35 results about "Nucleoside diphosphate" patented technology

The invention discloses a co-catalytic reaction system for preparing rare ginsenosides. The co-catalytic reaction system comprises glycosyltransferase GT (a); sucrose synthase SUS (b); uridine diphosphate UDP (c); and saccharose (d). Experiments show that in the presence of the saccharose and little UDP, an enzyme combination composed of the glycosyltransferase and the saccharose is capable of replacing an in-vitro reaction system to synthesize UDP sugar, one of expensive materials for the rare ginsenosides, and efficiently and economically converting substrates such as protopanoxadiol or protopanaxatriol into the rare ginsenosides, wherein the UDP is about 1/4 of the UDP sugar in price and is 1% of its original dosage; thus, preparation cost of the rare ginsenosides is greatly saved, and large-scale commercial preparation of the ginsenosides is better facilitated.

The invention belongs to the field of chemical synthesis and in particular relates to a method for synthesizing dinucleoside diphosphate and dinucleoside triphosphote. The method comprises the following steps: reacting nucleoside phosphoryl piperidine as a nucleoside phosphoryl donor, nucleoside phosphate or nucleoside diphosphate as a nucleoside phosphoryl receptor and 4,5-dicyanoimidazole (DCI) as an activating reagent in a high-polarity non-protonized solvent; purifying crude products through ion exchange chromatography to obtain dinucleoside diphosphate and dinucleoside triphosphote respectively. According to the method, the selected and used nucleoside phosphoryl piperidine raw material is simple and easily available, the coupling reaction speed is high in the presence of the activating reagent, the yield is high, few byproducts are generated, and the products are easily separated.

The invention discloses a synthesis method of nucleoside phosphate, which mainly comprises the following steps: mixing lithium diphosphate, nucleoside and a condensing agent in a solvent, stirring atroom temperature, adding a citric acid solution for washing after the reaction is finished, drying with anhydrous sodium sulfate to obtain a crude product, and carrying out column chromatography separation to obtain the product. According to the method disclosed by the invention, ion exchange resin does not need to be used for treatment in the process of synthesizing the raw material phosphodiester, the reaction can be carried out by directly using the lithium diphosphate, the method disclosed by the invention is simple to operate and low in equipment requirement, and the same meaningful method is provided for synthesizing nucleoside diphosphate.

The invention discloses a nucleoside diphosphate glucosyltransferase gene (Ta-UGT) and coded protein thereof, and belongs to the gene engineering field. The cDNA sequence of the nucleoside diphosphate glucosyltransferase gene (Ta-UGT) is SEQ ID NO.1, the sequence of coded amino acid thereof is SEQ ID NO.2, and the gene is from wheat scab (triticum asetivum L.), which is reported for the first time in the wheat and induced by deoxynivalenol (DON) to strengthen expression. An experiment shows that the over expression of the gene can improve the resistibility of plants to DON. The Ta-UGT is introduced into the wheat variety easily infected by gibberellic disease, so as to expect to reduce the content of the DON in the wheat grain.

The invention discloses a method for synthesizing nucleoside triphosphate and nucleoside diphosphate from an all-protected nucleoside phosphoramidite intermediate through acid catalysis. The method comprises the following steps of: generating a phosphoramidite intermediate crude product under an alkaline condition by taking carbobenzoxy protected nucleoside and benzyloxydiisopropylamino phosphorochloridite as raw materials, subjecting the phosphoramidite intermediate crude product to weak acid catalytic hydrolysis to obtain a nucleoside- H-phosphonate intermediate crude product, subjecting the nucleoside-H-phosphonate intermediate crude product and alkylamine to oxidative coupling, carrying out column chromatography on the resulting product to obtain an all-protected nucleoside phosphoramidite precursor, subjecting the nucleoside phosphoramidite precursor to catalysis and hydrogenation in an N-N-dimethylfomamide solution to remove all protective groups, filtering the resulting product to remove palladium on an activated carbon, and reacting the resulting product with pyrophosphate or monophosphoric acid alkyl ammonium salt under a condition in the presence of a weak acid catalyst to obtain a nucleoside triphosphate product and a nucleoside diphosphate product. By adopting the method disclosed by the invention, reaction velocity and yield of the pyrophosphoric acid and monophosphoric acid reagents with the phosphoramidite intermediate are greatly improved, and an efficient, universal and novel method for the chemical synthesis of the nucleoside 5'- triphosphate and the nucleoside 5'-diphosphate is established.

