15 results about "Uridylic Acids" patented technology

The invention provides a method for catalytically producing uridine monophosphate and application, and belongs to the technical field of genetic engineering and microbial engineering. The uridine kinase CaUDK-F gene with the nucleotide sequence as shown in SEQ ID NO.1 is adopted, the uridine kinase CaUDK-F gene is overexpressed through escherichia coli and then combined with polyphosphate kinase PPK to conduct biological catalysis on a substrate uridine, uridylic acid can be efficiently produced, the concentration of uridylic acid in a catalysis system is high, separation and purification are convenient, and the technology is simple. Meanwhile, the uridine kinase CaUDK-F is subjected to site-specific mutagenesis by adopting a site-specific mutagenesis technology, and three key amino acid residue sites S85, L87 and E88 which influence enzyme activity are subjected to site-specific mutagenesis, so that the catalytic efficiency and the conversion rate of the uridine kinase are further improved; wherein the enzyme activity of the mutant S85A is increased to 2.6 times that of a wild enzyme, and when the mutant S85A catalyzes uridine to produce uridylic acid, the conversion rate is increased to 92% from 80% of the wild enzyme.

The invention provides uridine kinase, a mutant thereof, a method for catalytically producing uridine monophosphate and application, and relates to the field of genetic engineering and microbial engineering. The uridine kinase CaUDK-T gene with the nucleotide sequence as shown in SEQ ID NO.1 is adopted, after escherichia coli is over-expressed, a substrate uridine is biologically catalyzed in combination with polyphosphate kinase PPK, and uridylic acid can be efficiently produced. Meanwhile, amino acid residues E148, S152 and V153 of uridine kinase CaUDK-T are replaced with glycine in a site-specific mutagenesis mode, mutants E148G, S152G and V153G are obtained through construction, the catalytic efficiency and conversion rate of uridine kinase are further improved, the enzyme activity of the uridine kinase mutant V153G is improved to 1.97 times that of wild enzyme, the conversion rate of 50 g / L uridine can reach 90%, efficient production of UMP is achieved, and the application prospect is wide. Therefore, the industrial production is promoted.

This application relates to the field of metabolic engineering and genetic engineering technology for production, specifically providing a uracil-producing strain, its construction method, and its application. This strain is... E. coli Knockout of the uridine kinase gene in the W3110 genome udk Simultaneously integrates the T7 RNA polymerase gene; weakens the uridine kinase gene. pyrH Overexpression of carbamoyl phosphate synthase gene carAB ribose-phosphokinase gene prsA pyrimidine-5'-nucleotide nuclease gene ppnN UMP phosphohydrolase gene umpH and umpG Ribonucleotide hydrolase 1 gene rihA and glutamine synthase gene glnA Heterogeneous introduction of wild-type Bacillus subtilis B. subtilis 168 orotate nucleoside monophosphate decarboxylase gene pyrF and orotic acid ribosyltransferase gene pyrE To achieve safe, efficient, and low-cost production of uracil.

This invention provides a kit for the simultaneous detection of nine flavor nucleotides in livestock and poultry products. The flavor nucleotides are cytidine monophosphate, uridine monophosphate, guanylic acid, inosine monophosphate, adenosine monophosphate, hypoxanthine, inosine, adenosine diphosphate, and adenosine triphosphate. The kit includes: a high-concentration extraction reagent, a standard curve solution, a high-concentration phosphate buffer solution, pH adjustment solution A, and pH adjustment solution B. The high-concentration extraction reagent is perchloric acid; the standard curve solution is a mixed solution of the nine flavor nucleotides; the high-concentration phosphate buffer solution is a solution of potassium dihydrogen phosphate and dipotassium hydrogen phosphate; pH adjustment solution A is sodium hydroxide; and pH adjustment solution B is phosphate. This invention also provides a method for the simultaneous detection of the nine flavor nucleotides using the above kit. This invention enables the simultaneous determination of the content of nine flavor nucleotides in livestock and poultry products. The method is simple, rapid, accurate, and sensitive, and can be used for batch determination, showing good prospects for practical application.

