CN109097409A - The preparation method of D- amino acid and alpha keto acid - Google Patents
The preparation method of D- amino acid and alpha keto acid Download PDFInfo
- Publication number
- CN109097409A CN109097409A CN201810905471.5A CN201810905471A CN109097409A CN 109097409 A CN109097409 A CN 109097409A CN 201810905471 A CN201810905471 A CN 201810905471A CN 109097409 A CN109097409 A CN 109097409A
- Authority
- CN
- China
- Prior art keywords
- amino acid
- acid
- preparation
- aar
- alpha keto
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P13/00—Preparation of nitrogen-containing organic compounds
- C12P13/04—Alpha- or beta- amino acids
- C12P13/08—Lysine; Diaminopimelic acid; Threonine; Valine
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0012—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.6, 1.7)
- C12N9/0014—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.6, 1.7) acting on the CH-NH2 group of donors (1.4)
- C12N9/0022—Oxidoreductases (1.) acting on nitrogen containing compounds as donors (1.4, 1.5, 1.6, 1.7) acting on the CH-NH2 group of donors (1.4) with oxygen as acceptor (1.4.3)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/90—Isomerases (5.)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P13/00—Preparation of nitrogen-containing organic compounds
- C12P13/04—Alpha- or beta- amino acids
- C12P13/06—Alanine; Leucine; Isoleucine; Serine; Homoserine
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P13/00—Preparation of nitrogen-containing organic compounds
- C12P13/04—Alpha- or beta- amino acids
- C12P13/22—Tryptophan; Tyrosine; Phenylalanine; 3,4-Dihydroxyphenylalanine
- C12P13/222—Phenylalanine
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y104/00—Oxidoreductases acting on the CH-NH2 group of donors (1.4)
- C12Y104/03—Oxidoreductases acting on the CH-NH2 group of donors (1.4) with oxygen as acceptor (1.4.3)
- C12Y104/03002—L-Amino-acid oxidase (1.4.3.2)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y501/00—Racemaces and epimerases (5.1)
- C12Y501/01—Racemaces and epimerases (5.1) acting on amino acids and derivatives (5.1.1)
- C12Y501/0101—Amino-acid racemase (5.1.1.10)
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention belongs to field of biotechnology, it is related to the preparation method of D- amino acid and alpha keto acid, include the following steps: that 1. recombinant bacteriums construct: full genome synthesizes l-amino acid deaminase gene LAAD, is connected into expression vector, conversion constructs recombinant bacterium PMLAAD to expression bacterial strain；Full genome synthesizing amino acid racemase gene AAR, is connected into expression vector, and conversion constructs recombinant bacterium AAR to expression bacterial strain；2. fermentation；3. enzymatic；4. isolating and purifying.Enzymatic production alpha-calcium picrolonate of the invention and D- amino acid, two products all have economic value, and product quality is high, thus greatly reduces production cost.
Description
Technical field
The invention belongs to field of biotechnology, are related to the preparation method of D- amino acid and alpha keto acid.
Background technique
There are 20 kinds of amino acid for nature, are L-type amino acid, they are the basic structural units of constitutive protein matter.Institute
With, it is considered that D- amino acid seldom exists in nature, in recent years, with going deep into for scientific research, it has been found that D- ammonia
Although base acid is not the basic structure nitrogen source for constituting protein, there is D- amino in many plants, microorganism and higher plant
The presence of acid.
The application field of D- type amino acid is mainly at present: 1. medicine aspects.For synthetic polypeptide medicaments, such as polypeptide antibiosis
Plain (aspoxicillin ASPOXICILLIN), medicine for enterogastritis (OXTREOTIDE), diuretics, diarrhea medicine (Sandostatin LAR Depot SANDOSTATIN
ACETATE), enzyme inhibitor, corticotropin(ACTH) analog relieve pain antalgesic, slimming drugs, type II diabetes medicine (Nateglinide
NATEGLINIDE) etc..2. in terms of food.Aspartame (Aspartame), the novel sweeteners such as third dipeptides of day (alitame).D-
Type amino acid can also be used in the products such as food flavor, antistaling agent, preservative, food coupler, food flavor.3. pesticide side
Face.Agrochemicals of Amino Acids is used without large area, can not only reduce manpower and material resources and survival dose, and have toxicity it is low, efficiently without public affairs
The features such as evil, be easily degraded utilization, and raw material sources are extensive, become new pesticide research hotspot.4. scientific research.Pass through postgraduate
The change of object vivo acid D, L- content of isomer can be used as year band timer and carry out weather, hydrologic research.D- amino acid is also
The research that can be used in terms of the Structure-function analysis of enzyme.
