CN107739733A - aspartate transaminase and preparation method thereof - Google Patents

aspartate transaminase and preparation method thereof Download PDF

Info

Publication number
CN107739733A
CN107739733A CN201710916374.1A CN201710916374A CN107739733A CN 107739733 A CN107739733 A CN 107739733A CN 201710916374 A CN201710916374 A CN 201710916374A CN 107739733 A CN107739733 A CN 107739733A
Authority
CN
China
Prior art keywords
aspartate transaminase
thalline
seq
preparation
ala
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.)
Granted
Application number
CN201710916374.1A
Other languages
Chinese (zh)
Other versions
CN107739733B (en
Inventor
宫春杰
张抒杨
陶冶
胡征
杨波
王毅
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Beijing Yongbo Technology Co ltd
Original Assignee
Hubei University of Technology
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Hubei University of Technology filed Critical Hubei University of Technology
Priority to CN201710916374.1A priority Critical patent/CN107739733B/en
Publication of CN107739733A publication Critical patent/CN107739733A/en
Application granted granted Critical
Publication of CN107739733B publication Critical patent/CN107739733B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • 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
    • C12N9/10—Transferases (2.)
    • C12N9/1096—Transferases (2.) transferring nitrogenous groups (2.6)
    • 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
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/70—Vectors or expression systems specially adapted for E. coli
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y206/00—Transferases transferring nitrogenous groups (2.6)
    • C12Y206/01—Transaminases (2.6.1)
    • C12Y206/01001—Aspartate transaminase (2.6.1.1), i.e. aspartate-aminotransferase

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • Biomedical Technology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Plant Pathology (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The present invention relates to biology field, more particularly to a kind of aspartate transaminase and preparation method thereof.The preparation method of aspartate transaminase, comprises the following steps：1) base sequence shown in SEQ ID NO.2 is expanded using round pcr；2) base sequence shown in SEQ ID NO.2 is cloned into vector plasmid, builds the expression vector of aspartate transaminase；3) expression vector is transformed into E. coli competent thalline and obtains recombinating thalline, obtain aspartate transaminase albumen through expanding culture, induced expression, collecting thalline, broken thalline and purifying protein process successively.Method using Ecoli BL21 pET21a AT recombinant bacterial strain induced expression bacterium glutamine synthetase proteins is simple, it is easy to operate, and test proves that the aspartate transaminase of the present invention is cold-adapted enzyme, there is the tolerance of preferable low temperature, can be that industrial production saves energy consumption.

