CN113234607A - Method for synthesizing geraniol by utilizing aspergillus oryzae - Google Patents
Method for synthesizing geraniol by utilizing aspergillus oryzae Download PDFInfo
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- CN113234607A CN113234607A CN202110576712.8A CN202110576712A CN113234607A CN 113234607 A CN113234607 A CN 113234607A CN 202110576712 A CN202110576712 A CN 202110576712A CN 113234607 A CN113234607 A CN 113234607A
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
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- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/10—Transferases (2.)
- C12N9/1085—Transferases (2.) transferring alkyl or aryl groups other than methyl groups (2.5)
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- C12P5/00—Preparation of hydrocarbons or halogenated hydrocarbons
- C12P5/02—Preparation of hydrocarbons or halogenated hydrocarbons acyclic
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- C12Y101/00—Oxidoreductases acting on the CH-OH group of donors (1.1)
- C12Y101/01—Oxidoreductases acting on the CH-OH group of donors (1.1) with NAD+ or NADP+ as acceptor (1.1.1)
- C12Y101/01034—Hydroxymethylglutaryl-CoA reductase (NADPH) (1.1.1.34)
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- C12Y—ENZYMES
- C12Y205/00—Transferases transferring alkyl or aryl groups, other than methyl groups (2.5)
- C12Y205/01—Transferases transferring alkyl or aryl groups, other than methyl groups (2.5) transferring alkyl or aryl groups, other than methyl groups (2.5.1)
- C12Y205/0101—(2E,6E)-Farnesyl diphosphate synthase (2.5.1.10), i.e. geranyltranstransferase
Abstract
The invention belongs to the field of microbial fermentation, and particularly relates to a method for synthesizing geraniol by utilizing aspergillus oryzae. The specific technical scheme is as follows: an Aspergillus oryzae capable of synthesizing geraniol, wherein the Aspergillus oryzae contains and can simultaneously express a GES gene, a tHMG1 gene and a mFPS gene. According to the invention, a gene engineering method is adopted, so that a specific Aspergillus oryzae can simultaneously express three genes of tHMG1, mFPS and GES for the first time, and thus the Aspergillus oryzae is endowed with the capacity of high-yield geraniol production; and the produced geraniol is easy to separate, has high purity and is suitable for industrial popularization and use.
Description
Technical Field
The invention belongs to the field of microbial fermentation, and particularly relates to a method for synthesizing geraniol by utilizing aspergillus oryzae.
Background
Geraniol (Geraniol), also known as Geraniol, 2, 6-dimethyl-2, 6-octadien-8-ol, is an acyclic monoterpene enol. The molecular formula is as follows: c10H18OMolecular weight: 154.25. geraniol is colorless or yellow oil at room temperatureLiquid, with a mild rose odour. And is therefore an important aroma component. Geraniol is not only used in the cosmetic industry, the food industry, and the like, but also has a wide range of pharmacological effects, such as antibacterial, anti-inflammatory, antioxidant, antiulcer, and nerve protection functions, and is clinically used for treating chronic bronchitis. Therefore, the geraniol has higher economic value.
Although geraniol has important applications in a number of industrial areas, its production currently relies on plant extraction. The production method is influenced by various factors such as plant varieties, planting areas, weather and climate, the quantity and quality of products fluctuate greatly, and the downstream application is not facilitated. Thus, microbial fermentative production becomes a possible alternative route. The biosynthesis genes of the aromatic compounds are transferred into microorganisms through synthetic biology, and then the microorganisms are utilized for fermentation production. The method can overcome the disadvantages of the plant extraction method, and has the advantages of low production cost and high product purity.
Disclosure of Invention
The invention aims to provide a method for synthesizing geraniol by using aspergillus oryzae.
In order to achieve the purpose of the invention, the technical scheme adopted by the invention is as follows: an Aspergillus oryzae capable of synthesizing geraniol, wherein the Aspergillus oryzae contains and can express GES gene.
Accordingly, an Aspergillus oryzae capable of synthesizing geraniol, which contains and can simultaneously express a GES gene, a tHMG1 gene and a mFPS gene.
Preferably, the aspergillus oryzae is from China general microbiological culture Collection center, and the preservation number is as follows: CGMCC No. 20748.
