyr Thr Asn Ala Arg Thr His Gln Gln Asn Gly Pro Leu

20 25 30

Val Ile Glu Arg Gly Glu Gly Pro Tyr Val Phe Asp Ala Ala Gly Lys

35 40 45

Arg Tyr Phe Glu Ala Met Ala Gly Leu Trp Ser Val Gly Leu Gly Phe

50 55 60

Asn Glu Lys Arg Leu Val Glu Ala Ala Tyr Arg Gln Met Gln Ala Leu

65 70 75 80

Pro Phe Tyr His Thr Phe Ser Ala Lys Ser His Gly Pro Ser Ile Asp

85 90 95

Leu Ala Glu Lys Leu Val Ala Leu Ala Pro Val Pro Met Ser Lys Val

100 105 110

Phe Phe Thr Asn Ser Gly Ser Glu Ala Asn Asp Thr Val Leu Lys Leu

115 120 125

Ile Ala Tyr Arg Ser Asn Ala Leu Gly Gln Pro Gln Arg Lys Lys Val

130 135 140

Ile Ser Arg Met Arg Ala Tyr His Gly Val Thr Ile Ala Ser Ala Ser

145 150 155 160

Leu Thr Gly Leu Pro Asn Asn His Arg Ser Phe Asp Leu Pro Leu Pro

165 170 175

Asn Ile Leu His Thr Gly Ser Pro His Phe Tyr Lys Asp Gly Gln Ala

180 185 190

Gly Glu Ser Glu Glu Ala Phe Ala Thr Arg Arg Ala Glu Glu Leu Glu

195 200 205

Ala Leu Ile Val Lys Glu Gly Pro Asp Thr Ile Ala Ala Phe Phe Gly

210 215 220

Glu Pro Val Met Gly Ala Gly Gly Val Ile Val Pro Pro Ala Thr Tyr

225 230 235 240

Trp Asp Lys Val Gln Gln Val Leu Lys Lys Tyr Asp Ile Leu Leu Val

245 250 255

Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Lys Met Phe Gly

260 265 270

Cys Asp Thr Tyr Asn Ile Lys Pro Asp Ala Ile Val Val Ser Lys Gln

275 280 285

Ile Thr Ser Ser Tyr Phe Pro Phe Ser Ala Val Leu Met Asn Asp Arg

290 295 300

Met Phe Glu Pro Ile Ala Asp Glu Ser Asn Lys Ile Gly Val Leu Gly

305 310 315 320

His Gly Phe Thr Ala Gly Gly His Pro Val Gly Ala Ala Val Ala Leu

325 330 335

Glu Asn Leu Lys Ile Ile Glu Glu Arg Gly Leu Val Ala His Ala Ala

340 345 350

Glu Thr Gly Ala His Leu Gln Ala Arg Leu Arg Glu Leu Ser Ser His

355 360 365

Pro Leu Val Gly Glu Val Arg Gly Val Gly Met Ile Ala Ala Ile Glu

370 375 380

Leu Val Leu Asp Lys Gln Arg Lys Thr Ala Ala Ala Thr Pro Gly Ala

385 390 395 400

Val Gly Gly Ile Ala Ala Arg Met Leu Gln Glu Arg Gly Val Ile Ser

405 410 415

Arg Asn Met Leu Glu Ala Ile Ala Ile Cys Pro Pro Leu Ile Val Thr

420 425 430

Lys Ser Gln Ile Asp Glu Leu Val Gly Thr Ile Ala Gly Val Leu Asp

435 440 445

Asp Met Lys Thr Glu Ala Ala Lys Leu Thr Pro Ala

450 455 460

<210> 3

<211> 456

<212> PRT

<213> Brucella neotomae

<400> 3

Met Thr Ala Gln Pro Asn Ser Leu Glu Ala Arg Asp Ile Arg Tyr His

1 5 10 15

Leu His Ser Tyr Thr Asp Ala Val Arg Leu Glu Ala Glu Gly Pro Leu

20 25 30

Val Ile Glu Arg Gly Asp Gly Ile Tyr Val Glu Asp Ile Ala Gly Lys

35 40 45

Arg Tyr Ile Glu Ala Met Ser Gly Leu Trp Ser Val Gly Val Gly Phe

50 55 60

Ser Glu Gln Arg Leu Ala Glu Ala Ala Ala Arg Gln Met Lys Lys Leu

