32 results about "Terpene synthase" patented technology

The present invention pertains to novel modified terpene synthases and their use for a preparation method for pseudopterosin intermediates and / or pseudopterosins. The method is based on the use of a modified terpene synthase comprising at least one modified amino acid residue, which enables a terpene synthase-catalyzed increased production of pseudopterosin intermediates and / or pseudopterosins from Geranylgeranyl pyrophosphate as starting material. The new modified terpene synthases of this invention allow the production of pseudopterosin intermediates, such as Isoelisabethatriene A, Isoelisabethatriene B, Erogorgiaene, or Seco-Pseudopterosin and / or the production of pseudopterosins, such as Pseudopterosin A, in a cost-efficient, economical, and sustainable manner. Also provided are nucleic acids, encoding for the modified terpene synthases of this invention, as well as expression vectors capable of expressing such nucleic acids and host cells comprising the same.

The present invention discloses a recombinant bacterium for synthesizing key components of plant essential oil, a construction method thereof and applications. The recombinant bacterium contains exogenous genes; the exogenous genes include genes of terpene synthase, cytochrome P450 and short-chain dehydrogenase; the key components of the plant essential oil include thymol and carvacrol. By introducing the genes of terpene synthase, cytochrome P450 and short-chain dehydrogenase, the present invention successfully constructs a recombinant bacterium capable of synthesizing thymol and carvacrol, realizing the heterologous synthesis of thymol derivatives. Moreover, by using the recombinant bacterium of the present invention to transform and produce thymol and carvacrol, the production efficiency is high, it is green and environmentally friendly, and the cost is low, having good industrial application prospects.

The present invention relates to a bifunctional terpene synthase and its mutants, as well as the catalytic products. The amino acid sequence of the bifunctional terpene synthase is shown in SEQ ID NO.1. The bifunctional terpene synthase mutant is a protein composed of a new amino acid sequence formed by mutating the leucine at the 89th position of the amino acid sequence shown in SEQ ID NO.1 to glutamine or asparagine. The bifunctional terpene synthase FoFS to be modified in the present invention simultaneously has an isopentenyl transferase domain and a terpene cyclase domain, and has the dual functions of catalyzing the head-to-tail connection and cyclization of the terpene synthesis precursors DMAPP and IPP. FoFS has sesquiterpene products with multiple different ring systems. By modifying FoFS, the mutant protein can catalyze sesquiterpene compounds with other complex ring systems, which is of great significance for expanding the library of terpene natural products and adding more potential drug-value terpene compounds to the reserve force.

This invention discloses a mango terpene synthase gene TPS4 and its applications, the nucleotide sequence of which is shown in SEQ ID NO:1. The amino acid sequence of the protein it encodes is shown in SEQ ID NO:2. The invention also discloses a recombinant expression vector containing the mango terpene synthase gene TPS4, a recombinant engineered strain, and the recombinant mango terpene synthase TPS4; and the application of the mango terpene synthase gene TPS4, its encoded protein, the recombinant expression vector, the recombinant engineered strain, or the recombinant TPS4 in regulating the synthesis of terpenoid compounds from geraniol diphosphate (GPP), neroli diphosphate (NPP), and farnesyl diphosphate (FPP). This invention is the first to clone and verify a terpene synthase gene TPS4 from the Sacred Heart mango genome, capable of simultaneously catalyzing the synthesis of monoterpenes and sesquiterpenes from three substrates: GPP, NPP, and FPP. It has broad application prospects and significant economic value.

The invention relates to the technical field of gene engineering, in particular to an Atractylodes lancea terpene synthase gene AlTPS42, a product coded by the Atractylodes lancea terpene synthase gene AlTPS42 and application of the Atractylodes lancea terpene synthase gene AlTPS42, and the Atractylodes lancea terpene synthase gene AlTPS42 has a nucleotide sequence as shown in SEQ ID NO.1 or a nucleotide sequence which is obtained by substituting, deleting or adding one or more nucleotides to the nucleotide sequence as shown in SEQ ID NO.1 and expresses protein with the same function. The enzyme can be applied to a pathway prepared by taking farnesyl pyrophosphate (FPP) as a substrate. By utilizing the gene provided by the invention, the content of the terpenoid substances of the atractylodes lancea can be increased through a genetic engineering technology.

