47 results about "Protein stability" patented technology

The present disclosure provides drug responsive domains derived from human carbonic anhydrase 2 that can modulate protein stability for human interleukin 15 (IL15) payloads, as well as compositions and methods of use thereof.

The present disclosure relates to the use of a maltose dependent degron to control stability of a protein of interest fused thereto at the post-translational level. The present disclosure also relates to the use of a maltose dependent degron in combination with a maltose-responsive promoter to control gene expression at the transcriptional level and to control protein stability at the post-translational level. The present disclosure also relates to the use of a stabilization construct that couples expression of a cell-growth-affecting protein with the production of non-catabolic compounds. The present disclosure further relates to the use of a synthetic maltose-responsive promoter. The present disclosure further provides compositions and methods for using a maltose dependent degron, a maltose-responsive promoter, and a stabilization construct, either alone or in various combinations, for the production of non-catabolic compounds in genetically modified host cells.

Disclosed herein are peptide hydrogel-based formulations. In some embodiments, the peptide hydrogel-based formulations encapsulate antibodies. In some embodiments, the peptide hydrogel-based formulations prevent formation of high molecular weight protein species.

This invention discloses a mutant, gene, and recombinant bacteria of a highly thermostable single-chain sweet protein. The mutant is characterized by the following amino acid sequences shown in SEQ ID NO.1: isoleucine (I) at position 5 is mutated to glutamic acid (E), glutamic acid (E) at position 23 is mutated to alanine (A), isoleucine (I) at position 26 is mutated to arginine (R), cysteine ​​(C) at position 41 is mutated to alanine (A), tyrosine (Y) at position 65 is mutated to isoleucine (I), glycine (G) at position 83 is mutated to arginine (R), and asparagine (N) at position 90 is mutated to glutamic acid (E). Based on structural analysis and computationally assisted design for protein stability, this invention achieves site-directed mutagenesis of the single-chain sweet protein, resulting in a highly thermostable mutant. Compared to the original single-chain sweet protein, the thermostable mutant of this invention exhibits higher T... m It was heated by more than 20.4°C while maintaining its sweetness.

The application discloses a methionine adenosyltransferase mutant and application of the mutant in immobilized reaction, the mutant is obtained by mutating an amino acid sequence of wild-type Escherichia coli methionine adenosyltransferase, the mutant enhances the interaction force between subunits by introducing a disulfide bond at a specific position; and / or, the protein stability is enhanced by constructing a connecting peptide between adjacent subunits. The mutant provided in the application enhances the thermal stability and catalytic efficiency of the enzyme, reduces the optimum reaction pH value, reduces the generation of by-products and the precipitation of a reaction solution. The mutant enzyme can maintain high activity at a temperature of about 40 DEG C, and is particularly suitable for industrial production. In addition, the immobilized form of the mutant enzyme exhibits good stability and reusability in continuous use, significantly reduces production cost, simultaneously reduces environmental pollution, improves the yield of SAM, is more suitable for large-scale industrial production, and has more market competitiveness.

The application discloses a mutant and gene of single-chain sweet protein with high thermal stability, wherein the mutant is obtained by mutating isoleucine (I) at the 5th position, glutamic acid (E) at the 23rd position, isoleucine (I) at the 26th position, tyrosine (Y) at the 65th position, glycine (G) at the 83rd position and asparagine (N) at the 90th position in the amino acid sequence shown in SEQ ID NO. 1 to alanine (A), arginine (R), isoleucine (I), glutamic acid (E) and aspartic acid (D) respectively; the application is based on structural analysis and protein stability calculation aided design, and the single-chain sweet protein is subjected to site-directed mutagenesis, so that the mutant of single-chain sweet protein with high thermal stability is obtained; compared with the single-chain sweet protein, the mutant of single-chain sweet protein with high thermal stability has a T m increase of 18.4 DEG C or more and maintains sweetness.