65 70 75 80

Pro Phe Tyr His Thr Phe Ser Tyr Arg Ser His Gly Pro Val Ile Asp

85 90 95

Leu Ala Glu Lys Leu Val Ala Met Ala Pro Val Pro Met Ser Lys Ala

100 105 110

Tyr Phe Thr Asn Ser Gly Ser Glu Ala Asn Asp Thr Val Val Lys Leu

115 120 125

Ile Trp Tyr Arg Ser Asn Ala Leu Gly Glu Pro Glu Arg Lys Lys Ile

130 135 140

Ile Ser Arg Lys Arg Gly Tyr His Gly Val Thr Ile Ala Ser Ala Ser

145 150 155 160

Leu Thr Gly Leu Pro Asn Asn His Arg Ser Phe Asp Leu Pro Ile Asp

165 170 175

Arg Ile Leu His Thr Gly Cys Pro His His Tyr His Asp Ala Leu Pro

180 185 190

Gly Glu Ser Glu Glu Gln Phe Thr Thr Arg Leu Ala Asn Glu Leu Glu

195 200 205

Gln Leu Ile Leu Ala Glu Gly Pro His Thr Ile Ala Ala Phe Ile Gly

210 215 220

Glu Pro Val Met Gly Ala Gly Gly Val Val Val Pro Pro Lys Thr Tyr

225 230 235 240

Trp Glu Lys Val Gln Ala Val Leu Gly Arg Tyr Asn Ile Leu Leu Val

245 250 255

Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Asn Leu Phe Gly

260 265 270

Cys Gln Thr Phe Asp Ile Lys Pro Asp Ile Leu Val Met Ser Lys Gln

275 280 285

Leu Ser Ser Ser Tyr Leu Pro Ile Ser Ala Phe Leu Ile Asn Glu Arg

290 295 300

Val Tyr Ala Pro Ile Ala Glu Glu Ser His Lys Ile Gly Thr Leu Gly

305 310 315 320

Thr Gly Phe Thr Ala Ser Gly His Pro Val Ala Ala Ala Val Ala Leu

325 330 335

Glu Asn Leu Ala Ile Ile Glu Glu Arg Asp Leu Val Ala Asn Ala Arg

340 345 350

Glu Arg Gly Ala Arg Met Gln Lys Arg Leu Arg Glu Leu Gln Asp His

355 360 365

Pro Leu Val Gly Glu Val Arg Gly Val Gly Leu Ile Ala Gly Val Glu

370 375 380

Leu Val Ile Asp Lys Glu Ala Lys Thr Gly Leu Glu Gln Pro Gly Ala

385 390 395 400

Leu Gly Ala Arg Ala Asn Ala Ala Leu Gln Glu Arg Gly Val Ile Ser

405 410 415

Arg Ala Met Gly Asp Thr Leu Ala Phe Cys Pro Pro Leu Ile Ile Asn

420 425 430

Asp Gln Gln Val Asp Thr Met Val Ser Ala Leu Glu Ala Ala Leu Asn

435 440 445

Asp Val Gln Ala Ser Leu Gly Lys

450 455

<210> 4

<211> 460

<212> PRT

<213> Novosphingobium acidiphilum

<400> 4

Met Leu Asp Ala Pro Asn Ser Ala Glu Ala Arg Asp Ile Arg Phe His

1 5 10 15

Leu His Ser Tyr Thr Asn Ala Arg Leu His Glu Gln Val Gly Pro Leu

20 25 30

Val Ile Glu Gly Gly Asp Gly Ile Tyr Val Ile Asp Ser Glu Gly Arg

35 40 45

Arg Tyr Ile Glu Ala Met Ser Gly Leu Trp Ser Val Gly Val Gly Phe

50 55 60

Ser Glu Lys Arg Leu Val Ala Ala Ala Thr Glu Gln Met Asn Lys Leu

65 70 75 80

Pro Phe Tyr His Thr Phe Thr His Lys Ser His Gly Pro Ala Ile Asp

85 90 95

Leu Ala Glu Lys Leu Val Thr Leu Ala Ser Pro Leu Thr Pro Gly Gly

100 105 110

Met Ser Lys Ala Phe Phe Thr Asn Ser Gly Ser Glu Ala Asn Asp Thr

