CN114774438A - Osmanthus gene OfTPS380.1 and application thereof - Google Patents
Osmanthus gene OfTPS380.1 and application thereof Download PDFInfo
- Publication number
- CN114774438A CN114774438A CN202210420147.0A CN202210420147A CN114774438A CN 114774438 A CN114774438 A CN 114774438A CN 202210420147 A CN202210420147 A CN 202210420147A CN 114774438 A CN114774438 A CN 114774438A
- Authority
- CN
- China
- Prior art keywords
- gene
- geraniol
- saccharomyces cerevisiae
- osmanthus
- expression vector
- 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
Links
Images
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
- C12N9/14—Hydrolases (3)
- C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
-
- 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
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
- C12N15/81—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
-
- 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
- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
- C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y301/00—Hydrolases acting on ester bonds (3.1)
- C12Y301/07—Diphosphoric monoester hydrolases (3.1.7)
- C12Y301/07011—Geranyl diphosphate diphosphatase (3.1.7.11)
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Mycology (AREA)
- Biophysics (AREA)
- Plant Pathology (AREA)
- Physics & Mathematics (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention belongs to the technical field of molecular biology, and discloses an osmanthus geneOfTPS380.1And the application thereof, the geraniol synthetase gene highly expressed in the osmanthus petals is screened through the bioinformatics analysis of genome dataOfTPS380.1，OfTPS380.1The full-length sequence of gene CDS is shown as SEQ ID NO.1 and contains geneOfTPS380.1The expression vector of the method is used for transforming saccharomyces cerevisiae and fermenting and culturing, the yield of the geraniol reaches 21.67mg/L, and the method has industrial production value.
Description
Technical Field
The invention belongs to the technical field of molecular biology, and particularly relates to an osmanthus gene OfTPS380.1 and application thereof in production of geraniol
Background
Geraniol is an acyclic monoterpene alcohol compound extracted from plants. Colorless to yellow oily liquid at normal temperature, mild and sweet rose smell, and bitter taste. It is one of the main components of essential oils such as rose oil, clove oil and citronella oil, and is also present in small amounts in geranium and lemon. Geraniol and geranyl ester thereof are widely used as daily essence and edible essence, are main agents of rose essence, and are used for preparing daily products and foods. Meanwhile, the geraniol is used for antibiosis and expelling parasites, has good effect of clinically treating chronic bronchitis, can improve the lung ventilation function and reduce the airway resistance, and is also beneficial to improving the body immunity.
The osmanthus fragrans is a famous sweet plant and contains abundant terpenoids, and the osmanthus fragrans not only can bring pleasant mental feelings to people, but also has health care values of sterilization, inflammation diminishing, oxidation resistance, aging resistance and the like. The genome of the osmanthus fragrans contains a large number of aroma genes, but related reports of geraniol synthase genes do not exist in the osmanthus fragrans. The invention screens the geraniol synthase gene in osmanthus fragrans by using bioinformatics technology, and performs function verification in saccharomyces cerevisiae.
Disclosure of Invention
In order to overcome the defects of the prior art, the geraniol synthase gene OfTPS380.1 highly expressed in the osmanthus fragrans petals is screened from the osmanthus fragrans genome, and the function verification is carried out in the saccharomyces cerevisiae.
In order to achieve the purpose, the invention adopts the following technical scheme:
sweet-scented osmanthus leaf alcohol synthetase gene OfTPS380.1: through the analysis of genome data bioinformatics, a terpenoid gene OfTPS380.1 highly expressed in the petals of the osmanthus fragrans is screened, and the CDS sequence of the gene is shown as SEQ ID NO. 1.
An expression vector contains geraniol synthetase gene OfTPS380.1.
The recombinant saccharomyces cerevisiae expressing the OfTPS380.1 gene and the application thereof are as follows: transforming a yeast expression vector containing the gene OfTPS380.1 into saccharomyces cerevisiae, and obtaining recombinant saccharomyces cerevisiae for expressing the gene OfTPS380.1 through a resistant plate and sequencing screening; and (3) inoculating the recombinant saccharomyces cerevisiae into a YPD culture medium, adding isopropyl myristate for covering, and extracting geraniol after fermentation is finished.