Description

Aspartate transaminase and preparation method thereof
Technical field
The present invention relates to biology field, more particularly to a kind of aspartate transaminase and preparation method thereof.
Background technology
Aspartate transaminase, alias：Glutamic-oxaloacetic transaminase, English name：Aspartate transaminase(AST).Aspartate transaminase rises in the combined deamination in human body in amino acid catabolic Important function, it is catalyzed the transamination of aspartic acid and a-ketoglutaric acid, forms glutamic acid and oxaloacetic acid.Divide extensively It is distributed in human body respectively to organize, it is predominantly located at liver, kidney and skeletal muscle, with levels and myocardial contents highest.Aspartate transaminase contains The important indicator being scheduled on medically as liver function test is measured, for judging whether liver function suffers damage.
Aspartate transaminase in liver has two kinds of isodynamic enzymes, be respectively be present in liver cell mitochondria m-AST and Intracytoplasmic s-AST.AST normal value is 0~40U/L in serum.When liver cell has slight lesion, s-AST is released Into blood, when lesion aggravates, m-AST can also be discharged in succession, and AST content and hepatocellular damage degree are into positive in serum Close, therefore can infer whether liver function is normal by calculating in serum AST contents.
There is great application prospect using aspartate transaminase production aromatic amino acid, under optimum conditions, with Aspartic acid and several ketone propylhomoserins are l-amino acid corresponding to substrate synthesis.Utilizing aspartate transaminase synthetic styrene-acrylic ketone There is preferable effect when acid, p-hydroxyphenylpyruvic acid and indole-3-pyruvic acid.
It is poor to the tolerance of low temperature that obtained aspartate transaminase is produced at present, can hardly play work at low temperature Property, in industrial production, the requirement to temperature is higher.
The content of the invention
To solve problem above, the present invention provides a kind of aspartate transaminase and preparation method thereof, obtained asparagus fern ammonia Sour transaminase has preferable low temperature tolerant.
An object of the present invention is to provide a kind of amino acid sequence of aspartate transaminase；The aspartic acid turns ammonia The amino acid sequence of enzyme is as shown in SEQ ID NO.1.
The second object of the present invention is to provide a kind of base sequence of codes for aspartate aminotransferase gene；The asparagus fern ammonia The base sequence of sour aminotransferase gene is as shown in SEQ ID NO.2.
The third object of the present invention is to provide a kind of preparation method of aspartate transaminase.
The preparation method of aspartate transaminase, comprises the following steps：
1) base sequence shown in SEQ ID NO.2 is expanded using round pcr；
2) base sequence shown in SEQ ID NO.2 is cloned into vector plasmid, builds the table of aspartate transaminase Up to carrier；
3) expression vector is transformed into E. coli competent thalline and obtains recombinating thalline, successively through expanding culture, luring Lead expression, collection thalline, broken thalline and purifying protein process and obtain bacterial aspartate apotransminase.
Preferably, in the step 1), round pcr amplification primer used is：Forward primer such as SEQ ID NO.3 institutes Show, reverse primer is as shown in SEQ ID NO.4.
Preferably, vector plasmid is pET21a (+) in the step 2).
Preferably, in the step 3), E. coli competent thalline is e. coli bl21 competence thalline.
Preferably, in the step 3), carried out during the purifying protein using Ni-NTA affinity chromatography chromatographic columns Protein purification.
The beneficial effects of the present invention are：By round pcr, the amplification in cold Bacillus is liked obtains aspartic acid to the present invention Aminotransferase gene, and prove that the aspartate aminotransferase gene base sequence belongs to by the comparison of NCBI Blast base sequences Like the new aspartate aminotransferase gene sequence of cold Bacillus.Obtain expressing aspartate transaminase by technique for gene engineering The Ecoli BL21-pET21a-AT recombinant bacterial strains of albumen, induced expression recombinant bacterial strain obtain aspartate transaminase albumen.Profit Method with Ecoli BL21-pET21a-AT recombinant bacterial strain induced expression aspartate transaminase albumen is simple, easy to operate, and And test proves that the aspartate transaminase of the present invention is cold-adapted enzyme, i.e., performance is active under cryogenic, have preferable The tolerance of low temperature, can be that industrial production saves energy consumption.
Brief description of the drawings