Accordingly, a method for synthesizing geraniol using Aspergillus oryzae expressing a GES gene.
Preferably, the Aspergillus oryzae expresses the GES gene, the tHMG1 gene and the mFPS gene simultaneously.
Preferably, the aspergillus oryzae is from China general microbiological culture Collection center, and the preservation number is as follows: CGMCC No. 20748.
Preferably, the method comprises the following steps:
(1) constructing a recombinant plasmid containing the GES gene;
(2) transferring the recombinant plasmid into aspergillus oryzae to obtain recombinant aspergillus oryzae;
(3) culturing recombinant Aspergillus oryzae to obtain geraniol.
Preferably, the recombinant plasmid in step (1) contains the GES gene, the tHMG1 gene and the mFPS gene at the same time.
Preferably, in step (3), the recombinant Aspergillus oryzae is cultured at 28 ℃.
Preferably, geraniol is obtained and extracted with ethyl acetate.
The invention has the following beneficial effects: the invention firstly enables a specific Aspergillus oryzae to simultaneously express three genes of tHMG1, mFPS and GES through a genetic engineering method, thereby endowing the Aspergillus oryzae with the capability of high-yield geraniol. And the produced geraniol is easy to separate, has high purity and is suitable for industrial popularization and use. The inventors found that, when the production is carried out using Aspergillus oryzae which the inventors previously studied and deposited, the gene expression level and geraniol yield are superior to those of Aspergillus oryzae which is generally commercially available, and the geraniol yield can be increased by 35% or more. In addition, the inventor tries various promoters, and finds that when other promoters are used besides the promoter PamyB, the expression amount is not ideal, and about 1/5-1/3 is obtained when only the promoter PamyB is used, so that the final yield of the geraniol is not ideal, and even the yield is too low to extract.
Detailed Description
The invention provides a method for synthesizing geraniol by using aspergillus oryzae, which comprises the following steps: three genes were expressed simultaneously in aspergillus oryzae: tHMG1, mFPS and GES, to obtain recombinant Aspergillus oryzae strains. Wherein the gene sequence of tHMG1 is shown as SEQ ID NO: 1, and the gene sequence of mFPS is shown as SEQ ID NO: 2, the gene sequence of GES is shown as SEQ ID NO: 3, respectively.
Fermenting and filtering the recombinant aspergillus oryzae strain to obtain a recombinant aspergillus oryzae strain mycelium, extracting a product, and separating and purifying to obtain the high-purity geraniol.
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art.
The first embodiment is as follows: construction of Aspergillus oryzae Strain capable of expressing GES Gene
1. A pTYGSa1 recombinant plasmid was constructed. The DNA fragment of the GES gene is obtained by chemical synthesis from Shanghai bioengineering Co., Ltd, and the correctness of the gene sequence is verified by sequencing. The DNA fragment is taken as a template, sequence amplification is carried out by utilizing a specific primer through a PCR technology, and Hind III and EcoRI enzyme cutting sites are respectively introduced into the 5 'end and the 3' end of an amplification product fragment. The amplified fragment and pTYGS plasmid were digested with HindIII and EcoRI, respectively, to obtain two digestion products. The two enzyme products were measured for concentration with a DNA concentration measuring instrument, mixed at a DNA concentration of 1:1, and enzymatically ligated with T4 DNA ligase at 16 ℃. The E.coli DH 5. alpha. strain was transformed with the enzyme-linked product, and the transformed strain was selected on a kanamycin-resistant plate (LB medium plate containing 50mg/L kanamycin) (37 ℃ C., 24 hours) to grow an E.coli strain containing the correct recombinant plasmid (designated as pTYGSa 1). The screened Escherichia coli is picked into a fresh LB medium plate to be continuously cultured for 24h, the Escherichia coli thallus is collected by centrifugation, and the recombinant plasmid pTYGSa1 is extracted by a plasmid extraction kit.
