CN117683756A - Tobacco sesquiterpene synthase NtTPS143 and application thereof - Google Patents

Abstract

The invention belongs to the technical fields of plant molecular biology and plant genetic engineering, and particularly relates to tobacco sesquiterpene synthase NtTPS143 and application thereof. The amino acid sequence of the tobacco sesquiterpene synthase NtTPS143 is shown in SEQ ID NO. 1. The invention discovers that the NtTPS143 can be expressed in a large quantity through escherichia coli, and can efficiently catalyze farnesyl pyrophosphate to generate 4 sesquiterpene compounds and 1 compound with unknown structure. The invention lays a foundation for the research of tobacco terpene secondary metabolism, provides important candidate genes for developing tobacco genetic engineering research and stress-tolerant breeding, and has important significance for producing sesquiterpene compounds by applying genetic engineering technology.

Description

Tobacco sesquiterpene synthase NtTPS143 and application thereof

Technical Field

The invention belongs to the technical fields of plant molecular biology and plant genetic engineering, and particularly relates to tobacco sesquiterpene synthase NtTPS143 and application thereof.

Background

Terpenoids are natural products that are widely available in nature and most diverse in chemical structure. The volatile terpene substances have unique odor and biological activity, and have important biological functions in the processes of plant growth, insect resistance, disease resistance and the like. According to the structure in which isoprene unit (C) ₅ ) In different amounts, terpenoids are divided into monoterpenes (C ₁₀ ) Sesquiterpenes (C) ₁₅ ) Diterpene (C) ₂₀ ) Triterpenes (C) ₃₀ ) Etc. In plant cells, geranyl pyrophosphate (GPP, C), a precursor compound synthesized by the mevalonate pathway in the cytoplasm (MVA pathway) and the methyl erythritol-4-phosphate pathway in the plastid (MEP pathway) ₁₀ ) Farnesyl pyrophosphate (FPS, C) ₁₅ ) And geranylgeranyl pyrophosphate (GGPP, C) ₂₀ ) The Terpene synthase (TPS) catalyzes to generate monoterpenes, sesquiterpenes and diterpenoids with different structures, and the monoterpenes, the sesquiterpenes and the diterpenoids are directly released into the environment or stored in specific organs, tissues or cells, or are oxidized, reduced and glycosidated by cytochrome P450, alcohol dehydrogenase, glycosidase and the like to form derivatives with various structures. MVA pathway and MEP pathway are relatively conserved in higher plants, whereas TPS has diverse catalytic activity and expression characteristics, determining structural diversity, distribution characteristics and environmental response patterns of plant terpenoids. Most TPS synthesizes only one or a few products, such as gin synthase of cotton, amorpha-4,11-diene synthase (ADS) of sweet wormwood all synthesize a single compound, whereas caryophyllene synthase of arabidopsis, rice, cotton, tomato can synthesize caryophyllene and lupulin simultaneously, but TPS which synthesizes multiple sesquiterpenes simultaneously with a single substrate FPP is less common.

alpha-Guaiene (formula C) ₁₅ H ₂₄ Molecular weight 204.35, CAS number 3691-12-1) is presentThe grape and the patchouli are important components of patchouli essential oil, are oxidized into (-) -rotundane by CYP71BE5 in the grape, and have close relation with the flavor of the wine. Compound (3S, 3aR,3bR,4S,7R,7 aR) -4-Isopropyl-3, 7-dimethylgctahydro-1H-cyclopena [1,3]]cyclopropa[1,2]Benzen-3-ol (formula C ₁₅ H ₂₆ O, molecular weight 222.37, CAS No. 23445-2-5), naphthalene,1,2,3,5,6,7, 8a-octahydro-1,8a-dimethyl-7- (1-methyl-phenyl) -, [1S- (1. Alpha., 7. Alpha., 8. Alpha.)]- (formula C) ₁₅ H ₂₄ Molecular weights 204.35, CAS number 10219-75-7) and 2- (4 a,8-Dimethyl-1,2,3, 4a,5,6, 7-octahydro-naphthalen-2-yl) -prop-2-en-1-ol (formula C) ₁₅ H ₂₄ O, molecular weight 220.35) has not been reported in plants. Because of the rare and low content of the compounds in the plant kingdom, the compounds are difficult to separate and purify, the chemical properties and the physiological functions of the compounds are rarely known at present, and the development and the utilization of the compounds are limited. The key TPS genes are separated and identified, and the compounds are efficiently produced by a synthetic biological method, so that the method has important significance for researching the physiological activity of the compounds and for industrial development and application in the fields of medicines, fragrances, cosmetics and the like.

