33 results about "Isopentenyl pyrophosphate" patented technology

Methods for synthesizing isopentenyl pyrophosphate are provided. A first method comprises introducing into a host microorganism a plurality of heterologous nucleic acid sequences, each coding for a different enzyme in the mevalonate pathway for producing isopentenyl pyrophosphate. A related method comprises introducing into a host microorganism an intermediate in the mevalonate pathway and at least one heterologous nucleic acid sequence, each sequence coding for an enzyme in the mevalonate pathway necessary for converting the intermediate into isopentenyl pyrophosphate. The invention also provides nucleic acid sequences, enzymes, expression vectors, and transformed host cells for carrying out the methods.

Methods for synthesizing amorpha-4,11-diene synthase from isopentenyl pyrophosphate are provided. A first method comprises introducing into a host microorganism a plurality of heterologous nucleic acid sequences, each coding for a different enzyme in the mevalonate pathway for producing isopentenyl pyrophosphate. Amorpha-4,11-diene synthase is then produced using an optimized amorpha-4,11-diene synthase gene. The invention also provides nucleic acid sequences, enzymes, expression vectors, and transformed host cells for carrying out the methods.

Methods for synthesizing isopentenyl pyrophosphate are provided. A first method comprises introducing into a host microorganism a plurality of heterologous nucleic acid sequences, each coding for a different enzyme in the mevalonate pathway for producing isopentenyl pyrophosphate. A related method comprises introducing into a host microorganism an intermediate in the mevalonate pathway and at least one heterologous nucleic acid sequence, each sequence coding for an enzyme in the mevalonate pathway necessary for converting the intermediate into isopentenyl pyrophosphate. The invention also provides nucleic acid sequences, enzymes, expression vectors, and transformed host cells for carrying out the methods.

A unique carotenogenic biosynthetic gene cluster has been isolated from Panteoa agglomerans strain DC404, wherein the genetic organization of the cluster is crtE-idi-crtY-crtI-crtB-crtZ. The genes contained within this cluster encode geranylgeranyl pyrophosphate (GGPP) synthetase (CrtE), isopentenyl pyrophosphate isomerase (Idi), lycopene cyclase (CrtY), phytoene desaturase (CrtI), phytoene synthase (CrtB), and β-carotene hydroxylase (CrtZ). The gene cluster, genes and their products are useful for the conversion of farnesyl pyrophosphate to carotenoids. Vectors containing those DNA segments, host cells containing the vectors and methods for producing those enzymes by recombinant DNA technology in transformed host organisms are disclosed.

Methods of treatment and prophylaxis of various diseases and disorders, and in particular diseases and disorders of lipid and bone metabolism, involving the administration of prenyl transferase (farnesyl pyrophosphate synthase) and / or isopentenyl pyrophosphate isomerase inhibitor compounds are disclosed.

Nucleic acid sequences encoding ∈-cyclase, isopentenyl pyrophosphate isomerase and beta-carotene hydroxylase as well as vectors containing the same and hosts transformed with the vectors. Methods for controlling the ratio of various carotenoids in a host and for the production of novel carotenoid pigments. The present invention also provides a method for screening for eukaryotic genes encoding carotenoid biosynthesis, and for modifying the disclosed enzymes.

The present invention provides recombinant DNA comprising a transcription promoter and a downstream sequence to be expressed, in operable linkage therewith, wherein the transcription promoter comprises a region found upstream of the open reading frame of a highly expressed Phaffia gene, preferably a glycolytic pathway gene, more preferably the gene coding for Glyceraldehyde-3-Phosphate Dehydrogenase. Further preferred recombinant DNAs according to the invention contain promoters of ribosomal protein encoding genes, more preferably wherein the transcription promoter comprises a region found upstream of the open reading frame encoding a protein as represented by one of the amino acid sequences depicted in any one of SEQIDNOs: 24 to 50. According to a further aspect of the invention an isolated DNA sequence coding for an enzyme involved in the carotenoid biosynthetic pathway of Phaffia rhodozyma is provided, preferably wherein said enzyme has an activity selected from isopentenyl pyrophosphate isomerase activity, geranylgeranyl pyrophosphate synthase activity, phytoene synthase activity, phytoene desaturase activity and lycopene cyclase activity, still more preferably those coding for an enzyme having an amino acid sequence selected from the one represented by SEQIDNO: 13, SEQIDNO: 15, SEQIDNO: 17, SEQIDNO: 19, SEQIDNO: 21 or SEQIDNO: 23. Further embodiments concern vectors, transformed host organisms, methods for making proteins and / or carotenoids, such as astaxanthin, and methods for isolating highly expressed promoters from Phaffa.

