43 results about "Prenyltransferase" patented technology

The invention provides a method for biosynthesizing menadione by using yarrowia lipolytica. The method is realized by constructing recombinant yarrowia lipolytica, and the construction method of the recombinant yarrowia lipolytica for synthesizing menadione comprises the following steps: intensively expressing a mevalonic acid pathway in the yarrowia lipolytica, and synthesizing menadione by introducing a synthesis pathway of heterologous geranyl and geranyl pyrophosphate. A sufficient isoprene side chain donor is provided for biosynthesis of menadione; on the basis, an aromatic isopentenyl transferase coding gene which is optimized by codons and has menadione substrate specificity is over-expressed in yarrowia lipolytica. The construction method of the yarrowia lipolytica for biosynthesis of menadione provided by the invention is simple to operate, the constructed yarrowia lipolytica can obtain a menadione product with relatively high concentration by taking commercially available menadione as a substrate, the biosynthesis steps are relatively short, and the yarrowia lipolytica has relatively high production and application values.

The invention relates to the discovery of a biosynthetic gene cluster of an alkaloid, namely, exopoid C, and the biosynthetic gene cluster of the exopoid. A biosynthetic gene cluster exo of an exopoid compound is obtained from a fungus Penicillium sp.HDN14-431 through genome mining, the gene cluster comprises a heme-dependent oxidase gene exoA which is composed of indole dioxygenase and a cytochrome b5 structural domain, a cytochrome P450 enzyme gene exoB and an isopentenyl transferase gene exoC which is composed of indole dioxygenase and a cytochrome b5 structural domain, and the cytochrome P450 enzyme gene exoB and the isopentenyl transferase gene exoC are used as biosynthetic genes of the exopoid compound. All genes in a gene cluster are expressed through an aspergillus nidulans heterologous expression system, two isopentenyl indole alkaloids are obtained in total, and a compound I is a new compound exopoid C. And the compound II, namely the exopoid, is a known compound. The biosynthesis pathway of the compound is reported for the first time.

The invention discloses a biosynthesis method and application of cannabinoid compounds. The specific method comprises the following steps: constructing a non-reductive polyketide synthase (NRPKS) AtnG from a fungus Arthrinum sp.NF2194, an isopentenyl transferase NphBV49W / Y288P from an actinomycetes Streptomyces sp.CL190, a cyclase Svz9 from an actinomycetes Streptoviensis NA431, a GPP synthase AgGPPS from a plant Abis grandis and a terpene synthesis enhancement path MEV into a heterologous expression vector, and sequentially introducing the heterologous expression vector into escherichia coli, so as to successfully realize the production of the cannabinoid compounds OA, CBGA and CBCA. The cannabinoid compound disclosed by the invention has important medical and industrial values.

This invention provides a chalcone isopentenyltransferase gene. GiPT16 This invention relates to the GiPT16 protein, amplification primer set, and applications, belonging to the field of biogenetics technology. It is based on *Glycyrrhiza inflata* (GiPT16 protein, amplification primer set, and applications). Glycyrrhiza inflata Using whole-genome sequencing data and a reverse genetics strategy, the key aromatic isopentenyltransferase GiPT16, involved in the isopentenylation modification of chalcone active ingredients, was successfully identified and functionally characterized. This enzyme was confirmed to specifically catalyze the biosynthesis of psoralen and glycyrrhizin C using DMAPP as a donor. It is the first chalcone isopentenyltransferase characterized in *Glycyrrhiza inflata*, filling a gap in the study of key enzymes in the chalcone isopentenylation metabolic pathway of this species and providing important evidence for further elucidating the molecular mechanisms of quality formation in *Glycyrrhiza inflata* medicinal materials.

The invention relates to the field of molecular biology, in particular to an enzyme for cannabinoid synthesis and a method for preparing cannabinoid. More specifically, the present invention relates to a method for producing a cannabinoid phenolic acid (CBGA) or an analog thereof, the method comprising the steps of: reacting a heterologous expressed isoprenyltransferase with a geranyl diphosphate (GPP) and an acid; wherein the amino acid sequence of the isoprenyltransferase is SEQ ID No: 1, or the isoprenyltransferase is expressed by the nucleotide sequence of SEQ ID No: 2.

The application provides a kind of aromatic isopentenyl transferase mutant, which is based on the aromatic isopentenyl transferase gene MenA derived from Bacillus subtilis ( Bacillus subtilis ) and six aromatic isopentenyl transferase mutants MenA-1 to MenA-6 are obtained by site-directed mutagenesis PCR technology, and the recombinant bacteria Bacillus subtilis expressing the above mutants are constructed, respectively. The yield of heptamethylnaphthoquinone fermented by the recombinant bacteria is 64.49 mg / L, 69.30 mg / L, 83.86 mg / L, 105.84 mg / L, 119.23 mg / L and 125.45 mg / L, respectively, which is 133%, 143%, 173%, 219%, 246% and 259% of the yield of heptamethylnaphthoquinone fermented by the Bacillus subtilis expressing the wild-type aromatic isopentenyl transferase MenA.

