54 results about "Prenyltransferase" patented technology

Many GTPases such as Ras, Ral and Rho require post-translational farnestylation or geranylgeranylation for mediating malignant transformation. Dual farnesyltransferase (FT) (FTI) and geranylgeranyltransferase-I (GGT-1) inhibitors (GGTI) were developed as anticancer agents from based on an ethylenediamine scaffold. On the basis of a 4-fold substituted ethylenediamine scaffold, the inhibitors are structurally simple and readily derivatized, facilitating extensive structure-activity relationship studies. The most potent inhibitor is compound exhibited an in vitro hFTase IC50 value of 25 nM and a whole cell H-Ras processing IC50 value of 90 nM. Several of the inhibitors proved highly selective for hFTase over the related prenyltransferase enzyme geranylgeranyltransferase-I (GGTase-I). A crystal structure of an inhibitor cocrystallized with farnesyl pyrophosphate in the active site of rat FTase illustrates that the para-benzonitrile moiety is stabilized by a π-π stacking interaction with the Y361β residue, suggesting an importance of this component of the inhibitors.

The invention discloses recombinant pichia pastoris (GS115-ppic9-novQ) preserved in CCTCC (China center for type culture collection) on March 11, 2016 with the preservation number of CCTCC M 2016105. The invention further discloses a construction method of the recombinant pichia pastoris and an application in production of aromatic prenyltransferase. By means of induction culture of the engineering bacterium, soluble recombinant aromatic prenyltransferase NovQ with biological activity can be directly obtained in supernatant of a fermentation liquid, and processes such as cell collection, cell disruption and the like are omitted, accordingly, the production process is simplified. Compared with enzymes from other sources, the aromatic prenyltransferase has the advantages that the activity does not depend on structures such as a living cell or a biological membrane, the substrate specificity is low, prenylation of multiple substrates can be catalyzed, the catalytic activity is high and the like.

Methods and compositions for modulating plant development are provided. Polynucleotide sequences and amino acid sequences encoding isopentenyl transferase (IPT) polypeptides are provided. The sequences can be used in a variety of methods including modulating root development, modulating floral development, modulating leaf and / or shoot development, modulating senescence, modulating seed size and / or weight, and modulating tolerance of plants to abiotic stress. Polynucleotides comprising an IPT promoter are also provided. The promoter can be used to regulate expression of a sequence of interest. Transformed plants, plant cell, tissues, and seed are also provided.

A microorganism having the reduction or loss of a 4-hydroxybenzoate-polyprenyltransferase activity or a 2-octapernylphenol 2-octaprenyl-6-methoxyphenol;flavin reductase activity and capable of producing lactic acid, particularly a microorganism having chromosomal DNA having the deletion of a part or the entirety of a gene encoding a protein having a 4-hydroxybenzoate-polyprenyltransferase activity or a protein having a 2-octapernylphenol 2-octaprenyl-6-methoxyphenol;flavin reductase activity; and a process for production of lactic acid using the microorganism.

Provided are a transformant produced by introducing a gene coding for a cis-prenyltransferase and a gene coding for a Nogo-B receptor, which are considered to be involved in polyisoprenoid biosynthesis, into a host to allow the host to express the cis-prenyltransferase and the Nogo-B receptor, and a method for producing a polyisoprenoid using the transformant. The present invention relates to a transformant produced by introducing a gene coding for a cis-prenyltransferase and a gene coding for a Nogo-B receptor into a host to allow the host to express the cis-prenyltransferase and the Nogo-B receptor.

Provided is a method for producing a polyisoprenoid, which can increase natural rubber production by enhancing the rubber synthesis activity of rubber particles. The present invention provides methods for producing a polyisoprenoid using a gene coding for a cis-prenyltransferase (CPT) family protein, a gene coding for a Nogo-B receptor (NgBR) family protein and a gene coding for a rubber elongation factor (REF) family protein, specifically a method for producing a polyisoprenoid in vitro using rubber particles bound to proteins coded for by these genes, and a method for producing a polyisoprenoid in vivo using a recombinant organism (plant) having these genes introduced therein.

The invention relates to a construction method and application of a genetically-engineered bacterium for expressing prenyltransferase ComQ, and belongs to the technical field of gene engineering and protein engineering. The method is mainly characterized by including the steps of designing a primer of the prenyltransferase ComQ, amplifying target genes of the prenyltransferase ComQ, constructing recombinant plasmid through an enzyme digestion and enzyme linking method, identifying the recombinant plasmid and constructing a genetically-engineered bacterial strain through chemical transformation. By means of the method, the genetically-engineered bacterial strain for expressing the prenyltransferase is constructed for biosynthesis of various aromatic compounds with the pharmacological activity and isopentenylated peptides compounds by means of the prenyltransferase, and the genetically-engineered bacteria are used for heavily expressing, separating and purifying the prenyltransferase; bymeans of the prenyltransferase which is obtained through separation and purification, isopentenylated treatment can be conducted on aromatic radicals in peptide compounds so that the method can be used for isopentenylated treatment of the aromatic compounds.

A novel PSL family prenyltransferase has relaxed substrate specificity, which can use a variety of cyclic dipeptides and prenyl donors as substrates to produce various terpenylated diketopiperazines. An amino acid sequence of the prenyltransferase is SEQ ID NO:1. An application of the prenyltransferase is transferring different prenyl groups to Trp-containing cyclic dipeptides. The prenyltransferase catalyzes the formation of terpenylated diketopiperazines by assembling prenyl groups onto cyclic di peptides, which provides a new strategy for drug development of diketopiperazines.