The present invention includes modified Arabidopsis Nucleoside Di-Phosphate Kinase 2 (NDPK2) nucleic acid molecules whose enzymatic activity have been increased (i.e. hyperactive). NDPKs are ubiquitous housekeeping enzymes that catalyze the transfer of γ-phosphoryl group from a nucleoside triphosphate (NTP) to a nucleoside diphosphate (NDP), and also multifunctional proteins that regulate a variety of eukaryotic cellular activities, including cell proliferation, development, and differentiation. In plants, NDPKs are reported to play a key role in the signaling of both stress and light. Among three NDPKs (NDPK1, NDPK2, NDPK3) in a model plant, Arabidopsis thaliana, NDPK2 was reported as a positive signal transducer of phytochrome-mediated plant light signaling and to regulate cellular redox state, which enhances multiple stress tolerance in transgenic plants. Thus, the plants with the hyperactive NDPK2 are expected to possess higher efficiency of light utilization and enhanced tolerance to various environmental stresses such as cold, salt, and oxidative stresses. Since abiotic stress is one of the most important factors to limit the productivity of many crops, the hyperactive NDPK2 can be used for the development of high-yielding multiple stress tolerant plants with higher efficiency of light utilization. In this invention, several hyperactive NDPK2 were generated by C-terminal deletion and site-directed mutagenesis. Therefore, the present invention can be utilized to develop multiple stress tolerant and efficiently light-utilizing plants, which can eventually increase crop yields. The invention also includes plants having at least one cell expressing the modified NDPK2, vectors comprising at least one portion of the modified NDPK2 nucleic acids, and methods using such vectors for producing plants with enhanced light sensitivity and stress tolerance.

This invention relates to one method to filter liver cell cancer organism and side organism difference expression protein, which finds out one liver cell cancer organism expression protein, wherein, the immune experiment further proves G protein beta2 subunit in liver cell cancer organism different expression; The protein can be used as protein molecule label for liver cancer by the relative property.

The present invention relates to a method of increasing or facilitating the clearance of pulmonary mucus secretions in a subject. The present invention also relates to a method of facilitating hydration of pulmonary mucus secretions in a subject. The invention also relates to a method of preventing or treating a disease or condition associated with impaired lung or airway function in a human or other mammal. The method comprises administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of a P2Y6 receptor agonist compound, wherein said amount is effective to activate P2Y6 receptors on the luminal surface of the lung epithelium. The P2Y6 receptor agonist compounds used in the present invention include mononucleoside 5'-diphosphate, dinucleoside monophosphate, dinucleoside diphosphate or dinucleoside triphosphate of general formula I, or salts and solvates thereof , Hydrate.

The invention belongs to the field of chemical synthesis, and relates to a synthesis method of nucleoside diphosphate saccharide. The synthesis method comprises the following steps: 1) taking fully-protected nucleoside phosphoryl piperidine, removing all protecting groups through palladium carbon catalytic hydrogenation in an N,N-dimethylformamide or dimethylsulfoxide solution, and filtering to remove palladium and carbon to obtain corresponding deprotected nucleoside phosphoryl piperidine; 2) adding glycosyl-1-monophosphate and 4, 5-dicyanoimidazole (DCI) into the nucleoside phosphoryl piperidine, and reacting to obtain a nucleoside diphosphate saccharide product; 3) performing ethanol precipitation, reverse phase C18HPLC (high performance liquid chromatography) purification and final ion exchange on a crude product to obtain corresponding nucleoside diphosphate 6-deoxy-L-pyranose with high purity. By the synthesis method, the yield is high, the reaction time is short and the product separation is simple; therefore, the synthesis method has a high application value.

The invention discloses a method to get nucleic diphosphokinase A (NDPK-A) oxidation-reduction isomer through gene engineering in genetic engineering domain, which is characterized by the following: producing a amino acid abrupt change in NDPK-A; generating the oxidation-reduction isomer; abrupt-changing forth position cysteine to serine; getting amino acid abrupt change; setting the amino acid sequence as SEQ ID NO. 2. This isomer possesses stronger activity, which can increase activity of phosphate group transferase of NDPK and DNase (inhibit cancer).