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.

The invention relates to application of isopentenyl transferase FnPT1, and belongs to the technical field of enzymology. The isopentenyl transferase FnPT1 screened and verified in the invention is different from most of reported flavonoid isopentenyl transferases, the isopentenyl transferase FnPT1 can catalyze the isopentenyl modification reaction of a larger substrate of flavone glycoside, and the isopentenyl transferase FnPT1 has the potential to be applied to the synthetic biology of the flavone glycoside. The important application value is realized.

To provide a genetically engineered microorganism capable of producing a cannabinoid.SOLUTION: Provided is a genetically engineered microorganism that is a photosynthetic microalgae or cyanobacterium and does not contain an exogenous nucleic acid molecule encoding an aromatic prenyltransferase. In one aspect, the genetically engineered microorganism is capable of producing tetrahydrocannabinolic acid or tetrahydrocannabinol and does not contain an exogenous nucleic acid molecule encoding a tetrahydrocannabinolic acid synthase.SELECTED DRAWING: Figure 1

To provide a polyisoprenoid production method capable of suitably producing polyisoprenoid even when using a callus.SOLUTION: A method for producing polyisoprenoid comprises using a transformed callus into which a gene encoding a cis-prenyltransferase (CPT) family protein and a gene encoding a Rubber Elongation Factor (REF) family protein are introduced, to produce polyisoprenoid.SELECTED DRAWING: Figure 2

The present application relates to the field of synthetic biology technology, and specifically discloses a cannabinol-producing microbial cell, its construction method, and application. A cannabinol-producing microbial cell includes a host cell, and the host cell includes a synthetic pathway for geranyl pyrophosphate, a synthetic pathway for olivate, and an isopentenyl transferase mutant. The construction method is as follows: based on the geranyl pyrophosphate-producing host cell yCAN10, the olivate synthesis pathway is constructed to form yCG02; a copy gene of the olivate synthase OLS is integrated into yCG02 to obtain yCG03; an acetyl-CoA carboxylase mutant ACC1 (S659A, S1157A) is integrated into yCG03 to obtain yCG04; and an isopentenyl transferase mutant is integrated into yCG04 to obtain yCG05, i.e., a cannabinol-producing microbial cell. The application is to produce cannabinol from a carbon source, which alleviates the problem of a large gap in cannabinol resources in the current market.

The invention discloses an isopentenyl transferase and an application of the isopentenyl transferase in synthesis of bakuchiol. Specifically, recombinant saccharomyces cerevisiae for expressing the isopentenyl transferase is constructed, the isopentenyl transferase CcPT6 from fructus psoraleae is obtained through screening, the isopentenyl transferase CcPT6 can catalyze an isopentenylation reaction of a coumaric acid substrate to generate bakuchiol, and the activity of the isopentenyl transferase CcPT6 is higher than that of a reported sequence BAK36. The CcPT7 is obtained by truncating 54 amino acids at the N end of the CcPT6, the catalytic activity is improved by 74%, and the activity of the CcPT7 is higher than that of a reported BAK36 mutant. In addition, the invention also provides a recombinant saccharomyces cerevisiae and yarrowia lipolytica engineering strain for integrally expressing the isopentenyl transferase CcPT6 or CcPT7, so that the artificial biosynthesis of the bakuchiol is realized. The discovery of the isopentenyl transferase and the construction of the recombinant engineering strain have important meanings for promoting the biosynthesis of the bakuchiol.

The present disclosure refers to therapeutic combinations and pharmaceutical compositions comprising glycolysis inhibitors / modulators and prenyltransferase inhibitors for treating brain tumors. In particular, the disclosure describes therapeutic combinations and pharmaceutical compositions of dichloroacetate (DCA) and perillyl alcohol (POH) useful for treating brain cancers such as glioblastoma multiforme (GBM). Additionally, the disclosure provides methods for treating GBM and other forms of brain cancer using the same.

The invention discloses an oxygen isopentenyl transferase participating in synthesis of sesquiterpene coumarin and an application of the oxygen isopentenyl transferase. Belongs to the technical field of gene engineering. The invention discloses an open reading frame sequence of a Xinjiang ferula oxygen isopentenyl transferase FsPT9 gene and an amino acid sequence coded by the Xinjiang ferula oxygen isopentenyl transferase FsPT9 gene. The FsPT9 gene is obtained by screening and cloning based on combined analysis of multi-omics data of ferula sinkiang, a yeast expression vector is constructed, and a catalytic mechanism of the FsPT9 is identified in vitro by taking 7-hydroxycoumarin and farnesyl pyrophosphate as substrates. A yeast heterologous expression system proves that the FsPT9 can catalyze 7-hydroxycoumarin and farnesyl pyrophosphoric acid to generate umbelliferone. The invention discloses a key enzyme for initial synthesis of sesquiterpene coumarin for the first time, and the key enzyme is an oxygen isopentenyl transferase which is reported for the first time and is used for catalyzing three isopentenyl units at the same time, so that theoretical guidance is provided for synthesis and transformation of pharmacodynamic substances of asafetida sesquiterpene coumarin.