Methods and compositions for modulating plant development are provided. Polynucleotide sequences encoding isopentenyl transferase (IPT) polypeptides are provided, as are the amino acid sequences of the encoded polypeptides. The sequences can be used in a variety of methods including modulating root development, modulating floral development, modulating leaf and / or shoot development, modulating senescence, modulating seed size and / or weight, and modulating tolerance of plants to abiotic stress. Transformed plants, plant cells, tissues, and seed are also provided.

The invention discloses a method for catalytically synthesizing vitamin K2 through NovQ aromatic isopentenyl transferase. The method comprises the steps that 1, vitamin K3 is dissolved in an ethanol aqueous solution, NaBH4 is added, under the protection of N2, the temperature is controlled to be not more than 50 DEG C, stirring reaction is conducted till the system becomes light green transparent liquid, after neutralization is conducted till the pH is 4.0, extraction is conducted on a product with ethyl acetate, after concentration is conducted, freezing crystallization is conducted, clean liquid is filtered, and vitamin K3 hydroquinone is obtained; 2, vitamin K3 hydroquinone and DMAPP are added into an enzymatic reaction system, reaction is conducted under the closed and N2 protection conditions, after the reaction is completed, Fe<3+> is added, N2 protection is removed, after adequate oxidation is conducted, extraction is conducted with ethyl acetate, and the product is obtained. According to the method for catalytically synthesizing the vitamin K2 through the NovQ aromatic isopentenyl transferase, the core link adopts a biological enzyme catalytic engineering technology, and the cheap vitamin K3 is utilized for catalytically synthesizing high-value vitamin K2; the production technology is simple, the production cost is low, and the production process is green and environmentally friendly.

According to the isopentenyl transferase mutant and the method for producing cannabinoid phenol, existing isopentenyl transferase is subjected to site-directed mutagenesis, so that olive alcohol and geranyl pyrophosphate can be helped to catalyze to generate cannabinoid phenol with relatively high catalytic activity and purity, and the isopentenyl transferase mutant has relatively high production and application prospects.

The invention discloses degenerate primers for identifying hybrid isoprenoid-producing bacteria and application thereof, belonging to the technical field of biology. The sequences of the degenerate primers are disclosed as SEQ ID NO.1-SEQ ID NO.2. The invention also discloses a method for identifying hybrid isoprenoid-producing bacteria by using the degenerate primers. A direct PCR (polymerase chain reaction) method is utilized to identify hybrid isoprenoid-producing bacteria, thereby avoiding the defects of high complexity, time consuming, blindness, high leakage possibility and the like in the strain fermentation, fermentation product purification and identification and other processes in the traditional identification method, greatly enhancing the work efficiency, and having higher purposiveness. The degenerate primers have important meanings in discovering hybrid isoprenoid microbial natural product drugs and exploring novel prenyltransferase.

The invention discloses a plant expression vector without antibiotic marker genes and a construction method thereof. The T-DNA region of the vector contains a gene cassette which comprises CaMV 35S promoter, Xba I, Bam HI and XmaI restriction endonuclease enzyme sites, isopentenyl transferase (ipt) gene and nos terminator sequence. The construction method of the vector includes: using a binary vector pBI121 as a base, replacing gus gene with ipt gene, deleting npt II gene through Bsu 36 I and Xba I enzyme sites, reconnecting the CaMV 35S promoter to the T-DNA region. With the technical scheme provided by the invention, transgenic plants can be induced and regenerated on a culture medium with a proper amount of low-concentration cytokinin growth regulators, and the obtained transgenic plants are the transgenic plants expressed by the target genes, which can reduce steps for identifying whether the target gene is expressed, and can avoid potential risks of antibiotic marker genes.

A method for producing natural rubber by using a recombinant microorganism is disclosed, the method comprising: (a) manufacturing an expression vector capable of expressing a gene encoding a cis-prenyltransferase, which is a guayule-derived natural rubber synthetase represented by the amino acid sequence of SEQ ID NO: 2 or a Hevea brasiliensis-derived natural rubber synthetase represented by the amino acid sequence of SEQ ID NO: 6, and an expression vector capable of expressing a gene coding for a natural rubber precursor synthetase; (b) transforming a host microorganism with the expression vector; (c) culturing the transformed host microorganism; and (d) separating natural rubber from the cultured transformed host microorganism. The natural rubber obtained by the method has a white powder form identical to that of natural rubber, and shows an FT-IR spectrum pattern extremely similar to that of natural rubber.

The invention discloses a fermentation and one-step purification method for obtaining aromatic prenyl transferase. Through plasmid construction and the use of fusion tags, a large amount of soluble prenyltransferase NovQ can be obtained in the cytosol of cells, and then a highly active protein can be obtained after one-step purification, which improves the efficiency of protein acquisition. Co-expression of the fusion tag and the NovQ protein can obtain a high-purity enzyme protein without destroying the enzyme result. Compared with other prenyltransferase acquisition methods, the present invention has the advantages of high yield, simple enzyme extraction and purification methods, etc., and has wide application prospects in the in vitro and in vivo production processes of terpenoids.