2. Preparation of recombinant aspergillus oryzae strain a 1: a pre-cultured Aspergillus oryzae strain (classified and named as Aspergillus oryzae, which is preserved in China general microbiological culture Collection center of China Committee for culture Collection of microorganisms, the preservation date is 11 months and 5 days in 2020, the preservation number is CGMCC No.20748, the preservation address is No. 3 Hospital No. 1 of North Chen West Lu of the sunny district in Beijing, and the first publication is carried out in patent CN 112430547A) is inoculated on a fresh PDA slant and cultured for 7 days at 28 ℃. Then, the mycelium was washed with sterile water to obtain Aspergillus oryzae spore liquid. Diluting spore liquid to 1 × 10 with sterile water5one/mL. 2mL of the spore solution was inoculated into 50mL of fresh PDB medium at 120rp at 28 ℃And culturing for 3 days under m. The mycelia were then obtained by filtration through a nylon filter cloth, and 0.3mL of a cell-wall breaking enzyme solution (10U/mL) was added to 5mL of a phosphate buffer (50mM, pH 6.0), and 0.25g of wet Aspergillus oryzae mycelia was further added to remove cell walls from the Aspergillus oryzae mycelia, thereby obtaining Aspergillus oryzae cell protoplasts. To the centrifuge tube containing the prepared protoplast, 20. mu.L of the recombinant plasmid pTYGSa1, 1mL of a filter-sterilized PEG 4000 (60%, w/v) solution was added, and the mixture was allowed to stand at 25 ℃ for 20 min. And (3) coating the mixed solution in the centrifugal tube on a PDA culture medium plate, placing the plate on a PDA culture medium for culture at the temperature of 30 ℃, picking out a single colony after the recombinant aspergillus oryzae grows on the PDA culture medium for 5 days, and amplifying the GES gene by using a PCR technology to verify that the correct recombinant aspergillus oryzae strain A1 is obtained.
3. Preparing geraniol: inoculating the recombinant Aspergillus oryzae strain A1 into a fresh PDB culture medium, culturing for 7 days at 28 ℃ and 120rpm, filtering and collecting 1.0g of mycelium, quickly adding liquid nitrogen, cooling, grinding into fine powder, adding 10mL of ethyl acetate at room temperature, violently mixing, placing the mixture in a shaking table, shaking for 2 hours at 40 ℃, extracting geraniol into an ethyl acetate organic phase, and finally separating by using a separating funnel to obtain an organic phase containing the geraniol. The organic solvent was removed by using a vacuum distillation apparatus to obtain a solid sample containing geraniol, which was then dissolved in methanol at a concentration of 1 mg/mL. The comparison of geraniol in the methanol solution with the standard geraniol species using LC-MS/MS techniques confirmed that geraniol was indeed present in the extracted sample. Then, the content of geraniol in the extract was measured by HPLC method, and the geraniol yield was calculated to be 1.4mg/L and the purity was calculated to be 88.4% by measuring a standard curve using a commercially available geraniol standard.
Example two: construction of Aspergillus oryzae Strain that can simultaneously express GES Gene, tHMG1 Gene and mFPS Gene
1. According to the method of step 1 of the example, DNA fragments of tHMG1 gene and mFPS gene were synthesized by Shanghai bioengineering Co., Ltd, and then amplification products were obtained by PCR, assembled into plasmid pTYGS together with the GES gene fragment, and promoter PamyB was added to 5' ends of 3 DNA fragments, respectively, to obtain recombinant plasmid pTYGSa 2. Further, according to the method of step 2 in the example, protoplasts of Aspergillus oryzae cells were transformed by PEG-mediated transformation to obtain recombinant Aspergillus oryzae strain A2 (which can express GES gene, tHMG1 gene and mFPS gene simultaneously).
2. The fermentation of A.oryzae strain A2 was carried out as in step 3 of example, and the mycelia of A.oryzae strain A2 were obtained by filtration, followed by extraction of the product as shown in step 3 of example 1 and determination of the product concentration by HPLC. And finally, calculating to obtain the geraniol with the yield of 53.4mg/L, and obtaining a high-purity geraniol product, wherein the purity of the geraniol is 95% after the organic solution is evaporated and removed.
The above-described embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various changes, modifications, alterations, and substitutions which may be made by those skilled in the art without departing from the spirit of the present invention shall fall within the protection scope defined by the claims of the present invention.