Fig. 1 is the agarose gel electrophoresis qualification figure that PCR expands aspartate aminotransferase gene；Wherein M swimming lanes are DNA Marker, No. 1 swimming lane are the product that PCR expands aspartate aminotransferase gene；
Fig. 2 is that the polyacrylamide of recombinant bacterial strain Ecoli BL21-pET21a-AT expression aspartate transaminase albumen coagulates Gel electrophoresis qualification figure；Wherein marker swimming lanes are protein marker；AT swimming lanes are aspartate transaminase albumen.
Fig. 3 is expression vector pET21a-AT structure schematic diagram；
Embodiment
Embodiment 1
The acquisition of aspartate aminotransferase gene
1) bacterial strain containing target gene is obtained
The present invention likes cold Bacillus genus strain screened from Changbai Mountain soil, specifically screened using cold Bacillus genus strain is liked The Chinese invention patent of journey such as Patent No. 201710034491.5, the bacterial strain are named as：Cryobacterium Baishanse 02, it is stored in China typical culture collection center, deposit number CCTCC NO：M2016604.Preservation date For on October 31st, 2016, preservation address was Wuhan, China, Wuhan University.
2) extraction of genome
Utilize the bacterial strains of DNA extraction kit (TAKARA Dalian) extraction Cryobacterium baishanse 02 Genome is stand-by in -20 DEG C of preservations as template, the genomic samples of extraction.
3) PCR amplifying target genes
Design primer：Forward primer is as shown in SEQ ID NO.3, and reverse primer is as shown in SEQ ID NO.4；PCR reacts System is as shown in table 1 below：
Table 1
PCR reaction conditions are：95 DEG C of pre-degeneration 5min；95 DEG C of 30s, 55 DEG C of 30s, 68 DEG C extension 1.30min, 30 altogether Circulation；68 DEG C of extension 10min；15 DEG C of holding 10min.
By pcr amplification product through 1% agarose gel electrophoresis, as shown in figure 1, the size of pcr amplification product with it is expected that 1140bp is approached, and by pcr amplification product gel extraction, send biotech firm's sequencing with the base sequence of accurate judgement pcr amplification product Row, sequencing commission Bo Shang Bioisystech Co., Ltd completes.
Sequencing result is as shown in SEQ ID NO.2, by the base sequence shown in SEQ ID NO.2 in NCBI Blast websites (https://blast.ncbi.nlm.nih.gov/Blast.cgi) on carry out base sequence comparison, comparison result with Cryobacterium arcticum strain PAMC 27867 aspartate transaminase similitude is 93%, you can is judged Base sequence shown in SEQ ID NO.2 belongs to the new aspartate transaminase sequence of Cryobacterium category.
Embodiment 2
Build the expression vector of aspartate transaminase
The pcr amplification product obtained in embodiment 1 is subjected to glue reclaim, with restriction enzyme EcoRI and NdeI to PCR Amplified production carries out double digestion reaction；Double digestion is carried out to carrier pET21a (+) with restriction enzyme EcoRI and NdeI simultaneously Reaction, then through efficient DNA ligase High Ligation (TOYOBO) catalysis connections through the reacted pET21a of double digestion (+) and pcr amplification product；Structure obtains the expression vector pET21a-AT of aspartate transaminase, expression vector pET21a-AT Structure it is as shown in Figure 3.
Embodiment 3
Expression and purifying aspartate transaminase albumen
The expression vector pET21a-AT that embodiment 2 obtains is transformed into the competence thalline of e. coli bl21, large intestine The preparation of bacillus BL21 competence uses CaCl2Method, CaCl2The competence that method prepares e. coli bl21 is conventional for laboratory Laboratory facilities, it will not be repeated here.Recombinant bacterium is obtained after obtained expression vector pET21a-AT is transformed into e. coli bl21 Strain Ecoli BL21-pET21a-AT.Ecoli BL21-pET21a-AT are expanded into culture to OD6001mM is added after=0.6 IPTG, 28 DEG C of Fiber differentiations express aspartate transaminase albumen, and Fiber differentiation collects thalline after terminating, successively through bacterial cell disruption Aspartate transaminase albumen after purification is obtained after purification with Ni-NTA affinity chromatography chromatographic columns, as shown in Fig. 2 obtaining asparagus fern The molecular weight of albumen of propylhomoserin apotransminase is consistent with theoretical predicted value 40.7KD between 29.0-44.3, shows that purifying obtains Albumen be aspartate transaminase albumen.
Embodiment 4
Reaction temperature is tested the effect of vigor of aspartate transaminase
Determined using Mavrides methods；
The preparation of substrate：50mmol/L L-Aspartic acid and the reaction solution of α-ketoglutaric acid are prepared, reaction solution PH is 7.0；
Enzyme activity unit defines：Aspartate transaminase at certain temperature and pH condition, each second catalytic conversion 1mol substrate amino acids generate corresponding product and are defined as a unit of activity.As a result it see the table below 1：