Asymmetry conversion is the main method for preparing D- amino acid, can be divided into chemistry not according to preparation means and approach difference
Symmetrical conversion and the asymmetric conversion of biology.Can be divided into according to initial feed difference: method of asymmetric synthesis is directly prepared；Synthesis is outer
It is split after raceme；Asymmetric conversion preparation is passed through by l-amino acid or dl aminoadipic acid.Wherein, it generally uses both at home and abroad now
Method be first with chemical method synthesize racemic modification, then carry out split preparation D- amino acid.
Chemical method produce the reaction condition of D- amino acid relatively acutely, pollution is big, low yield and costly, therefore not
It is suitble to large-scale production.And developing very fast and most application potential is biological preparation method.In biological preparation method,
According to initial feed difference, study at present it is more be divided into it is following several:
1, it using the DL- amino acid of racemization or derivative as raw material, is extracted with complete microbial cell or from microbial cell
Enzyme as catalyst, hydrolyze amino acid or derivatives thereof.It, can only be selective since the stereoselectivity of biocatalyst is strong
The a certain optically-active amino acid of hydrolysis or derivatives thereof, it is another not hydrolyzed or hydrolyze seldom.Therefore, two kinds of optically-active amino acid tools
There is different combining forms, also just there is different physicochemical properties, it may be convenient to be divided by means of physics, chemical means
From.N- acetyl-amino-acylase (N-acyl-D-amio acid amidohydrolase) is to split the preparation of DL- type amino acid
One of main enzyme of D- amino acid.From the point of view of document report, most of most suitable substrates of amino-acylase are mostly neutral ammonia
Base acid.Liao Benren etc. is split to obtain L-type tryptophan with amino-acylase as substrate using DL- acetyltryptophan.Do not digested
N- acetyl-D-trp, then D-trp is obtained with hydrochloric acid water solution.The yield and optical purity of D-trp and L-Trp are all
It can achieve 98% or more.
2, a certain racemic intermediate is prepared first, and the hydrolase and racemase generated by microbial body is in certain condition
Under keep racemic intermediate amino acid converting to D-.The more of this aspect research is that 5- replaces glycolylurea, and 5- replaces glycolylurea
Under the action of glycolylurea hydrolase and carbamoylase, corresponding D- amino acid is generated.Kyriakos etc. is using containing the sea D-
Because one step of microorganism of enzyme and N- carbamyl-D- aminoacidase conversion DL- 4-Hydroxyphenyl hydantoin is that D- para hydroxybenzene is sweet
Propylhomoserin, the theoretical yield of two step enzyme reaction processes is up to 100%.Japan Ajinomoto, Germany Bayer company, Holland
The companies such as DSM are all producing D-pHPG with hydantoinase, and scale has reached thousands of tons/year.In addition to this, extra large
Because enzyme process can be also used for preparing D-trp, D-Val etc..
3, not right under the catalytic action of enzyme using the relatively cheap L-type amino acid of from a wealth of sources, price as starting material
Title is converted to D- type amino acid.Yagasaki etc. [18] constructs two plants of weights with Racemase activity and decarboxylase respectively
Group bacterial strain, l-amino acid obtain DL- amino acid after racemization, then split to obtain D- amino acid again, be obtained by the method
D-Glu and D-PROLINE ee value reach 99% or more, but yield is less than 50%.Ian etc. is the study found that l-amino acid is passed through
Deamination forms ketone acid, and ketone acid produces D- amino acid under the action of D- amino acid aminotransferase, according to this principle, structure
It builds with l-amino acid deaminase active and the active genetic engineering bacterium of D- amino acid transaminase, asymmetric can convert a variety of L-
Type amino acid, wherein leucine, tyrosine, tryptophan, the yield of methionine and ee value are 90% or more.
Above several method, it is all pure to be not suitable in certain disadvantage or to partial amino-acid, or be that production is uneconomical.