Disclosure of Invention

The invention discovers that the sesquiterpene synthase from tobacco can be expressed in a large amount by using escherichia coli genetic engineering bacteria, and has good effect of catalyzing and synthesizing sesquiterpene compounds.

In a first aspect of the invention, there is provided a tobacco sesquiterpene synthase NtTPS143, the amino acid sequence of which is shown in SEQ ID NO. 1.

In a second aspect of the invention there is provided the use of the tobacco sesquiterpene synthase NtTPS143 in catalysing the synthesis of a sesquiterpene compound from a substrate.

Further, the substrate is farnesyl pyrophosphate and the sesquiterpene compound includes (3S, 3aR,3bR,4S,7R,7 aR) -4-Isopropyl-3, 7-dimethylopyra hydro-1H-cyclopara [1,3] cyclopara [1,2] benzen-3-ol, alpha-Guaiene, naphthalene,1,2,3,5,6,7, 8a-octahydro-1,8 a-dimethylol-7- (1-methyl-thenyl) -, [1S- (1 alpha, 7 alpha, 8a alpha) ] -and 2- (4 a, 8-dimethylol-1, 2,3, 4a,5,6, 7-octahydro-naphthalen-2-yl) -prop-2-en-1-ol.

In a third aspect of the invention, there is provided a gene encoding said tobacco sesquiterpene synthase NtTPS 143.

Further, the gene has a nucleotide sequence shown in any one of a1-a 2:

a1, a nucleotide sequence shown as SEQ ID NO. 2;

a2, a nucleotide sequence encoding said tobacco sesquiterpene synthase NtTPS143, but which differs from that shown in SEQ ID No.2 due to the degeneracy of the genetic code.

In a fourth aspect of the present invention, there is provided a recombinant expression vector comprising the gene.

Further, the recombinant expression vector is obtained by inserting the gene between BamHI and SacI sites of pET28a plasmid in forward direction.

In a fifth aspect of the present invention, there is provided a genetically engineered bacterium comprising the recombinant expression vector.

Furthermore, the host bacteria used by the genetically engineered bacteria are escherichia coli.

In a fifth aspect of the present invention, there is provided a method for preparing a sesquiterpene compound using the tobacco sesquiterpene synthase NtTPS143 as a catalyst and farnesyl pyrophosphate as a substrate in Dithiothreitol (DTT) and MgCl ₂ Catalytic synthesis of sesquiterpene compounds under the action of (2); the sesquiterpene compound comprises (3S, 3aR,3bR,4S,7R,7 aR) -4-isopopyl-3, 7-dimethyloctahydro-1H-cyclopena [1,3]]cyclopropa[1,2]benzen-3-ol、α-Guaiene、Naphthalene,1,2,3,5,6,7,8,8a-octahydro-1,8a-dimethyl-7-(1-methylethenyl)-,[1S-(1α,7α,8aα)]-and 2- (4 a,8-Dimethyl-1,2,3, 4a,5,6, 7-octahydro-naphthalen-2-yl) -prop-2-en-1-ol.

The invention has the following beneficial effects:

the invention expresses the tobacco-derived sesquiterpene synthase NtTPS143 in escherichia coli, and performs functional verification on the synthesis of the sesquiterpene compound, discovers that the NtTPS143 can be expressed in a large amount through escherichia coli, and can catalyze farnesyl pyrophosphate to generate the sesquiterpene compound (3S, 3aR,3bR,4S,7R,7 aR) -4-Isopropyl-3, 7-dimethylosta-1H-cyclopena [1,3] cyclopepaa [1,2] benzen-3-ol, alpha-Guaiene, naphthalene,1,2,3,5,6,7, 8a-octahydro-1,8a-Dimethyl-7- (1-methylpentanyl) - [1S- (1 alpha, 7 alpha, 8a alpha) ] -and 2- (4 a,8-Dimethyl-1,2,3,4, 7, 6-octahydro-1, 4 a-1-4 a-2-propanediol) and unknown structures of the sesqui-1, 2-propanediol-2-1, 4 a-2-propanediol.

The invention gives consideration to prokaryotic soluble expression of the NtTPS143 and specific and efficient catalytic activity on target products, obviously exceeds the level of the prior art, and has wide industrial practical prospect and potential for large-scale development.