Sequence listing
<110> university of agriculture in Huazhong
<120> method for synthesizing geraniol using Aspergillus oryzae
<160> 3
<170> SIPOSequenceListing 1.0
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<211> 2760
<212> DNA
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atgatcggca ggctgttcag ggcccacggc gagttctgcg ccagccaccc ctgggaggtg 60
atcgtggccc tgctgaccat caccgcctgc atgctgaccg tggacaagaa caacaccctg 120
gacgccagca gcggcctggg caccgccacc gccagcgccg ccgccgccgg cggcagcggc 180
agcggcgccg gcagcggcgc cagcggcacc atccccccca gcagcatggg cggcagcgcc 240
accagcagca ggcacaggcc ctgccacggc tggagccaga gctgcgacgg cctggaggcc 300
gagtacaacg ccgccgacgt gatcctgatg accatcgtga ggtgcaccgc cgtgctgtac 360
tgctactacc agttctgcag cctgcacagg ctgggcagca agtacgtgct gggcatcgcc 420
ggcctgttca ccgtgttcag cagcttcatc ttcaccaccg ccatcatcaa gttcctgggc 480
agcgacatca gcgagctgaa ggacgccctg ttcttcctgc tgctggtgat cgacctgagc 540
aacagcggca ggctggccca gctggccctg agcggcagca accaggccga ggtgacccag 600
aacatcgcca ggggcctgga gctgctgggc cccgccatca gcctggacac catcgtggag 660
gtgctgctgg tgggcgtggg caccctgagc ggcgtgcaga ggctggaggt gctgtgcatg 720
ttcgccgtgc tgagcgtgct ggtgaactac gtggtgttca tgaccttcta ccccgcctgc 780
ctgagcctga tcttcgacct gagcaggagc ggcgtggaca tgagcgtggt gagggagaag 840
gccaagggca gcctgctgct gaagagcctg accgaggagg agcagaaggc caaccccgtg 900
ctgcagaggg tgaagctgat catgaccacc ggcctgatgg ccgtgcacat ctacagcagg 960
gtggccttca gcggcagcga ctacgacgcc gtggacaaga ccctgacccc caccctgagc 1020
ctgaacgtga gcaacaacag gaccgagagc ggcgagatcg ccgacatcat catcaagtgg 1080
ctgaccatga gcgccgacca catcgtgatc agcatcgtgc tgatcgccct ggtggtgaag 1140
ttcatctgct tcgacaacag ggaccccctg cccgaccagc tgaggcagag cggccccgtg 1200
gccatcgccg ccaaggccag ccagaccacc cccatcgacg aggagcacgt ggagcaggag 1260
aaggacaccg agaacagcgc cgccgtgagg accctgctgt tcaccatcga ggaccagagc 1320
agcgccaacg ccagcaccca gaccgacctg ctgcccctga ggcacaggct ggtgggcccc 1380
atcaagcccc ccaggcccgt gcaggagtgc ctggacatcc tgaacagcac cgaggagggc 1440
agcggccccg ccgccctgag cgacgaggag atcgtgagca tcgtgcacgc cggcggcacc 1500
cactgccccc tgcacaagat cgagagcgtg ctggacgacc ccgagagggg cgtgaggatc 1560
aggaggcaga tcatcggcag cagggccaag atgcccgtgg gcaggctgga cgtgctgccc 1620
tacgagcact tcgactacag gaaggtgctg aacgcctgct gcgagaacgt gctgggctac 1680
gtgcccatcc ccgtgggcta cgccggcccc ctgctgctgg acggcgagac ctactacgtg 1740
cccatggcca ccaccgaggg cgccctggtg gccagcacca acaggggctg caaggccctg 1800
agcgtgaggg gcgtgaggag cgtggtggag gacgtgggca tgaccagggc cccctgcgtg 1860
aggttcccca gcgtggccag ggccgccgag gccaagagct ggatcgagaa cgacgagaac 1920
tacagggtgg tgaagaccga gttcgacagc accagcaggt tcggcaggct gaaggactgc 1980
cacatcgcca tggacggccc ccagctgtac atcaggttcg tggccatcac cggcgacgcc 2040
atgggcatga acatggtgag caagggcgcc gagatggccc tgaggaggat ccagctgcag 2100
ttccccgaca tgcagatcat cagcctgagc ggcaacttct gctgcgacaa gaagcccgcc 2160
gccatcaact ggatcaaggg caggggcaag agggtggtga ccgagtgcac catcagcgcc 2220
gccaccctga ggagcgtgct gaagaccgac gccaagaccc tggtggagtg caacaagctg 2280
aagaacatgg gcggcagcgc catggccggc agcatcggcg gcaacaacgc ccacgccgcc 2340