Toxic chemical substance is used when formerly synthesizing substrate sea mattress such as extra large mattress enzyme process, is polluted the environment, and extra large mattress enzyme process is to major part
D- amino acid is not applicable.Transaminase method: transaminase enzyme activity is generally relatively low, and needs amino group donor, needs first to prepare ketone acid.Also
There is some other method all to have some limitations.
Summary of the invention
The present invention provides the preparation method of a kind of D- amino acid and alpha keto acid, and enzymatic produces α-calcium picrolonate and D-
Amino acid, two products all have economic value, and product quality is high, thus greatly reduces production cost.
The present invention adopts the following technical scheme that:
A kind of preparation method of D- amino acid and alpha keto acid, includes the following steps:
1. recombinant bacterium constructs
Full genome synthesis source in proteus mirabilis (Proteus mirabilis) l-amino acid deaminase (L-amino
Acid deaminase, LAAD) gene (genbank accession number EU669819.1) LAAD, and be connected into pET24a carrier (NdeⅠ/Hind), plasmid LAAD is constructed, conversion to E.Coli BL21(DE3) constructs recombinant bacterium PMLAAD.
Full genome synthesis source in lactic acid bacteria (Lactobacillus buchneri Strain JCM 1115) amino
Sour racemase (amino acid racemase, AAR) gene (genbank accession number KC413940.1) AAR, and be connected into
PET24a carrier (NdeⅠ/Hind), plasmid AAR is constructed, conversion to E.Coli BL21(DE3) constructs recombinant bacterium AAR.
2. fermentation
Recombinant bacterium PMLAAD and AAR are fermented using TB.
1) culture medium
(1) LB:
(2) agar LB-Kan plate (g/L):
Note: medium sterilization is cooled to 50-60 DEG C, and 100ml LB adds 100 μ l Kan solution (100mg/ml), mixes, inverted plate
(25-30ml/6cm plate).
(3) TB (g/L):
2) fermentation process
(1) actication of culture: bacterium is connect to Kan LB agar plate, 37 DEG C of culture 12-14h from glycerol tube or milk pipe.
(2) first order seed: picking from the plate single bacterium to 4ml LB test tube, adds Kan to final concentration of 100mg/L, and 37 DEG C,
220rpm overnight incubation (16h).
(3) secondary seed: fresh bacterium solution is taken to be inoculated in LB(100ml/500ml by 2% inoculum concentration) shaking flask, add Kan dense to end
Degree is 100mg/L, and 37 DEG C, 220rpm cultivates 3.0-3.5h, OD600It is 1.0 or so.
(4) the TB fermentation of 1L/5L: the inoculum concentration of secondary seed 2%-10% accesses TB culture medium (1L/5L fermentation shake flask), adds
Kan to 25mg/L 37 DEG C, 220rpm, cultivates 3h, starts 25 DEG C of cooling；Add IPTG to final concentration (0.1mM), 220rpm, 25
Degree, overnight incubation.
(5) ferment 20-24h, and thalline were collected by centrifugation by 3500rpm*25min, and bacterium sets -30 DEG C of refrigerator freezings for 24 hours.
3. enzymatic
Using 5 liters of fermentors, the l-amino acid (amino acid such as valine, phenylalanine, leucine, isoleucine of 100g are weighed
One of), add 3 liters of water, 300rpm stirring and dissolving, sodium hydroxide solution tune pH6-8 controls 35 DEG C of temperature, the AAR bacterium of freezing is added
Body 10g adds phosphopyridoxal pyridoxal phosphate (PLP) 150mg.It is stirred to react 6h, centrifugation recycling AAR, centrifugation supernatant is heated to 80 DEG C, 10min
Inactivate remaining Racemase activity.Supernatant is cooled to 35 DEG C, and PMLAAD thallus 20g is added, commodity-hydrogen peroxide of 3g is added
Enzyme is passed through the air of 3L/min, and 300rpm is stirred to react to amino acid and no longer reduces, and stops reaction, bacteria recovered by centrifugation.
4. isolating and purifying
Conversion centrifugation supernatant crosses ceramic membrane, and ultrafiltration membrane, decolourize film, adjusts pH3-5, and upper cation exchange resin adsorbs D- amino acid,
It collects α-ketone acid and flows through liquid, calcium chloride is added.Ammonium hydroxide elutes lower D- amino acid.Condensing crystallizing α-calcium picrolonate and D- ammonia respectively
Base acid.Filter dry acquisition α-ketone acid calcium salt 40g and D- amino acid 38g.
