998 results about "Phosphorylation" 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.

The invention discloses a deoxyribozyme probe for identifying carbapenem-resistant klebsiella pneumoniae and application of the deoxyribozyme probe, and belongs to the technical field of deoxyribozyme probes. The method comprises the following steps: firstly, mixing a phosphorylated library chain, a substrate chain and a connecting chain, then, carrying out covalent linkage to construct a DNA library, firstly, carrying out reverse screening, separating and purifying a DNA band of an uncut substrate RNA site, constructing a chain-shaped DNA library by using the obtained DNA band, carrying out forward screening, separating and purifying the DNA band of the cut substrate RNA site, carrying out PCR (Polymerase Chain Reaction) amplification, and recovering a positive-sense chain, so as to obtain the DNA library. The deoxyribozyme probe is obtained by performing repeated screening and enriching, and the probe can recognize carbapenem-resistant klebsiella pneumoniae with high specificity and has the potential of exerting advantages in the aspect of biosensing application.

The method for preparing D-mannose through catalysis can effectively reduce the amount of by-products glucose and fructose, and greatly improves the conversion rate of D-mannose. Specifically, engineering bacteria for expressing isoamylase genes and engineering bacteria for expressing alpha-glucan phosphorylase genes, glucophosphate mutase genes, difunctional enzyme glucophosphate isomerase / mannose 6-phosphate isomerase genes and mannose 6-phosphate phosphatase genes are utilized, starch or starch derivatives are used as substrates, and the starch or starch derivatives are used as substrates. A phosphate buffer solution and Mg < 2 + > are added, a preliminary catalysis system is constructed, after the reaction is completed, an incompletely-reacted substrate and maltodisaccharide exist in the reaction system, the incompletely-reacted substrate and maltodisaccharide can be hydrolyzed into glucose by glucoamylase, and the polyphosphoglucokinase can be used for catalyzing the reaction of the polyphosphoglucokinase under the assistance of sodium hexametaphosphate. All glucose is converted into phosphorylated glucose, then D-mannose is generated, and by-products in a system are removed while the yield is increased.

The invention provides application of scEMC10 in treatment and diagnosis of cerebral vascular endothelial cell senescence, and relates to the technical field of biomedicine. The research of the inventor finds that brain microvascular endothelial cell senescence is caused by the lack of scEMC10; after the brain microvascular endothelial cells are treated by scEMC10, beta-galactosidase, SASP and classic senescence markers related to senescence can be down-regulated; after scEmc10 is supplemented to old mice and Alzheimer's disease 5XFAD model mice, the motion coordination ability and the spatial cognitive ability of the mice can be obviously improved, the relative content of a marker phosphorylated alpha-synuclein of the Parkinson's disease of the old mice can be obviously reduced, and the content of tyrosine hydroxylase phosphorylated active forms can be increased. Therefore, the scEMC10 can be used for treating brain microvascular endothelial cell senescence, improving Alzheimer's disease and Parkinson's disease and detecting brain microvascular endothelial cell senescence as a marker.

The invention relates to the technical field of biological information, and discloses a multi-mode phosphorylation site prediction method and device, equipment and a medium, and the method comprises the steps: extracting a subsequence of a candidate phosphorylation site from a to-be-predicted protein sequence, and generating a sequence feature of the subsequence through a deep learning model; constructing a graph structure based on the three-dimensional structure information of the candidate phosphorylation sites, and extracting structural features of the candidate sites through a graph neural network according to the graph structure; performing multi-modal feature fusion on the sequence features and the structural features to generate fusion features; and predicting the phosphorylation probability of the candidate site based on the fusion feature to obtain a phosphorylation probability prediction result. According to the method, the protein sequence language model and the multi-modal features of the three-dimensional structure diagram convolutional network are fused, so that the accuracy and comprehensiveness of phosphorylation site prediction can be effectively improved, and meanwhile, the calculation efficiency and the bioinformatics interpretation are considered.