The invention discloses a method for preparing farnesene by using a biodiesel by-product. The method comprises the steps of purification of the biodiesel by-product, wherein the purity of the purified glycerin reaches 63.8% (w / w), building of a recombinant host cell for preparing farnesene from the purified glycerin as the raw material, wherein the recombinant host cell comprises related genes in a mevalonate pathway or deoxymutulose-5-hexose monophosphate pathway, an isopentene pyrophosphate isomerase gene, a farnesene isopentenyl pyrophosphate synthetase gene and a farnesene synthetase gene. The method also comprises fermentation of the purified glycerin as the raw material in the recombinant host cell to prepare the farnesene, wherein the concentration of the obtained farnesene is 0.025g / L; and optimum construction of the recombinant host cell capable of improving the yield of the farnesene by employing the purified glycerin as the raw material, wherein the concentration of the obtained farnesene reaches 2.9g / L, and the concentration is improved by 115 times in comparison with that before optimization.

The invention relates to a wolfberry lycopene epsilon-cyclase gene and a recombinant vector comprising the gene. The gene is selected from one of the following nucleotide sequences: 1) a nucleotide sequence shown in a sequence SEQ ID NO.1; and 2) a homologous sequence or an allelic gene thereof and a derivative nucleotide sequence thereof in which 70% of one or more nucleotides are added, replaced, inserted or deleted in the nucleotide sequence shown in the sequence SEQ ID NO. 1. By extracting the total RNA of fresh wolfberry leaves, a wolfberry lycopene epsilon-cyclase gene Lm LcyE is cloned by using a 3'RACE technology, and an obtained complete genome sequence is 1569. An escherichia coli expression vector pET28a-LmLcyE is constructed, the enzymatic activity of cloned wolfberry isopentenyl pyrophosphate isomerase gene Lm LcyE encoding is identified by using an escherichia coli heterologous expression system, and the wolfberry lycopene epsilon-cyclase gene Lm LcyE can catalyze lycopene for cyclizing the lycopene into delta carotene, so that metabolism flows downstream, thereby improving the content of alpha-carotenoid.

The invention provides for a method for producing a 5-carbon alcohol in a genetically modified host cell. In one embodiment, the method comprises culturing a genetically modified host cell which expresses a first enzyme capable of catalyzing the dephosphorylation of an isopentenyl pyrophosphate (IPP) or dimethylallyl diphosphate (DMAPP), such as a Bacillus subtilis phosphatase (YhfR), under a suitable condition so that 5-carbon alcohol is 3-methyl-2-buten-1-ol and / or 3-methyl-3-buten-1-ol is produced. Optionally, the host cell may further comprise a second enzyme capable of reducing a 3-methyl-2-buten-1-ol to 3-methyl-butan-1-ol, such as a reductase.

The invention discloses a genetically engineered bacterium with gamma-terpinene synthesis capacity and a constructing method and application thereof, and belongs to the technical field of genetic engineering. According to engineered escherichia coli, a path for synthesizing MVA from mevalonic acid is reconstructed by heterogeneous expression of hydroxymethylglutaryl coenzyme A synthetase mvaS, hydroxymethylglutaryl coenzyme A reductase mvaE, mevalonate kinase Erg12, phosphomevalonate kinase Erg8, mevalonate pyrophosphate decarboxylase Erg19 and Isopentenyl diphosphate isomerase Idi1, meanwhile, geranyl pyrophosphate synthetase GPPS2 and gamma-terpinene TPS2 are led into a bacterium body, and by utilizing the biotransformation capacity of the genetically engineered bacterium, gamma-terpinene can be efficiently produced in an environment-friendly mode. Through fermentation with the genetically engineered bacterium, the output of gamma-terpinene can reach 80 mg / L. The genetically engineered bacterium and the gamma-terpinene synthesis method are suitable for practical industrial production.