The application provides a kind of aromatic isopentenyl transferase mutant, to be derived from Bacillus subtilis ( Bacillus subtilis )SEPHCHC synthase gene MenD is based, six aromatic isopentenyl transferase mutants MenD-1, MenD-2, MenD-3, MenD-4, MenD-5 and MenD-6 are obtained by site-directed mutagenesis PCR technology, and the recombinant bacteria Bacillus subtilis expressing the above mutants is respectively constructed, and the yield of heptamethylnaphthoquinone fermentation is 70.77 mg / L, 75.48 mg / L, 81.72 mg / L, 88.41 mg / L, 99.88 mg / L and 133.40 mg / L respectively, which is 116%, 123%, 134%, 144%, 162% and 218% of the yield of heptamethylnaphthoquinone of Bacillus subtilis expressing wild-type SEPHCHC synthase MenD respectively.

A prenyltransferase capable of producing bakuchiol or its analogues from geranyl pyrophosphate (GPP) and cinnamic acid, coumaric acid, caffeic acid and / or ferulic acid is disclosed herein. Manipulated cells expressing a prenyltransferase capable of producing bakuchiol or its analogues, as well as methods of using the prenyltransferase and cells to produce bakuchiol or its analogues, are also disclosed herein.

The application provides a prenyltransferase mutant and application thereof in production of icariin, and belongs to the technical field of bioengineering. The application solves the problem of low icariin yield in the prior art. The prenyltransferase mutant is obtained by mutating any one of the 52th, 131th, 205th and 303th positions of the amino acid shown in SEQ ID NO. 2. The prenyltransferase mutant is mainly used for efficient prenyltransferase and application thereof in synthesis of icariin.

The present disclosure refers to therapeutic combinations and pharmaceutical compositions comprising glycolysis inhibitors / modulators and prenyltransferase inhibitors for treating brain tumors. In particular, the disclosure describes therapeutic combinations and pharmaceutical compositions of dichloroacetate (DCA) and perillyl alcohol (POH) useful for treating brain cancers such as glioblastoma multiforme (GBM). Additionally, the disclosure provides methods for treating GBM and other forms of brain cancer using the same.

This invention discloses a protein with O-isopentenyltransferase activity, related biomaterials, and applications, belonging to the field of biotechnology. This invention screened and cloned an O-isopentenyltransferase gene (GlOPT), which can be used to prepare O-isopentenyltransferase. Experiments have confirmed that this O-isopentenyltransferase can catalyze the in vitro conversion of 8-hydroxybergamot lactone to arugulatin; catalyze the conversion of bergamotol to isoimperatorin; and catalyze the conversion of xanthotoxin to imperatorin. This provides technical support for the production of isopentenyl furanocoumarin-like active ingredients from arugula, lays the foundation for research on the biosynthetic pathways of coumarin compounds, and has significant theoretical and practical implications.

The application relates to application of an isopentenyl transferase FnPT1 and belongs to the technical field of enzymology. The isopentenyl transferase FnPT1 screened and verified by the application is different from most reported flavonoid isopentenyl transferases, the isopentenyl transferase FnPT1 can catalyze isopentenyl modification reaction of flavonoid glycoside which is a relatively large substrate, the isopentenyl transferase FnPT1 has potential application in flavonoid glycoside synthetic biology, and has important application value.

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 provides a method for heterologous synthesis of a compound Angulotetrone for resisting phytophthora parasitica var nicotianae, which is characterized in that non-reduced polyketide synthase Atn1, UbiA type isopentenyl transferase Atn2, flavin dependent monooxygenase Atn3 and terpene cyclase Atn4 are co-expressed in an aspergillus oryzae host, so that a silent biosynthesis process is successfully activated; according to the invention, a novel antibacterial compound Angulotetrone is found and synthesized. The compound cannot be obtained in the original strain. Structural identification and activity tests show that the compound Angulotetrone shows remarkable inhibitory activity on phytophthora parasitica var nicotianae, and the minimum inhibitory concentration (MIC) is 9.01 g / mL. The invention not only provides a new candidate drug for prevention and treatment of tobacco black shank, but also provides an efficient technical platform for mining dark substances of microbial genomes and discovering active natural products with novel structures.