The invention provides application of an expression inhibitor of KMT5A in preparation of a medicine for preventing or treating esophageal squamous carcinoma. In-vitro experiments prove that ICA can inhibit proliferation of esophageal squamous carcinoma cells in a concentration-dependent manner and remarkably promote cell apoptosis. The action mechanism of the ICA is closely related to the direct targeting KMT5A protein, and experiments show that the ICA can reduce the expression level of the KMT5A and influence the protein stability of the KMT5A. Clinical sample analysis further shows that KMT5A is highly expressed in esophageal cancer tissues, and ICA treatment in animal experiments significantly reduces the tumor volume. The invention provides a new targeted treatment strategy for esophageal squamous carcinoma, and has important clinical transformation value.

The invention relates to the technical field of protease mutation, and discloses a bisabolol synthase mutant with high catalytic activity and application of the bisabolol synthase mutant. According to the method, alpha-bisabolol synthase derived from compositae herbaceous plant globe artichoke is taken as wild enzyme, mutants are virtually screened by applying a protein language model SaProt, a deep learning model Catapro and a protein stability prediction tool Dynaamt, and the effect of producing bisabolol is verified through experiments; according to the present invention, the mutants obtained by mutation of 11 specific amino acid sites such as D125N, F159Y, F159K, N346D, T372S, Y432F, N346S, D184E, N157E, E374A and T258S are obtained through the mutation of the above 11 specific amino acid sites; wherein through 96-hour shake flask fermentation and gas chromatography detection, the bisabolol yields of the obtained single-point mutation T258S and double-point mutation T258S / F159K are respectively increased by 72.6% and 90.2% compared with the wild type yield; the Kcat / Km value of the two-point mutation T258S / F159K is 4.57 times that of a wild type.

The application discloses a mutant of single-chain sweet protein with high thermal stability and a gene of the mutant, wherein the mutant is obtained by mutating isoleucine (I) at the 5th position, glutamic acid (E) at the 23rd position, isoleucine (I) at the 26th position, tyrosine (Y) at the 65th position, glycine (G) at the 83rd position and asparagine (N) at the 90th position in the amino acid sequence shown in SEQ ID NO. 1 to alanine (A), arginine (R) and aspartic acid (D) respectively. m The Tm of the mutant of single-chain sweet protein with high thermal stability is increased by more than 19.4 DEG C, and the sweetness is maintained.

This invention belongs to the field of plant genetic engineering and relates to the application and method of reducing the expression of the Zm7649 gene in improving plant stress resistance. This invention utilizes an EMS mutant with premature termination of expression. Zm7649-ems Heterologous overexpression with Arabidopsis thaliana Zm7649-OE Verified Zm7649 The function of negatively regulating salt stress and high temperature stress was studied, and its preliminary analysis was performed. Zm00001d027649 The molecular mechanism regulating stress involves the interaction between Zm7649 and a maize catalase (Zm4848), with ubiquitination degrading Zm4848 and reducing its protein stability. This invention provides new genetic resources and theoretical basis for maize stress-resistance breeding or germplasm innovation.

The present application relates to the construction method of the ophthalmic protein drug sustained-release preparation, the ophthalmic protein drug is contacted with the weakly interacting excipient of pharmaceutically acceptable, and the protein pre-holding complex is formed;The first hydration excipient and the second hydration excipient are sequentially added to the protein pre-holding complex, and the layered hydrated protein complex is formed;The layered hydrated protein complex is dispersed in the first continuous phase of the ionizable group-containing pharmaceutically high polymer, and the ion pairing release blocking layer is formed in the periphery by adjusting the ion environment, and the intermediate is obtained;The intermediate is dispersed in the second continuous phase of the ophthalmic pharmaceutically acceptable, and the ophthalmic protein drug sustained-release preparation is prepared.The ophthalmic protein drug is pre-held with the weakly interacting excipient, the differentiated occupation area and the layered hydration structure are constructed, the protein stability is improved;Through the stage ion regulation, the pharmaceutically high polymer forms the non-closed ion pairing release blocking layer, the protein release fine control is realized, the drug efficacy is prolonged and the curative effect is improved.