115 120 125

Ala Ile Lys Leu Ile Trp Tyr Arg Ser Asn Ala Leu Gly Lys Pro Glu

130 135 140

Lys Lys Lys Leu Ile Ser Arg Val Arg Ala Tyr His Gly Val Thr Leu

145 150 155 160

Ala Ala Ala Ser Leu Thr Gly Leu Pro Asn Asn His Arg Ser Phe Asp

165 170 175

Leu Pro Met Pro Gly Val Leu His Thr Gly Ser Pro His Tyr Trp Arg

180 185 190

Glu Gly Leu Pro Gly Glu Ser Glu Glu Ala Phe Ala Thr Arg Arg Ala

195 200 205

Glu Glu Leu Asp Ala Leu Ile Gln Ala Glu Gly Pro Asp Thr Ile Ala

210 215 220

Ala Phe Phe Ala Glu Pro Val Met Gly Ala Gly Gly Val Val Val Pro

225 230 235 240

Pro Ala Thr Tyr Trp Asp Lys Ile Gln Ala Val Leu Ala Lys Tyr Asp

245 250 255

Ile Leu Phe Val Ala Asp Glu Val Ile Asn Gly Phe Gly Arg Thr Gly

260 265 270

Thr Met Phe Ala Cys Glu Thr Tyr Gly Ile Arg Pro Asp Ile Leu Ile

275 280 285

Val Ser Lys Gln Leu Ser Ser Ser Tyr Met Pro Ile Ala Ala Leu Ile

290 295 300

Ala Asn Asp Arg Val Leu Gly Pro Val Met Asp Glu Ser Ala Arg Ile

305 310 315 320

Gly Thr Leu Gly His Gly Tyr Thr Ala Gly Gly His Pro Val Ala Thr

325 330 335

Ala Val Ser Leu Glu Asn Leu Arg Ile Ile Glu Glu Asp Gly Leu Val

340 345 350

Ala Asn Cys Ala Ile Gln Gly Ala Arg Leu Arg Ala Gly Leu Ala Ala

355 360 365

Leu Ser His His Pro Leu Val Gly Glu Val Arg Gly Val Gly Met Ile

370 375 380

Ala Ala Ile Glu Leu Val Thr Asp Lys Ala Gly Lys Val Ala Leu Asp

385 390 395 400

Val Pro Gly Lys Leu Gly Gly Met Val Asn Lys Ala Leu Gln Asp Asn

405 410 415

Gly Val Ile Cys Arg Ala Val Val Asp Ala Ile Cys Phe Cys Pro Pro

420 425 430

Met Ile Ile Thr Ala Asp Gln Ile Asp Asp Leu Leu Ala Ala Val Ser

435 440 445

Ala Ser Leu Asp Arg Val Ala Ala Asp Leu Gly Leu

450 455 460

<210> 5

<211> 456

<212> PRT

<213> Ochrobactrum intermedium

<400> 5

Met Thr Ala Gln Pro Asn Ser Leu Glu Ala Arg Asp Ile Arg Tyr His

1 5 10 15

Leu His Ser Tyr Thr Asp Ala Val Arg Leu Glu Ala Glu Gly Pro Leu

20 25 30

Val Ile Glu Arg Gly Asp Gly Ile Tyr Val Glu Asp Val Thr Gly Lys

35 40 45

Arg Tyr Ile Glu Ala Met Ser Gly Leu Trp Ser Val Gly Val Gly Phe

50 55 60

Ser Glu Pro Arg Leu Ala Glu Ala Ala Ala Arg Gln Met Lys Lys Leu

65 70 75 80

Pro Phe Tyr His Thr Phe Ser Tyr Arg Ser His Gly Pro Val Ile Asp

85 90 95

Leu Ala Glu Lys Leu Val Ser Met Ala Pro Val Pro Met Ser Lys Ala

100 105 110

Tyr Phe Thr Asn Ser Gly Ser Glu Ala Asn Asp Thr Val Ile Lys Leu

115 120 125

Ile Trp Tyr Arg Ser Asn Ala Leu Gly Glu Pro Glu Arg Lys Lys Ile

130 135 140

Ile Ser Arg Lys Arg Gly Tyr His Gly Val Thr Ile Ala Ser Ala Ser

145 150 155 160

Leu Thr Gly Leu Pro Asn Asn His Arg Ser Phe Asp Leu Pro Ile Asp

165 170 175