The invention discloses a kit for jointly detecting phosphorylated Tau-217 and non-phosphorylated Tau-217. The kit comprises a magnetic microsphere reagent, a detection antibody reagent and a fluorescent microsphere reagent, the magnetic microsphere reagent comprises a magnetic microsphere compound, the detection antibody reagent comprises a first detection antibody compound and a second detection antibody compound, and the fluorescent microsphere reagent comprises a first fluorescent microsphere compound and a second fluorescent microsphere compound. The invention provides a kit for jointly detecting phosphorylated Tau-217 and non-phosphorylated Tau-217, which is based on a single molecule detection technology, and realizes accurate distinguishing and high-sensitivity detection on the phosphorylated Tau-217 and the non-phosphorylated Tau-217 at the same time through a double-target synchronous detection design and a double-color fluorescence report system.

The present application relates to a pyridoxal kinase mutant, a recombinant expression vector and a microbial cell and their applications, and belongs to the technical field of biological catalysis. In order to solve the problem of low product concentration in existing enzyme catalysis, a pyridoxal kinase mutant is provided, the amino acid sequence is selected from the amino acid sequence shown in SEQ ID NO. 1, the lysine at position 229 is mutated to alanine, phenylalanine, methionine, arginine, threonine, histidine, serine, tyrosine, valine, leucine, isoleucine, proline, asparagine, aspartic acid or glutamic acid; the pyridoxal kinase mutant is used for catalyzing pyridoxal to synthesize pyridoxal phosphate, and a recombinant expression vector and a microbial cell can be further formed. The present application has good enzyme activity, high product conversion rate, high concentration of pyridoxal phosphate obtained, and the concentration of the product catalyzed by the wild-type pyridoxal kinase to phosphorylate pyridoxal is obviously improved.

The invention relates to the technical field of biological tissue adhesives, in particular to a biological tissue adhesive composition. The biological tissue adhesive composition comprises a powder A, a powder B and a powder C, wherein the powder A is obtained by curing calcium silicate and acidic calcium phosphate in a curing liquid, curing, drying and crushing; powder B: bivalent metal salt; powder C: phosphorylated amino acid; and an aqueous solvent; the biological tissue adhesive is used for bone adhesion and bone repair. According to the biological tissue adhesive composition provided by the invention, a silicate / hydroxyapatite composite material with a bone induction function is synthesized through optimized bone cement reaction, a curing liquid is added to react with phosphorylated amino acid, and a bone adhesive material is obtained through organic and inorganic material coordination reaction; the bone repairing and regenerating capability and the bone bonding capability are good.

The invention discloses a method for enhancing squalene synthesis through oil tea HMGR gene editing. According to the method, phosphorylation regulation and control sites Ser47 and / or Thr208 recognized by MAPK are targeted, and enzymatic activity inhibition is relieved and metabolic flux is enhanced by constructing phosphorylation resistance mutants (such as Ser-> Ala and Thr-> Val). The constructed editing vector can be introduced into camellia oleifera tissues in manners of PEG-mediated protoplast transient transfer, agrobacterium infection of calluses and the like, so that stable expression is realized. Experiments prove that the enzyme activity of HMGR in the editor is remarkably improved, the squalene accumulation amount is increased by 50% or above, the editing efficiency is high, expression is stable, and the method is suitable for high-value development of plant triterpenoid functional components and excellent strain breeding. The method also has the potential of popularization and application in other triterpenoid synthetic crops.

The invention relates to the technical field of biology, and particularly discloses a polypeptide and application thereof in enhancing immunity and / or resisting inflammation. The invention provides a polypeptide. The amino acid sequence of the polypeptide is shown as SEQ ID NO. 1. The compound has the dual functions of regulating immunity and resisting inflammation, on one hand, the immune function of cells under the normal condition can be enhanced, and on the other hand, cell inflammation can be relieved by reducing the phosphorylation level of key protein in an LTR4-NF-kappa B signal channel in the inflammatory state. The polypeptide is low in cytotoxicity, high in safety, simple in preparation method and low in cost, can be used as an ideal immune and anti-inflammatory regulator, and has good application potential and value.

