100 results about "Dephosphorylation" patented technology

The invention discloses a method and application for enhancing cytidine production, and belongs to the field of genetic engineering and microbial engineering. The present invention knocks out cytidine deaminase gene cdd, uridine kinase gene udk, pyrimidine-specific ribonucleoside hydrolase gene rihB, nucleotide 5'-monophosphate nucleosidase gene ppnN, pyruvate oxidase gene poxB and integrates and expresses 5'-CTP diphosphohydrolase, orotate phosphoribosyltransferase, uridylate kinase mutant gene PyrH (R92G / D93G), as well as phosphoribosyl pyrophosphate synthetase gene prs, glucose-6-phosphate dehydrogenase gene zwf and 6-phosphogluconate dehydrogenase gene gnd, and constructs a recombinant Escherichia coli with enhanced cytidine synthesis ability, and the cytidine yield reaches 61.9g / L in a 30L fermenter without adding antibiotics and inducers, which can be used to realize the industrial production of cytidine and realize green biomanufacturing.

The invention discloses application of hairyvein agrimony in preparation of a weight-reducing medicine. The weight-losing medicine is a hairyvein agrimony decoction, extract or monomer agrimonin, the effective components in the medicine are agrimonin and derivatives thereof, and STAT3 activity and SOCS3 expression can be inhibited, so that the STAT3-SOCS3 activity is kept in a proper range, and the weight-losing effect is achieved. It is found through experiments that the agrimonin can target SET protein, relieve inhibition of SET on PP2A and promote dephosphorylation of PP2A-mediated STAT3, NF-kB and other signals, so that the activity of the STAT3, the NF-kB and the other signals is inhibited, SOCS3 expression is reduced, and the purpose of weight reduction is achieved. The weight-reducing medicine comprises a hairyvein agrimony decoction, a hairyvein agrimony extract and agrimonin, and human body experiments prove that the hairyvein agrimony decoction and the hairyvein agrimony extract have a remarkable weight-reducing effect.

The invention relates to an alkaline phosphatase mutant as well as a preparation method and application thereof. The alkaline phosphatase mutant comprises polypeptide obtained by mutating an amino acid sequence of parent alkaline phosphatase; the mutation comprises the following steps: replacing threonine at the 124 site on the amino acid sequence of the parent alkaline phosphatase with leucine; the nucleotide sequence of the parent alkaline phosphatase is as shown in SEQ ID NO.1, and / or the amino acid sequence of the parent alkaline phosphatase is as shown in SEQ ID NO.3. The alkaline phosphatase mutant has relatively high expression quantity, can be applied to dephosphorylation of an enzymolysis phosphoric acid substrate, and particularly can be used for labeling an antibody and / or catalyzing chemiluminescence of a luminescent substrate.

The invention relates to a process for dephosphorylation of skim milk, ultra-filtered milk (UF milk) or micellar casein isolate (MCI) comprising (i) acidification of a skim milk, UF milk or MCI preferably not lower than pH 6.0, preferably between 6.0 and 6.7, (ii) cooling the acidified skim milk, UF milk or MCI to the temperature between 0° C. and 15° C., (iii) adding gluconate and / or maleate to the cooled skim milk, UF milk or MCI and, (iv) washing the skim milk, UF milk or MCI, to remove phosphorus, thus dephosphorylating the skim milk, UF milk or MCI, preferably to an extent that the total phosphorous content of the skim milk, UF milk or MCI is reduced with at least 20%, preferably 20-40%, more preferably 30-40% compared to the material provided to step (i). The invention also relates to dephosphorylated MPC, MPI or MCI obtainable by the process of the invention, and to a liquid heat-sterilized enteral nutritional composition comprising 2.0-3.0 kcal / ml wherein 16-35 en % is provided by protein, the combination of caloric content and relative protein caloric content selected such that there is 10-18 g / 100 ml protein, preferably 12-18 g / 100 ml protein in the composition, wherein the protein comprises micellar casein (MC), whey protein (WP) and optionally caseinate (CAS), wherein there is at least 70 wt % MC and less than 15 wt % WP, based on total protein content, and wherein the composition has a total amount of phosphorous less than 192 mg / 100 ml and / or 30-80 mg / 100 kcal, preferably at least 30-80 mg / 100 kcal, most preferably less than 192 mg / 100 ml and 30-80 mg / 100 kcal.

