38 results about "Nucleotidase" patented technology

The invention discloses a genetically engineered bacterium for producing uridine diphosphate glucose as well as a construction method and application of the genetically engineered bacterium, and belongs to the technical field of genetic engineering. The following genes are overexpressed in the genetically engineered bacterium: a pyrimidine nucleoside operon gene, a carbamyl phosphate synthetase gene, an aspartate carbamyl transaminase gene, an orotic acid phosphoribose transferase gene, a phosphoribose pyrophosphate synthase gene, a uridylate kinase gene and a glucophosphate mutase gene. A uridine triphosphate glucose-1-phosphate uridine group transferase gene; the genetically engineered bacterium does not express the following genes: a purine nucleotide repressor protein gene, a deoxyribonucleoside triphosphate reductase gene, a nucleoside triphosphate pyrophosphohydrolase gene, a cytidine triphosphate synthase gene, a pyrimidine nucleotidase gene, a nucleotidase gene, an ornithine carbamoyltransferase gene, a UDPG dehydrogenase gene and a lacI repressor protein gene. The genetically engineered bacterium can be used for efficiently synthesizing UDPG by taking glucose as a substrate.

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.

The invention relates to the technical production field of metabolic engineering and genetic engineering, and particularly provides a genetically engineered bacterium for producing 2 '-deoxyuridine as well as a construction method and application of the genetically engineered bacterium. According to the strain, deoxyuridine kinase tdk is knocked out on an E.coli W3110 genome, and T7 RNA polymerase is integrated; an aspartate transaminase gene aspC, a glutamine synthetase gene glnA, a 5 '-deoxynucleotidase gene yfbR and a uridine monophosphate kinase gene pyrH are subjected to overexpression; the method comprises the following steps: introducing a ribonucleoside diphosphate reductase gene nrdBCA from Escherichia phage T4 in a heterologous series connection manner; the CTP synthase gene pyrG * of Corynebacterium glutamicum is introduced in a heterologous mode, and the CTP synthase gene pyrG * of Corynebacterium glutamicum is introduced in a heterologous mode. The safe, efficient and low-cost production of the 2 '-deoxyuridine is realized.

Compounds that modulate the conversion of AMP to adenosine by 5′-nucleotidase, ecto, and compositions containing the compounds and methods for synthesizing the compounds, are described herein. The use of such compounds and compositions for the treatment and / or prevention of a diverse array of diseases, disorders and conditions, including cancer- and immune-related disorders, that are mediated by 5′-nucleotidase, ecto is also provided.

Compounds that modulate the conversion of AMP to adenosine by extracellular 5'-nucleotidase, compositions containing the compounds, and methods for synthesizing the compounds are described herein. Also provided are uses of such compounds and compositions for treating and / or preventing a variety of diseases, disorders, and conditions (including cancer and immune-related disorders) mediated by extracellular 5'-nucleotidase.

The invention discloses an escherichia coli recombinant bacterium for whole-cell catalytic synthesis of rebaudioside M as well as a construction method and application of the escherichia coli recombinant bacterium. The Escherichia coli recombinant bacterium for the whole-cell catalytic synthesis of the rebaudioside M, which is constructed by the invention, contains a glycosyl transferase UGT76G1 coding gene mutant, a glycosyl transferase UGT91C1 coding gene mutant, a sucrose synthase atSuS1 gene and a polyphosphoglucokinase cgPPGK gene; a glucose-1-phosphate uridine transferase galU gene, a nucleoside diphosphate kinase ndk gene and a nucleoside diphosphate kinase ndk gene are over-expressed; meanwhile, an encoding gene araA gene of L-arabinose isomerase and an encoding gene ushA gene of 5 '-nucleotidase / UDG hydrolase are knocked out. According to the recombinant escherichia coli constructed by the invention, Reb D is prevented from being directly used as a synthetic substrate, so that the raw materials are easier to obtain, and the conversion rate of the substrate and the synthetic rate and yield of rebaudioside M are remarkably improved.