The present invention relates to a TRPV3 (transient receptor potential vanilloid 3) activity inhibitor, more precisely to a method for inhibiting TRPV3 activity including the step of treating isopentenyl pyrophosphate and a method for treating skin disease containing the step of administering isopentenyl pyrophosphate to a subject with skin disease or applying the same on the skin of the subject. Isopentenyl pyrophosphate of the present invention controls increase of sensory cell reactivity to current or migration and proliferation of skin cells induced by TRPV3, so that it can be effectively used for the development of a pain reliever or a therapeutic agent for skin disease.

Novel proteins of microorganism E-396 (FERM BP-4283) and the DNA sequences which encode these proteins have been discovered to provide an improved biosynthetic pathway from farnesyl pyrophosphate and isopentyl pyrophosphate to various carotenoids, especially zeaxanthin, astaxanthin, adonixanthin and canthaxanthin.

The production of carotenoid is accomplished using a DNA molecule that encodes a polypeptide as obtained from Haematococcus pluvialis, Phaffia rhodozyma, or Saccharomyces cerevisiae, having isopentonyl pyrophosphate (IPP) isomerase activity, or DNA molecule having a nucleotide sequence that hybridizes thereto. In particular, one can introduce such a DNA molecule into a carotenoid-producing microorganism, culture the microorganism thus transformed, and then obtain carotenoids in the culture broth and cells.

The invention relates to a saffron, a saffron endophytic fungi GGPPS (geranylgeranyl pyrophosphate synthase) gene, a gene cloning method and application, and belongs to the technical field of gene engineering. The invention discloses two genes including the saffron and GGPPS in metabolic pathway of the saffron endophytic fungi; the two genes have nucleotide sequences shown as SEQ ID NO.4 and SEQ ID NO.3, have the identical open reading frames shown as SEQ ID NO.1 and code amino acid sequences shown as code SEQ ID NO.2. The color genetic complementary reaction verifies that the GGPPS gene coding protein has a typical GGPPS enzyme function; FPP (farnesyl pyrophosphate) and IPP (isopentenyl pyrophosphate) can be catalyzed to be synthesized into the GGPPS (C<20>).

The invention discloses a genetically engineered bacterium for synthesizing D-limonene, and a construction method and application of the genetically engineered bacterium, and relates to the field of genetic engineering and fermentation engineering. In the genetically engineered bacterium, an HMG-CoA synthetase gene, an acetyl CoA acetylase mvaE, a GPPS (geranyl pyrophosphate synthase) gene, a D-limonene synthetase gene C1L2, an MVA kinase ERG12 gene, an MVAP kinase ERG8 gene, a mevalonic acid decarboxylase ERG19 gene and an isopentenyl pyrophosphate isomerase IDI gene are subjected to overexpression. In an LB culture medium, after flask shaking fermentation is carried out, the D-limonene can be subjected to heterologous synthesis, and the yield of the D-limonene is 27.3mg / L of fermentationliquor. The genetically engineered bacterium can be used for the industrial production of the D-limonene.

The invention relates to a construction method and a process of engineering bacteria for producing amorpha-4,11-diene. According to the strain for efficiently producing an artemisinin precursor amorpha-4,11-diene, recombinant bacteria are obtained by regulating the expression / or the activity of escherichia coli and an exogenous gene, wherein the gene comprises an acetoacetyl coenzyme A thiolase atoB gene, an HMG-CoA synthase hmgS gene, an HMG-CoA reductase hmgR gene, a mevalonate kinase mevK gene, a phosphohydroxyproline kinase pmk gene, a mevalonic acid-5-pyrophosphoric acid decarboxylase pmdgene, an isopentenyl pyrophosphate isomerase idi gene, a farnesyl pyrophosphate synthase ispA gene and an amorpha-4,11-diene synthase ads gene.