The Machilus rubescens TPS synthetase gene MnTPS12 provided by the invention is a key gene for synthesis of gamma-Eudesmol and Linalool in plant genetic engineering, a novel terpene synthetase gene MnTPS12 is obtained by cloning in Machilus rubescens for the first time, and the gene can catalyze FPP and GPP to generate gamma-Eudesmol and Linalool, and plays an important role in improving the content and resistance of plant terpene components. The terpene synthase gene MnTPS12 researched by the invention can be used for preparing gamma-Eudesmol and Linalool, and can be further used for preparing essential oil, essence and medicines containing the components of the gamma-Eudesmol and the Linalool. The gene segment of the terpene synthase gene MnTPS12 is constructed on a plant expression vector, other plant materials can be transformed through an external source, so that a transgenic material containing the terpene aroma gene is obtained, and an effective method is provided for cultivating aroma and medicinal plants.

The present invention relates to a protein, comprising the following 1)-3): 1) an amino acid sequence as shown in SEQ ID NO.1; 2) a sequence having the same function as the sequence as shown in SEQ ID NO.1 and obtained by replacing, deleting or adding one or more amino acids; 3) a fusion protein having the same function as the protein as shown in SEQ ID NO.1 and connected to a tag at the N-terminus or C-terminus; and also discloses the use of the AarTPS89 gene in catalyzing the precursor substance geranyl pyrophosphate GPP to generate borneol diphosphate, and further generating borneol through the catalytic action of phosphatase; wherein the AarTPS89 gene sequence is as shown in SEQ ID NO.2.

The invention relates to a difunctional terpene synthase mutant and a sesterterpenoid compound generated by catalysis of the difunctional terpene synthase mutant, the difunctional terpene synthase mutant is a protein composed of a new amino acid sequence formed by mutating the 92nd isoleucine of the amino acid sequence as shown in SEQ ID NO.1 into phenylalanine, and the amino acid sequence of the difunctional terpene synthase mutant is as shown in SEQ ID NO.3. The invention also relates to a preparation method of the difunctional terpene synthase mutant. The structures of the sesterterpenoids generated by catalysis are as shown in formulas 2-8. Compared with the prior art, the mutant protein ZbSS-I92F disclosed by the invention has the advantages that the 92 isoleucine of the wild difunctional terpene synthase ZbSS is transformed into phenylalanine, so that the mutant protein ZbSS-I92F is catalyzed to obtain sesterterpene compounds of other complex ring systems, such as 5 / 6 / 5 / 5 / 5 pentacyclic, 5 / 15 bicyclic and 14-membered ring, instead of being catalyzed to generate tetracyclic sesterterpene originally. The method has important significance in expanding a terpenoid natural product library and adding more terpenoid compound reserve with potential drug value. Therefore, the function of the difunctional terpene synthase is expanded, and a foundation is further laid for analysis of an enzyme catalysis mechanism.

Described herein are plant-derived haloacid dehalogenase-like (HAD-like) polypeptides having cyclic terpene synthase (TPS) activity, in particular suitable for use in biochemical methods of producing drimane-type sesquiterpenes, encompassing drimenol and / or albicanol and related compounds, like phosphorylated drimane-type sesquiterpene alcohols, in particular phosphorylated drimenol and / or albicanol compounds and derivatives. Also described herein are the coding nucleotide sequences of the TPS activity, corresponding expression constructs, recombinant hosts, and methods of preparing such polypeptides and mutants and variants thereof. Also described herein is a method of using such polypeptides, the method including using the polypeptides in the production of odorants, flavours and fragrance ingredients.

The invention discloses a tripterygium wilfordii transcription factor MYB92 and application thereof. The full length of the disclosed TwMYB92 coding gene is 1002bp, and 333 amino acids are coded. The invention finds that the transcription factor can be specifically combined with a promoter region of tripterygium wilfordii terpenoid synthetase and has remarkable transcriptional regulation activity. By regulating and controlling the expression of the TwMYB92 transcription factor, the expression of the terpene synthetase gene can be effectively activated, so that the accumulation of terpene substances is promoted. By identifying the function of the TwMYB92 transcription factor, a foundation is laid for improving the biosynthesis of tripterygium wilfordii terpenoids through a genetic engineering means, a feasible scheme is provided for protecting wild tripterygium wilfordii resources, and the application prospect is wide and the economic value is important.