Arg Ile Leu His Ala Gly Cys Pro His His Tyr Arg Glu Gly Gln Ala

180 185 190

Gly Glu Thr Glu Glu Gln Phe Ala Thr Arg Leu Ala Asp Glu Leu Glu

195 200 205

Gln Leu Ile Leu Ala Glu Gly Pro His Thr Ile Ala Ala Phe Ile Gly

210 215 220

Glu Pro Val Met Gly Ala Gly Gly Val Val Val Pro Pro Arg Thr Tyr

225 230 235 240

Trp Glu Lys Val Gln Ala Val Leu Lys Arg Tyr Asp Ile Leu Leu Val

245 250 255

Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Asn Leu Phe Gly

260 265 270

Ser Gln Thr Phe Asp Ile Lys Pro Asp Ile Leu Val Met Ser Lys Gln

275 280 285

Leu Ser Ser Ser Tyr Leu Pro Ile Ser Ala Phe Leu Ile Asn Glu Arg

290 295 300

Val Tyr Ala Pro Ile Ala Glu Glu Ser His Lys Ile Gly Thr Leu Gly

305 310 315 320

Thr Gly Phe Thr Ala Ser Gly His Pro Val Ala Ala Ala Val Ala Leu

325 330 335

Glu Asn Leu Ala Ile Ile Glu Glu Arg Gln Leu Val Ala Asn Ala Arg

340 345 350

Asp Arg Gly Ala Tyr Met Gln Lys Arg Leu Arg Asp Leu Lys Asp His

355 360 365

Pro Leu Val Gly Glu Val Arg Gly Val Gly Leu Ile Ala Gly Val Glu

370 375 380

Leu Val Thr Asp Lys Gln Ala Lys Thr Gly Leu Glu Gln Ala Gly Ala

385 390 395 400

Leu Gly Ala Gln Ala Asn Ala Ala Leu Gln Asp Arg Gly Val Ile Ser

405 410 415

Arg Ala Met Gly Asp Thr Leu Ala Phe Cys Pro Pro Leu Ile Ile Asn

420 425 430

Asp Gln Gln Ile Asp Thr Met Val Ser Ala Leu Glu Ala Ala Leu Asn

435 440 445

Asp Val Gln Ala Ser Leu Ala Arg

450 455

<210> 6

<211> 456

<212> PRT

<213> Pseudaminobacter salicylatoxidans

<400> 6

Met Asp Thr Gln Pro Asn Ser Pro Glu Ala Arg Asp Ile Arg Tyr Asn

1 5 10 15

Leu His Ala Tyr Thr Asn Ala Arg Arg His Gln Glu Ala Gly Pro Leu

20 25 30

Ile Ile Glu Lys Gly Glu Gly Ile Tyr Val Glu Asp Asn Ser Gly Lys

35 40 45

Arg Tyr Ile Glu Ala Met Ala Gly Leu Trp Ser Val Ala Val Gly Phe

50 55 60

Ser Glu Gln Arg Leu Val Asp Ala Ala Thr Lys Gln Met Ser Lys Leu

65 70 75 80

Pro Tyr Tyr His Ser Phe Thr His Lys Ala His Ser Pro Leu Ile Asp

85 90 95

Leu Ala Glu Lys Leu Val Gln Met Ala Pro Val Pro Met Ser Lys Ala

100 105 110

Phe Phe Thr Asn Ser Gly Ser Glu Ala Asn Asp Thr Ala Met Lys Met

115 120 125

Ile Trp Tyr Arg Ser Asn Ala Leu Gly Gln Pro Gln Arg Lys Lys Ile

130 135 140

Ile Ser Arg Gln Arg Gly Tyr His Gly Val Thr Ile Ala Ser Ala Ser

145 150 155 160

Leu Thr Gly Leu Pro Asn Asn Gln Ile Ser Phe Asp Leu Pro Ile Ala

165 170 175

Asn Val Leu His Thr Ala Ser Pro His Tyr Trp Arg Glu Ala Arg Pro

180 185 190

Gly Glu Thr Glu Glu Gln Phe Ser Thr Arg Leu Ala Glu Glu Leu Glu

195 200 205

Lys Leu Ile Leu Ala Glu Gly Pro Glu Thr Val Ala Ala Phe Ile Gly

210 215 220