The invention discloses a method for producing isoprene by in vitro enzymatic reaction and an application of isoprene and belongs to the technical field of biological engineering. The method disclosed by the invention comprises the following steps of respectively introducing an obtained mevalonate kinase gene, a phosphomevalonate kinase gene, a mevalonate-pyrophosphate decarboxylase gene, an isopentenyl pyrophosphate isomerase gene and an isoprene synthase into host bacteria, carrying out fermental cultivation to obtain five recombinant bacteria, respectively carrying out ultrasonic disruption on each thallus to obtain a crude enzyme solution, mixing five crude enzyme solutions to obtain a reaction enzyme solution and adding a reaction substrate solution into the reaction enzyme solution to in vitro synthesize isoprene. By the method disclosed by the invention, the in-vitro synthesis of isoprene from mevalonic acid by virtue of enzymatic reaction is achieved and the synthesis of isoprene by precisely in vitro controlling the content of metabolic enzyme is realized.

The present invention provides recombinant DNA comprising a transcription promoter and a downstream sequence to be expressed, in operable linkage therewith, wherein the transcription promoter comprises a region found upstream of the open reading frame of a highly expressed Phaffia gene, preferably a glycolytic pathway gene, more preferably the gene coding for Glyceraldehyde-3-Phosphate Dehydrogenase. Further preferred recombinant DNAs according to the invention contain promoters of ribosomal protein encoding genes, more preferably wherein the transcription promoter comprises a region found upstream of the open reading frame encoding a protein as represented by one of the amino acid sequences depicted in any one of SEQIDNOs: 24 to 50. According to a further aspect of the invention an isolated DNA sequence coding for an enzyme involved in the carotenoid biosynthetic pathway of Phaffia rhodozyma is provided, preferably wherein said enzyme has an activity selected from isopentenyl pyrophosphate isomerase activity, geranylgeranyl pyrophosphate synthase activity, phytoene synthase activity, phytoene desaturase activity and lycopene cyclase activity, still more preferably those coding for an enzyme having an amino acid sequence selected from the one represented by SEQIDNO: 13, SEQIDNO: 15, SEQIDNO: 17, SEQIDNO: 19, SEQIDNO: 21 or SEQIDNO: 23. Further embodiments concern vectors, transformed host organisms, methods for making proteins and / or carotenoids, such as astaxanthin, and methods for isolating highly expressed promoters from Phaffia.

The present invention relates to an isolated DNA sequence encoding an enzyme in the mevalonate pathway or the pathway from isopentenyl pyrophosphate to farnesyl pyrophosphate. Vectors and plasmids including such DNA, are also set forth. The invention also includes host cells transformed by such DNAs, or vectors or plasmids containing such DNAs. A process for the production of isoprenoids and carotenoids using such transformed host cells is also provided.

The invention discloses recombinant saccharomyces cerevisiae for producing alpha-cubene and application of the recombinant saccharomyces cerevisiae, and belongs to the field of metabolic engineering and bioengineering. According to the recombinant saccharomyces cerevisiae strain for producing alpha-cubene, saccharomyces cerevisiae is taken as a host, alpha-cubene synthase is subjected to heterologous expression, and farnesyl pyrophosphate synthase and isopentenyl pyrophosphate isomerase are subjected to overexpression. The recombinant saccharomyces cerevisiae strain for producing the alpha-cubene is obtained by transferring recombinant plasmids capable of expressing an alpha-cubene synthase gene, a farnesyl pyrophosphate synthase gene and an isopentenyl pyrophosphate isomerase gene into saccharomyces cerevisiae. According to the invention, the exogenous gene is introduced into the saccharomyces cerevisiae to produce the alpha-cubene, and petroleum resources are not required to be used for chemical synthesis, so that the consumption of the petroleum resources and the pollution to the environment are reduced, and the sustainable development is realized. The alpha-cubene can be efficiently produced, and the application of the alpha-cubene in the fields of food, medicine, agriculture, tobacco and the like is promoted.