Drawings

FIG. 1 is a total ion chromatogram of GC-MS analysis of the catalytic activity of the NtTPS143 recombinant protein in vitro (Total Ion Chromatography, TIC). The retention time (min) is on the abscissa and the peak height of the chromatographic response value is on the ordinate. 1-5 are products of the catalysis of FPP by NtTPS 143.

FIG. 2 is a mass spectrum comparison of the sesquiterpene compound 1 synthesized by the catalysis of NtTPS143 with (3S, 3aR,3bR,4S,7R,7 aR) -4-Isopropyl-3, 7-dimethyloctahydro-1H-cyclopena [1,3]cy clopropa[1,2]benzen-3-ol. The mass spectrum of the NtTPS143 catalytic product 1 is on the abscissa and the mass spectrum of (3S, 3aR,3bR,4S,7R,7 aR) -4-Isopropyl-3, 7-dimethylfectahydro-1H-cyclopena [1,3]cy clopropa[1,2]benzen-3-ol is on the abscissa.

FIG. 3 is a mass spectrum and a structural diagram of sesquiterpene compound (3S, 3aR,3bR,4S,7R,7 aR) -4-Isopropyl-3, 7-dimethyloctahydro-1H-cyclopena [1,3]cy clopropa[1,2]benzen-3-ol). The abscissa is the ion charge to mass ratio and the ordinate is the relative ion fragment intensity.

FIG. 4 is a mass spectrum comparison of sesquiterpene compound 2 synthesized catalytically by NtTPS143 with α -guanene. The mass spectrum of the NtTPS143 catalytic product 2 is on the abscissa and the mass spectrum of the alpha-guanene is on the abscissa.

FIG. 5 is a mass spectrum and a structure diagram of the sesquiterpene compound α -guanene. The abscissa is the ion charge to mass ratio and the ordinate is the relative ion fragment intensity.

FIG. 6 is a chart comparing the mass spectra of sesquiterpene compound 3 synthesized by the catalysis of NtTPS143 with Naphthalene,1,2,3,5,6,7, 8a-octahydro-1,8a-dimethyl-7- (1-methyl-phenyl) - [1S- (1. Alpha., 7. Alpha., 8. Alpha.) ] -mass. The mass spectrum of the NtTPS143 catalytic product 3 is on the abscissa, and the mass spectrum of Naphthalene,1,2,3,5,6,7, 8a-octahydro-1,8a-dimethyl-7- (1-methyl-phenyl) -, [1S- (1 alpha, 7 alpha, 8 alpha) ] -is on the abscissa.

FIG. 7 is a mass spectrum and a structural diagram of sesquiterpene compounds Naphthalene,1,2,3,5,6,7, 8a-octahydro-1,8a-dimethyl-7- (1-methyl-phenyl) -, [1S- (1. Alpha., 7. Alpha., 8. Alpha.) ]. The abscissa is the ion charge to mass ratio and the ordinate is the relative ion fragment intensity.

FIG. 8 is a mass spectrum comparison of the sesquiterpene compound 4 synthesized catalytically by NtTPS143 with 2- (4 a,8-Dimethyl-1,2,3, 4a,5,6, 7-octahydro-naphthalen-2-yl) -prop-2-en-1-ol. The mass spectrum of the NtTPS143 catalytic product 4 is on the abscissa and the mass spectrum of 2- (4 a,8-Dimethyl-1,2,3, 4a,5,6, 7-octahydro-naphthalen-2-yl) -prop-2-en-1-ol is on the abscissa.

FIG. 9 is a mass spectrum and a structural diagram of the sesquiterpene compound 2- (4 a,8-Dimethyl-1,2,3, 4a,5,6, 7-octahydro-naphthalen-2-yl) -prop-2-en-1-ol. The abscissa is the ion charge to mass ratio and the ordinate is the relative ion fragment intensity.

Fig. 10 is an expression pattern of NtTPS143 in each tissue of tobacco.

Detailed Description

The present invention will now be described in detail with reference to the drawings and specific examples, which should not be construed as limiting the invention. Unless otherwise indicated, the technical means used in the following examples are conventional means well known to those skilled in the art, and the materials, reagents, etc. used in the following examples are commercially available unless otherwise indicated.