Glu Pro Val Met Gly Ala Gly Gly Val Val Val Pro Pro Ala Gly Tyr

225 230 235 240

Trp Glu Lys Ile Gln Ala Val Leu Lys Lys Tyr Asp Ile Leu Leu Ile

245 250 255

Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Glu Met Phe Gly

260 265 270

Ser Gln Thr Tyr Gly Leu Gln Pro Asp Ile Met Val Met Ser Lys Gln

275 280 285

Leu Ser Ser Ser Tyr Leu Pro Ile Ser Ala Leu Leu Ile Asn Asp Lys

290 295 300

Val Phe Glu Pro Leu Ala Asp Glu Ser Asn Arg Ile Gly Thr Phe Gly

305 310 315 320

His Gly Phe Thr Ala Gly Gly His Pro Val Ala Ala Ala Val Ala Leu

325 330 335

Glu Asn Leu Arg Leu Ile Glu Glu Arg Asp Leu Val Gly Asn Ala Arg

340 345 350

Glu Val Gly Ala Tyr Met Gln Gln Arg Leu Arg Glu Leu Ser Ser His

355 360 365

Glu Leu Val Gly Glu Val Arg Gly Ile Gly Leu Ile Ala Ala Ile Glu

370 375 380

Leu Val Ala Asp Lys Ala Thr Lys Ala Pro Trp Gly Gln Pro Gly Ala

385 390 395 400

Leu Gly Ala Leu Thr Asn Gly Leu Leu Gln Gln Asn Gly Val Ile Ser

405 410 415

Arg Asn Met Gly Asp Ala Leu Ala Phe Cys Pro Pro Leu Ile Ile Thr

420 425 430

Arg Ala Gln Val Asp Glu Ile Ile Ser Ala Leu Lys Thr Ser Leu Asp

435 440 445

Glu Ala Leu Lys Gln Val Arg Ala

450 455

<210> 7

<211> 456

<212> PRT

<213> Brucella abortus

<400> 7

Met Thr Ala Gln Pro Asn Ser Leu Glu Ala Arg Asp Ile Arg Tyr His

1 5 10 15

Leu His Ser Tyr Thr Asp Ala Val Arg Leu Glu Ala Glu Gly Pro Leu

20 25 30

Val Ile Glu Arg Gly Asp Gly Ile Tyr Val Glu Asp Ile Ala Gly Lys

35 40 45

Arg Tyr Ile Glu Ala Met Ser Gly Leu Trp Ser Val Gly Val Gly Phe

50 55 60

Ser Glu Gln Arg Leu Ala Glu Ala Ala Ala Arg Gln Met Lys Lys Leu

65 70 75 80

Pro Phe Tyr His Thr Phe Ser Tyr Arg Ser His Gly Pro Val Ile Asp

85 90 95

Leu Ala Glu Lys Leu Val Ala Met Ala Pro Val Pro Met Ser Lys Ala

100 105 110

Tyr Phe Thr Asn Ser Gly Ser Glu Ala Asn Asp Thr Val Val Lys Leu

115 120 125

Ile Trp Tyr Arg Ser Asn Ala Leu Gly Glu Pro Glu Arg Lys Lys Ile

130 135 140

Ile Ser Arg Lys Arg Gly Tyr His Gly Val Thr Ile Ala Ser Ala Ser

145 150 155 160

Leu Thr Gly Leu Pro Asn Asn His Arg Ser Phe Asp Leu Pro Ile Asp

165 170 175

Arg Ile Leu His Thr Gly Cys Pro His His Tyr His Asp Ala Leu Pro

180 185 190

Gly Glu Ser Glu Glu Gln Phe Ala Thr Arg Leu Ala Asn Glu Leu Glu

195 200 205

Gln Leu Ile Leu Ala Glu Gly Pro His Thr Ile Ala Ala Phe Ile Gly

210 215 220

Glu Pro Val Met Gly Ala Gly Gly Val Val Val Pro Pro Lys Thr Tyr

225 230 235 240

Trp Glu Lys Val Gln Ala Val Leu Gly Arg Tyr Asn Ile Leu Leu Val

245 250 255

Ala Asp Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Asn Leu Phe Gly

260 265 270

Cys Gln Thr Phe Asp Ile Lys Pro Asp Ile Leu Val Met Ser Lys Gln