Compared with the prior art, the invention has the following advantages and beneficial effects:
the research of the geraniol synthase gene in plants is very little, the geraniol synthase gene is excavated in the osmanthus fragrans for the first time and is subjected to functional verification in saccharomyces cerevisiae, the geraniol yield reaches 21.67mg/L, and the method has the value of industrial production.
Drawings
FIG. 1 is a gel map of the OfTPS380.1 clone in example 1
FIG. 2 is a map of the expression vector plasmid in example 2.
FIG. 3 shows the GC-MS detection results of geraniol.
Fig. 4 is a graph of geraniol ion fragmentation.
Detailed Description
The technical solutions of the present invention are described below with reference to the drawings and the specific embodiments, which are only used for explaining the present invention and are not used for limiting the scope of the present invention.
Example 1 Gene screening and cloning
Gene screening and cloning: the analysis of genome data bioinformatics shows that geraniol synthase gene (OfTPS380.1) highly expressed in osmanthus petals is screened, specific primers OfTPS380.1-CDS-F and OfTPS380.1-CDS-R (shown in table 1) for amplifying the full length of OfTPS380.1 gene CDS are designed and amplified by Primer5.0 software, and amplification primers are synthesized by Beijing Openkogaceae biotechnology. Taking the osmanthus fragrans cDNA as a template, amplifying a target gene CDS by referring to Phanta high-fidelity enzyme specifications, and setting a PCR reaction system and a program as shown in tables 2 and 3.
TABLE 1 primer List
TABLE 2PCR reaction System
TABLE 3PCR reaction procedure
Gel electrophoresis detection and gene sequencing: 30ml TAE added 0.45g agarose powder, microwave oven boiling after adding 3 u l 10000X nucleic acid dye, pouring into the rubber plate, after solidification sample. After 30min of 120V, 150mA electrophoresis, the bands were observed on a Gel-Logie200 Gel scanning imager (as shown in FIG. 1). And recovering the target band, connecting the recovered target band with a T-vector, transforming escherichia coli for sequencing, wherein the sequence of the OfTPS380.1 gene CDS is shown as SEQ ID No.1, extracting a Plasmid from a bacterial solution with correct sequencing, and using the Plasmid as a template for later vector construction, wherein the Plasmid is named as OfTPS380.1-CDS-Plasmid.
Example 2 vector construction
Construction of a yeast vector: OfTPS380.1-CDS-Plasmid is used as a template, an upstream primer and a downstream primer of OfTPS380.1-BsaI-F/R (table 1) carrying BsaI enzyme cutting sites are designed to clone OfTPS380.1 gene fragments, and the obtained PCR product and an expression vector Plasmid (a Plasmid map is shown in figure 2 and is presented to the master of Liutiangang teacher) required to be constructed are subjected to Goldengate ligation. The PCR reaction system and the procedure are shown in tables 2 and 3, the product is directly transformed into escherichia coli, monoclonal positive detection is carried out, the positive single colony is shaken to extract plasmids, then enzyme digestion verification is carried out, the correct plasmids are sequenced through enzyme digestion verification, and the correctly sequenced plasmids are named as follows: OfTPS380.1-Pkz762-Plasmid for the subsequent transformation of s.cerevisiae.
Example 3 Yeast transformation and plate screening
YPD solid medium preparation: 2% of peptone, 1% of yeast extract and 10g/L of agar, adding distilled water, heating and dissolving, and fixing the volume to 0.9 time of the final volume; sterilizing at 115 deg.C for 30min, and adding 0.1 volume of 20% glucose solution to obtain YPD culture medium. SC-URA screening culture medium: 0.67% yeast nitrogen source basic culture medium YNB, weighing the uracil-deficient amino acid mixture according to the following table 4, adjusting the pH to 6.5 by using 2M NaOH solution, adding distilled water, heating for dissolving, fixing the volume to 0.9 time, and sterilizing at 115 ℃ for 30 min; after sterilization, a 0.1-fold volume of 20% glucose solution was added.
TABLE 4 amino acid mixture formula
10 × TE configuration: 100mM Tris and 10mM EDTA, adjusted to pH 7.5 with hydrochloric acid, and finally autoclaved at 115 ℃ for 30min, and stored at room temperature.
10 × LiAc (lithium acetate) configuration: 1M LiAc, adjusted to pH 7.5 with glacial acetic acid (acetic acid), then sterilized by filtration through a 0.22. mu.M sterile filter and kept in a freezer at 4 ℃ until use.