By implementing above-mentioned technical proposal, enzymatic production α-calcium picrolonate of the invention and D- amino acid, two products are all
With economic value, product quality is high, thus greatly reduces production cost.
Specific embodiment
A kind of preparation method of D- amino acid and alpha keto acid, includes the following steps:
1. recombinant bacterium constructs:
Full genome synthesis source in proteus mirabilis (Proteus mirabilis) l-amino acid deaminase (L-amino
Acid deaminase, LAAD) gene (genbank accession number EU669819.1) LAAD, and be connected into pET24a carrier (NdeⅠ/Hind), plasmid LAAD is constructed, conversion to E.Coli BL21(DE3) constructs recombinant bacterium PMLAAD.
Full genome synthesis source in lactic acid bacteria (Lactobacillus buchneri Strain JCM 1115) amino
Sour racemase (amino acid racemase, AAR) gene (genbank accession number KC413940.1) AAR, and be connected into
PET24a carrier (NdeⅠ/Hind), plasmid AAR is constructed, conversion to E.Coli BL21(DE3) constructs recombinant bacterium AAR.
2. fermentation
Recombinant bacterium PMLAAD and AAR are fermented using TB.
3) culture medium
(1) LB:
(2) agar LB-Kan plate (g/L):
Note: medium sterilization is cooled to 50-60 DEG C, and 100ml LB adds 100 μ l Kan solution (100mg/ml), mixes, inverted plate
(25-30ml/6cm plate).
(3) TB (g/L):
4) fermentation process
(1) actication of culture: bacterium is connect to Kan LB agar plate, 37 DEG C of culture 12-14h from glycerol tube or milk pipe.
(2) first order seed: picking from the plate single bacterium to 4ml LB test tube, adds Kan to final concentration of 100mg/L, and 37 DEG C,
220rpm overnight incubation (16h).
(3) secondary seed: fresh bacterium solution is taken to be inoculated in LB(100ml/500ml by 2% inoculum concentration) shaking flask, add Kan dense to end
Degree is 100mg/L, and 37 DEG C, 220rpm cultivates 3.0-3.5h, OD600It is 1.0 or so.
(4) the TB fermentation of 1L/5L: the inoculum concentration of secondary seed 2%-10% accesses TB culture medium (1L/5L fermentation shake flask), adds
Kan to 25mg/L 37 DEG C, 220rpm, cultivates 3h, starts 25 DEG C of cooling；Add IPTG to final concentration (0.1mM), 220rpm, 25
Degree, overnight incubation.
(5) ferment 20-24h, and thalline were collected by centrifugation by 3500rpm*25min, and bacterium sets -30 DEG C of refrigerator freezings for 24 hours.
3. enzymatic
Using 5 liters of fermentors, the l-amino acid (amino acid such as valine, phenylalanine, leucine, isoleucine of 100g are weighed
One of), add 3 liters of water, 300rpm stirring and dissolving, sodium hydroxide solution tune pH6-8 controls 35 DEG C of temperature, the AAR bacterium of freezing is added
Body 10g adds phosphopyridoxal pyridoxal phosphate (PLP) 150mg.It is stirred to react 6h, centrifugation recycling AAR, centrifugation supernatant is heated to 80 DEG C, 10min
Inactivate remaining Racemase activity.Supernatant is cooled to 35 DEG C, and PMLAAD thallus 20g is added, commodity-hydrogen peroxide of 3g is added
Enzyme is passed through the air of 3L/min, and 300rpm is stirred to react to amino acid and no longer reduces, and stops reaction, bacteria recovered by centrifugation.
4. isolating and purifying
Conversion centrifugation supernatant crosses ceramic membrane, and ultrafiltration membrane, decolourize film, adjusts pH3-5, and upper cation exchange resin adsorbs D- amino acid,
It collects α-ketone acid and flows through liquid, calcium chloride is added.Ammonium hydroxide elutes lower D- amino acid.Condensing crystallizing α-calcium picrolonate and D- ammonia respectively
Base acid.Filter dry acquisition α-ketone acid calcium salt 40g and D- amino acid 38g.