The invention relates to ribonucleotide reductase from 'Bailing' producing strain cordyceps sinensis hirsutella sinensis for anabolism of deoxidized pyrimidine nucleoside diphosphate from pyrimidine nucleoside diphosphate, a gene coding the ribonucleotide reductase and application of the ribonucleotide reductase. The ribonucleotide reductase has over 90% of homology to the amino acid sequence shown by SEQ ID No.1 or SEQ ID No.3; and the coding gene has over 90% of homology to the nucleotide sequence shown by SEQ ID No.2 or SEQ ID No.4. According to the invention, the metabolic pathway for synthesizing deoxidized pyrimidine nucleoside diphosphate from pyrimidine nucleoside diphosphate is studied in details in principle, the cloning DNA (Deoxyribonucleic Acid) comprising the nucleotide sequence provided by the invention can be transferred into engineering bacteria through transduction, transformation and combined transfer, high expressivity of the host deoxidized pyrimidine nucleoside diphosphate is obtained by adjusting the expression of the deoxidized pyrimidine nucleoside diphosphate biosynthesis gene, an effective means of increasing output of the deoxidized pyrimidine nucleoside diphosphate is provided, and a significant application prospect is realized.

The present invention aims to provide a method for producing a polysaccharide with high efficiency using a polysaccharide synthase. The present invention provides a method for producing a polysaccharide, including allowing polysaccharide synthase (B) to act on ribonucleoside diphosphate-monosaccharide (A) shown below to produce a polysaccharide, wherein in 10 to 100% of the duration in which (B) acts on (A), the concentration of ribonucleoside diphosphate in a reaction solution is lower than 100 times an inhibitory concentration IC₅₀ described below against polysaccharide synthase (B).

Inhibitory concentration IC₅₀: a concentration of ribonucleoside diphosphate at which an enzyme activity of polysaccharide synthase (B) is reduced by half while under a condition where (B) has a concentration at which (B) acts on ribonucleoside diphosphate-monosaccharide (A), wherein ribonucleoside diphosphate-monosaccharide (A) is used as a substrate and ribonucleoside diphosphate is used as an inhibitor.

Ribonucleoside diphosphate-monosaccharide (A): a sugar nucleotide in which a proton of at least one hydroxyl group of at least one monosaccharide (a) selected from the group consisting of triose (a-1), tetrose (a-2), pentose (a-3), hexose (a-4), heptose (a-5), and monosaccharide (a-6) described below is substituted with a functional group represented by any one of chemical formulae (1) to (5) below.

Monosaccharide (a-6): a monosaccharide selected from the group consisting of (a-1), (a-2), (a-3), (a-4), and (a-5), in which at least one member selected from the group consisting of a proton, a hydroxyl group, and a hydroxymethyl group of the monosaccharide is substituted with substituent (E) described below.

Substituent (E) is at least one substituent selected from the group consisting of carboxyl, amino, N-acetylamino, sulfate, methylester, N-glycolyl, methyl, 1,2,3-trihydroxypropyl, phosphate, and 2-carboxy-2-hydroxyethyl groups.

The invention relates to an enzyme from Bailing production bacterium Cordyceps Chinese Hirsutella for synthesizing metabolic (desoxy) pyrimidine nucleoside triphosphate from (desoxy) pyrimidine nucleoside diphosphate, a gene for coding the enzyme and application thereof. The enzyme is nucleoside diphosphokinase which has more than 90% of homology with amino acid sequence disclosed as SEQ ID NO.1; and the coding gene has more than 90% of homology with nucleotide sequence disclosed as SEQ ID NO.2. The invention researches the metabolic pathway of (desoxy) pyrimidine nucleoside diphosphate synthesized (desoxy) pyrimidine nucleoside triphosphate in details in principle. The cloned DNA (deoxyribonucleic acid) comprising the nucleotide sequence provided by the invention can be transformed into engineering bacterium by transduction, conversion and conjugal transfer. By adjusting the expression of the (desoxy) pyrimidine nucleoside triphosphate biosynthesized gene, the host (desoxy) pyrimidine nucleoside triphosphate is endowed with high expressivity, thereby providing an effective way for enhancing the yield of the (desoxy) pyrimidine nucleoside triphosphate and having great application prospects.