The invention relates to an isopentenyl diphosphate isomerase gene and applications thereof. A technical problem that efficiency of synthesis of terpenoids using engineering bacteria is low is solved. The invention provides the isopentenyl diphosphate isomerase gene, proteins obtained via expression of the isopentenyl diphosphate isomerase gene, and preparation method and applications of a prokaryotic expression vector containing the isopentenyl diphosphate isomerase gene and engineering bacteria capable of producing terpenoids. The isopentenyl diphosphate isomerase gene can be widely applied in the field of terpenoid preparation.

The invention discloses a method for constructing high-yield coenzyme Q10 engineering bacteria and an application of the method, belongs to the technical field of genetic engineering, and solves the problem of low yield by an existing coenzyme Q10 production method. The method disclosed by the invention comprises the following steps of performing extraction to obtain genome DNA from a gene parentstrain containing coding isopentenyl diphosphate isomerase IDI; amplifying out a homologous gene of the isopentenyl diphosphate isomerase IDI through a polymerase chain reaction; enabling the amplified homologous gene to be connected with a plasmid vector, and constructing a recombinant carrier; and guiding the recombinant carrier into a host cell so as to obtain target engineering bacteria. The invention provides an application of the method for constructing the high-yield coenzyme Q10 engineering bacteria to production of a coenzyme Q10. According to the method disclosed by the invention, through constructing the isopentenyl diphosphate isomerase IDI overexpressed by engineering bacteria, precursor substances synthesized by the coenzyme Q10 are increased, the synthesis capacity of the coenzyme Q10 can be greatly increased, the yield of the coenzyme Q10 is notably increased, and the method can be used for large-scale industrial production of the coenzyme Q10.

The invention relates to a Platycodon grandiflorum geranyl geranyl pyrophosphate synthase gene PgGGPPS and an encoding product and application thereof. The encoding gene (PgGGPPS) of the Platycodon grandiflorum geranyl geranyl pyrophosphate synthase is cloned from the root of Platycodon grandiflorum, and the encoding gene (PgGGPPS) can be applied to a path for preparing geranyl geranyl pyrophosphate by taking DMAPP and IPP as substrates. The technology can be used for subsequently producing a large amount of geranyl and geranyl pyrophosphate through a bacterial system, and an effective method is provided for meeting the huge market demand faced by terpenoid components. By utilizing the gene, the content of the platycodon grandiflorum terpenoids can be increased through a genetic engineering technology.

The invention discloses a genetically engineered bacterium for co-producing geraniol and nerol, its construction method and application, and belongs to the technical field of genetic engineering. The present invention expresses acetyl-CoA acyltransferase / hydroxymethylglutaryl-CoA reductase, 3-hydroxy-3-methylglutaryl-CoA synthase, mevalonate kinase, formazan Valonate-5-phosphate kinase, mevalonate-5-diphosphate decarboxylase, isopentenyl pyrophosphate isomerase, geranyl diphosphate synthase, and geraniol synthase or phosphatase. The invention utilizes genetic engineering means to successfully establish a metabolic pathway for synthesizing geraniol and nerol in Escherichia coli, and can directly bioconvert glucose into geraniol and nerol.

The invention discloses a method for preparing farnesene by using a biodiesel by-product. The method comprises the steps of purification of the biodiesel by-product, wherein the purity of the purified glycerin reaches 63.8% (w / w), building of a recombinant host cell for preparing farnesene from the purified glycerin as the raw material, wherein the recombinant host cell comprises related genes in a mevalonate pathway or deoxymutulose-5-hexose monophosphate pathway, an isopentene pyrophosphate isomerase gene, a farnesene isopentenyl pyrophosphate synthetase gene and a farnesene synthetase gene. The method also comprises fermentation of the purified glycerin as the raw material in the recombinant host cell to prepare the farnesene, wherein the concentration of the obtained farnesene is 0.025g / L; and optimum construction of the recombinant host cell capable of improving the yield of the farnesene by employing the purified glycerin as the raw material, wherein the concentration of the obtained farnesene reaches 2.9g / L, and the concentration is improved by 115 times in comparison with that before optimization.