The invention relates to a method for producing hyaluronic acid through cell-free catalysis, and belongs to the technical field of biological catalysis. According to the invention, a system capable of being applied to cell-free catalytic production of hyaluronic acid is constructed, and genetically engineered bacteria are used for expressing N-acetyl hexosamine 1-kinase, glucosamine-1-phosphate acetyl transferase, glucuronide kinase, GlcA-1-P uridinetransferase, hyaluronic acid synthase, polyphosphate kinase, uridine monophosphate kinase and inorganic pyrophosphatase; according to the present invention, N-acetylglucosamine GlcNAc, glucuronic acid GlcA, ATP and UMP are adopted as substrates to perform catalytic production, UDP and ADP produced during the reaction process are converted into ATP and UTP, such that nucleotide circulation is achieved, raw material consumption is reduced, the final hyaluronic acid yield reaches 1.28 g / L, and an economic and efficient brand new path is provided for hyaluronic acid industrial production.

The application discloses a method for preparing 2'-methoxy-uridine triphosphate, which comprises the following steps: taking 2'-methoxy-uridine acid as a substrate, taking adenosine triphosphate as an initial phosphate donor, constructing a phosphate circulation system, adding an enzyme catalyst, and adopting one-pot enzyme method to perform enzyme catalysis reaction, so as to obtain the end product 2'-methoxy-uridine triphosphate. The phosphate circulation system is composed of adenosine triphosphate, adenosine diphosphate and phosphate compounds. The enzyme catalyst catalyzes the dephosphorylation of the phosphate compounds to provide inorganic phosphate to the adenosine diphosphate, and the adenosine triphosphate is regenerated to realize the circulation energy supply. The method adopts high-activity enzyme combination of specific sources, has high conversion rate of the substrate 2'-methoxy-uridine acid, and has high conversion rate of the final 2'-methoxy-uridine triphosphate up to 92.6%, and has the advantages of high synthesis efficiency, simple steps, green environmental protection and controllable cost.

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.

The invention discloses a method for preparing 2 '-methoxy-uridine triphosphate, which comprises the following steps: in a liquid reaction system, constructing a phosphoric acid circulating system by taking 2'-methoxy-uridine monophosphate as a substrate and adenosine triphosphate as an initial phosphoric acid donor, adding an enzyme catalyst, and carrying out an enzyme catalysis reaction by adopting a one-pot enzyme method to obtain the 2 '-methoxy-uridine triphosphate. The phosphoric acid circulating system is composed of adenosine triphosphate, adenosine diphosphate and a phosphate compound, inorganic phosphoric acid is provided for adenosine diphosphate by catalyzing dephosphorizing of the phosphate compound through the enzyme catalyst, and adenosine triphosphate is regenerated to achieve circulating energy supply. According to the method, a high-activity enzyme combination with a specific source is adopted, the conversion rate of the substrate 2 '-methoxyl-uridine monophosphate is high, the final conversion rate of 2'-methoxyl-uridine triphosphate reaches up to 92.6%, and the method has the advantages of being high in synthesis efficiency, simple in step, environmentally friendly and controllable in cost.

The invention discloses a UTP-glucose-1-phosphate uridine monophosphate transferase mutant and application thereof, and belongs to the technical field of biological engineering. The problem of low salidroside yield in the prior art is solved. The invention provides a UTP-glucose-1-phosphate uridylic acid transferase mutant, which is obtained by taking an amino acid sequence as shown in SEQ ID NO.2 as a starting sequence and mutating any one of the 6th site, the 126 site, the 134 site, the 140th site, the 177th site, the 180th site, the 214th site, the 249th site, the 284th site, the 289th site, the 296th site, the 298th site and the 300th site. The invention is mainly used for high-efficiency UTP-glucose-1-uridine phosphate transferase and application of the UTP-glucose-1-uridine phosphate transferase in de novo synthesis of salidroside.