The invention discloses a modular fluorescent RNA aptamer biosensor system, and relates to the field of medicine. Comprising a probe module, an enzyme system, a dye and a reaction buffer solution, the probe module comprises a promoter probe P and a reporter probe R, the enzyme system comprises SplintR ligase and T7RNA polymerase, and the reaction buffer solution comprises a transcription buffer solution and a SplintR ligase buffer solution. The promoter probe P comprises a T7 promoter sequence and a 5 '-phosphorylated upstream recognition region, and the length of the promoter probe P is 10nt. According to the present invention, the ligase-assisted probe assembly and the T7RNA polymerase-mediated fluorescent RNA aptamer transcription are integrated, the simultaneous detection of the multiple circRNA can be achieved without the complex probe labeling, the femtomole-level sensitivity and the excellent single base mutation distinguishing ability in the complex sample are provided, and the real-time fluorescence instrument and the portable device are adapted.

The invention relates to isolated synthetic or recombinant peptides comprising an epitope of human tau 2N4R, wherein the tau peptide sequence comprising the epitope is not phosphorylated. The invention also relates to use of such peptides to generate binding molecules, such as antibodies, specific for the non-phosphorylated tau epitopes and to such peptides and binding molecules, such as antibodies, for use in investigation, diagnosis and treatment of tauopathies, such as Alzheimer's disease.

Provided in the present invention are an oxamide compound having a structure as shown in formula (I), or a pharmaceutically acceptable salt, isotope derivative or solvate thereof, or a stereoisomer, geometric isomer or tautomer thereof, or a prodrug molecule or metabolite thereof, and a pharmaceutical composition thereof and the use thereof. The compound involved in the present invention can efficiently inhibit STAT6 phosphorylation, can be used for preparing drugs for preventing and treating inflammatory diseases, and can be used for preparing anti-tumor drugs.

The invention discloses a phosphorylated peptidomimetic compound and a medical application thereof. The invention provides a phosphorylated peptidomimetic compound with a novel structure. Activity research shows that the phosphorylated peptidomimetic compound has excellent Cbl-b inhibitory activity and is an effective Cbl-b inhibitor. Technicians in the field know that Cbl-b overexpression can inhibit T cell immune response, and inhibition of Cbl-b can activate killing of an immune system on tumor cells. Therefore, the phosphorylated peptidomimetic compound or pharmaceutically acceptable salts and solvates thereof provided by the invention can be developed and prepared into drugs for treating diseases (such as non-small cell lung cancer, breast cancer, prostate cancer, head and neck squamous cell carcinoma, liver cancer, pancreatic cancer, colorectal cancer, ovarian cancer and other tumors) treated or relieved by inhibiting Cbl-b (such as non-small cell lung cancer, breast cancer, prostate cancer, head and neck squamous cell carcinoma, liver cancer, pancreatic cancer, colorectal cancer, ovarian cancer and the like) the medicine prospect is promising.

The invention belongs to the field of biological medicine, and particularly relates to PGK1 K17 site succinylation modification and application. Experiments show that the 17-site lysine of the PGK1 protein has succinylation modification (PGK1 K17su), and the existence of the modification is confirmed through a specific succinylation modified antibody for the first time. Further, it is found that PGK1 K17su can weaken lung cancer cell oxidative phosphorylation and promote glycolysis metabolism by inhibiting lung cancer cell mitochondrial generation, and functional experiments prove that PGK1 K17 succinylation modification promotes proliferation and migration of lung cancer cells.

The peptide complex of the present invention promotes the expression of the SIRT1 gene, referred to as a longevity gene, promotes the expression of the FGF21 gene, and increases the phosphorylation of AMPK. In addition, the peptide complex of the present invention exhibits an anti-inflammatory effect. Therefore, the peptide complex of the present invention inhibits aging, promotes the expression of the longevity gene SIRT1 gene, and can be used for treating or preventing diseases associated with SIRT1.

The invention belongs to the field of antibodies, and particularly relates to a p-Tau217 phosphorylated protein binding antibody or an antigen binding part thereof and application thereof. The invention provides the amino acid sequences of the heavy chain CDR and the light chain CDR of the antibody specifically bound with the p-Tau217 phosphorylated protein, and the antibody can specifically recognize and bind the p-Tau217 phosphorylated protein and has the advantages of high affinity and good specificity to the protein. Based on the advantages, the antibody can be applied to diagnosis of diseases (such as nervous system degenerative diseases) related to abnormal p-Tau217 phosphorylated protein level, and especially can be applied to clinical diagnosis of Alzheimer's disease.