The invention discloses a tobacco terpenoid synthase gene NtTPS2184 and application thereof, and belongs to the technical field of plant molecular biology, the nucleotide sequence of the tobacco terpenoid synthase gene NtTPS2184 is shown as SEQ ID NO.1, the invention provides a recombinant expression vector, the recombinant expression vector is obtained by positively inserting the tobacco terpenoid synthase gene NtTPS2184 between BamHI and SacI sites of pGEX4.0 plasmid, and the recombinant expression vector is used for expressing the tobacco terpenoid synthase gene NtTPS2184. The invention also provides an application of the tobacco terpene synthase gene NtTPS2184 or the recombinant expression vector in the synthesis of linalool and trans-nerolidone by a catalytic substrate, and the terpene synthase gene NtTPS2184 provided by the invention can be massively expressed by using escherichia coli genetically engineered bacteria, so that the yield of the tobacco terpene synthase gene NtTPS2184 is increased, and the yield of the tobacco terpene synthase gene NtTPS2184 is increased. In the presence of a substrate geranyl pyrophosphate, the monoterpene synthase exerts the activity of the monoterpene synthase to catalytically synthesize linalool; and in the presence of a substrate farnesyl pyrophosphate, the activity of the sesquiterpene synthase is exerted, and the trans-nerolidinol is catalytically synthesized.

The invention relates to the technical field of biology and natural medicines, in particular to an Atractylodes lancea terpene synthase gene AlTPS21, a product coded by the gene AlTPS21 and application of the gene AlTPS21, and the gene AlTPS21 has a nucleotide sequence as shown in SEQ ID NO.1 or a nucleotide sequence which is obtained by substituting, deleting or adding one or more nucleotides to the nucleotide sequence as shown in SEQ ID NO.1 and expresses protein with the same function. According to the invention, a coding gene of the Atractylodes lancea terpene synthase gene (AmTPS) is successfully cloned from the rhizome of Atractylodes lancea, and the synthase can be applied to a pathway prepared by using farnesyl pyrophosphate (FPP) as a substrate. By utilizing the gene provided by the invention, the content of the terpenoid substances of the atractylodes lancea can be increased through a genetic engineering technology.

This invention relates to the terpene synthase AaTPS48 from Artemisia argyi, its encoding gene, and its applications. It provides an important foundation for utilizing genetic engineering techniques to increase the content of the active components myrcene and (trans)-β-farnesene in Artemisia argyi, or to directly produce myrcene and (trans)-β-farnesene. The technical solution is that the terpene synthase AaTPS48 is a protein of the following a) or b) or c): a) the amino acid sequence of the protein shown in SEQ ID No. 1; b) a fusion protein obtained by attaching a tag to the N-terminus and / or C-terminus of the protein shown in SEQ ID No. 1; c) a protein with the same function obtained by substituting and / or deleting and / or adding one or more amino acid residues of the amino acid sequence shown in SEQ ID No. 1. The AaTPS48 protein of this invention can catalyze both GPP to form myrcene and FPP to form (trans)-β-farnesene, playing an important role in the biosynthesis of myrcene, (trans)-β-farnesene, and other terpene compounds in Artemisia argyi.

The application provides a machilus thunbergii TPS synthetase gene MnTPS12 The application is a key gene for synthesis of gamma-eudesmol and linalool in plant genetic engineering, and a new terpene synthase gene is cloned from machilus thunbergii for the first time MnTPS12 The gene can catalyze FPP and GPP to generate gamma-eudesmol and linalool, and plays an important role in improving the content and resistance of plant terpene components. MnTPS12 The terpene synthase gene studied in the application can be used for preparing gamma-eudesmol and linalool, and for further preparing essential oil, essence and medicine containing gamma-eudesmol and linalool. MnTPS12 The gene fragment of the terpene synthase gene is constructed on a plant expression vector, and can be transformed into other plant materials by exogenous transformation, so as to obtain transgenic materials containing the terpene aroma gene, and provide an effective method for cultivating aroma and medicinal plants.