Provided herein, inter alia, are methods for treating, delaying and / or reducing the severity of conditions associated with age-related decline of various physiological pathways, including mitochondrial health and associated physical states, by administration of compositions that can degrade extracellular ATP (eATP), including NTPDase enzymes, nucleotidases such as apyrases and members of the GDA1_CD39 superfamily, phosphatases and ATP degrading enzyme producing yeast strains.

The present invention discloses the use of a nonspecific 5'-nucleotidase in the preparation of nicotinamide riboside. The nonspecific 5'-nucleotidase is selected from a nonspecific 5'-nucleotidase derived from Shewanella sp. HN-41. The nonspecific 5'-nucleotidase of the present invention can efficiently catalyze the hydrolysis of high-concentration nicotinamide mononucleotide (NMN) to produce nicotinamide riboside (NR), exhibits high enzymatic activity, and has value for industrial application.

The present invention relates to radio- and fluorescence-labeled compounds, as well as their use as Diagnostics for Detecting Ecto-5′-Nucleotidase (CD73). The invention is further directed to a pharmaceutical composition comprising said compounds as well as to the compounds and the pharmaceutical composition for use in a method of diagnosis of a disease associated with increased or decreased CD73-expression as well as in the treatment of a disease associated with increased CD73-expression.

Compounds that modulate the conversion of AMP to adenosine by 5′-nucleotidase, ecto, and compositions containing the compounds and methods for synthesizing the compounds, are described herein. The use of such compounds and compositions for the treatment and / or prevention of a diverse array of diseases, disorders and conditions, including cancer- and immune-related disorders, that are mediated by 5′-nucleotidase, ecto is also provided.

The present invention relates to an Escherichia coli that efficiently produces 5'-inosinic acid and its application, belonging to the field of bioengineering technology. In order to reduce the degradation of 5'-inosinic acid in Escherichia coli, four phosphatase encoding genes were knocked out using the CRISPR / Cas9 system, and it was found that knocking out the UMP phosphatase encoding gene nagD and the 5'-nucleotidase encoding gene ushA respectively can increase the production of 5'-inosinic acid. Subsequently, in order to inhibit the further conversion of 5'-inosinic acid into guanylate and adenylate, the 5'-inosinic acid dehydrogenase encoding gene guaB was knocked out alone. Finally, in order to enhance the supply and utilization of formyltetrahydrofolate required in the purine synthesis process, the promoter of the serine hydroxymethyltransferase encoding gene glyA was replaced with P tac The recombinant Escherichia coli produced 2113.12 mg / L of 5'-inosinic acid in 48 hours of fermentation, and no antibiotics were required during the fermentation process, indicating that the product has broad application prospects.

Provided are CD73 (also known as ecto-5'-nucleotidase) inhibitor compounds, and compositions and uses thereof for treating and / or preventing CD73-associated diseases, disorders, and conditions, including cancer-associated and immune-associated disorders. CD73 inhibitor compounds include compounds having a structure according to Formula I': (I') and pharmaceutically acceptable esters or salts thereof.

The invention belongs to the technical field of biological catalytic synthesis, and particularly relates to a method for synthesizing beta-nicotinamide mononucleotide (NMN) by enzyme catalysis, which comprises the following steps: by taking D-ribose as a starting substrate, and utilizing a three-step enzyme catalysis reaction system which is synergistically completed by an S-methyl-5-thioribose kinase mutant (SMTK), purine nucleotidase (PNP) and nicotinamide ribose kinase (NRK), synthesizing beta-nicotinamide mononucleotide (NMN) by enzyme catalysis. And efficient synthesis of the NMN is realized. In order to further optimize the cost, an ATP (adenosine triphosphate) regeneration system is innovatively introduced in the process, so that the consumption of ATP in the reaction process is effectively reduced. On the basis, by combining an enzyme directed evolution technology, an immobilized enzyme technology and a continuous reactor design, the catalytic reaction efficiency and the product purity level are remarkably improved. The whole process has the characteristics of greenness and environmental protection, the reaction conditions are mild and controllable, the product yield stably reaches more than or equal to 85%, and the requirements of industrial large-scale production are completely met.

