14 results about "Uridine phosphorylase" patented technology

The invention provides a uracil production strain and a construction method and application thereof.According to the strain, on an E.coli UR14 genome by means of a CRIPSR / Cas9 gene editing technology, firstly, psuG genes, preTA genes, rutA genes and upp genes are knocked out, so that decomposition of uracil is blocked; then, a uridine phosphorylase gene udp and a pyrimidine-5 '-nucleotide nucleotidase gene ppnN are subjected to overexpression, and synthesis and accumulation of uracil are synergistically enhanced; and finally, overexpression of the ribose phosphate mutase gene pgm further enhances the conversion of a by-product ribose phosphate 1-precursor 5-ribose phosphate 1-pyrophosphate and improves the carbon utilization rate, and the obtained strain has good genetic stability and high fermentation yield, can stably produce uracil, and has wide application prospects.

Methods of treating a subject for a liver condition, e.g., NAFLD, NASH, and / or DILI, are provided. Aspects of the methods include administering to the subject an effective amount of a UPase inhibitor, optionally in combination with a uridine active agent (e.g., uridine (UR), a UR pro-drug or a UR mimetic), such as supplemental uridine, to treat the subject for the liver condition. Also provided are compositions for use in practicing the subject methods.

The present invention provides a method and composition for the enzyme-catalyzed synthesis of nucleosides containing protecting groups. The method includes using pyrimidine nucleoside phosphorylase or uridine phosphorylase and purine nucleoside phosphorylase to catalyze the synthesis of nucleosides containing protecting groups from a substrate, the substrate including a substrate nucleoside, a substrate base, and a substrate phosphate, the substrate base containing a protecting group, the pyrimidine nucleoside phosphorylase including proteins having 80% or more identity to proteins P and NP shown in SEQ ID NO: 1, the uridine phosphorylase including proteins having 80% or more identity to protein UP shown in SEQ ID NO: 2, and the purine nucleoside phosphorylase including proteins having 80% or more identity to protein PNP shown in SEQ ID NO: 3, 7, or 8. This method solves the problem in the prior art that there is no biosynthetic method for producing nucleosides containing protecting groups, and is suitable for the field of enzyme catalysis.

The invention provides application of uridine phosphorylase UP in screening of drugs for preventing and / or treating brain tumors. The invention also provides a reagent for reducing the expression of uridine phosphorylase UP. And / or a reagent for reducing the activity or function of uridine phosphorylase UP is used for preparing a medicine for preventing and / or treating brain tumors.

The invention relates to a nucleotide PET molecular probe targeting uridine phosphorylase as well as a preparation method and application of the nucleotide PET molecular probe. The PET molecular probe comprises a compound [< 18 > F] Uridine or [< 18 > F] Tripolyridine which is marked by uracil nucleosides < 18 > F. Compared with the prior art, the PET molecular probe prepared by the invention has a positioning diagnosis effect on tumor uridine metabolism change, and simultaneously realizes tumor metabolism image typing and pathological evaluation after tumor treatment.

The invention discloses an enzyme composition, an immobilized enzyme and application. The enzyme composition consists of uridine phosphorylase and thymine phosphorylase. The enzyme composition and the immobilized enzyme are simple to prepare, have the characteristics of high conversion efficiency, high yield and simplicity and convenience in operation when being used for synthesizing 2 '-fluoro-2'-deoxycytidine, and can effectively reduce the production cost. Meanwhile, synthetic reaction conditions of the 2 '-fluoro-2'-deoxycytidine based on the enzyme composition and the immobilized enzyme are mild, green and environment-friendly, and the environmental problem caused by chemical synthesis can be avoided.

This application discloses the application of uridine phosphorylase 1 (UPP1) as an inhibitory target in the preparation of drugs for treating systemic lupus erythematosus (SLE), relating to the field of biomedical technology. This application is the first to discover abnormally elevated UPP1 expression in peripheral blood mononuclear cells of SLE patients, which is positively correlated with disease activity. Validation experiments demonstrate that the specific inhibitor of UPP1, BAU, can effectively inhibit UPP1 enzyme activity. In cell models, it reduces inflammatory factors and type I interferon responses induced by STING pathway activation. In SLE animal models, BAU significantly reduces splenomegaly, proteinuria, and pathological damage to the kidneys and lungs, decreases the proportion of pathogenic immune cells such as M1 macrophages, Th1 cells, and plasma cells in the spleen, and inhibits the deposition of immune complexes in the glomeruli. This invention reveals for the first time the potential of UPP1 as a novel therapeutic target for SLE and provides BAU as a candidate drug for treating SLE and related organ damage, offering a novel strategy for clinical intervention.