Example 1: tobacco NtTPS143 Gene cloning

Tobacco material (100 mg) was thoroughly ground in liquid nitrogen, transferred to a 1.5mL centrifuge tube, 1mL Trizol (Invitrogen, cat. 15596-018) was added, mixed well, left at room temperature for 5min, centrifuged at 12000rpm for 10min, and the pellet was discarded. 0.2mL of chloroform was added to the supernatant, the mixture was homogenized, centrifuged at 12000rpm for 10min, and 0.5mL of isopropanol was added to the supernatant to precipitate RNA. Centrifuge at 12000rpm for 10min and the pellet was dissolved in 100. Mu.L water.

Reverse transcription was performed using an RNAPCR system (TaKaRa, cat DRR 019A), the reaction system being: 10 Xbuffer 1μL，dNTP 1μL，MgCl ₂ 2. Mu.L Oligo dT 1. Mu.L, RNAinhibitor 0.5. Mu.L, AMV RTase 0.5. Mu.L, RNA 2. Mu.L. After 30min of reaction at 42 ℃, ice bath is carried out.

Performing PCR amplification by using cDNA obtained by reverse transcription reaction as a template, wherein forward primers are as follows: ntTPS143-F:5'-ATGGCCTCAGCAGCAGCAGT-3' (SEQ ID NO. 3), reverse primer: ntTPS143-R:5'-TCAAATTTTGATGGATTCCA-3' (SEQ ID NO. 4). PCR amplification was performed using Takara PrimeSTAR Max DNA polymerase. The reaction system is as follows: 10 XBuffer 5. Mu.L, 10mM dNTP 2. Mu.L, forward primer 2. Mu.L, reverse primer 2. Mu.L, DNA polymerase 1. Mu.L, cDNA 1. Mu.L. The PCR conditions were: 95 ℃ for 5min;95 ℃ 30s,55 ℃ 30s,72 ℃ 120s,35 cycles; extending at 72℃for 5min. The PCR product was detected by 1% agarose gel electrophoresis, and the size of the NtTPS143 gene fragment was 1653bp, which corresponds to the expected size.

The nucleotide sequence of the NtTPS143 gene is shown as SEQ ID NO.2, and the amino acid sequence of the encoded protein is shown as SEQ ID NO. 1. Sequence homology alignment analysis shows that the protein is a typical sesquiterpene synthase.

Example 2: expression vector construction and E.coli transformation

Recovering target gene fragment by agarose gel electrophoresis gel recovery kit method, and connecting target fragment to carrierCloning vector (full gold: CB 101-01) and then transforming it into E.coli DH 5. Alpha. Clone under the following conditions: adding 5 mu L of the ligation product into 100 mu L of competent cells, lightly mixing, and then carrying out ice bath for 30min; rapidly placing into a water bath at a temperature of 42 ℃ for heat shock for 90s, and immediately placing on ice for 2-3min; adding 800 mu L of LB liquid medium, and slowly shaking and culturing for 1h at 37 ℃; the bacterial liquid was centrifuged at 6000rpm for 1min, 700. Mu.L of the supernatant was discarded, the bacterial cells were suspended, and the suspension was spread on LB plates containing ampicillin (Amp, 100 mg/L) and subjected to inverted dark culture at 37℃for 12 to 16 hours. Positive clone screening is carried out by colony PCR, positive monoclonal colonies are selected, plasmids are extracted, and then the plasmids are subjected to delivery sequencing.

Through sequencing analysis, the sesquiterpene synthetase gene NtTPS143 of tobacco is cloned and obtained, and the nucleotide thereofThe sequence is shown as SEQ ID NO.1, the encoded protein contains 1653 bases, the encoded protein is named as sesquiterpene synthase NtTPS143, total 550 amino acids, and the specific amino acid sequence is shown as SEQ ID NO. 2. Thus, the sesquiterpene synthase gene NtTPS143 was insertedThe recombinant plasmid of the cloning vector was designated pEASY-TPS143.

The plasmid pEASY-TPS143 is used as a template, and the TakaRa company PrimeSTAR Max DNA polymerase is used for amplifying the NtTPS143 gene, and the primers are as follows: ORF-NtTPS143-F-BamHI:5'-GCGGATCCATGGCCTCAGCAGCAGCAGT-3' (SEQ ID NO. 5) and ORF-NtTPS143-R-SacI:5'-GCGAGCTCTCAAATTTTGATGGATTCCA-3' (SEQ ID NO. 6). The reaction system is as follows: 10 XBuffer 5. Mu.L, 10mM dNTP 2. Mu.L, forward primer 2. Mu.L, reverse primer 2. Mu.L, DNA polymerase 1. Mu.L, cDNA 1. Mu.L. The PCR conditions were: 95 ℃ for 5min;95 ℃ 30s,55 ℃ 30s,72 ℃ 120s,35 cycles; extending at 72℃for 5min. The PCR product was detected by 1% agarose gel electrophoresis, and the target gene fragment was recovered by the method of agarose gel electrophoresis gel recovery kit, and double-digested with BamH I and Sac I was used to ligate with plasmid pET28a (Novagen: 69864-3) digested with BamH I and Sac I. E.coli is transformed by the connection product, clone culture is selected, and plasmid sequencing is extracted. The positive plasmid was designated pET28-TPS143.