50% PEG 4000: weighing 40g of PEG4000, adding a proper amount of water, heating until the PEG4000 is completely dissolved, and adding water to a constant volume of 80 mL; sterilizing at 115 deg.C for 30min, packaging with 600 μ l/tube, and storing in refrigerator at 4 deg.C.
DMSO, DMSO: filtering with 0.22 μ M sterile filter membrane, sterilizing, and storing in refrigerator at 4 deg.C.
Single-stranded milt dna (ssdna): purchased from kulai technology limited, beijing.
The saccharomyces cerevisiae strain YZL141 transformation and plate screening method comprises the following specific steps:
1. selecting about 5 single clones of saccharomyces cerevisiae strain YZL141 to contain 5 single clonesCulturing in PA bottle of ml YPD liquid culture medium at 30 deg.C overnight with shaking table; 50 μ l of the bacterial solution was diluted 20 times with sterile water, and OD was measured600Value (20 times dilution, not able to make OD value greater than 1, otherwise would be inaccurate); by OD600The volume x is 10 (e.g. 5ml if OD 2, typically between 500 and 1000. mu.l) is transferred to 50ml fresh YPD medium and incubated at 30 ℃ and 220rpm for 3.5 h; when the OD600 is 0.5-0.8, centrifuging at 3000rpm for 5min to collect thallus; adding 20ml of 1 × TE solution to resuspend the cells, centrifuging at 3000rpm for 5min, and discarding the supernatant; 2ml of 1 XLiAc/0.5 XTE (1700. mu. l H) was added to the precipitated cells2O + 200. mu.l of 10 XLiAc + 100. mu.l of 10 XTE), left at room temperature for 10min (preferably 25 ℃);
2. boiling the single-chain milt DNA in water for 10min, quickly putting back to ice for cooling to ensure that the milt DNA is in a single-chain state, taking 100 mu l of yeast strain YZL141 suspension, adding 10 mu l of denatured single-chain milt DNA, adding 300ng of OfTPS380.1-Pkz762-Plasmid which is diluted for later use in advance, slightly flicking and uniformly mixing, and finally adding 700 mu l of 1 XLiAc/40% PEG4000/1 XTE solution (560 mu l of PEG4000+70 mu l of 10 XLiAc +70 mu l of 10 XTE) for uniformly mixing; culturing at 30 deg.C for 30 min; adding 88 mul DMSO, mixing, and performing heat shock at 42 deg.C for 9 min; centrifuging at 10000rpm for 30s, and discarding the supernatant; adding 1ml YPD culture medium to resuspend the thallus, centrifuging at 8000rpm for 30s, and discarding the supernatant; adding 1ml YPD culture medium to suspend thallus, and shake-culturing at 30 deg.C for 30 min; centrifuging at 10000rpm for 30s, and discarding the supernatant; 1ml of 1 × TE solution is added to resuspend the thalli, centrifugation is carried out for 30s at 10000rpm, and the supernatant is discarded;
3. and coating the transformed YZL141 on an SC-URA screening plate, drying the liquid, and culturing in an incubator at 30 ℃ until obvious single colonies are visible for about 2-3 days.
EXAMPLE 4 Positive detection of Yeast transformed colonies
Single colonies of the transformed yeast were picked into 10. mu.l of sterile water, 2. mu.l was taken out, 10. mu.l of 20mM NaOH solution was added, treated with PCR instrument at 99 ℃ for 20min, placed on ice for template application, and the template was vortexed for 2s to prevent sedimentation before use.
PCR System (10. mu.l): 2 XTaq Master Mix 5. mu.l, OfTPS380.1-Yeast transformation Positive detection-F/R0.5. mu.l each, template 1. mu.l, water 3. mu.l, PCR conditions: pre-denaturation at 94 ℃ for 2min, denaturation at 94 ℃ for 20S, annealing at 55 ℃ for 20S, extension at 72 ℃ for 1min/kb, setting 30 cycles, and final extension at 72 ℃ for 5 min. And (3) observing a band after electrophoresis of the PCR product, and performing yeast fermentation and geraniol GC-MS detection if an obvious positive band (a target band 1752bp) exists.