Provided herein are genetic circuits and cell state classifiers for detecting the microRNA profile of a cell. The cell state classifiers of the present disclosure utilize phosphorylation state of a transcription factor to control classifier output. Kinases and phosphatase pairs that function in phosphorylating or dephosphorylating the transcription factor are integrated into the circuit, their expression tuned by the presence of microRNAs of interest (e.g., in a cell). The genetic circuits and cell state classifiers may be used in various applications (e.g., therapeutic or diagnostic applications).

The invention belongs to the field of feed protein processing, and particularly relates to a method for improving the bioavailability of animal feed protein through chemical enzymolysis. The method comprises the following steps: mixing a compound protein raw material with water to form a suspension; sodium tripolyphosphate is added for phosphorylation pretreatment, so that protein is subjected to specific modification; after the pH is adjusted to be acidic, pepsase and acid phosphatase are added at the same time for an enzymolysis reaction; and after the reaction is finished, performing enzyme deactivation and drying to obtain the modified feed protein. According to the method, a specific enzyme cutting site is created on a protein molecule through chemical phosphorylation, then directional hydrolysis is achieved through efficient recognition of pepsin on the site, and meanwhile dynamic regulation and control of the hydrolysis process are achieved by means of the dephosphorylation effect of acid phosphatase. The obtained product has the outstanding advantages of high content of small peptides, good digestive absorbability, low bitterness, strong biological activity and the like, and the bioavailability of feed protein is remarkably improved.

This invention discloses a polypeptide that inhibits the desimilarization activity of tau protein and its applications, belonging to the field of biomedical technology. The polypeptide of this invention is tau. 438 The peptide is obtained through endogenous tau protein synthesis. The peptide provided by this invention can inhibit tau protein desimulation and increase tau mimicry, exhibiting high specificity and safely and effectively improving cognitive and memory function in AD model mice.

The invention relates to application of lentinan in preparation of a medicine for enhancing Kukuzhou cell immune function, and belongs to the technical field of immunopharmacology. The invention discloses a mechanism and application of lentinan LNT (Lentinan) based on a TLR2 (Toll-Liver Receptor 2) signal channel for enhancing the immune function of liver cumness cells and promoting lysosome maturation. By constructing an LNT fluorescence labeling system, a Kuptake cell in-vivo tracing model and a TLR2 function intervention model, it is proved that LNT can be directly combined with TLR2 and activate a TLR2 / MyD88 / NF-kappa B signal axis, and therefore the phagocytosis and immune response capacity of Kuptake cells is enhanced. Meanwhile, TLR2 signal-induced reactive oxygen species (ROS) up-regulation can promote TFEB dephosphorylation and nuclear translocation, lysosome maturation is driven, and degradation metabolism of LNT is accelerated. The invention provides a TLR2-based immune enhancement mechanism of the LNT, and provides a theoretical basis and an application basis for developing immunomodulatory targeted drugs.

The present invention relates to a process for the dephosphorylation of skimmed milk, ultra-filtered milk (UF milk) or micellar casein isolate (MCI) comprising (i) acidifying the skimmed milk, UF milk or MCI, preferably not below pH 6.0, preferably between 6.0 and 6.7, (ii) cooling the acidified skimmed milk, UF milk or MCI to a temperature between 0 DEG C and 15 DEG C, (iii) adding gluconate and / or maleate to the cooled skimmed milk, UF milk or MCI, and (iv) washing the skimmed milk, UF milk or MCI to remove phosphorus, thereby dephosphorylating the skimmed milk, UF milk or MCI, preferably to the extent that the total phosphorus content of the skimmed milk, UF milk or MCI is reduced by at least 20%, preferably 20-40%, more preferably 30-40%, compared to the material provided to step (i). The invention also relates to a dephosphorylated MPC, MPI or MCI obtainable by the process of the invention and to a liquid enteral nutritional composition comprising 2.0-3.0 kcal / ml of heat sterilization wherein 16 en%-35 en% is provided by a protein, the combination of the calorie content and the relative protein calorie content being selected such that 10-18 g / 100 ml protein, preferably 12-18 g / 100 ml protein is present in the composition, preferably 12-18 g / 100 ml protein, preferably 12-18 g / 100 ml protein, preferably 12-18 g / 100 ml protein, preferably 12-18 g / 100 ml protein, preferably 12-18 g / 100 ml protein, preferably 12-18 g / 100 ml protein, preferably 12-18 g / 100 ml protein, preferably 12-18 g / 100 ml protein. Wherein the protein comprises micellar casein (MC), whey protein (WP) and optionally caseinate (CAS) wherein at least 70 wt% MC and less than 15 wt% WP are present, based on the total protein content, and wherein the composition has a total amount of phosphorus of less than 192 mg / 100 ml and / or 30-80 mg / 100 kcal, preferably at least 30-80 mg / 100 kcal, most preferably less than 192 mg / 100 ml and 30-80 mg / 100 kcal.