Claims (8)
- The preparation method of 1.D- amino acid and alpha keto acid, which comprises the steps of:1, recombinant bacterium constructsFull genome synthesizes l-amino acid deaminase gene LAAD, is connected into expression vector, and conversion constructs recombinant bacterium to expression bacterial strain PMLAAD；Full genome synthesizing amino acid racemase gene AAR, is connected into expression vector, and conversion constructs recombinant bacterium AAR to expression bacterial strain；2, it ferments；3, enzymatic；4, it isolates and purifies.
- 2. the preparation method of D- amino acid and alpha keto acid according to claim 1, which is characterized in that the l-amino acid Deaminase gene LAAD derives from proteus mirabilis.
- 3. the preparation method of D- amino acid and alpha keto acid according to claim 1 or claim 2, which is characterized in that the amino Sour racemase gene AAR derives from lactic acid bacteria.
- 4. the preparation method of D- amino acid and alpha keto acid according to claim 3, which is characterized in that the expression vector For pET24a carrier.
- 5. the preparation method of D- amino acid and alpha keto acid according to claim 3, which is characterized in that the expression bacterial strain For E.Coli BL21(DE3).
- 6. the preparation method of D- amino acid and alpha keto acid according to claim 1, which is characterized in that step 3 enzymatic Concrete operations are as follows: take l-amino acid, be dissolved in water, sodium hydroxide solution tune pH6-8, control 35 DEG C of temperature, be added AAR bacterium Body, phosphopyridoxal pyridoxal phosphate are stirred to react 6h, centrifugation recycling AAR, and centrifugation supernatant is heated to 80 DEG C, and 10min inactivates remaining racemase Activity, supernatant are cooled to 35 DEG C, and PMLAAD thallus is added, and catalase is added, is passed through air, 300rpm is stirred to react It is no longer reduced to amino acid, stops reaction, bacteria recovered by centrifugation, supernatant is used for step 4.
- 7. the preparation method of D- amino acid and alpha keto acid according to claim 6, which is characterized in that the l-amino acid For one of valine, phenylalanine, leucine, isoleucine.
- 8. the preparation method of D- amino acid and alpha keto acid according to claim 1, which is characterized in that separated in step 4 The concrete operations of purifying are as follows: the last resulting supernatant of step 3 passes sequentially through ceramic membrane, ultrafiltration membrane, decoloration film, adjusts pH3-5, on Cation exchange resin adsorbs D- amino acid, collects α-ketone acid and flows through liquid, and calcium chloride is added, and ammonium hydroxide elutes lower D- amino Acid, condensing crystallizing α-calcium picrolonate and D- amino acid, filter dry acquisition α-ketone acid calcium salt and D- amino acid respectively.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810905471.5A CN109097409A (en) | 2018-08-10 | 2018-08-10 | The preparation method of D- amino acid and alpha keto acid |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810905471.5A CN109097409A (en) | 2018-08-10 | 2018-08-10 | The preparation method of D- amino acid and alpha keto acid |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109097409A true CN109097409A (en) | 2018-12-28 |
Family
ID=64849112
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810905471.5A Pending CN109097409A (en) | 2018-08-10 | 2018-08-10 | The preparation method of D- amino acid and alpha keto acid |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109097409A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109097383A (en) * | 2018-08-10 | 2018-12-28 | 浙江正硕生物科技有限公司 | A kind of method of high flux screening l-amino acid deamination enzyme mutant recombinant bacterial strain |
| CN110938580A (en) * | 2019-12-26 | 2020-03-31 | 江南大学 | Method for improving production efficiency of D-tyrosine |
| JP2020188753A (en) * | 2019-05-14 | 2020-11-26 | 大洋香料株式会社 | Beverage flavor-improver |
| CN112626100A (en) * | 2020-12-29 | 2021-04-09 | 江南大学 | Method for high-throughput screening of alpha-keto acid high-yield strains |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013082542A2 (en) * | 2011-12-02 | 2013-06-06 | Invista North America S.A.R.L. | Methods for biosynthesizing 1,3butadiene |