Table 1
As can be seen from Table 1, the aspartate transaminase optimum temperature that the present invention obtains is 50 DEG C, and aspartic acid turns Ammonia enzyme has 11% relative enzyme activity at 4 DEG C, and compared with existing aspartate transaminase, the aspartic acid that the present invention obtains turns Ammonia enzyme has preferable low temperature tolerant, because the base sequence of codes for aspartate transaminase comes from Cryobacterium baishanse 02, Cryobacterium baishanse 02 is pyschrophile, so as to get asparagus fern ammonia Sour transaminase has low temperature tolerant characteristic, and aromatic series can be produced under cryogenic using the aspartate transaminase of the present invention Amino acid, there is great application prospect.
Embodiment described above only expresses the several embodiments of the present invention, and its description is more specific and detailed, but simultaneously Therefore the limitation to the scope of the claims of the present invention can not be interpreted as.It should be pointed out that for one of ordinary skill in the art For, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to the guarantor of the present invention Protect scope.Therefore, the protection domain of patent of the present invention should be determined by the appended claims.
Sequence table
<110>Hubei University Of Technology
<120>Aspartate transaminase and preparation method thereof
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 379
<212> PRT
<213> Cryobacterium
<400> 1
Met Pro Gln Ile Ser Ser Thr Ala Arg Ser Val Pro Gly Ser Gly Ile
1 5 10 15
Arg Arg Ile Phe Glu Leu Ala Ala Asp Leu Pro Asp Val Ile Gln Leu
20 25 30
Ser Val Gly Glu Pro Glu Glu Arg Val Ala Pro His Ile Leu Asp Ala
35 40 45
Gly Ala Arg Ala Trp Arg Asn Asp Ala Thr Asn Tyr Thr Pro Asn Ser
50 55 60
Gly Leu Leu Leu Leu Arg Ser Ala Ile Val Ser Lys Leu Gly Glu Phe
65 70 75 80
Asn Gly Tyr Ala Val Asp Glu Asp Gln Val His Ile Thr Ala Gly Gly
85 90 95
Ser Gln Ala Leu His Met Ala Met Leu Leu Thr Leu Asp Ala Gly Asp
100 105 110
Glu Ile Leu Ile Pro Asp Pro Gly Tyr Ala Thr Phe Ser Met Ala Ser
115 120 125
Arg Leu Val Gly Ala Ile Pro Val Pro Tyr Arg Leu Thr Ala Glu His
130 135 140
Ala Phe Arg Pro Arg Ile Asp Asp Leu Glu Gln Leu Val Ser Pro Arg
145 150 155 160
Thr Arg Val Leu Leu Ile Asn Ser Pro Ser Asn Pro Leu Gly Val Val
165 170 175
Phe Glu Pro Glu Val Leu Arg Glu Leu Leu Asp Phe Ala Ala Arg His
180 185 190
Asp Leu Trp Val Ile Ser Asp Glu Val Tyr Glu Tyr Phe Thr Phe His
195 200 205
Gly Gly Tyr Thr Ser Val Ala Ser Leu Asp Pro Asn Asp Arg Val Phe
210 215 220
Ser Val Tyr Ser Leu Ser Lys Thr Tyr Gly Leu Thr Gly Gly Arg Ile
225 230 235 240
Gly Tyr Leu Val Thr Pro Pro Gly Val Ala Gly Thr Phe Arg Ala Ala
245 250 255
Gln Glu Ala Ile Val Ser Cys Val Asn Thr Pro Ala Gln Leu Ala Ala
260 265 270
Leu Ala Ala Ile Glu Gly Asp Gln Ser Ala Val Ala Asp Ala Arg Glu
275 280 285
His Tyr Ser Arg Asn Leu Ala Ala Ala Gly Ala Ala Leu Asp Ala Arg
290 295 300
Gly Ile Glu Tyr Tyr Arg Pro Asp Gly Ala Phe Tyr Leu Trp Ile Asn
305 310 315 320
Val Ser Tyr Cys Ser Asn Gly Asn Val Ala Gln Trp Ala Glu Glu Phe
325 330 335
Leu Leu Ser Gln Arg Val Ala Val Ala Pro Gly Ser Ala Phe Gly Ala
340 345 350
Gly Gly Glu Gly Trp Ile Arg Ile Cys Ala Ala Gly Gln Arg Lys Pro
355 360 365
Leu Leu Thr Ala Leu Ser Arg Leu Pro Ala Val
370 375
<210> 2
<211> 1140
<212> DNA
<213> Cryobacterium
<400> 2
atgccccaga tttcgtccac ggcgcggtcg gttcccgggt cgggcatccg ccgcatcttc 60
gagctcgccg ccgacctgcc cgacgtgatc cagctcagcg tgggcgagcc cgaggaacgc 120
gtcgcgccgc acattctcga tgcgggcgcc cgagcgtggc ggaacgacgc caccaactac 180
acccccaaca gcggtctgct gctgctccgg agcgctatcg tgagcaagct cggcgagttc 240
aacgggtatg ccgtggacga agaccaggtg cacatcacgg ccggcgggtc gcaggccctg 300
cacatggcca tgctcctcac cctcgacgcg ggcgacgaga tcctcattcc cgacccgggt 360
tacgccacct tctccatggc ttcgaggctc gtcggcgcca ttccggtgcc gtaccggctc 420
acggccgagc acgccttccg gccgcggatc gatgacctcg aacagctggt gagcccacgc 480
acccgggtgc tgctgatcaa cagcccgtcc aatccgctcg gcgtcgtctt cgagccggag 540
gtgctgcggg aactgctcga cttcgccgcc cggcacgacc tctgggtgat cagcgacgag 600
gtctacgagt acttcacctt ccacggcggc tacacgagcg tggcgagcct ggacccgaac 660
gaccgggtgt tcagcgtcta ctcgctctcc aagacctatg gcctcacggg cggccggatc 720
ggctatctgg tgaccccgcc cggcgtggcc ggcaccttcc gggcggcgca agaggcgatc 780
gtcagctgcg tgaacacccc ggcgcagctc gccgccctgg ccgcgatcga gggcgaccag 840
tcggcggtcg cggatgcccg ggagcactac agcagaaacc tcgcggcggc cggtgctgcg 900
ctggatgccc gcggcatcga gtactaccgc cccgacgggg cgttctacct ctggatcaac 960
gtctcgtact gttcgaacgg gaacgtcgcc cagtgggccg aggagttcct gctcagccag 1020
cgggtggcgg tggcgcccgg ttcggccttc ggcgccggcg gggagggctg gatcaggatc 1080
tgcgccgctg gccagcgcaa accgctgctc accgcgctgt cccggctgcc tgccgtctaa 1140
<210> 3
<211> 31
<212> DNA
<213> Artificial Sequence
<400> 3
ggaattccat atgccccaga tttcgtccac g 31
<210> 4
<211> 28
<212> DNA
<213> Artificial Sequence
<400> 4
ggaattcgac ggcaggcagc cgggacag 28