A system and method for identifying a nucleobase in a template polynucleotide is disclosed. In an embodiment, such a method may include providing a substrate comprising a plurality of double-stranded template polynucleotides in a cluster. Each double-stranded template polynucleotide may include a first strand and a second strand. The method may further include contacting the plurality of double-stranded template polynucleotides with a first primer that binds to the first strand and a second primer that binds to the second strand. The method may further include extending the first primer and the second primer by contacting the cluster with a labeled nucleobase to form a first labeled primer and a second labeled primer. The method may further include stimulating light emission from the first and second labeled primers, where the amplitude of the signal generated by the first labeled primer is greater than the amplitude of the signal generated by the second labeled primer. The method may further include identifying the labeled nucleobases added to the first and second primers based on the amplitude of the signal generated by the labeled nucleobases.

Provided is a fish in which accumulation of inosinic acid is enhanced or accelerated during ripening. In a fish of the present disclosure, the function of the ecto-5′-nucleotidase (nt5e) gene is lost.

The invention provides application of non-specific 5 '-nucleotidase in production of beta-nicotinamide ribose (NR), and belongs to the technical field of biology. The invention discovers that the non-specific 5 '-nucleotidase derived from specific bacteria has obviously better catalytic activity for catalyzing the hydrolysis of nicotinamide nucleotide (NMN) to generate beta-nicotinamide ribose (NR) compared with UshA derived from escherichia coli, and the 5'-nucleotidase can be used for large-scale industrial production of beta-nicotinamide ribose.

Methods of identifying compounds that modulate the conversion of AMP to adenosine by 5′-nucleotidase, ecto, and that possess particular pharmacokinetic characteristics are described herein. Methods of such compounds, and compositions comprising same, for the treatment and / or prevention of a diverse array of diseases, disorders and conditions, including cancer- and immune-related disorders, are also provided.

CD73 (also known as ecto-5′-nucleotidase) inhibitor compounds are provided, as well as compositions and uses thereof for treating or preventing CD73-associated or related diseases, disorders and conditions, including cancer- and immune-related disorders. CD73 inhibitor compounds include compounds having the structure set forth in Formula I′ and pharmaceutically acceptable esters or salts thereof.

The invention relates to the technical field of in-vitro detection reagents, in particular to a 5 '-nucleotidase calibrator and a preparation method thereof.The calibrator comprises 5'-nucleotidase, a borate buffering agent and animal serum, and the animal serum is selected from at least one of newborn calf serum, fetal calf serum, calf serum or sheep serum. The calibrator disclosed by the invention shows good stability and interchangeability, and ensures the detection accuracy of 5 '-nucleotidase.

The present invention provides a compound which is, as an antisense oligonucleotide that controls the gene expression of RasGRP4 and is capable of treating myositis and rheumatoid arthritis, an antisense oligonucleotide having a nucleobase sequence that contains 8-80 consecutive nucleosides and has at least eight continuous nucleobases which are complementary to a transcript of RasGRP4, or a pharmaceutically acceptable salt thereof.

The application provides a cytosine nucleoside producing strain and a construction method and application thereof, and the strain is constructed by using CRIPSR / Cas9 gene editing technology E. coli The cdd gene and cmk gene are knocked out on the UR12 genome, the nucleoside triphosphate reductase gene nrdD is weakened, and the nucleoside triphosphate pyrophosphatase gene nudG, the nucleoside diphosphate kinase gene ndk and the uridine acid kinase gene pyrH carrying a mutation point are overexpressed (D93A) The cytosine triphosphate synthase gene pyrG carrying a mutation point is introduced into the Corynebacterium glutamicum heterologously and in multiple copies (D160E、E162A、E168K，cgl) The bifunctional nucleotidase gene of Saccharomyces cerevisiae PHM8 (sce) The obtained strain has good genetic stability, high fermentation yield and can stably produce cytosine nucleoside.

Described herein are compounds that modulate the conversion of AMP to adenosine by an extracellular 5 '-nucleotidase, as well as compositions containing the compounds and methods of synthesizing the compounds. Also provided are uses of such compounds and compositions for the treatment and / or prevention of a variety of diseases, disorders and conditions, including cancer and immune-related disorders, mediated by extracellular 5 '-nucleotidase.