The invention provides a genetically engineered bacterium for efficiently producing pseudouridine and application of the genetically engineered bacterium. The genetically engineered bacterium is based on a high-yield uridylic acid strain E. coli MB219 (the patent application number is 202410153359.6), and the high-efficiency pseudouridine engineering bacterium is constructed through systemic gene modification. The genetically engineered bacterium is characterized in that (1) gene knockout: 10 genes such as pbs1, psuK and the like are knocked out to block a metabolic pathway of byproducts; (2) metabolism regulation: replacing a J23110 strong promoter, and overexpressing zwf / gnd to enhance a phosphopentose pathway; (3) heterologous gene integration: integrating a specific dephosphorylase gene HDHD1, and introducing RjpsuG, NmygdH and thyA to construct a co-expression plasmid pRL06; the fermentation yield of the finally obtained engineering bacterium CY20 (pRL06) reaches 45.3 g / L and is increased by 6 times compared with that of a traditional method, the production efficiency is greatly improved, the product purity is high, and by-products are few.

The present invention is a composition for inhibiting PTEN dephosphorylation enzymes, and specifically, the present invention is characterized by presenting a composition in which a CopA3 peptide derived from the small horned dung beetle binds to a PTEN enzyme via a covalent bond, thereby enabling inhibition of PTEN dephosphorylation enzymes.

This invention discloses a tandem-responsive self-assembled molecular probe and its applications. The HPO-CYFF-TPP of this invention is converted to HO-CYFF-TPP under the dephosphorylation of overexpressed ALP, restoring the intramolecular charge transfer effect. It then self-assembles into nanoparticles in situ through π-π stacking, exhibiting fluorescence signal quenching and photoacoustic signal enhancement at low viscosity. Upon laser irradiation, the NPs generate a large amount of singlet oxygen in the mitochondria, thereby achieving efficient photodynamic therapy (PDT) and leading to increased viscosity. At high viscosity, due to the suppression of the distorted intramolecular charge transfer effect of HO-CYFF-TPP, the NPs activate the FL signal with a high signal-to-noise ratio and further enhance the PA signal. Therefore, the HPO-CYFF-TPP of this invention can achieve efficient PDT and enable real-time therapeutic effect assessment through FL / PA signals.

The present invention provides a method for directly detecting organophosphorus pesticides based on nano-biosensors, which realizes direct detection of organophosphorus pesticides OPs in one step by using a biosensor with carbon dots rich in phosphate groups on the surface, with high sensitivity and good specificity, and can avoid interference from components such as proteins, reducing sugars, amino acids, and metal ions, without the need for complex sample pre-treatment. The method comprises the following steps: using organic small molecules containing phosphate groups as raw materials, preparing carbon dots PCDs rich in phosphate groups on the surface by carbonization; mixing PCDs as fluorophores with dephosphorylase to construct a nano-biosensor; mixing organophosphorus pesticides OPs of known different concentrations with the nano-biosensor, measuring the fluorescence spectrum of the nano-biosensor after standing, and drawing a standard curve; mixing the OPs sample to be detected with the nano-biosensor, measuring the fluorescence spectrum of the nano-biosensor, and calculating the concentration of the OPs to be detected according to the standard curve.