Claims (7)

  1. A kind of 1. aspartate transaminase, it is characterised in that：The amino acid sequence of the aspartate transaminase such as SEQ ID Shown in NO.1.
  2. A kind of 2. gene for encoding the aspartate transaminase described in claim 1, it is characterised in that：The aspartic acid turns ammonia The base sequence of enzyme gene is as shown in SEQ ID NO.2.
  3. A kind of 3. method for preparing the aspartate transaminase described in claim 1, it is characterised in that：Comprise the following steps：
    1) base sequence shown in SEQ ID NO.2 is expanded using round pcr；
    2) base sequence shown in SEQ ID NO.2 is cloned into vector plasmid, the expression for building aspartate transaminase carries Body；
    3) expression vector is transformed into E. coli competent thalline and obtains recombinating thalline, successively through expanding culture, induction table Reach, collect thalline, broken thalline and purifying protein process and obtain aspartate transaminase albumen.
  4. 4. the preparation method of aspartate transaminase according to claim 3, it is characterised in that：In the step 1), PCR Technology expands primer used：Forward primer is as shown in SEQ ID NO.3, and reverse primer is as shown in SEQ ID NO.4.
  5. 5. the preparation method of aspartate transaminase according to claim 4, it is characterised in that：Carrier in the step 2) Plasmid is pET21a (+).
  6. 6. the preparation method of aspartate transaminase according to claim 5, it is characterised in that：In the step 3), greatly Enterobacteria competence thalline is e. coli bl21 competence thalline.
  7. 7. the preparation method of aspartate transaminase according to claim 6, it is characterised in that：In the step 3), institute Stating utilizes Ni-NTA affinity chromatography chromatographic columns to carry out protein purification during purifying protein.
CN201710916374.1A 2017-09-30 2017-09-30 Aspartate aminotransferase and preparation method thereof Active CN107739733B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201710916374.1A CN107739733B (en) 2017-09-30 2017-09-30 Aspartate aminotransferase and preparation method thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201710916374.1A CN107739733B (en) 2017-09-30 2017-09-30 Aspartate aminotransferase and preparation method thereof

Publications (2)

Publication Number Publication Date
CN107739733A true CN107739733A (en) 2018-02-27
CN107739733B CN107739733B (en) 2019-12-27

Family

ID=61236514

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201710916374.1A Active CN107739733B (en) 2017-09-30 2017-09-30 Aspartate aminotransferase and preparation method thereof

Country Status (1)