The invention discloses an ultrasonic activated liposome for regulating an ATP-ADO axis to relieve immunosuppression and application of the ultrasonic activated liposome, and belongs to the technical field of biological medicines. The liposome is formed by self-assembly of lipid, a sound-sensitive agent and an active oxygen response type CD39 inhibitor prodrug. According to the invention, the sonosensitizer capable of generating ROS and the ROS responsively released excision nucleotidase inhibitor prodrug are utilized to synthesize the ultrasonic activation type cascade drug release liposome for regulating the ATP-ADO axis to relieve immunosuppression, and only after the liposome is subjected to ultrasonic irradiation, the sonosensitizer can generate active oxygen and further activate the prodrug to release the excision nucleotidase inhibitor, so that the effect of relieving immunosuppression is achieved. According to the present invention, the tumor cell ICD is induced while the ATP level with immune activation is improved by adjusting the ATP-ADO axis, and the ADO accumulation with immunosuppression is reduced so as to achieve the dual anti-tumor immune effect; targeted external toxic and side effects caused by the excision nucleotidase inhibitor are avoided in an ultrasonic triggering mode.

This invention discloses a xanthine oxidase. It also discloses a mutant xanthine oxidase with good thermostability and high activity. Furthermore, this invention discloses the DNA sequence encoding xanthine oxidase, an expression vector containing the DNA sequence, a recombinant bacterial strain, and a method for preparing xanthine oxidase. This invention also discloses the application of xanthine oxidase in the preparation of clinical detection kits for 5'-nucleotidase and adenosine deaminase, demonstrating good reproducibility and high precision.

Compounds that modulate the conversion of AMP to adenosine by 5′-nucleotidase, ecto, and compositions containing the compounds and methods for synthesizing the compounds, are described herein. The use of such compounds and compositions for the treatment and / or prevention of a diverse array of diseases, disorders and conditions, including cancer- and immune-related disorders, that are mediated by 5′-nucleotidase, ecto is also provided.

According to the application of the purine nucleotidase inhibiting substance in preparation of the medicine for treating rheumatoid arthritis, a high-efficiency inhibitor Forodesine of PNP is selected to be subjected to preventive intragastric administration on CIA mice, and after administration, arthritis morbidity is relieved, the disease course is delayed, the clinical score is reduced, and the morbidity is reduced.

The present invention provides a cell including a recombinant DNA molecule including a first nucleic acid sequence encoding at least one first polypeptide being an enzyme selected from: inosine monophosphate dehydrogenase (Impdh), guanosine monophosphate synthetase (gmps), nucleotidase 5C (nt5c), polynucleotide phosphorylase (Pnp), or any combination thereof. Further provided is a method of using the cell of the invention, such as for synthesizing guanine.

The present invention provides a one-pot multi-enzyme method for synthesizing pseudouridine, comprising: weighing D-ribose, ATP and metal cofactor Mn 2+ Mg 2+ and K + Salt, deionized water dissolves to obtain the first solution;Uracil is weighed again, deionized water dissolves to obtain the second solution;The first solution and the second solution are mixed and stirred evenly, and the pH is adjusted to 7.0-8.0 for standby use;Ribokinase, pseudouridine 5-phosphoglycosidase, 5'-nucleotidase are added, and the reaction is sufficient to obtain a pseudouridine reaction solution;Pseudouridine reaction solution is purified and filtered and dried to obtain pseudouridine. This method greatly shortens the reaction process and reaction time, and the raw materials required are cheap, the reaction process is simple and rapid, and expensive and complex instruments and equipment are not required, the reaction conditions are mild, the yield is high and the output is stable, and the use of enzyme method is also environmentally friendly, and it is a green and efficient pseudouridine synthesis technology that can be used for industrial large-scale.

Compounds that modulate the conversion of AMP to adenosine by 5′-nucleotidase, ecto, and compositions containing the compounds and methods for synthesizing the compounds, are described herein. The use of such compounds and compositions for the treatment and / or prevention of a diverse array of diseases, disorders and conditions, including cancer- and immune-related disorders, that are mediated by 5′-nucleotidase, ecto is also provided.