The present disclosure in various aspects provides host cells and methods for making sesquiterpene components of cypriol oil or oud oil. In some embodiments, the host cells express one or more terpene synthase (TPS) enzymes. In some embodiments, the invention provides host cells producing oxidized sesquiterpene components of cypriol oil or oud oil. In these embodiments, the host cells further expresses one or more cytochrome P450 (CYP) enzymes.

The present invention discloses a method for utilizing OfTPS7 A method for in vitro synthesis of terpene aroma substances and screening of key terpene synthase genes from Osmanthus fragrans OfTPS7 The key terpene synthase OfTPS7 was expressed using a prokaryotic expression system. The purified OfTPS7 protein was subjected to in vitro enzymatic activity experiments using geranyl diphosphate (GPP) as a substrate. The products were detected using headspace solid-phase microextraction combined with GC-MS. The results showed that OfTPS7 can catalyze GPP to produce terpene aroma substances such as linalool, β-myrcene, and D-limonene.

This disclosure generally relates to the use of enzyme combinations or recombinant microbes comprising same to make isoprenoid precursors, isoprenoids and derivatives thereof including prenylated aromatic compounds. Novel metabolic pathways exploiting Claisen, aldol, and acyloin condensations are used instead of the natural mevalonate (MVA) pathway or 1-deoxy-d-xylulose 5-phosphate (DXP) pathways for generating isoprenoid precursors such as isopentenyl pyrophosphate (IPP), dimethylallyl pyrophosphate (DMAPP), and geranyl pyrophosphate (GPP). These pathways have the potential for better carbon and or energy efficiency than native pathways. Both decarboxylative and non-carboxylative condensations are utilized, enabling product synthesis from a number of different starting compounds. These condensation reactions serve as a platform for the synthesis of isoprenoid precursors when utilized in combination with a variety of metabolic pathways and enzymes for carbon rearrangement and the addition / removal of functional groups. Isoprenoid alcohols are key intermediary products for the production of isoprenoid precursors in these novel synthetic metabolic pathways. These precursors can be modified to various isoprenoid products through prenyl transferase, terpene synthase, or terpene cyclases. The production of prenylated aromatic compounds is achieved through prenyl transfer of the hydrocarbon units of isoprenoid precursors to polyketides.

The disclosure relates to a method of making at least one serrulatane comprising contacting a terpene or a terpenoid substrate with at least one of a cis-prenyl transferase, a terpene synthase, and a cytochrome P45Q. The disclosure also relates to an expression system comprising one or more expression cassettes, each expression cassette comprising a promoter operably linked to a nucleic acid segment encoding at least one of: a cis-prenyl transferase, a terpene synthase, and a cytochrome P450. The disclosure also relates to a host cell comprising an expression system comprising one or more expression cassettes, each expression cassette comprising a promoter operably linked to a nucleic acid segment encoding at least one of: a cis-prenyl transferase, a terpene synthase, and a cytochrome P450.

The invention relates to the technical field of plant genetic engineering, in particular to a ginkgo terpene synthase gene GbTPS13 and application of the ginkgo terpene synthase gene GbTPS13 in heterologous synthesis of a bilobalide precursor in tobacco. The nucleotide sequence of the ginkgo terpene synthase gene GbTPS13 is as shown in SEQ ID NO. 1, and the coded amino acid sequence is as shown in SEQ ID NO. 2; ginkgo biloba diterpene synthase GbTPS13 is identified in tobacco tissues, L-pimaric diene can be catalytically generated through overexpression of GbTPS13 in tobacco, the diterpene compound is a common precursor of bilobalide, and the method is used for constructing a tobacco chassis system capable of being used for synthesizing bilobalide. And a potential starting platform and basic framework are provided for complete and large-scale biosynthesis of bilobalide.

A terpenoid synthase for producing germacrene A and application thereof.The present application relates to a germacrene A synthase AarTPS34 gene and its coding product and application.The AarTPS34 gene is cloned from folium artemisiae argyi, and the AarTPS34 gene is a key enzyme gene for synthesizing germacrene A, a sesquiterpenoid compound, which is obtained from artemisia for the first time.The experiment proves that the AarTPS34 protein of the present application can catalyze the precursor material farnesyl pyrophosphate (FPP) to generate the sesquiterpenoid compound germacrene A, and germacrene A is a synthetic precursor of anticancer active ingredient beta-elemene, and the research on germacrene A has important theoretical and practical significance for improving the quality of folium artemisiae argyi and producing plant monoterpene compounds.