Example 5 Yeast fermentation and geraniol GC-MS detection
1. Shake flask fermentation culture
YPD liquid medium preparation: the final concentration of peptone (Angel brand) is 20g/L, the final concentration of yeast powder (Angel brand) is 10g/L, the final concentration of glucose is 10g/L, the final concentration of galactose is 10g/L (glucose and galactose are independently prepared and added into a culture medium after sterilization), and isopropyl myristate (IPM) is filtered by a sterile filter membrane before being used.
Inoculation: from the transformed plate, 5 single colonies that were positive in detection were picked up in 5ml of YPD liquid medium and cultured overnight at 30 ℃ at 220 rpm. The next day, overnight culture broth is extracted and added into 50ml liquid culture medium according to a certain proportion to make initial OD600The value was about 0.1, and 10% isopropyl myristate was added for covering, and the cells were incubated at 30 ℃ for 72 hours on a shaker at 220 rpm.
GC-MS quantitative detection of geraniol
After the fermentation is finished, centrifuging at 4000rpm of 4 ℃ for 10 minutes, taking 200 mu L of isopropyl myristate, putting the isopropyl myristate into a sample bottle, detecting the product by GC-MS, and simultaneously adding 1 mu L of methyl nonanoate with the concentration of 4.75mg/L as an internal standard for quantitative calculation, wherein the unconverted saccharomyces cerevisiae strain YZL141 is used as a negative control, and the quantitative calculation formula is as follows: C1/V1 ═ C2/V2(C1 is the peak area of methyl nonanoate, V1 is the concentration of methyl nonanoate, C2 is the peak area of geraniol, and V2 is the concentration of geraniol).
And (3) GC-MS detection:
GC setting: the injection port temperature is 250 ℃, the injection mode is no shunt, the initial temperature of the temperature rise program is 40 ℃, the temperature is kept for 3.5min, the temperature rise rate of 10 ℃/min is increased to 100 ℃, the temperature is kept for 3min, and then the temperature is increased to 280 ℃ at the rate of 5 ℃/min and is kept for 5 min. The total program time was 46.5 min.
Setting the MS: the interface temperature is 280 ℃, the loading gas is helium, the flow rate is 1.0ml/min, the ion source temperature is 220 ℃, the EI ionization mode is adopted, the electron energy is 70ev, and the scanning range is 50-500 amu.
The sample injection mode is a non-flow splitting mode, the temperature of the sample injection port is maintained at 230 ℃, and the temperature of the transmission line is 240 ℃. Electron energy 70eV, scan range 40-450 amu, ion source temperature 150 ℃, high purity helium (99.999%) as carrier gas, flow rate 0.8 mL/min. The initial column temperature was 40 deg.C, held for 3min, then raised to 80 deg.C at a rate of 1 deg.C/min, held for 3min, then raised to 220 deg.C at a rate of 10 deg.C/min, and held for 15 min.
GC-MS detection shows that the target product peak is geraniol, the target peak-off time is 7.58min, and the final concentration of the geraniol is 21.67 mg/L.
Sequence listing
<110> university of agriculture in Huazhong
Bingning high-new osmanthus fragrans industry and technology research institute
<120> osmanthus fragrans gene OfTPS380.1 and application thereof
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1752
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 1
atggagtgta caatgaggag tatttcctta ttttcacagt catctaatgg tatttcagga 60
actgcaagaa gtccatttca atggccgata aaccatcgat tttcatcggg tcaacgtgat 120
ttcgtttgca agtcgttgcc ggtgtcgttg cctagtgtaa cgccattgat tcctgctgaa 180
aatgatgctc tgtataatta tatacgccaa cctgtgatag ttactcccga agtcgatgag 240
ggcgcgaagc acagagaatt gctcgaaaaa actcgacgag aactgcaaag aagtacaaaa 300
ccagttgaga cgctaaaact tatagacaaa ctccaacgat taggaattgc gtattatttt 360
gaggatgata tcaatgcaat actggatcga ttctctgatg gcttgcctaa tgaagatctc 420
ttcacaacag ccttatgctt ccgcttgctc cgtgataacg gctacaaaac tgattctgat 480
gtctttctta aattcatgga aaagaacaag aaattcaaag aacatttggc tcaagacacc 540