The invention discloses a synthesis method of 2 '-fluoro-2'-deoxyguanosine. According to the method, thymine nucleoside phosphorylase and purine nucleoside phosphorylase are used as biocatalysts, and 2 '-fluoro-2'-deoxyuridine and guanine are used as substrates to synthesize the 2 '-fluoro-2'-deoxyguanosine. Compared with a chemical method, the production cost is greatly reduced; meanwhile, the synthesis method can effectively solve the problem that the product is difficult to separate.

The invention discloses recombinant escherichia coli with high yield of N-acetylneuraminic acid and application of the recombinant escherichia coli, and relates to the technical field of biological genetic engineering. The invention relates to a recombinant escherichia coli, which is characterized in that the escherichia coli is taken as a host, and free expression of an N-acetylmannosamine epimerase gene yihS from Streptomyces xiamenensis or an N-acetylmannosamine epimerase gene ce3 from Bacteroides polymorpha and an exogenous N-acetylneuraminic acid lyase gene nano A is carried out; and carrying out recombinant expression on N-acetyl hexosamine 1-kinase nahK, a UDP-N-acetyl glucosamine pyrophosphorylase gene glmU and a UDP-N-acetyl glucosamine-2-epimerase gene neuC in the other synthetic route of the ManNAc. According to the recombinant escherichia coli with high yield of N-acetylneuraminic acid, the yield of N-acetylneuraminic acid can reach 23.08 g / L under a shake flask fermentation condition; a two-stage batch feeding strategy is adopted, the yield of N-acetylneuraminic acid in a 5L fermentation tank reaches 71.25 g / L, the molar conversion rate of GlcNAc reaches up to 57.60%, and the method has the potential of industrial application.

The present invention relates to use of a JWA polypeptide in the preparation of a drug for resisting androgenetic alopecia. An amino acid sequence of the polypeptide is shown as I or II: I: FPGSDRF-Z; II: X-FPGSDRF-Z, wherein an amino acid S is subjected to phosphorylation modification, and X and Z are an amino acid or an amino acid sequence, respectively. The described JWA polypeptide can directly target integrin molecules onto hair follicle cells and enter the cells to play a role in regulating and controlling hair follicle proliferation, promoting hair growth and the like; the telogen phase of the hair follicles is significantly shortened, and the anagen phase of the hair follicles is prolonged; the hair shaft and follicle volume is increased; and the expression level of the target cell integrin molecule αvβ1 can be improved, and the hair follicle stem cell signal channel MEK / ERK / E2F1 / SP1 / Wnt10a / 10b / β-catenin can be accurately activated.

The invention relates to the field of konjak food, provides konjak gel food as well as a preparation method and application thereof, and aims to solve the problems of poor stability, insufficient brittleness and poor antibacterial property of existing konjak gel food. According to the gel food, phosphorylated konjac glucomannan serves as a main body, chitosan / microcrystalline cellulose composite filler, carrageenan, sodium alginate, calcium salt, citrate and other auxiliary materials are added, a multi-cross-linked network structure is constructed through the synergistic effect of multiple components, and the stability and mechanical performance of gel are remarkably enhanced. In the preparation process, a temperature gradient control strategy is adopted, and ordered construction and layer-by-layer curing of a gel network are achieved. The obtained product has high gel strength, excellent brittleness, low bleeding rate and excellent antibacterial performance, can be widely applied to functional food simulating the mouth feel of animal cartilage, and has good storage stability and sensory quality.