| CN103789247A (en) * | 2014-02-14 | 2014-05-14 | 江南大学 | Method for producing alpha-ketoisocaproate by whole-cell transformation |
| CN107129436A (en) * | 2016-06-27 | 2017-09-05 | 通辽梅花生物科技有限公司 | A kind of isoleucine extraction process |
-
2018
- 2018-08-10 CN CN201810905471.5A patent/CN109097409A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013082542A2 (en) * | 2011-12-02 | 2013-06-06 | Invista North America S.A.R.L. | Methods for biosynthesizing 1,3butadiene |
| CN103789247A (en) * | 2014-02-14 | 2014-05-14 | 江南大学 | Method for producing alpha-ketoisocaproate by whole-cell transformation |
| CN107129436A (en) * | 2016-06-27 | 2017-09-05 | 通辽梅花生物科技有限公司 | A kind of isoleucine extraction process |
Non-Patent Citations (4)
| Title |
|---|
| BAEK等: "Accession ID：EU669819.1，Proteus mirabilis L-amino acid deaminase gene， complete cds", 《GENEBANK DATABASE》 * |
| MUTAGUCHI等: "Accession ID：KC413940.1，Lactobacillus buchneri strain JCM 1115 putative 4-aminobutyrate aminotransferase gene， complete cds", 《GENEBANK DATABASE》 * |
| WOLF.DIETER FESSNER: "《现代生物催化 高立体选择及环境友好的反应》", 30 June 2016, 中国轻工业出版社 * |
| 陆阳等: "双酶级联生物合成 D-赖氨酸", 《精细化工》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109097383A (en) * | 2018-08-10 | 2018-12-28 | 浙江正硕生物科技有限公司 | A kind of method of high flux screening l-amino acid deamination enzyme mutant recombinant bacterial strain |
| JP2020188753A (en) * | 2019-05-14 | 2020-11-26 | 大洋香料株式会社 | Beverage flavor-improver |
| CN110938580A (en) * | 2019-12-26 | 2020-03-31 | 江南大学 | Method for improving production efficiency of D-tyrosine |
| CN112626100A (en) * | 2020-12-29 | 2021-04-09 | 江南大学 | Method for high-throughput screening of alpha-keto acid high-yield strains |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109097409A (en) | The preparation method of D- amino acid and alpha keto acid | |
| CN105296456B (en) | A kind of stability-enhanced glutamic acid decarboxylase enzyme mutant of pH and its application | |
| CN101240302B (en) | Method for producing L-phenylalanine by using bacillus subtilis to ferment feather | |
| CN109735559B (en) | Biological preparation method of gamma-aminobutyric acid | |
| CN107217048A (en) | It is a kind of to catalyze and synthesize aminopeptidase of carnosine and its preparation method and application | |
| CN111944856A (en) | Synthetic method of pregabalin intermediate | |
| CN108220352A (en) | A kind of method of raw material fermentation production gamma-polyglutamic acid | |
| CN109136298B (en) | Preparation method of D-amino acid | |
| KR101740968B1 (en) | Method for production of GABA with lactic acid bacteria through L-glutamic acid feeding | |
| CN102864190A (en) | Producing method of gamma-aminobutyric acid | |
| CN110055292A (en) | A kind of pyruvic acid and levodopa joint production process and application | |
| JP2018531602A (en) | Bio-based N-acetyl-L-methionine and uses thereof | |
| CN115725520A (en) | Preparation method of glutathione synthetase and method for catalytically producing glutathione | |
| CN105695427A (en) | Biological enzyme for catalyzing synthesis of glutathione and preparation and extraction methods of biological enzyme | |
| CN109576200A (en) | A kind of recombinant bacterium producing glutamate racemase and its construction method and application | |
| KR100614219B1 (en) | Fermentation of L-lysine using acid-treated product of soybean meal | |
| JPS61274690A (en) | Production of d-alpha-amino acid | |
| JPS62111693A (en) | Production of l-amino acid from alpha-ketocarboxilic acid byfermentation | |
| JP3635106B2 (en) | Biocatalytic continuous reaction method | |
| JPS6319158B2 (en) | ||
| JPS5816692A (en) | Preparation of l-tryptophan by enzyme | |
| CN112522171A (en) | Engineering bacterium, treatment method of ornithine-containing solution and kit | |
| CN117757862A (en) | Method for producing valine through fermentation and application | |
| JPS60164493A (en) | Preparation of l-phenylalanine | |
| CN117658697A (en) | Method for preparing fertilizer and culture medium by utilizing threonine fermentation waste liquid of phage |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20181228 |