275 280 285

Leu Ser Ser Ser Tyr Leu Pro Ile Ser Ala Phe Leu Ile Asn Glu Arg

290 295 300

Val Tyr Ala Pro Ile Ala Glu Glu Ser His Lys Ile Gly Thr Leu Gly

305 310 315 320

Thr Gly Phe Thr Ala Ser Gly His Pro Val Ala Ala Ala Val Ala Leu

325 330 335

Glu Asn Leu Ala Ile Ile Glu Glu Arg Asp Leu Val Ala Asn Ala Arg

340 345 350

Glu Arg Gly Ala Arg Met Gln Lys Arg Leu Arg Glu Leu Gln Asp His

355 360 365

Pro Leu Val Gly Glu Val Arg Gly Val Gly Leu Ile Ala Gly Val Glu

370 375 380

Leu Val Ile Asp Lys Glu Ala Lys Thr Gly Leu Glu Gln Pro Gly Ala

385 390 395 400

Leu Gly Ala Arg Ala Asn Ala Ala Leu Gln Glu Arg Gly Val Ile Ser

405 410 415

Arg Ala Met Gly Asp Thr Leu Ala Phe Cys Pro Pro Leu Ile Ile Asn

420 425 430

Asp Gln Gln Val Asp Thr Met Val Ser Ala Leu Glu Ala Ser Leu Asn

435 440 445

Asp Val Gln Ala Ser Leu Gly Lys

450 455

<210> 8

<211> 453

<212> PRT

<213> Paracoccus denitrificans PD1222

<400> 8

Met Asn Gln Pro Gln Ser Trp Glu Ala Arg Ala Glu Thr Tyr Ser Leu

1 5 10 15

Tyr Gly Phe Thr Asp Met Pro Ser Val His Gln Arg Gly Thr Val Val

20 25 30

Val Thr His Gly Glu Gly Pro Tyr Ile Val Asp Val His Gly Arg Arg

35 40 45

Tyr Leu Asp Ala Asn Ser Gly Leu Trp Asn Met Val Ala Gly Phe Asp

50 55 60

His Lys Gly Leu Ile Glu Ala Ala Lys Ala Gln Tyr Asp Arg Phe Pro

65 70 75 80

Gly Tyr His Ala Phe Phe Gly Arg Met Ser Asp Gln Thr Val Met Leu

85 90 95

Ser Glu Lys Leu Val Glu Val Ser Pro Phe Asp Asn Gly Arg Val Phe

100 105 110

Tyr Thr Asn Ser Gly Ser Glu Ala Asn Asp Thr Met Val Lys Met Leu

115 120 125

Trp Phe Leu His Ala Ala Glu Gly Lys Pro Gln Lys Arg Lys Ile Leu

130 135 140

Thr Arg Trp Asn Ala Tyr His Gly Val Thr Ala Val Ser Ala Ser Met

145 150 155 160

Thr Gly Lys Pro Tyr Asn Ser Val Phe Gly Leu Pro Leu Pro Gly Phe

165 170 175

Ile His Leu Thr Cys Pro His Tyr Trp Arg Tyr Gly Glu Glu Gly Glu

180 185 190

Thr Glu Ala Gln Phe Val Ala Arg Leu Ala Arg Glu Leu Glu Asp Thr

195 200 205

Ile Thr Arg Glu Gly Ala Asp Thr Ile Ala Gly Phe Phe Ala Glu Pro

210 215 220

Val Met Gly Ala Gly Gly Val Ile Pro Pro Ala Lys Gly Tyr Phe Gln

225 230 235 240

Ala Ile Leu Pro Ile Leu Arg Lys Tyr Asp Ile Pro Met Ile Ser Asp

245 250 255

Glu Val Ile Cys Gly Phe Gly Arg Thr Gly Asn Thr Trp Gly Cys Leu

260 265 270

Thr Tyr Asp Phe Met Pro Asp Ala Ile Ile Ser Ser Lys Asn Leu Thr

275 280 285

Ala Gly Phe Phe Pro Met Gly Ala Val Ile Leu Gly Pro Asp Leu Ala

290 295 300

Lys Arg Val Glu Ala Ala Val Glu Ala Ile Glu Glu Phe Pro His Gly

305 310 315 320

Phe Thr Ala Ser Gly His Pro Val Gly Cys Ala Ile Ala Leu Lys Ala

325 330 335