aacatggtga ccgccgtgtt cctggccacc ggccaggacc ccgcccagaa cgtgaccagc 2400
agcaactgca gcaccgccat ggagtgctgg gccgagaaca gcgaggacct gtacatgacc 2460
tgcaccatgc ccagcctgga ggtgggcacc gtgggcggcg gcaccggcct gcccggccag 2520
agcgcctgcc tggagatgct gggcgtgagg ggcgcccacg ccaccaggcc cggcgacaac 2580
gccaagaagc tggcccagat cgtgtgcgcc accgtgatgg ccggcgagct gagcctgatg 2640
gccgccctgg tgaacagcga cctggtgaag agccacatga ggcacaacag gagcagcatc 2700
gccgtgaaca gcgccaacaa ccccctgaac gtgaccgtga gcagctgcag caccatcagc 2760
<210> 2
<211> 1062
<212> DNA
<213> mFPS
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atgagcgccg acggcgccaa gaggaccgcc gccgagaggg agagggagga gttcgtgggc 60
ttcttccccc agatcgtgag ggacctgacc gaggacggca tcggccaccc cgaggtgggc 120
gacgccgtgg ccaggctgaa ggaggtgctg cagtacaacg cccccggcgg caagtgcaac 180
aggggcctga ccgtggtggc cgcctacagg gagctgagcg gccccggcca gaaggacgcc 240
gagagcctga ggtgcgccct ggccgtgggc tggtgcatcg agctgttcca ggccttcttc 300
ctggtggccg acgacatcat ggaccagagc ctgaccagga ggggccagct gtgctggtac 360
aagaaggagg gcgtgggcct ggacgccatc aacgacagct tcctgctgga gagcagcgtg 420
tacagggtgc tgaagaagta ctgcaggcag aggccctact acgtgcacct gctggagctg 480
ttcctgcaga ccgcctacca gaccgagctg ggccagatgc tggacctgat caccgccccc 540
gtgagcaagg tggacctgag ccacttcagc gaggagaggt acaaggccat cgtgaagtac 600
aagaccgcct tctacagctt ctacctgccc gtggccgccg ccatgtacat ggtgggcatc 660
gacagcaagg aggagcacga gaacgccaag gccatcctgc tggagatggg cgagtacttc 720
cagatccagg acgactacct ggactgcttc ggcgaccccg ccctgaccgg caaggtgggc 780
accgacatcc aggacaacaa gtgcagctgg ctggtggtgc agtgcctgca gagggtgacc 840
cccgagcaga ggcagctgct ggaggacaac tacggcagga aggagcccga gaaggtggcc 900
aaggtgaagg agctgtacga ggccgtgggc atgagggccg ccttccagca gtacgaggag 960
agcagctaca ggaggctgca ggagctgatc gagaagcaca gcaacaggct gcccaaggag 1020
atcttcctgg gcctggccca gaagatctac aagaggcaga ag 1062
<210> 3
<211> 1785
<212> DNA
<213> GES
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atgataactt catcgtcgtc cgttcgatct ttatgctgtc cgaaaaccag tatcatttca 60
ggcaaacttc ttccttcatt gctcctcact aacgtaataa atgtcagtaa tggtacgagt 120
agtcgcgctt gcgtttccat gtcgtcttta ccggtttcca agtccactgc ctcctccatt 180
gcagctccgt tggtccggga caacggctcc gcccttaact tttttcctca agctccacag 240
gttgaaatag atgaaagcag tagaataatg gagttggtag aagccactag aagaacactc 300
agaaacgaat cttctgattc tacggagaag atgaggttga tcgactccct tcaacgcctg 360
ggattgaacc accatttcga acaagacata aaggagatgc tacaagattt cgcaaatgag 420
cacaagaata caaaccaaga tctctttact acttcactac gttttcgatt gctaagacat 480
aatggcttta atgtcacgcc tgatgtattc aataaattca cggaggaaaa tggcaagttt 540
aaggaatcat taggcgagga cacaattggg attttaagtt tgtatgaagc gtcgtattta 600
gggggaaagg gggaagaaat attatcggag gcaatgaaat ttagcgaatc taaattaaga 660
gaatcttctg gacacgtggc acstcatatt cgtcgacaga ttttccagtc gttggagctt 720
ccgaggcatt tgagaatggc taggttagaa tcccgaagat acatcgaaga agattacagc 780
aatgaaattg gggctgattc ttccttgctt gaattagcaa aactcgattt caattcagta 840
caagcacttc accaaatgga attaaccgaa atttctaggt ggtggaaaca attgggtttg 900
tctgataagc taccatttgc acgcgatcga ccgttggaat gcttcttgtg gaccgtcgga 960