The invention discloses a vetiver grass terpene synthase gene VzTPS9 and an application of the vetiver grass terpene synthase gene VzTPS9 in synthesis of terpenoids. The invention belongs to the field of gene engineering, the terpene synthase gene VzTPS9 is cloned and separated from vetiver grass for the first time, and the gene function is explored through prokaryotic expression. It is found and proved that when FPP is used as a substrate, VzTPS9 can be used for synthesizing a sesquiterpene compound cedrol and a monoterpene compound linalool. The gene has important value for improving the insect trapping capability of vetiveria zizanioides.

According to the technical route of prokaryotic expression, protein purification and enzyme activity verification, novel synthase AaTPS19 in artemisia apiacea is found, GPP is catalyzed by the novel synthase AaTPS19 to generate 2-Pinanol and Geraniol, the novel synthase AaTPS19 is the first identified 2-PinanolSynthase, the product type of the monoterpene synthase is broadened, potential functions and evolution characteristics of the monoterpene synthase are analyzed, and the novel synthase AaTPS19 has the advantages that the novel synthase AaTPS19 has the advantages that the novel synthase AaTPS19 has the advantages that the novel synthase AaTPS19 is used for catalyzing GPP to generate the 2-Pinanol and the Geraniol; and a tissue expression mode of the gene is obtained through fluorescent quantitative PCR, so that a foundation is laid for prokaryotic expression and in-vitro functional verification of the TPS.

The invention discloses a plant-sensitive piercing-sucking pest related gene MED25 and application thereof, and relates to the technical field of gene engineering, the MED25 gene is derived from corn, the nucleotide sequence of the MED25 gene is shown as SEQ ID NO.1, and the amino acid sequence of encoded protein of the MED25 gene is shown as SEQ ID NO.2. It is proved for the first time that the corn MED25 gene mutation enables the gene function to be lost, and the corn MED25 gene mutation enables the gene function to be lost. The MED25 gene can improve the resistance of corn to piercing-sucking pests, and the MED25 gene participates in the piercing-sucking pest resistance through transcriptional regulation of the expression of a terpene synthase gene (TPSs gene), provides important theoretical technical support for cultivation of piercing-sucking pest resistant corn varieties, and has important production and application values.

The invention provides a masson pine alcohol dehydrogenase gene, a product and application thereof, masson pine alcohol dehydrogenase and application thereof, and relates to the technical field of biology. The pinus massoniana alcohol dehydrogenase gene is cloned from pinus massoniana for the first time, the nucleotide sequence and the amino acid sequence of the pinus massoniana alcohol dehydrogenase gene are determined, the blank that in the prior art, terpene synthase in pinus massoniana and biosynthetic genes of the terpene synthase are unknown is filled up, and theoretical and basic support is provided for related research of pinus massoniana. Besides, the masson pine alcohol dehydrogenase gene is recombined to obtain a recombinant vector and an engineering bacterium, the masson pine alcohol dehydrogenase is successfully purified to obtain the masson pine alcohol dehydrogenase, the masson pine alcohol dehydrogenase can catalyze cis-3-hexenal to synthesize cis-3-hexenol in an in-vitro enzyme activity experiment, and the masson pine alcohol dehydrogenase has important significance on prevention of phytophagous diseases and insect pests.

The invention provides a machilus TPS synthetase gene MnTPS10 and application thereof, the terpene synthetase gene MnTPS10 with a dual catalytic function is cloned from machilus, the gene can catalytically synthesize Hedycarylol and Geraniol at the same time, the research on the dual-function terpene synthetase gene in machilus plants is enriched, and the application of the gene in the machilus plants is developed. Important gene resources and theoretical basis are provided for deeply researching the diversity synthesis mechanism of terpenoids. The MnTPS10 gene disclosed by the invention has a unique dual catalytic function, two high-value terpenoids can be respectively synthesized by utilizing different terpenoid precursors (GPP and FPP), and compared with a synthetase gene with a single function, the MnTPS10 gene disclosed by the invention is wider in application range, the biosynthesis cost can be reduced, and the production efficiency can be improved.