The invention relates to the field of biochemical detection, and discloses a method for detecting the content of cytidine in fermentation liquor based on an enzyme method and high flux, and the method comprises the following steps: providing a first reagent containing uridine phosphorylase, dihydropyrimidine dehydrogenase, glucose-6-phosphate dehydrogenase and related coenzyme substrates, and a second reagent containing cytidine deaminase. During detection, a sample and a first reagent are mixed for background absorbance monitoring; then a second reagent is added to start a specific reaction, cytidine is catalyzed into uridine, the uridine consumes the reduced coenzyme through a cascade enzyme reaction, and meanwhile, the coenzyme is cyclically amplified by a regeneration system. The cytidine content is determined by monitoring the decline rate of the specific wavelength absorbance. By adopting a two-step sample adding method, the interference of a sample matrix is effectively deducted, and the detection accuracy is improved; by utilizing the design of coenzyme regeneration cycle, a detection signal is amplified, and the sensitivity is improved; by combining the automatic operation of the porous microplate, high-throughput detection is realized, and the efficiency is improved.

The present invention provides engineered uridine phosphorylase (UP) enzymes, polypeptides having UP activity, and polynucleotides encoding these enzymes, as well as vectors and host cells comprising these polynucleotides and polypeptides. Methods for producing UP enzymes are also provided. The present invention further provides compositions comprising the UP enzymes and methods of using the engineered UP enzymes. The present invention finds particular use in the production of pharmaceutical compounds.

The invention relates to a method of producing cladribine with a purity greater than 99.9 wt.% from 2-deoxyuridine and 2-chloroadenine by using the catalytic action of high concentrations (at least 20.000 U / L) of freely dissolved transglycosidases UP (uridine phosphorylase) and PNP (purine nucleoside phosphorylase) in an enzymatic synthesis in an aqueous environment without the presence of any organic solvent.

This invention provides a uracil-producing strain, its construction method, and its application. The strain utilizes CRIPSR / Cas9 gene editing technology to produce... E. coli On the UR14 genome, firstly by knocking out psuG Gene, black Gene, rutA Genes and upp Genes were introduced to block the breakdown of uracil; then, the uridine phosphorylase gene was overexpressed. udp and pyrimidine-5'-nucleotide nuclease gene ppnN This synergistically enhances the synthesis and accumulation of uracil; finally, overexpression of the phosphoribosyltransferase gene... pgm, Further enhancing the conversion of the byproduct 1-phosphate ribose into the precursor 5-phosphate ribose-1-pyrophosphate improves carbon utilization. The resulting strain exhibits good genetic stability and high fermentation yield, enabling stable production of uracil and demonstrating broad application prospects.

The invention describes a method of the enzymatic synthesis of cordycepin and its 2-halogen derivatives by reacting 3 -deoxyuridine with adenine or its 2-halogen derivatives in an aqueous phosphate reaction medium under the catalytic action of uridine phosphorylase (UP) and purine nucleoside phosphorylase (PNP). The synthetic method according to the invention is fast, technologically and energetically undemanding and environmentally friendly, and does not require the presence of organic solvents or chromatographic purification of pure cordycepin, while being easily transferable to laboratory and mass production conditions.

The application discloses a method for enhancing 5'-cytidylic acid accumulation and application, and belongs to the field of genetic engineering and microbial engineering. The application knocks out ppnN, ushA, yrfG, yjjG, umpH and umpG genes, overexpresses 5'-CTP diphosphohydrolase gene nudG and orotidine-5'-phosphate ribosyltransferase gene pyrE, integrates expression of uridine phosphorylase mutant gene PyrH(R92G / D93G), further knocks out cytidine deaminase gene cdd and ribonucleoside-triphosphate reductase gene nrdD, and integrates expression of uridine-cytidine kinase gene udk, phosphoribosyl pyrophosphate synthetase gene prs, glucose-6-phosphate dehydrogenase gene zwf and 6-phosphogluconate dehydrogenase gene, so that the 5'-cytidylic acid accumulation capacity of Escherichia coli is improved. Under the condition that no antibiotic and inducer are added, the 5'-cytidylic acid yield of the recombinant bacteria reaches 39.6 g / L in a 60L fermenter.

The invention describes a method of the enzymatic synthesis of cordycepin and its 2-halogen derivatives by reacting 3 -deoxyuridine with adenine or its 2-halogen derivatives in an aqueous phosphate reaction medium under the catalytic action of uridine phosphorylase (UP) and purine nucleoside phosphorylase (PNP). The synthetic method according to the invention is fast, technologically and energetically undemanding and environmentally friendly, and does not require the presence of organic solvents or chromatographic purification of pure cordycepin, while being easily transferable to laboratory and mass production conditions.