The invention discloses an application of an engineered exosome rich in SIRT3 in preparation of a medicine for improving mitochondrial dysfunction. The method comprises the following steps: screening to obtain a polypeptide molecular gel G12 targeting HSP60, forming a fusion protein SIRT3-G12 from the G12 and SIRT3, constructing a recombinant plasmid for expressing the fusion protein, transfecting cells, culturing, and extracting an exosome, thereby obtaining the engineering exosome SIRT3-Exo rich in SIRT3. The SIRT3-Exo is used for treating mitochondrial dysfunction, and the research shows that the SIRT3-Exo can remarkably promote mitochondrial oxidative phosphorylation, promote expression of cell antioxidant enzymes and deacetylation of FOXO3a, enhance the antioxidant capacity of cells and remove ROS in the cells, and has the effect of promoting mitochondrial function recovery. The problem that SIRT3 cannot be used for treating mitochondrial dysfunction through exosomes is solved, and good application prospects are achieved.

The invention discloses a self-assembled PROTAC nano material based on natural product mitochondria targeting, which is a nano particle prepared from mitochondria targeting molecules, a photosensitizer and PROTAC as raw materials through a nano precipitation method. The mitochondrial targeting molecule is berberine, the photosensitizer is hypericin, and the PROTAC is dBET57; the ratio of the amount of substance of the dBET57 to the amount of substance of the berberine is (1-10): (1-10), and the ratio of the amount of substance of the dBET57 to the amount of substance of the hypericin is (1-10): (1-10). The nano material can be used for blocking multiple energy metabolism pathways of tumor cells, including glycolysis and oxidative phosphorylation, meanwhile, mitochondria is damaged, ferroptosis is induced, and the anti-tumor capacity of pharmacodynamic molecules is improved; bRD4 in tumor cells and downstream carcinogenic protein c-Myc of the BRD4 can be efficiently degraded. The invention also discloses an application of the nano material in preparation of antitumor drugs.

The invention relates to the technical field of antibodies, in particular to antigen peptides of human PP1 alpha protein and Thr320 site phosphorylated protein of the human PP1 alpha protein, and a preparation method and application of an antibody of the antigen peptides of the human PP1 alpha protein and the Thr320 site phosphorylated protein. The invention provides an antigen peptide of human PP1 alpha protein and Thr320 site phosphorylated protein thereof, the antigen peptide has obvious advantages in the aspects of immunogenicity and the like, animals can be induced to generate high-level antibodies, and the generated antibodies have high specificity, affinity and titer. Based on the antigen peptide, the invention provides an anti-human PP1 alpha protein or a polyclonal antibody with phosphorylated Thr320 site thereof and a preparation method thereof, the polyclonal antibody has the characteristics of strong specificity, high affinity and high titer, and through immunoblotting and immunohistochemical verification, the polyclonal antibody can be used for preparing the anti-human PP1 alpha protein or the anti-human PP1 alpha protein or the anti-human PP1 alpha protein or the anti-human PP1 alpha protein or the anti-human PP1 alpha protein. The probe shows good specificity and sensitivity in detection of phosphorylation modification of human PP1 alpha protein and Thr320 site thereof, and has a good application prospect.

The invention discloses recombinant escherichia coli for producing sucrose phosphorylase as well as a construction method and application of the recombinant escherichia coli, and belongs to the technical field of biological engineering. The sucrose phosphorylase is constructed, the amino acid sequence of the sucrose phosphorylase is shown as SEQ ID NO.11, a recombinant escherichia coli BdSP-L341V / L343F / V346P for expressing the sucrose phosphorylase is constructed, the sucrose phosphorylase expressed by recombinant bacteria or whole cells of the recombinant bacteria are used for carrying out catalytic reaction, and efficient production of ascorbyl glucoside can be realized. The invention also provides a method for producing ascorbic acid glucoside, the catalytic reaction is carried out at normal temperature and normal pressure, toxic and harmful organic solvents are not used, and the pollution to the environment is reduced; the whole catalytic process is a one-step reaction, has the advantages of high conversion rate, simple purification step and simple process, can better save the production cost, and has wide industrial application prospects.