The invention provides an isopentenyl transferase mutant and application of the isopentenyl transferase mutant in production of icaritin, and belongs to the technical field of bioengineering. The problem of low icaritin yield in the prior art is solved. The isopentenyl transferase mutant comprises an isopentenyl transferase mutant, and the isopentenyl transferase mutant is obtained by taking amino acid shown in SEQ ID NO.2 as a starting sequence and mutating any one of the 52nd site, the 131 site, the 205th site and the 303th site of the isopentenyl transferase mutant. The isopentenyl transferase is mainly used for high-efficiency isopentenyl transferase and application thereof in synthesis of icaritin.

A recombinant cell of Saccharomyces cerevisiae that includes in its genome nucleic acids encoding cannabinoid biosynthetic pathway genes. A cannabinoid is produced by the recombinant cell in the presence of a cannabinoid precursor substrate and at least one of the cannabinoid biosynthetic pathway genes is from an organism other than Cannabis sativa, wherein the at least one of the cannabinoid biosynthetic pathway genes encodes a prenyltransferase. In an embodiment, the prenyltransferase is NphB from Streptomyces sp. having the amino acid sequence of any one of SEQ ID NOs: 8-11. Also disclosed is a method for producing a cannabinoid with the recombinant cell and the cannabinoid precursor substrate.

The present invention provides an isolated DNA molecule including at least a first nucleic acid sequence encoding a first protein and at least a second nucleic acid sequence encoding a second protein, wherein the first protein and the second protein are derived from Helichrysum umbraculigerum and belonging to an enzyme family selected from: acyl activating enzyme (AAE), polyketide synthase (PKS), polyketide cyclase (PKC), prenyltransferase (PT), or cannabichromenic acid synthase (CBCAS), and wherein the first protein and the second protein belong to different enzyme families. Further provided are an artificial nucleic acid molecule including the isolated DNA molecule, a transgenic cell, a tissue, or a plant including same. Further provided is a method for synthesizing a cannabinoid, a precursor thereof, or any combination thereof.

The application provides a recombinant yeast for producing bakuchiol, a construction method and application thereof. The recombinant yeast is Yarrowia lipolytica containing isopentenyl transferase, and a gene coding the isopentenyl transferase is derived from Psoralea corylifolia Linn. The recombinant yeast can be used to produce bakuchiol in a large yield through a biosynthetic method, and provides technical support for industrialization of the bakuchiol.

The invention discloses a recombinant expression vector, a recombinant bacterium, a cannabidiol analogue O-1821 synthesis method and application thereof, and relates to the technical field of bioengineering. According to the method, the synthesis efficiency is remarkably improved through multiple innovative strategies. Firstly, the catalytic efficiency of a key enzyme can be enhanced by optimizing the metabolic flux of endogenous malonyl-coenzyme A and geranyl diphosphate; and secondly, a condensed phase vesicular enzyme active concentrate-diisopentenyl transferase synergistic expression system is introduced, and diisopentenyl transferase is synergistically expressed, so that the efficiency of converting the serotinic acid into the cannabis rhizopus acid is remarkably improved, and the efficient synthesis of the O-1821 and the precursor compound thereof is realized. The invention has wide application potential in the fields of drug research and development, natural product biosynthesis and biotechnology, and provides a new way for sustainable production and industrial development of cannabinoid compounds.

The present invention belongs to the field of bioengineering technology and specifically relates to an isoprenyltransferase mutant and its use. To address the prior art problem of microbial inability to synthesize polyisoprene, the present invention utilizes semi-rational enzyme design. Without changing the amino acid sequence of the enzyme's catalytic domain, the present invention obtains a substrate-specific isoprenyltransferase mutant by truncating the N-terminal hydrophobic amino acid, among other methods. The amino acid sequence of the mutant is shown in SEQ ID NO: 5. The gene encoding the mutant is recombined into an expression vector to obtain a recombinant expression vector, which is then transformed into the host cell Escherichia coli to ultimately produce an engineered bacterium capable of producing polyisoprene, thereby achieving heterologous synthesis of polyisoprene.

The application belongs to the technical field of organic synthesis, and relates to a synthesis method of vitamin K2 or a derivative thereof. The synthesis method comprises the following steps: obtaining a compound shown in formula I according to a reaction route shown in the following compound 1 and compound 2; wherein the compound 1 and the compound 2 are subjected to enzyme catalysis reaction (enzymatic reaction) to obtain a compound 3; the enzyme used in the enzyme catalysis reaction is one or more of isopentenyl transferase, geranylgeranyl transferase and heptaprenyl pyrophosphate transferase; and n is a natural number from 0 to 7. The synthesis method has the advantages of shortening a synthesis route, simplifying post-treatment operation, less pollution, mild conditions, high reaction efficiency, low production cost, high product purity and the like.