The invention discloses a method for changing zeta activity of DNA polymerase, and belongs to the technical field of biological medicine. The method comprises the following step: changing the zeta activity of the DNA polymerase by changing phosphorylation / dephosphorylation modification of residues at positions S279, S282 and S286 of human REV3L-N70 protein, or changing the stability of the REV3L-N70, or changing the interaction between the REV3L-N70 and REV3L-C protein, or changing a Caspase-mediated REV3L-C cleavage event. The zeta activity of the DNA polymerase cannot be effectively improved through overexpression of the REV3L, but after the REV3L is knocked out, the problem, such as influence on normal cell growth, also exists, and the invention provides an effective method for inhibiting or enhancing the zeta activity of the DNA polymerase from the perspective of stabilizing or removing an REV3L activity self-inhibition mechanism.

The invention discloses a GlcNAc6P induction component and a method for promoting synthesis of acetylglucosamine. The existing GlcNAc6P sensing element has the problems of serious leakage and small response range, which results in that in the process of regulating and controlling the expression of phosphatase by using the GlcNAc6P sensing element, the phosphatase also has higher expression intensity when intracellular GlcNAc6P is not accumulated, and the supply of precursor substances is influenced. Besides, along with rapid increase of the intracellular GlcNAc6P concentration, the upper limit of the expression quantity of phosphatase is low, the intracellular dephosphorylation requirement cannot be met, and further increase of the GlcNAc yield is limited. Therefore, an existing GlcNAc6P sensing element is optimized, a nagC binding sequence is subjected to directed evolution, expression intensity of repressor protein nagC is optimized, the leakage expression level of the sensor is reduced, and the response range of the sensor is expanded, so that the expression intensity of phosphatase is more effectively and rigorously regulated and controlled to adapt to the change of the concentration of GlcNAc6P in cells in different fermentation periods; and a foundation is laid for producing acetylglucosamine by further transforming recombinant bacteria through metabolic engineering.

The invention relates to a method for regulating and controlling selective absorption of cadmium and manganese by plants through dephosphorylation. The STNP-mediated phosphorylation is crucial to the selective uptake of cadmium and manganese of solanaceae NRAMP5 (Nuclear Receptor Amplification Protein 5). Simulation of non-phosphorylated mutation in NRAMP5 reduces the absorption of plants to cadmium without affecting the absorption of plants to manganese.

The present invention relates to a blood treatment device configured to dephosphorylate extracellular adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP) and / or lipopolysaccharide (LPS) in the blood of a patient in need thereof in an extracorporeal blood circuit, wherein the device comprises a matrix on which alkaline phosphatase (AP) is immobilized. The present invention also relates to an extracorporeal blood circuit comprising a blood treatment device of the present invention and to the use of the blood treatment device as a medicament or a method for treating an infection, preferably a blood or systemic infection such as sepsis, and / or for treating sepsis-related acute kidney injury (AKI).

Protein phosphatase 5 (PP5) is a serine / threonine protein phosphatase involved in the maturation and activation of numerous signaling pathways essential for cancer growth. PP5 activity is essential for the survival of clear cell renal cell carcinoma (ccRCC), however the mechanism remains unclear. Data demonstrates that PP5 interacts with caspase-8, FADD, and RIPK1, components of extrinsic apoptotic pathway Complex II. Specifically, PP5 dephosphorylates and inactivates the death effector proteins RIPK1 and FADD, preserving Complex II integrity and regulating extrinsic apoptosis. Protein phosphatases are considered to be ‘undruggable,’ however we have developed a specific inhibitor of PP5 (P-053) that prevents substrate binding to the active site. Encouragingly, PP5 inhibition using P-53 in VHL-null ccRCC robustly induces extrinsic apoptosis. Taken together, the data suggests that PP5 promotes ccRCC survival by suppressing extrinsic apoptosis, and small molecule inhibition of PP5 presents a viable therapeutic strategy for ccRCC.

