29 results about "Chaperonin" patented technology

The invention discloses an aspergillus niger recombinant bacterium AnCT-XynB-K-agdA and application thereof, and belongs to the technical field of microbiology and bioengineering. According to the invention, Aspergillus niger AnCat is taken as an expression host, firstly, an acid protease regulatory factor prtT is knocked out to obtain a defective strain, and then different chaperonins are subjected to fusion expression by optimizing integration sites of a laccase lcc9 expression cassette, so that the high-expression laccase chaperonin is obtained. According to the method, an expression vector pC3-5 'agdA-PcitA-XynB-L-lcc9-hph-3' agdA is constructed, a recombinant bacterium AnCT-XynB-K-agdA is further obtained, the enzyme activity of the extracellular laccase subjected to shake flask fermentation reaches 1821.2 U / L, and finally, the recombinant efficient expression of the laccase Lcc9 in aspergillus niger is realized.

This article describes a method for preventing or treating severe asthma by administering peptides associated with chaperone protein 60.1.

The invention provides a composition comprising a Plasmodium species multimeric protein capable of forming a nanoparticle and at least one antigen of interest, wherein the multimeric protein comprises a monomer chosen from Plasmodium species pyridoxal 5'-phosphate synthase (PLP), chaperone 60 protein (Cpn60), and caseinolytic protease (Clp). In embodiments, the invention provides the composition wherein the monomers assemble to form a multimeric nanoparticle and / or which comprises an adjuvant. The invention further provides a nucleic acid encoding the multimeric protein and the at least one antigen of interest, E. coli comprising the nucleic acid, and the use of such in a method for producing the inventive multimeric protein and the at least one antigen of interest. Also provided are a method and use of the inventive multimeric protein and the at least one antigen of interest for immunizing a subject against Plasmodium species.

A transformed microorganism having an ability to produce a polyhydroxyalkanoate includes a polyhydroxyalkanoate synthase gene and a gene encoding a chaperone belonging to the ClpB family. The gene encoding the chaperone belonging to the ClpB family is a gene which has been introduced into the transformed microorganism or whose expression has been enhanced in the transformed microorganism. The chaperone belonging to the ClpB family may be derived from the genus Cupriavidus, Escherichia, or Saccharomyces. A polyhydroxyalkanoate can be produced by culturing the transformed microorganism.

Provided are a mutant of a small molecular chaperone protein MOG1-encoding gene and use thereof. The mutant is more effective than a wild-type MOG1 in enhancing myocardial sodium current, and the mutant of the small molecular chaperone protein MOG1-encoding gene and a mutant of the small molecular chaperone protein MOG1 are used in the preparation of medicaments for the treatment of diseases such as Brugada syndrome, arrhythmia, dilated cardiomyopathy, or heart failure.

The invention relates to double stranded ribonucleic acid (dsRNAi) agents and compositions targeting the SCAP gene, as well as methods of inhibiting expression of a SCAP gene and methods of treating subjects having a SCAP-associated disorder, such as nonalcoholic fatty liver disease (NAFLD) or nonalcoholic steatohepatitis (NASH), using such dsRNAi agents and compositions.

This invention relates to the field of biotechnology, and more particularly to the application of the wheat molecular chaperone protein TaBiP1 in the control of wheat scab. This invention is the first to discover that overexpression of the wheat molecular chaperone protein TaBiP1 can significantly reduce the infection severity of wheat scab. Compared with the transgenic recipient parent Fielder, TaBiP1-overexpressing plants significantly reduce wheat scab infection and can be used to create new scab-resistant germplasm. In this invention, the TaBiP1 gene in recipient wheat is edited using CRISPR / Cas9 gene editing technology to obtain transgenic plants. bip1 The resistance of wheat to Fusarium head blight was weaker than that of Fielder, which confirms the positive significance of TaBiP1 in enhancing wheat resistance to Fusarium head blight.

The invention belongs to the technical field of globular protein synthesis, and particularly relates to a chaperonin-assisted recombinant human hFGF-2 protein preparation method and application thereof, the chaperonin-assisted recombinant human hFGF-2 protein preparation method comprises the following steps: S1, constructing a recombinant vector TrxA-6H-DDDDK-hFGF2 containing an hFGF-2 fusion protein coding sequence; s2, transforming the recombinant vector into an escherichia coli host cell for induced expression to obtain an expression thallus; s3, crushing the expression thalli to obtain a cell lysis supernatant; s4, taking the supernate, carrying out nickel column affinity chromatography, carrying out desalination treatment on the fusion protein by adopting gel chromatography, and collecting a fusion protein eluent; s5, performing enterokinase cutting treatment on the fusion protein to obtain an enzyme-digested product; s6, performing heparin affinity chromatography on the enzyme digestion product, and collecting hFGF-2 protein eluent; and S7, carrying out dialysis treatment on the protein eluent to obtain the recombinant hFGF-2 protein.

The application discloses a method for removing chaperone and application thereof. The method for removing chaperone comprises the following steps: after a sample containing a target recombinant protein is loaded on an affinity chromatography column, a washing liquid containing urea and KCl is used to flush the affinity chromatography column, so as to remove the chaperone combined with the target recombinant protein. The method of the application can effectively remove the chaperone of about 60 kDa from an escherichia coli host bacterium by combined washing of urea and KCl, and has the advantages of simple operation, low cost, good chaperone removal effect for various recombinant proteins with different molecular weights, wide universality, and the like, and can significantly improve the purity and quality of the recombinant protein.