Example 3: prokaryotic expression and Activity determination

pET28-TPS143 transformed E.coli Rosetta (DE 3). The monoclonal was picked and inoculated into liquid LB medium containing 50mg/L kanamycin for overnight culture. The culture was grown up to 50ml until OD600 = 0.6, IPTG was added to a final concentration of 1mM and the induction culture was continued for 12 hours at 20 ℃. The bacterial solution was centrifuged at 1000rpm for 1min and the pellet resuspended in lysis buffer (50mM Tris,pH7.5,5mM DTT,5mM MgCl) ₂ ) Cells were sonicated, centrifuged at 10000rpm for 2min, and supernatants were collected for activity assays.

To 100. Mu.L of the above protein was added 2. Mu.L of FPP (Sigma-Aldrich, F6892), and the mixture was reacted at 37℃for 1 hour, and 1ml of n-hexane was added and shaken, and 1. Mu.L was taken for gas chromatography-mass spectrometry (GC-MS). Gas Chromatography (GC) instrument model: thermo Trace1300 (HP-5 ms:30 m.times.0.25 mm.times.0.25 μm). Mass spectrometry instrument model Thermo ITQ 900 (EI ion source; ion trap detector). Sample injection amount: 1 μl. Chromatographic conditions: the temperature is kept at 60 ℃ for 3min, the temperature is increased to 280 ℃ at 10 ℃/min, the temperature is kept for 5min, and the helium flow rate is 1ml/min. Mass spectrometry conditions: the ion source temperature is 250 ℃, the interface temperature is 250 ℃, and the scan mode acquisition m/z is 50-500.

GC-MS detection showed that 5 products were detected at 15.12 (1), 15.22 (2), 15.94 (3), 16.35 (4), 16.58 (5) minutes (FIG. 1). Comparison of the mass spectrum with the NIST pool revealed that compound 1 was (3S, 3aR,3bR,4S,7R,7 aR) -4-Isopropyl-3, 7-dimethylicotina hydro-1H-cyclopena [1,3]cy clopropa[1,2]benzen-3-ol (FIGS. 2, 3), compound 2 was α -guanene (FIGS. 4, 5), compound 3 was napthalene, 1,2,3,5,6,7, 8a-octahydro-1,8 a-dimethylol-7- (1-methyl-thenyl) - [1S- (1 α,7 α,8 a) ] - (FIGS. 6, 7), and compound 4 was 2- (4 a, 8-dimethylol-1, 2,3, 4a,5,6, 7-octahydro-2-en-1-8, 8 a-1).

Example 4: expression pattern of the NtTPS143 Gene

Tobacco material (100 mg) was thoroughly ground in liquid nitrogen, transferred to a 1.5mL centrifuge tube, 1mL Trizol (Invitrogen, cat. 15596-018) was added, mixed well, left at room temperature for 5min, centrifuged at 12000rpm for 10min, and the pellet was discarded. 0.2mL of chloroform was added to the supernatant, the mixture was homogenized, centrifuged at 12000rpm for 10min, and 0.5mL of isopropanol was added to the supernatant to precipitate RNA. Centrifuge at 12000rpm for 10min and the pellet was dissolved in 100. Mu.L water.

Reverse transcription was performed using an RNA PCR system (TaKaRa, cat DRR 019A) with the following reaction system: 10 Xbuffer 1. Mu.L, dNTP 1. Mu.L, mgCl ₂ 2. Mu.L of Oligo dT 1. Mu.L, RNA Inhibitor 0.5. Mu.L, AMV RTase 0.5. Mu.L, and RNA 2. Mu.L. After 30min of reaction at 42 ℃, ice bath is carried out.