The application provides a uridine monophosphate specific glycoside hydrolase and application thereof in biosynthetic synthesis of pseudouridine. Nmygdh The function of the gene is verified by in-vitro enzyme activity experiment Nmygdh The recombinant protein has broad-spectrum and high-efficiency catalytic activity, and can efficiently catalyze the hydrolysis of glycosidic bonds in various nucleotides, such as uridine monophosphate (UMP), guanosine monophosphate (GMP), adenosine monophosphate (AMP) and cytidine monophosphate (CMP), and the activity of uridine monophosphate is the strongest. The application uses the protein to efficiently hydrolyze uridine monophosphate to obtain two substrates of pseudouridine, i.e. uracil and 5'-phosphoribose, and realizes the atom-economical and efficient biosynthetic synthesis of pseudouridine by combining the cascade reaction of pseudouridine glycosidase EcPsuG and dephosphorylase EcYjjG.

The invention discloses an enzymatic self-anchored pre-targeting molecule P-TCO as well as a preparation method and application thereof.The enzymatic self-anchored pre-targeting molecule P-TCO provided by the invention is subjected to a dephosphorylation reaction under the action of high-expression alkaline phosphatase of a tumor cell membrane, and an active intermediate with a QM group is generated. The intermediate can be used for covalently labeling ALP and membrane proteins around the ALP so as to ensure that a TCO group is stably anchored on a cell membrane. Subsequently, the TCO group covalently labeled on the membrane protein can be subjected to an efficient biological orthogonal reaction with the tetrazine probe, so that a targeted delivery platform of the multi-modal imaging probe is constructed, high-sensitivity visualization of living tumors is realized, and huge application potential in the field of precise diagnosis of clinical tumors is shown.

According to the application of the reagent for detecting the phosphorylation level of the S202 and T205 sites of the MAPT protein in preparation of the renal fibrosis diagnosis product, the function research of the MAPT protein is expanded from the traditional field of glomerular podocytes to renal tubular epithelial cells and the fibrosis pathological process of the renal tubular epithelial cells for the first time, and a brand new direction of the MAPT protein in renal disease research is opened up. Different from the conventional general research means which only depends on overall gene knockout or overexpression, the method provided by the invention realizes accurate analysis of the function of the MAPT key phosphorylation site by using the site-specific mutant. Through a systematic function determination experiment, the invention discloses an anti-intuition biological rule with great theoretical significance: although phosphorylation of the sites S202 and T205 obviously drives the fibrosis process of the renal tubular epithelial cells, the fibrosis phenotype cannot be improved by simulating the dephosphorylation state (S202A / T205A mutant) of the sites.

Various aspects of this disclosure relate to the discoveries that (1) acids can catalyze the spontaneous dephosphorylation of phosphoryloxytryptamines into hydroxytryptamines such as psilocybin into psilocin, and (2) hydroxytryptamines are less stable to spontaneous oxidation than phosphoryloxytryptamines. The salt of a phosphoryloxytryptamine and an anion that is the conjugate base of a weak acid can buffer pH upon dissolution of the salt to inhibit the acid-catalyzed dephosphorylation of the phosphoryloxytryptamine and thereby protect it from spontaneous oxidation. Suitable anions include acetate, bicarbonate, dihydrogen phosphate, aspartate, and glutamate.

The invention provides a bifunctional nucleic acid aptamer-nano antibody conjugate, which comprises: a, a nucleic acid aptamer, which is targeted to phosphatase; b, a nano antibody of a target receptor; c, one or more connexons, wherein the connexons are used for connecting the nucleic acid aptamer and the nano antibody; wherein the phosphatase is used for carrying out dephosphorylation on a target receptor. The bifunctional aptamer-nano antibody conjugate disclosed by the invention can enhance affinity and reduce off-target effect, and can be used for resisting tumor proliferation and overcoming drug resistance.

The invention provides a preparation method and application of a multi-target enzymatic self-assembly fluorescence activated nanoprobe, and belongs to the field of pharmacy. According to the method, an Fmoc solid-phase peptide synthesis strategy is adopted, an N end is connected with a fluorescence quencher Dabcyl, a C end is marked with fluorescein, a near C end is a self-assembly sequence Y (pY) YG capable of generating self-assembly behavior through dephosphorylation of alkaline phosphatase, and a middle section is a cleavage sequence KGGFLGK capable of being cleaved into a fluorescent'switch 'by cathepsin B; the near N end is a functional polypeptide F-pY-LyP-1 of a targeting sequence CGNKRTRGC or LyP-1 which can be highly combined with a p32 receptor on the surface of a foam cell. The fluorescence activated nanoprobe accurately reaches a plaque part under the synergistic effect of multiple target points and is self-assembled into spherical nanoparticles, so that the aggregation and retention effects of the probe in the plaque are improved, the non-specific signal interference is reduced, and the early atherosclerotic plaque is more accurately identified.