ataggcttat tgagcttata cgaagcatcg tacatgggag caaacggcga agaaatattg 600
tcagaggcca aggaatttac tgaaattcac cttagacagt cgatgcctcg gttggctctg 660
caacttcgtc gacaagttgg ttctgcctta gagctcccga ggcacctccg gatggctagg 720
ttagaagcta ggcgttacat tgaagaatat gctacagaaa gtgaacacga tccagccctt 780
ttggaactgg caagattaga ttataacaaa gtccagttac aacaccaaat ggaattgtct 840
gaaatatcaa gatggtggaa acaattgggg cttgttgaga agctgagctt tgcccgggac 900
agacccttgg aatgcttttt atggactgtg ggacttcttc cagaacccaa atactctagc 960
tgcagaattg aactggctaa gaccatagcc attctattgg taattgacga tatttttgac 1020
acatatggca aaatggagga acttgttctt ttcactgaag caattcaaag atgggatctt 1080
gatgaattgg aaactcttcc accatacatg agaatatgtt acatggcatt atacaacact 1140
accaatgaaa tctgctacaa aatcctcaag gagtatggat tttgtgtcct tccctacctt 1200
aaatccacgt ggatagatat gattgagggg tttatggtag aggcgaattg gtttaatggt 1260
ggacatggac caaatttgga ggaatatata gagaatggtg tttcaacagc aggagcatat 1320
atggctttgg tgcacctatt ctttcttata ggggaaggag ttacaaatga gaatattact 1380
aaattgttga gaaaaccata tcctaaactc ttttccacag ctgggagaat tcttcgtctc 1440
tgggatgatc taggaactgc aaaggaggag gaagaacgag gcgatcttgc atcgtgcatg 1500
cagatattaa tgagagagaa gaacatagat tgtgaaaacg aaggcagaaa atatattctg 1560
aaagctataa atagcctatg gaaagatctg aacgatgaac tgatttcacc aaacgcaatg 1620
ccattagcca ttaccaaagt tgcattgaac atggctagag catccgaagt tgtctacaag 1680
catgaagagg attcatactt ctccagcgtc gataattatg tgcaggcttt gttcttcact 1740
cctattaatt ga 1752
Claims (5)
1. Osmanthus fragrans geraniol synthetase geneOfTPS380.1Characterized in that the geneOfTPS380.1The nucleotide sequence of (A) is shown in SEQ ID NO. 1.
2. An expression vector comprising the geraniol synthase gene according to claim 1OfTPS380.1。
3. A recombinant Saccharomyces cerevisiae is characterized in that it contains geneOfTPS380.1The yeast expression vector transforms saccharomyces cerevisiae, and expression is obtained through resistance plate and sequencing screeningOfTPS380.1Recombinant saccharomyces cerevisiae of genes.
4. The gene according to claim 1OfTPS380.1Or the expression vector of claim 2 or the recombinant Saccharomyces cerevisiae of claim 3 for use in the synthesis of geraniol.
5. A method for synthesizing geraniol by yeast is characterized by comprising the following steps: the recombinant saccharomyces cerevisiae of claim 3 is inoculated in a galactose-containing YPD medium, isopropyl myristate is added for covering, and geraniol is extracted after fermentation is finished.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210420147.0A CN114774438B (en) | 2022-04-20 | 2022-04-20 | Osmanthus gene OfTPS380.1 and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210420147.0A CN114774438B (en) | 2022-04-20 | 2022-04-20 | Osmanthus gene OfTPS380.1 and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114774438A true CN114774438A (en) | 2022-07-22 |
| CN114774438B CN114774438B (en) | 2023-04-28 |
Family
ID=82432065
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210420147.0A Active CN114774438B (en) | 2022-04-20 | 2022-04-20 | Osmanthus gene OfTPS380.1 and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114774438B (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2002199884A (en) * | 2000-12-28 | 2002-07-16 | Toyota Motor Corp | Method for producing geranyl geraniol and its analogues |
| CN102344915A (en) * | 2011-09-16 | 2012-02-08 | 中国科学院研究生院 | Protein with cinnamyl alcohol dehydrogenase activity and coding gene as well as application thereof |
| CN111286482A (en) * | 2020-05-13 | 2020-06-16 | 中国科学院烟台海岸带研究所 | Escherichia coli engineering bacterium capable of rapidly producing geraniol and construction method and application thereof |
| CN113025594A (en) * | 2021-03-04 | 2021-06-25 | 安徽农业大学 | Polypeptide, nucleic acid and application of polypeptide and nucleic acid in synthesis of geraniol |
| CN113774079A (en) * | 2021-08-13 | 2021-12-10 | 中国科学院天津工业生物技术研究所 | Recombinant saccharomyces cerevisiae and construction method and application thereof |
-
2022