Country Link
CN (1) CN107739733B (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108546698A (en) * 2018-04-25 2018-09-18 浙江华睿生物技术有限公司 A kind of aspartic acid enzyme mutant

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107058167A (en) * 2017-01-17 2017-08-18 湖北工业大学 The preparation method and applications of microbial bacterial agent

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107058167A (en) * 2017-01-17 2017-08-18 湖北工业大学 The preparation method and applications of microbial bacterial agent

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
LEE J. 等: "Cryobacterium arcticum strain PAMC 27867 chromosome 1, complete sequence", 《GENBANK DATABASE》 *
LEILA BIROLO 等: "Aspartate aminotransferase from the Antarctic bacterium Pseudoalteromonas haloplanktis TAC 125 Cloning, expression, properties, and molecular modelling", 《EUR. J. BIOCHEM.》 *
张抒杨 等: "耐冷细菌适应低温机制研究进展及应用", 《山东化工》 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108546698A (en) * 2018-04-25 2018-09-18 浙江华睿生物技术有限公司 A kind of aspartic acid enzyme mutant
CN108546698B (en) * 2018-04-25 2020-03-31 浙江华睿生物技术有限公司 Aspartic enzyme mutant

Also Published As

Publication number Publication date
CN107739733B (en) 2019-12-27

Similar Documents

Publication Publication Date Title
CN108424900B (en) Nitrilase mutant and construction method and application thereof
CN105331642B (en) Method for catalytically producing α -ketoglutaric acid by using L-glutamic acid oxidase
CN109055327A (en) Aldehyde Ketoreductase mutant and its application
CN105062985A (en) Carbonyl reductase mutant and application thereof
CN106591271A (en) Arginine deiminase mutant with improved enzyme activity and temperature stability and application of mutant
CN103421734B (en) The method of the soluble expression of a kind of recombination tyrosine decarboxylase and the application of this enzyme
CN107739733A (en) aspartate transaminase and preparation method thereof
CN103992992A (en) Coding gene of (+) gamma-lactamase in Sulfolobus solfataricus P2, and application thereof
CN112410356A (en) Resveratrol synthase gene RS derived from radix tetrastigme and application thereof
CN109897870B (en) Method for preparing 10-hydroxy-2-decenoic acid from capric acid as raw material by using escherichia coli engineering bacteria
CN101497863B (en) Method for preparing N-terminated acetylated thymosin alpha 1 and special engineering bacteria therefor
CN101942422B (en) Genes of D-lactic dehydrogenase from serratia marcescens and research of cloning and expressing recombinant strains and recombinant enzymes
CN106434611A (en) Method for preparing L-ornithine by means of double-enzyme coupling by taking L-arginine as raw material
CN102732538B (en) Novel high-temperature-resisting lipase gene and coding product thereof
CN103382464B (en) Derive from the Ntn hydrolase of the ancient bacterium of hyperthermophilic and encoding gene thereof and application
CN110195088A (en) A kind of new arginine hydrolase and its encoding gene and application
CN106929491B (en) (S)-carbonyl reductase heteromer and its application in the more benzene ring compounds of catalysis
CN106434710A (en) Heatproof arginase gene expression sequence and application
CN104131017A (en) Rhodotorula glutinis phenylalanine deaminase gene and application thereof
CN105132394A (en) LIPASE 6 as well as encoding gene and application thereof
CN107739732A (en) Bacterium glutamine synthelase and preparation method thereof
CN104178469B (en) Cyclopropane formate hydrolase, gene, mutant and application thereof
CN115074303B (en) Genetically engineered bacterium capable of degrading feathers, construction method and application thereof
CN108018266A (en) A kind of marine source superoxide dismutase and its encoding gene and application
CN112322643B (en) Porphyra haitanensis glutamic-oxalacetic transaminase gene and application thereof

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
GR01 Patent grant
GR01 Patent grant
TR01 Transfer of patent right

Effective date of registration: 20231226

Address after: 101200 room 205-211526, No. 40, Fuqian West Street, Pinggu town, Pinggu District, Beijing (cluster registration)

Patentee after: BEIJING YONGBO TECHNOLOGY CO.,LTD.

Address before: 430068 No. 28 Nanli Road, Hongshan District, Wuhan City, Hubei Province

Patentee before: HUBEI University OF TECHNOLOGY

TR01 Transfer of patent right