A transmucosal psychoactive alkaloid composition including a psychoactive alkaloid extract or synthetic psychoactive alkaloid. The alkaloids in the extract are predominantly dephosphorylated rather than phosphorylated. The transmucosal psychoactive alkaloid composition also includes a mucoadhesive polymer, a carrier, and optional further excipients. The non-ingestive composition may be taken orally through the mucosa. A process for obtaining an oral transmucosal psychoactive alkaloid composition includes dephosphorylating the alkaloid during extraction, purifying the extracted alkaloid and standardizing to a specific concentration by adding measured quantities of excipients.

The present invention belongs to the field of pharmaceutical chemistry, and specifically relates to a class of compounds shown in formula I or formula II as protein phosphatase 5 agonists, preparation methods thereof, and pharmaceutical compositions containing these compounds and applications of these compounds or compositions in drug preparation. The present invention provides a class of 2-imine-2H-chromene-3-thioamide protein phosphatase (PP5) agonists and derivatives thereof, by discovering active small molecules with novel structures and good physicochemical properties, and the small molecules have shown good activity in various experiments, directly targeting PP5 to produce agonistic effects, and provide a new solution for the treatment of related diseases caused by PP5 abnormalities, and is expected to be developed into specific drugs for the treatment of Alzheimer's disease, obesity caused by glucocorticoid treatment, and cancer caused by protease dephosphorylation dysfunction, and has clinical application value.

The present invention relates to a cell lysate wherein the lysis buffer comprises magnesium acetate, the phosphorylation inhibitor K3L, the agonist of dephosphorylation GADD34 and a caspase inhibitor for use in, or as, a cell-free system and a method for preparing a cell-free protein synthesis system. Further provided is a method for expressing a protein in the cell-free protein synthesis system, a method for screening for a candidate compound for an effect on a target molecule and a use of the cell-free protein synthesis system. The present invention also provides a kit for the above-mentioned cell-free systems.

Provided is a method for producing an olefin compound using reaction with a base, whereby a target product with a higher purity than the prior art can be obtained. Specifically, provided is a method for producing an olefin compound by reacting a phosphorus-containing olefin compound salt with a base to obtain a dephosphorized and hydrogenated olefin compound, wherein the temperature of the reaction is 50° C. or less.

It is herein demonstrated that an increase in Dox-induced miRNA-106b-5p (miR-106b) is able to down-regulate PR55α subunit, which leds to a decrease both in PP2A activity as well as an accumulation of the phosphorylated HDAC4 protein in cytosol. Anti-miRNA-106b-5p (miR-106b) therapy is thus able to prevent Dox-induced PR55α degradation, facilitate the HDAC4 dephosphorylation by PP2A, and increase HDAC4-YY1 interaction, resulting in a decreased in sST2 expression and release by the LV myocardium. Further, and for first time, it has identified the role that sST2 isoform plays in Dox-induced CTX.

The invention discloses a lung cancer cell strain with YY1S247A mutation and application of the lung cancer cell strain. According to the present invention, phosphorylation modification of serine at the 247th site of YY1 is detected through mass spectrometry, and in order to explore the influence of the phosphorylation modification at the site on the function of YY1 in non-small cell lung cancer cells, a lung cancer cell strain in which a YY1S247A mutant gene is knocked is constructed through a CRISPR / Cas9 gene editing technology; dephosphorylated YY1 (YY1S247A mutation) is detected in a constructed lung cancer cell line to block the promotion effect of third-generation EGFR-TKI on CD24 expression, and the cell strain can be widely applied to research on the function of YY1 in non-small cell lung cancer and research on the drug resistance mechanism of the third-generation EGFR-TKI, and can also be used for developing therapeutic drugs for targeted inhibition of phosphorylation of YY1S247 to sensitize EGFR-TKI.