- 2022-04-20 CN CN202210420147.0A patent/CN114774438B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2002199884A (en) * | 2000-12-28 | 2002-07-16 | Toyota Motor Corp | Method for producing geranyl geraniol and its analogues |
| CN102344915A (en) * | 2011-09-16 | 2012-02-08 | 中国科学院研究生院 | Protein with cinnamyl alcohol dehydrogenase activity and coding gene as well as application thereof |
| CN111286482A (en) * | 2020-05-13 | 2020-06-16 | 中国科学院烟台海岸带研究所 | Escherichia coli engineering bacterium capable of rapidly producing geraniol and construction method and application thereof |
| CN113025594A (en) * | 2021-03-04 | 2021-06-25 | 安徽农业大学 | Polypeptide, nucleic acid and application of polypeptide and nucleic acid in synthesis of geraniol |
| CN113774079A (en) * | 2021-08-13 | 2021-12-10 | 中国科学院天津工业生物技术研究所 | Recombinant saccharomyces cerevisiae and construction method and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| XIANGLING ZENG ET AL.: ""Emission and Accumulation of Monoterpene and the Key Terpene Synthase (TPS) Associated with Monoterpene Biosynthesis in Osmanthus fragrans Lour"" * |
| 滕林佐等: ""四季桂香叶醇合成酶基因 GES 的克隆及表达模式分析"" * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114774438B (en) | 2023-04-28 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112813013B (en) | Recombinant escherichia coli for producing hydroxytyrosol and application thereof | |
| WO2023143136A1 (en) | YEAST ENGINEERING BACTERIUM FOR FERMENTATIVE PRODUCTION OF α-SANTALENE AND USE THEREOF | |
| CN102168099A (en) | 3-ketosteroid -delta 1-dehydrogenase, engineering bacterium and application thereof | |
| CN109337932B (en) | Method for increasing yield of monascus pigment | |
| CN109628420B (en) | Glucosyltransferase and application thereof in producing vanillin-alpha-D-glucoside | |
| CN111334522B (en) | Recombinant saccharomyces cerevisiae for producing ambergris alcohol and construction method | |
| CN106244615B (en) | Engineering bacterium, construction method thereof and application of engineering bacterium in preparation of geraniol | |
| CN105647958B (en) | Saccharomyces cerevisiae engineering bacterium for producing 2-phenylethyl alcohol and preparation method and application thereof | |
| CN114774438A (en) | Osmanthus gene OfTPS380.1 and application thereof | |
| CN116064266B (en) | Recombinant saccharomyces cerevisiae with enhanced salt stress resistance, and construction method and application thereof | |
| CN111518711B (en) | Enterobacter strain and application thereof in coproduction of microbial exopolysaccharide and 2,3-butanediol | |
| CN114438109B (en) | Osmanthus gene OfTPS13.2 and application thereof | |
| CN109371053B (en) | Construction method of monascus pigment producing strain | |
| CN110628804B (en) | Method for constructing saccharomyces cerevisiae strain with high tolerance to isobutanol | |
| CN112940956A (en) | Pichia pastoris for enhancing lactoferrin expression by adopting double promoters and application thereof | |
| CN108203697B (en) | Yeast engineering bacterium of fish natural killer cell enhancement factor and application thereof | |
| CN106967663B (en) | Recombinant strain for preventing and treating crop diseases | |
| CN112048447B (en) | Recombinant rhodotorula mucilaginosa and application thereof in production of water-soluble neutral polysaccharide | |
| CN109897863A (en) | A kind of method for transformation of Lactobacillus brevis | |
| CN112175850B (en) | Industrial saccharomyces cerevisiae capable of highly producing nucleic acid and application thereof | |
| CN116769619B (en) | Candida B2, capsicum product and method | |
| CN112920959B (en) | Method for increasing yield of L-menthol in saccharomycetes | |
| CN106967627A (en) | A kind of fermenting and producing ALA recombinant bacterial strain and preparation method thereof | |
| CN109468254B (en) | Method for improving production efficiency of L-alanine and application thereof | |
| CN107142219B (en) | Tricholoma trichomonaum for producing triterpenoids 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 |