32 results about "Prenyltransferase" patented technology

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 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

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 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 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 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.

Provided are a mutant cis-prenyltransferase (CPT) family protein and a method for producing a polyisoprenoid, which enable the production of a high molecular weight polyisoprenoid. Included is a mutant cis-prenyltransferase (CPT) family protein obtained by mutating the amino acid sequence of a C-terminal region of a cis-prenyltransferase (CPT) family protein not found on rubber particles to be identical or similar to the amino acid sequence of a C-terminal region of a cis-prenyltransferase (CPT) family protein found on rubber particles.

The application discloses a brewing yeast engineering strain for producing bakuchiol as well as a construction method and application thereof. The engineering strain overexpresses a gene in a geranyl pyrophosphate synthesis pathway, a gene in a p-coumaric acid synthesis pathway and a mutant isopentenyl transferase PcPT07 coding gene; the engineering strain has the ability to synthesize bakuchiol from glucose as a single carbon source; the classification and naming of the engineering strain is Saccharomyces cerevisiae BAK07, and the preservation number is CGMCC NO.31081. The application realizes the de novo synthesis of bakuchiol in the brewing yeast through overexpression of the gene in the geranyl pyrophosphate synthesis pathway, the gene in the p-coumaric acid synthesis pathway and the mutant isopentenyl transferase PcPT07 and combining metabolic engineering and other strategies, and lays a foundation for producing bakuchiol through microbial fermentation.

The invention discloses a protein with O-isopentenyl transferase activity, a related biological material and application, and belongs to the technical field of biology. According to the invention, an O-isopentenyl transferase gene (GlOPT) is screened and cloned, and the O-isopentenyl transferase can be prepared by using the gene. Experiments prove that the O-isopentenyl transferase can be used for catalyzing 8-hydroxybergamolactone in vitro to generate glehnia littoralis; the bergamot phenol is catalyzed to generate isoimperatorin; and the xanthotoxol is catalyzed to generate imperatorin. Technical support is provided for production of glehnia littoralis isopentenyl furocoumarin active ingredients, a foundation is laid for research on a biosynthesis route of coumarin compounds, and the method has important theoretical and practical significance.

The application provides a method for biosynthesis of tetraenemenaquinone by Yarrowia lipolytica. The method is achieved by constructing recombinant Yarrowia lipolytica, and the construction method of the recombinant Yarrowia lipolytica for synthesizing tetraenemenaquinone comprises the following steps: intensively expressing a mevalonate pathway in Yarrowia lipolytica, and introducing a heterologous synthesis pathway of geranylgeranyl pyrophosphate to provide sufficient isoprene side chain donors for the biosynthesis of tetraenemenaquinone; on the basis, overexpressing a codon-optimized aromatic isopentenyltransferase gene with menaquinone substrate specificity in Yarrowia lipolytica. The construction method of Yarrowia lipolytica for biosynthesis of tetraenemenaquinone provided by the application is simple in operation, the constructed Yarrowia lipolytica can obtain a high-concentration tetraenemenaquinone product by using commercially available menaquinone as a substrate, the biosynthesis steps are short, and the method has high production application value.

The disclosure relates to the biosynthesis of cannabinoids and related prenylated phenolic compounds using recombinant enzymes. In particular, the disclosure provides recombinant prenyltransferase enzymes engineered to produce a greater amount of a desired product, or to have a greater ability to catalyze a reaction using a desired substrate, as compared to the wild type prenyltransferase. The disclosure also provides methods of preparing such recombinant enzymes; as well as methods of use thereof in improving the biosynthesis of cannabinoids and related prenylated phenolic compounds.

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 belongs to the technical field of genetic engineering and enzymology, and particularly relates to isopentenyl transferase and application thereof to preparation of isopentenyl isoflavone. The isopentenyl transferase FoPT2 with the amino acid sequence shown as SEQ ID NO.2 is verified for the first time to show catalytic activity on 6 isoflavone compounds (genistein, dihydrogenistein, biochanin A, calycosin, genistin and 3 ', 4', 7-trihydroxy isoflavone), 21 isopentenyl products are generated in total, the structures of 10 products are analyzed, and 3 products are synthesized for the first time. The isopentenylation modification comprises 13 kinds of mono-isopentenylation (C- / O-connection) and 8 kinds of diisopentenylation (C-O connection / C-C connection), modification sites comprise C-6, 7-O, C-8, 4 '-O and the like, and wide regioselectivity is shown. By optimizing reaction conditions, the catalytic efficiency of FoPT2 is improved, the preparation of isopentenyl isoflavone with various structures is facilitated, and a new possibility is provided for isopentenyl isoflavone products.