The disclosure relates to a method of making at least one serrulatane comprising contacting a terpene or a terpenoid substrate with at least one of a cis-prenyl transferase, a terpene synthase, and a cytochrome P450. The disclosure also relates to an expression system comprising one or more expression cassettes, each expression cassette comprising a promoter operably linked to a nucleic acid segment encoding at least one of: a cis-prenyl transferase, a terpene synthase, and a cytochrome P450. The disclosure also relates to a host cell comprising an expression system comprising one or more expression cassettes, each expression cassette comprising a promoter operably linked to a nucleic acid segment encoding at least one of: a cis-prenyl transferase, a terpene synthase, and a cytochrome P450.

The invention relates to a difunctional terpene synthase mutant and a sesterterpenoid compound generated by catalysis of the difunctional terpene synthase mutant, the difunctional terpene synthase mutant is a protein composed of a new amino acid sequence formed by mutating 195th valine of an amino acid sequence as shown in SEQ ID NO.1 into alanine, and the amino acid sequence of the difunctional terpene synthase mutant is as shown in SEQ ID NO.3. The invention also relates to a preparation method of the difunctional terpene synthase mutant. Compared with the prior art, the method has the advantages that the difunctional terpene synthase FoFS is subjected to semi-rational transformation, so that the mutant FoFS-V195A of the difunctional terpene synthase FoFS can catalyze the generation of novel skeleton sesterterpenoids with different stereo chirality. The strategy not only is beneficial to expanding the structural diversity of terpenoid natural products, but also provides important molecular resources for developing novel terpenoid compounds with potential drug value, and has important scientific significance and application prospects.

The present invention relates to novel plant-derived halogenase-like (HAD-like) polypeptides having cyclic terpene synthase (TPS) activity, particularly suitable for use in biochemical methods for the production of drimane sesquiterpenes, including drimenol and / or selinol and / or related compounds, such as phosphorylated drimane sesquiterpene alcohols, particularly phosphorylated drimenol and / or selinol compounds and derivatives. The present invention also provides the novel TPS-activity-encoding nucleotide sequences, corresponding expression constructs, recombinant hosts, methods for the production of such novel polypeptides and mutants and variants thereof. The present invention also relates to the use of such novel polypeptides in the production of odorants, flavorants and fragrance ingredients.

The application relates to the field of biotechnology, and particularly discloses a Saccharomyces cerevisiae engineering strain LSc274 with high yield of sirenol and a construction method and application thereof, aiming to solve the problems of low content of sirenol in a natural host, difficulty in chemical synthesis, and low yield of biosynthesis. ROX1 、 EXG1 、 DPP1 、 TRP1 , the GAL80 gene is knocked out in a Saccharomyces cerevisiae host, and meanwhile, sirenol synthase coding genes, tHMG1, EGR20, TRP1, URA3, LEU2 and HIS3 genes are highly expressed. The yield of the constructed engineering strain reaches 905.83+33.41 mg / L in a flask fermentation, and the yield reaches 16.29+0.12 g / L in a 5 L fermenter, which is the highest yield reported at present, and the engineering strain has the application prospect of large-scale production of sirenol.

The application discloses a kind of for synthesizing β - terpene synthase CcTPS6 of eucalyptol and its encoding gene and its product β - the application of eucalyptol as anti-aging drug belongs to synthetic biology and natural medicine chemistry technical field. The application starts from Labiatae plant torch flower ( Colquhounia coccinea var. mollis ) Cloning and functional identification of a synthetic β - eucalyptol single-function terpene synthase CcTPS6 encoding gene, its nucleotide sequence is as shown in Seq ID No.2, after codon optimization and site-directed mutagenesis, connect with expression vector pCold-TF, construct into the recombinant plasmid capable of expressing in escherichia coli, again the recombinant plasmid is transformed into escherichia coli and is constructed into engineering cell, realizes the heterologous high-efficiency synthesis compound β - eucalyptol in escherichia coli. The gene engineering cell constructed by the application is safe and stable, and the production cycle is short, which shows its great value in application development. The CcTPS6 product provided by the application β - eucalyptol has anti-aging activity and can be applied in the preparation of anti-aging drugs.