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 provides a yeast hydrolysate as well as a preparation method and application thereof, the yeast hydrolysate comprises the following components in percentage by mass: 35-65% of true protein, aromatic amino acid and sulfur-containing amino acid, and the content ratio of the aromatic amino acid to the sulfur-containing amino acid is 2-5; the ratio of the content of the components with the weight-average molecular weight of 200-2000Da to the content of the components with the weight-average molecular weight of more than 2000Da is 4-10, and the ratio of the content of the components with the weight-average molecular weight of 200-2000Da to the content of the components with the weight-average molecular weight of less than 200Da is 4-9; the yeast hydrolysate further comprises disodium nucleotide, the disodium nucleotide comprises disodium inosinate, disodium guanylate, disodium cytidine monophosphate and disodium uridinemonophosphate, and the ratio of the mass sum of the disodium inosinate and the disodium guanylate to the mass sum of the disodium cytidine monophosphate and the disodium uridinemonophosphate to the disodium nucleotide is 1: (0.6-1.2): (1.6-2.2). The feed additive is beneficial for further improving the feed intake of the piglets and reducing the disease incidence of the piglets.

A nucleotide that encodes for a destabilized RNA that codes for ERBB2 (erythroblastic oncogene B) or MYC oncogenes. The destabilization occurs in an adenylate-uridylate rich element (ARE) stabilizing motif. Without wishing to be bound to any particular theory, changing the stabilizing motifs of ERBB2 and MYC to destabilization motifs is believed to lead to the rapid degradation of their transcripts. Thus, in turn, renders the composition useful as a therapeutic for cancer.

The present disclosure relates to relates to ribonucleic acid (RNA) molecules having a 3' UTRthat encodes for a destabilized TEAD1 or YAP1 protein. The destabilization occurs in an untranslated adenylate-uridylate rich element (ARE) stabilizing motif. The RNA molecule can be used to reverse transcribe a deoxyribonucleic acid (DNA) molecule, which can then be transfected into a vector and formulated as a pharmaceutical composition. Such vectors and pharmaceutical compositions comprising the vectors can be useful in the treatment of a variety of diseases and disorders for which dysregulation of the Hippo pathway is implicated including, but not limited to, cancer, obesity, liver diseases, and cardiac diseases. This abstract is intended as a scanning tool for purposes of searching in the particular art and is not intended to be limiting of the present invention.

The present invention is directed to methods and compositions for template-free enzymatic synthesis of a polyribonucleotide of a predetermined sequence from 3′-O-reversibly blocked nucleoside triphosphates using poly(A) and poly(U) polymerases.

The application discloses application of a 5'-monophosphate nucleotide composition in preparation of anti-aging and longevity drugs and / or functional foods, and belongs to the technical field of medicines.The composition is composed of 5'-monophosphate adenosine, 5'-monophosphate cytidine, 5'-guanylic acid disodium, 5'-uridylic acid disodium and hypoxanthine nucleotide, and the mass ratio of the acid types of CMP, AMP, UMP, GMP and IMP is respectively as follows: CMP: 15-78%, AMP: 6-44%, UMP: 7-40%, GMP: 7-51%, and IMP is 0 or greater than 0 and not higher than 2.5%. The composition can synergistically enhance various mechanisms such as significantly reducing an insulin resistance index, enhancing body anti-coagulation function, improving blood circulation, regulating metabolism of essential minerals such as lithium and copper and regulating intestinal flora ecology, delays aging and prolongs healthy life span. Animal experiment data show that, compared with a control group, the composition prolongs the median survival time of mice by 9.21-12.6%, and the equivalent human life span is 8.76-12.01 years. Human experiment data show that, compared with the control group, the composition intervention reduces the median age of DNA methylation by 3.08 years, and provides a new idea for developing anti-aging and longevity drugs and / or functional foods.