The invention discloses a mutant of HPV53 E6 protein and application of the mutant. Compared with wild type E6, the mutant has the following three mutation sites: K79W, S133P and N8K, the K79W mutation design enhances the stability of a hydrophobic space and forms new hydrophobic interaction with Y108 / H130, S133P enables the flexibility of a loop region to be more stable, N8K mutation forms a salt bridge with E10 / E11, and the relation between the loop region and helix is enhanced. The mutation designs can maintain the stability of the whole structure, significantly improve the protein stability and immunogenicity, break through the technical bottlenecks of low in-vivo expression quantity and weak immunogenicity of the natural E6 protein, and provide a new way for immunotherapy of HPV53 related lesions.

The application discloses a hair loss prevention shampoo targeting CD133 positive hair follicle dermal papilla cells and a preparation method thereof. The shampoo comprises a hair follicle targeting single-domain antibody-active peptide fusion, a liposome-encapsulated compound plant extract, a biological surfactant, a protein stability surfactant and an auxiliary material. The fusion is formed by fusing an anti-CD133 VHH single-domain antibody and a 5alpha-reductase inhibiting peptide DK-HP-7 through a flexible connecting peptide. Based on the discovery that CD133 positive dermal papilla cells are a key cell group driving hair follicle formation, the application uses single-domain antibodies to achieve precise targeting of active ingredients. The DK-HP-7 designed through molecular docking has an IC 50 of 4.28 μM. The rhamnolipid and Kolliphor HS 15 complex system can make the fusion protein activity retention rate higher than 89% after being stored at 40 ℃ for 3 months. The shampoo can significantly antagonize dihydrotestosterone-induced hair follicle growth inhibition, promote hair follicles to enter the growth phase and increase hair follicle density, and has good skin safety and stability.

This application provides a protein stability detection probe based on ascorbate peroxidase 2 and its application. The detection probe includes ascorbate peroxidase 2 displayed on the cell surface and an anchoring protein that anchors ascorbate peroxidase 2 to the cell. Ascorbate peroxidase 2 has an insertion site for the target protein, into which the target protein inserts. Ascorbate peroxidase 2 is divided into N-terminal and C-terminal ascorbate peroxidase 2, and the target protein is linked to both ends of the C-terminal ascorbate peroxidase 2 via flexible linkers. The intensity of the fluorescence signal generated by the cascade reaction catalyzed by the probe in this application has a good linear relationship with the stability of the inserted test protein mutant, enabling high-throughput identification of protein mutant stability and showing promising application prospects.

The application provides a hepatitis C antibody detection antigen composition and application, kit and detection method thereof, relates to the antibody detection technical field, and the hepatitis C antibody detection antigen composition comprises a labeled recombinant antigen and a coated recombinant antigen, the amino acid sequence of the labeled recombinant antigen includes Trx and hepatitis C Core region, NS3 protein, NS4b protein dominant epitope and His tag, and the full-length sequence is shown as SEQ ID NO. 1. The labeled recombinant antigen used in the application is obtained by mutating 7 cysteines (C) into alanine (A) by amino acid mutation on the NS3 protein, reducing the aggregation and precipitation caused by disulfide bond of the recombinant protein, and improving the protein stability. When colloidal gold is used for antigen labeling, a secondary indirect labeling method is adopted, which is beneficial to the exposure of antigen epitopes, can improve the reaction sensitivity, and reduces the antibody missed detection.