ttacttcctg aaccaaaata ttctggttgt aggattgagt tggccaaaac cattgccgtt 1020
ttgctcgtta ttgatgacat attcgacact tatggctcgt atgatcaact tattctcttc 1080
acaaacgcaa tcagaagatg ggatctagat gcaatggatg aacttcccga gtacatgaaa 1140
atatgctaca tggcattgta caacacgact aacgaaatat gttacaaagt acttaaagaa 1200
aacggatgga gtgttcttcc ttacctcgaa cgaacgtgga tagacatggt tgaaggattc 1260
atgctagaag ccaagtggtt aaacagcggc gaacaaccga atttggaagc atacatagaa 1320
aacggcgtta ccacagccgg ctcttacatg gctttagtac acttgttttt cctcatagga 1380
gacggcgtta acgatgagaa cgtgaaatta ctattagacc cttaccctaa actgttctca 1440
tcggctgggc gcattcttcg actgtgggac gatttgggta ctgctaagga ggaacaagag 1500
agaggggatg tatcttcaag catacaattg tacatgaagg agaagaatgt gagatcagag 1560
agtgaaggaa gggagggaat agtagagata atatacaact tgtggaaaga catgaatgga 1620
gaattgattg gttcaaacgc attgccgcaa gcaatcatag agacctcatt caacatggca 1680
agaacatcac aagttgtgta tcaacacgaa gatgatactt atttctctag tgtcgataac 1740
tatgtacaat ctctcttctt cactcctgtt tctgtttctg tttaa 1785
Claims (10)
1. An Aspergillus oryzae capable of synthesizing geraniol, which is characterized in that: the Aspergillus oryzae contains and can express a GES gene.
2. An Aspergillus oryzae capable of synthesizing geraniol, which is characterized in that: the Aspergillus oryzae contains and can express GES gene, tHMG1 gene and mFPS gene at the same time.
3. An aspergillus oryzae according to claim 1 or 2, characterized in that: the aspergillus oryzae is from China general microbiological culture Collection center, and the preservation number is as follows: CGMCC No. 20748.
4. A method for synthesizing geraniol by using Aspergillus oryzae is characterized in that: the Aspergillus oryzae expresses a GES gene.
5. The method for synthesizing geraniol according to claim 4, wherein: the Aspergillus oryzae expresses GES gene, tHMG1 gene and mFPS gene simultaneously.
6. The method for synthesizing geraniol according to claim 4, wherein: the aspergillus oryzae is from China general microbiological culture Collection center, and the preservation number is as follows: CGMCC No. 20748.
7. The method for synthesizing geraniol according to any one of claims 3 to 6, wherein: the method comprises the following steps:
(1) constructing a recombinant plasmid containing the GES gene;
(2) transferring the recombinant plasmid into aspergillus oryzae to obtain recombinant aspergillus oryzae;
(3) culturing recombinant Aspergillus oryzae to obtain geraniol.
8. The method for synthesizing geraniol using aspergillus oryzae according to claim 7, wherein: the recombinant plasmid in the step (1) simultaneously contains a GES gene, a tHMG1 gene and an mFPS gene.
9. The method for synthesizing geraniol using aspergillus oryzae according to claim 7, wherein: in step (3), recombinant Aspergillus oryzae was cultured at 28 ℃.
10. The method for synthesizing geraniol using aspergillus oryzae according to claim 7, wherein: after geraniol was obtained, it was extracted with ethyl acetate.
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