Ile Asp Val Val Met Asn Glu Gly Leu Ala Glu Asn Val Arg Arg Leu

340 345 350

Ala Pro Arg Phe Glu Ala Gly Leu Lys Arg Ile Ala Asp Arg Pro Asn

355 360 365

Ile Gly Glu Tyr Arg Gly Ile Gly Phe Met Trp Ala Leu Glu Ala Val

370 375 380

Lys Asp Lys Pro Thr Lys Thr Pro Phe Asp Ala Asn Leu Ser Val Ser

385 390 395 400

Glu Arg Ile Ala Asn Thr Cys Thr Asp Leu Gly Leu Ile Cys Arg Pro

405 410 415

Leu Gly Gln Ser Ile Val Leu Cys Pro Pro Phe Ile Leu Thr Glu Ala

420 425 430

Gln Met Asp Glu Met Phe Glu Lys Leu Glu Lys Ala Leu Asp Lys Val

435 440 445

Phe Ala Glu Val Ala

450

Claims

1. a kind of method for preparing R-3- amino n-butanol, includes the following steps:

Using 3- amino n-butanol racemic modification and pyruvic acid as substrate, it is catalyst with ω-transaminase, is catalyzed to stereospecificity S-3- amino n-butanol is reacted with pyruvic acid, and the positive fourth of enantiomer R-3- amino for having neither part nor lot in reaction is then recycled from reaction system Alcohol.

2. the method as described in claim 1, which is characterized in that the ω-transaminase derives from microorganism selected from the group below: Ochrobactrum anthropic、Bradyrhizobium sp.Ec3.3、Brucella neotomae、 Novosphingobium acidiphilum、Ochrobactrum intermedium、Pseudaminobacter salicylatoxidans、Brucella abortus、Paracoccus denitrificans PD1222。

3. the method as described in claim 1, which is characterized in that the ω-transaminase is selected from the group: SEQ ID NO:1, SEQ ID NO:3、SEQ ID NO:4、SEQ ID NO:7。

4. the method as described in claim 1, which is characterized in that be also added with phosphopyridoxal pyridoxal phosphate in reaction system.

5. method as claimed in claim 4, which is characterized in that the additive amount of phosphopyridoxal pyridoxal phosphate in the reaction system is 0.1- 1mmol/L。

6. the method as described in claim 1, which is characterized in that the ω-transaminase is in enzyme form or expression microorganism shape Formula.

7. method as claimed in claim 6, which is characterized in that the expression microorganism is that expression ω-transaminase recombination is big Enterobacteria.

8. the method as described in claim 1, which is characterized in that reaction solution used in the reaction system is aqueous solution, phosphorus Phthalate buffer, Tirs buffer or bicarbonate buffer.

9. the method as described in claim 1, which is characterized in that reaction system pH6.5-pH9.0.

10. the method as described in claim 1, which is characterized in that reaction temperature is 25-45 DEG C.