To provide a novel stereoselective method for producing a cyclic dinucleotide derivative that can be used for an antibody-immunostimulator conjugate, and a production intermediate thereof, and to provide methods for producing a cyclic dinucleotide-linker and antibody-immunostimulator conjugate using the production method.SOLUTION: The present invention relates to a method that makes it possible to produce, in large amounts and with high yield, a cyclic dinucleotide derivative having a desired steric configuration, by using an optically active phosphitylation agent twice, during coupling and cyclization.SELECTED DRAWING: None

The invention discloses recombinant bacillus subtilis with high yield of surfactin as well as a construction method and application of the recombinant bacillus subtilis, and belongs to the technical field of genetic engineering. According to the invention, non-essential genes are determined through analysis and identification, a simplified engineering strain is obtained, and the highest fermentation yield reaches 11.2 g / L; then, a PTS part sugar phosphorylase gene obtained through screening is transferred into the simplified engineering strain, the metabolism efficiency of carbohydrates is improved, transportation and utilization of the carbohydrates are optimized, a fatty acid transmembrane transporter gene is further transferred, fatty acid uptake is assisted, synthesis of surfactin is facilitated, and the yield of the surfactin is improved. The combination of the two realizes optimization of synthesis of surfactin through a composite carbon source. Compared with a simplified engineering strain, the genetically engineered bacterium obtained by the invention has the advantages that the yield of surfactin is increased by 57.1%, the yield is increased by 2.3 times compared with that of an unsimplified engineering strain EX, and the final shake-flask fermentation yield can reach 17.6 g / L. As an industrial chassis cell of high-yield lipopeptide, the cell has a good industrial application prospect.

The invention belongs to the technical field of biological materials and biological information, and provides a space phosphorylation modification omics detection method. According to the detection method disclosed by the invention, proteomics analysis can be carried out on trace sample phosphorylation, 4259 phosphorylation sites can be identified by 5 micrograms of peptide fragments, and the credibility of 3506 sites is greater than 0.75.

The invention discloses a preparation method of crocodile liver peptide for improving liver injury, and relates to the technical field of bioactive peptide extraction, and the preparation method comprises the following steps: crocodile liver pretreatment: cutting fresh crocodile liver into blocks, mixing with deionized water, homogenizing, and adding a zirconium phosphate molecular sieve to obtain crocodile liver homogenate; enzymatic hydrolysis reaction: adjusting the pH value of the homogenate, and adding a compound enzyme preparation for enzymatic hydrolysis; carrying out enzyme deactivation treatment, namely heating, cooling, centrifuging and taking supernate; performing ultrafiltration separation, namely filtering the supernate through an ultrafiltration membrane, and collecting a small molecule peptide solution; purifying and refining: purifying through sephadex column chromatography, and freeze-drying to obtain a crocodile liver peptide product; a double-particle-size zirconium phosphate molecular sieve is constructed to cooperate with a compound enzyme preparation for enzymolysis, electron transfer is enhanced by combining phosphorylation modification, the defects of metal inhibition enzyme activity, insufficient concealed peptide fragment cutting and oxidative damage are overcome, meanwhile, the application of the crocodile liver peptide in the aspect of improving the liver function is researched, and the crocodile liver peptide has a good application prospect. And a new choice is provided for prevention and treatment of liver diseases and liver health care.

The invention discloses an anti-human MerTK antibody and an application of the anti-human MerTK antibody. The anti-human MerTK antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3. The invention also discloses nucleic acid for coding the antibody, a recombinant expression vector, a transformant, a preparation method of the antibody, a chimeric antigen receptor, an antibody drug conjugate, a pharmaceutical composition or a kit containing the antibody, and applications of the nucleic acid, the recombinant expression vector, the transformant, the chimeric antigen receptor, the antibody drug conjugate, the pharmaceutical composition or the kit in preparation of drugs for preventing and / or treating tumors. The antibody disclosed by the invention is high in affinity, can specifically inhibit the interaction of the ligand Gas6-MerTK and the ligand protein S-MerTK at the same time, and does not inhibit the interaction of the ligand Tulp1-MerTK. And potential toxicity to the retina can be avoided when inhibition activity is exerted in vivo. In addition, the anti-human MerTK antibody provided by the invention can specifically and effectively inhibit phosphorylation of AKT in a Gas6-MerTK signal, and shows certain tumor inhibition activity in a mouse in-vivo model test.