The invention relates to a group of N-glycosylation mutants of type I alpha-mannosidase and application of the N-glycosylation mutants. According to the invention, N-glycosylation modification of tomato-sourced I-type alpha-mannosidase SlMNSI1 at amino acid residues N288 and N334 is analyzed for the first time, and the modification sites are located in a conserved structural domain of a GH47 glycoside hydrolase family. N288A, N334A and N288A / N334A mutants are further constructed, and the N288A, N334A and N288A / N334A mutants are transiently expressed in mesophyll cells of the nicotiana benthamiana. Positioning analysis shows that the mutant can still be partially positioned in the Golgi apparatus, but protein aggregates appear and are transferred to the periphery of the endoplasmic reticulum. Protein stability detection shows that the N-glycosylation modified mutant is obviously accelerated to degrade after being treated by the cycloheximide, which indicates that the glycosylation modification of the N288 and N334 sites has an important effect on the stability and subcellular localization of the SlMNSI1. The functional characteristics of key glycosylation sites of the type I alpha-mannosidase are disclosed for the first time, a group of N-glycosylation mutants with structure and function differences are provided, and the N-glycosylation mutants can be used as important tools for researching plant glycosylation regulation, protein engineering modification and gene function identification and have wide application prospects.

The invention relates to the technical field of protein engineering and metabolic engineering, in particular to a mutant for improving the activity and stability of AceA and application of the mutant in biosynthesis. The mutant is obtained by mutating serine at the 335th site and / or the 398th site into glutamine on the basis of an escherichia coli wild type AceA. The mutant has enzyme activity obviously higher than that of a wild type and enhanced protein stability. The gene encoding the mutant is introduced into engineering bacteria for producing acetyl coenzyme A derivatives (such as phloroglucinol and 3-hydracrylic acid) or glyoxylic acid pathway derivatives (such as glycolic acid), so that glyoxylic acid branches can be effectively strengthened, and the yield and carbon conversion efficiency of target products are remarkably improved. The invention provides a novel key enzyme element and a metabolic engineering strategy for efficiently producing various chemicals with high added values.

The invention discloses an application of Irisin autocrine probiotics in preparation of a medicine for preventing / treating salt-sensitive hypertension. According to the application disclosed by the invention, it is found for the first time that the probiotic B.Subtills-WB600-Irisin capable of automatically secreting Irisin protein simultaneously has the effects of preventing and treating salt-sensitive hypertension, can effectively inhibit the increase of systolic pressure and diastolic pressure induced by rats fed with high-salt diet, and can maintain the systolic pressure and diastolic pressure at a basic blood pressure level; in a Dahl-S rat model, the autotaxis Irisin probiotic treatment can improve the urine sodium excretion, so that the blood pressure rise induced by high-salt diet reverses 26 + / -8 mmHg (plt; 0.001). Compared with the traditional probiotics, the invention solves the key problems of protein stability, targeting and multi-mechanism synergy through a synthetic biological strategy, and can be used as a medicine for preventing and treating hypertension and related diseases thereof.

This invention discloses an ultrafiltration system with real-time protein concentration control, belonging to the field of biopharmaceutical technology. The system includes a replenishment tank, a recirculation tank, an ultrafiltration unit, and a controller. The recirculation tank is connected to the replenishment tank via a replenishment pipe, which is equipped with a replenishment pump and a valve, with the valve located between the replenishment pump and the recirculation tank. A protein concentration detection component A is located at the bottom of the recirculation tank. The ultrafiltration unit is connected to the recirculation tank via a delivery pipe and a return pipe. A circulation pump is installed on the delivery pipe, and a back pressure component and a protein concentration detection component B are installed on the return pipe. This system can improve the consistency of protein concentration in the recirculation tank during constant-volume dialysis, reduce batch-to-batch differences in protein concentration, and thus improve the uniformity of product quality. It can also prevent concentration polarization caused by excessively high protein concentration from affecting protein stability.