Using cDNA obtained by reverse transcription reaction as a template, the expression level of NtTPS143 was detected using Applied Biosystems 7500Real-Time PCR System (Applied Biosystems, waltham, mass., USA). The forward primer is qRT-NtTPS143-F:5'-CACATGTAAGGACTCATGCTGACG-3' (SEQ ID NO. 7), reverse primer qRT-NtTPS143-R:5'-GCACTCAACTGCTCTATCTCTAGC-3' (SEQ ID NO. 8). Tobacco EF-1a is used as an internal reference gene, and the primers are as follows: qRT-EF-1a-F:5'-TGAGATGCACCACGAAGCTC-3' (SEQ ID NO. 9) and qRT-EF-1a-R:5'-CCAACATTGTCACCAGGAAGTG-3' (SEQ ID NO. 10). Detection was performed using ChamQ SYBR qPCR Master Mix (novzan) reagent. The PCR reaction system is as follows: 2X ChamQ SYBR qPCR Master Mix. Mu.L, 1. Mu.L of forward primer, 1. Mu.L of reverse primer and 0.5. Mu.L of cDNA0. The PCR conditions were: 95 ℃ for 5min;95℃15s,58℃30s,72℃30s;40 cycles.

The results showed that the expression level of NtTPS143 was high in roots, and also some in senescent leaves, but not in organs such as stems, young leaves, flowers, etc. (fig. 10).

While preferred embodiments of the present invention have been described, additional variations and modifications in those embodiments may occur to those skilled in the art once they learn of the basic inventive concepts. It is therefore intended that the following claims be interpreted as including the preferred embodiments and all such alterations and modifications as fall within the scope of the invention.

It will be apparent to those skilled in the art that various modifications and variations can be made to the present invention without departing from the spirit or scope of the invention. Thus, it is intended that the present invention also include such modifications and alterations insofar as they come within the scope of the appended claims or the equivalents thereof.

Claims (10)

1. A tobacco sesquiterpene synthase NtTPS143 is characterized in that the amino acid sequence is shown in SEQ ID NO. 1.

2. Use of the tobacco sesquiterpene synthase NtTPS143 according to claim 1 for catalyzing the synthesis of sesquiterpene compounds from a substrate.

3. The use according to claim 2, wherein the substrate is farnesyl pyrophosphate and the sesquiterpene compound comprises (3 s,3ar,3br,4s,7r,7 ar) -4-isopopyyl-3, 7-dimethyloctahydro-

1H-cyclipta [1,3] cyclipta [1,2] benzen-3-ol, alpha-Guaiene, naphthalene,1,2,3,5,6,7, 8a-octahydro-1,8a-Dimethyl-7- (1-methyl-phenyl) -, [1S- (1 alpha, 7 alpha, 8 alpha) ] -and 2- (4 a,8-Dimethyl-1,2,3, 4a,5,6, 7-octahydro-naphthalen-2-yl) -prop-2-en-1-ol.

4. A gene encoding the tobacco sesquiterpene synthase NtTPS143 of claim 1.

5. The gene according to claim 4, wherein the gene has a nucleotide sequence shown in any one of a1 to a 2:

a1, a nucleotide sequence shown as SEQ ID NO. 2;

a2, a nucleotide sequence encoding said tobacco sesquiterpene synthase NtTPS143, but which differs from that shown in SEQ ID No.2 due to the degeneracy of the genetic code.

6. A recombinant expression vector comprising the gene of claim 4.

7. The recombinant expression vector of claim 6, wherein said gene is inserted forward between BamHI and SacI sites in pET28a plasmid.

8. A genetically engineered bacterium comprising the recombinant expression vector of claim 6.

9. The genetically engineered bacterium of claim 8, wherein the host bacterium used is e.

10. A process for producing a sesquiterpene compound, characterized in that the tobacco sesquiterpene synthase NtTPS143 according to claim 1 is used as a catalyst and farnesyl pyrophosphate is used as a substrate in dithiothreitol and MgCl ₂ Catalytic synthesis of sesquiterpene compounds under the action of (2); the sesquiterpene compound comprises (3S, 3aR,3bR,4S,7R,7 aR) -4-isopopyyl-3, 7-dimethyloctaHydro-

1H-cyclipta [1,3] cyclipta [1,2] benzen-3-ol, alpha-Guaiene, naphthalene,1,2,3,5,6,7, 8a-octahydro-1,8a-Dimethyl-7- (1-methyl-phenyl) -, [1S- (1 alpha, 7 alpha, 8 alpha) ] -and 2- (4 a,8-Dimethyl-1,2,3, 4a,5,6, 7-octahydro-naphthalen-2-yl) -prop-2-en-1-ol.