This invention discloses a combination formulation of a BRD7 stabilizer and a HER2-targeting drug, and its application, belonging to the field of pharmaceutical molecular biology. This invention is the first to discover that BRD7 can upregulate HER2 protein stability in triple-negative breast cancer, promoting the transformation of triple-negative breast cancer cells to HER2-positive cells, thereby increasing the sensitivity of triple-negative breast cancer cells to HER2-targeting antibody drugs. Further research demonstrates that BRD7 increases HER2 protein stability by inhibiting HER2 protein degradation mediated by the autophagy-lysosomal pathway. Therefore, the combination of a BRD7 stabilizer drug and a HER2-targeting monoclonal antibody drug exhibits significant anti-tumor effects. This invention provides a potential treatment strategy for the clinical treatment of triple-negative breast cancer and has broad application prospects.

The invention belongs to the technical field of bioengineering, and particularly relates to high-stability recombinant elastin as well as a preparation method and application thereof. The amino acid sequence comprises a repeating unit with a general formula of (VPGXG) n, X is one of V, L, A and G, and n is an integer from 20 to 40; a cell adhesion peptide RGD sequence is inserted behind the penultimate VPGXG repetitive unit; and non-natural amino acid p-acetyl phenylalanine is introduced into the sequence as a photo-crosslinking site. According to the high-stability recombinant elastin as well as the preparation method and the application thereof, unnatural amino acid and a specific functional domain are introduced, so that unification of protein stability, biological activity and controllable crosslinking property is realized, and a foundation is laid for constructing intelligent biological materials. The pichia pastoris expression and inverse phase change purification technology is adopted, the bottleneck of a traditional process is broken through, and efficient, low-cost and large-scale production of the recombinant elastin is achieved.

The invention discloses an anti-tumor medicine composition and application thereof, the anti-tumor medicine composition has a synergistic effect, the inhibition effect on lung cancer cells PC-9 or NCI-H1975 is remarkably improved, and the anti-tumor medicine composition can be used for inhibiting the growth of source organs of lung adenocarcinoma patients. The anti-tumor pharmaceutical composition comprises an SPTLC1 inhibitor and a targeted drug, and the SPTLC1 inhibitor is a compound for inhibiting the enzyme activity, transcription, translation or protein stability of SPTLC1.

An isolated polynucleotide encoding a modified luciferase polypeptide and substrates. The OgLuc variant polypeptide has at least 60% amino acid sequence identity to SEQ ID NO: 1 and at least one amino acid substitution at a position corresponding to an amino acid in SEQ ID NO: 1. The OgLuc variant polypeptide has at least one of enhanced luminescence, enhanced signal stability, and enhanced protein stability relative to the corresponding polypeptide of the wild-type Oplophorus luciferase.

The invention discloses a pig STING protein K61R mutant and application thereof, the mutant is obtained by mutating the 61th lysine of the pig STING protein into arginine, and the amino acid sequence of the pig STING protein is as shown in SEQ ID NO. 1. According to the invention, the species-specific key site K61 of the pig STING protein is identified for the first time, the site is unique to pigs and is different from other species, and by regulating the protein stability and downstream signal transduction, a key negative regulation effect on virus replication of ASFV, PRV, HSV-1 and the like is achieved; meanwhile, the K61 site can be directly used as a core molecular marker and a targeting site for pig antiviral breeding, and important support is provided for improving the research efficiency of disease-resistant breeding.

The present disclosure relates to modified proteins, methods, compositions and kits for mapping the methylation status of nucleic acids comprising 5-methylcytosine and 5-hydroxymethylcytosine. In some embodiments, the modified proteins have been altered to increase protein stability. In some embodiments, a protein selectively acts on certain modified cytosines of a target nucleic acid and includes one or more substitution mutations that enhance the protein's selectivity for certain modified cytosines, optionally enhance the stability of the protein, or optionally enhance both selectivity and stability. Also provided are compositions and kits comprising one or more of the proteins, as well as methods of using one or more of the proteins.

The invention provides recombinant camel chymosin and application thereof, and belongs to the technical field of gene recombination. The amino acid sequences of the recombinant camel chymosin disclosed by the invention are as shown in SEQ ID NO. 1 to SEQ ID NO. 5. The physicochemical properties, the amino acid composition and the structure of the bovine chymosin and the bactrian camel chymosin are compared and analyzed by utilizing a bioinformatics method. Analyzing a three-dimensional conformation of an enzyme-substrate compound through molecular docking, and identifying key residues influencing the catalytic efficiency and the protein stability by combining charge analysis, flexible analysis and alanine scanning; rosetta is used for site-specific saturation mutation library construction, and preliminary screening of mutants is realized through energy minimization and conformation enumeration; based on quantum mechanics / molecular mechanics simulation and machine learning prediction, the catalytic efficiency change rule of the mutant is quantified, and the optimal bactrian camel chymosin mutant can be screened.

The invention provides an application of a stress protein USP1 (University Stress Protein 1) in tomato bacterial diseases and a method thereof, according to the invention, through a real-time fluorescent quantitative PCR (Polymerase Chain Reaction) (qRT-PCR) technology, it is found that SlUSP1 is expressed in various tissues and is obviously induced by a pathogenic bacterium PstDC3000. SlUSP1 knockout and overexpression plants are constructed, and forward regulation and control of disease resistance are proved: disease spots of the knockout plants are increased, defensive gene expression and antioxidant enzyme activity are reduced, and resistance is enhanced. The screening and verification of the interaction protein show that the SlUSP1 and the E3 ubiquitin ligase component SlWD1 / SlWD3 interact and exist in a dimer form, and the protein stability is regulated and controlled by a 26S ubiquitin-proteasome pathway. The overexpression of the SlUSP1 promotes stem diameter and branch increase, plant height reduction and growth and development related gene expression improvement, and provides a key molecular target for disease-resistant high-yield breeding in lodging resistance and growth regulation.

The application belongs to the field of anti-solid tumor drugs, and particularly relates to application of GSK-F1 in preparation of a drug for reducing NSUN2 protein stability, solid tumor targeted treatment and / or radiotherapy sensitization. Researches of the application show that GSK-F1 has the effect of inhibiting NSUN2 protein stability, and can realize targeted treatment of solid tumors and radiotherapy sensitization based on inhibition of NSUN2 tumor protein stability.

The invention relates to the field of dairy product processing, in particular to a screening and pretreatment method and application of high-stability raw milk. The method comprises the following steps: sequentially carrying out a gradient alcohol test and an improved boiling test on raw milk to screen a high-protein-stability raw material; and further performing a thermal stability test on the raw milk to be used for the ultra-high-temperature instantaneous sterilization product. And sequentially carrying out degassing treatment and pre-pasteurization treatment on the qualified raw milk, and then rapidly cooling and storing. According to the method, the detection sensitivity and prediction accuracy of the stability of the raw milk protein are remarkably improved through multiple detection, special raw materials suitable for producing high-end products such as high-calcium milk, coffee / tea milk and concentrated milk can be effectively screened out, and the excellent flavor and storage stability of the raw materials are guaranteed through a matched pretreatment process; the technical problem that an existing conventional method cannot meet the requirement for accurate raw material screening of high-end dairy products is solved.

The application discloses application of USP33 in stabilizing HDAC3 in tumors, and relates to the technical field of biological medicines; the USP33 improves the protein stability of HDAC3 and promotes the progress of liver cancer. The application finds and clarifies that in hepatocellular carcinoma, the deubiquitinating enzyme USP33 promotes the proliferation of liver cancer by stabilizing the protein level of HDAC3; the application discloses a pathway for the synergistic regulation of liver cancer progress by USP33-HDAC3; the application proposes a strategy for targeting the signal shaft to treat hepatocellular carcinoma; by inhibiting the expression of USP33 or directly inhibiting the activity of HDAC3, the degradation of HDAC3 or the blocking of the function of HDAC3 can be promoted, so that the purpose of inhibiting the growth of tumors is achieved; and the application provides a new candidate target point for the drug research and development of hepatocellular carcinoma.