This invention discloses a sucrose isomerase site mutant, a genetically engineered bacterium, and a method for catalyzing the production of isomaltulose. This invention utilizes PROSS online analysis combined with sequence alignment analysis to screen key amino acid sites related to catalytic activity in sucrose isomerases. Through site-directed mutagenesis, the mutant WT-Gro7-Q474T was obtained, with a maximum specific enzyme activity of 683 U / mg. k cat (s ‑1 The concentration reached 718. By optimizing protein expression conditions, including temperature, IPTG concentration, bacterial concentration, and time, and combining this with the method of co-expression of molecular chaperone proteins, a highly efficient heterologous expression system for sucrose isomerase was established. This invention established an optimal isomaltulose-catalyzed reaction system: in a 2 L reactor, with 1 L of reaction solution and a substrate concentration of 800 g / L sucrose, after 6 hours of reaction, all sucrose was converted, yielding isomaltulose 792.50 g / L, with a conversion rate >99.5%.

Methods and compositions for enhancing heterologous protein expression in plants are provided. The invention is directed to the co-expression of a heterologous protein of interest with a specific endoplasmic reticulum (ER)-resident molecular chaperone protein. Co-expressing chaperones such as a native soybean protein disulfide isomerase-like protein (Glyma.01G003700, Glyma.04g247900, Glyma.03G218300, or Glyma.18G204000), or a heterologous mammalian chaperone (bovine DNAJB12) with casein proteins in soybean cells resulted in a significant increase in casein accumulation, with up to 4-fold enhancement observed. The methods are broadly applicable for improving the yield of complex recombinant proteins, including milk proteins, therapeutic proteins, and industrial enzymes, in plant-based production systems, thereby overcoming common limitations related to protein folding and stability.

A siRNA characterised by having a sequence selected from the group comprised of: - SEQ ID NO. 1 : 5 ' CUCUUUCCCUUGAAUAAGC 3 '; - SEQ ID NO. 2 : 5 ' UGAUUCCCAAAGCUCACAG 3 '; and - a sequence including at least 16 contiguous nucleotides differing by no more than 3 nucleotides from SEQ ID No. 1 or SEQ ID No. 2, to silence TRAPl-low SEQ ID N°3. The use of the 5 'UTR SEQ ID N°4 region of the TRAPl-low transcript SEQ ID N°3 or a portion of that 5 'UTR region selected from the group comprised of: - the SEQ ID No. 6 portion of said 5 ' UTR region comprising nucleotide positions 1 through 644; - a portion of said region 5 ' UTR comprising the portion of SEQ ID No. 6 extended by a predetermined number of nucleotide positions beyond nucleotide position 644 along said region 5 'UTR.

Provided are tumor-targeted protein degradation chimeras, termed chaperone-mediated protein degraders (CHAMPs) comprising a first moiety that is capable of binding to a target protein (e.g., KRAS(G12D) or proteins and a second moiety that is capable of binding a chaperone protein or proteins or protein component of chaperone complexes (e.g., HSP90). Pharmaceutical compositions comprising the disclosed CHAMPs and their uses for treating thereof, which are useful for treating cancers and related conditions are also provided.

This disclosure provides expression systems comprising secretion signals that promote production and / or secretion of proteins of interest, as well as one or more polynucleotides encoding chaperone proteins (e.g., CRT and / or PDIA3) which, as demonstrate herein, enhance production and / or secretion of proteins. Moreover, genetically modified host cells comprising these expression systems are capable of producing high levels of protein of interest, such as bovine lactoferrin (bLF), bovine lactoglobulin (bLG), or ovalbumin (Ova).

Disclosed are fusion polypeptides that are substrates for white Kunitz's transglutaminase. The fusion polypeptides comprise one or more FKBP chaperone proteins and a target polypeptide. Each of these elements is separated from adjacent elements by a linker amino acid sequence. It has been found that it is advantageous to insert a glutamic acid-containing transglutaminase recognition motif into the linker amino acid chain. The subsequent labeling reaction catalyzed by the transglutaminase surprisingly provides a labeled fusion polypeptide with superior properties compared to chemically randomly labeled fusion polypeptides of similar design. Assays and kits for detecting a target antibody in a sample in vitro are provided.

The invention discloses a redox chaperonin PDX mutant for improving steroid hydroxylation catalysis activity of cytochrome P450 and application, according to the PDX mutant provided by the invention, the efficiency of OlePM for catalyzing LCA to generate UDCA is remarkably improved, mutants such as Y34I, Y34S, Y34E, Y34H, R67G, R67D, R67A, R67S, I90L, I90M, I90E, I90V and I90S are excellent in performance, and the UDCA concentration can reach 0.13-0.21 g / L and is improved by 5-8 times compared with that reported in literatures. By improving the electron transfer efficiency of the redox chaperonin, the steroid hydroxylation performance of the cytochrome P450 is remarkably enhanced, a new biocatalyst is provided for efficient and green biosynthesis of UDCA, and the method has important industrial application value.

The application discloses a kind of RNA chaperone, coding gene and high stress resistance recombinant bacteria expressing the RNA chaperone and its preparation method, it is related to the field of bioengineering.RNA chaperone PyCsp has the activity of binding RNA.By constructing the recombinant plasmid of expression RNA molecule PyCsp chaperone, the recombinant plasmid is transferred into target bacteria to prepare recombinant bacteria, i.e., the recombinant bacteria with high stress resistance, the stress resistance is low-temperature tolerance, high-pressure tolerance or high-temperature tolerance.The application digs out a heat-resistant RNA chaperone from super-thermophilic pressure-loving microorganism (optimum growth temperature 95 DEG C).Such high-temperature-resistant RNA chaperone as stress resistance element has important application value in improving the stress resistance of microorganism to high temperature, low temperature and high hydrostatic pressure and other environmental stress conditions, and has unique advantages in improving the stress resistance of strain used in fermentation in the application scene of bioengineering related to high temperature and high pressure.

Genetically modified cells (e.g., recombinant mammalian cells) and methods for improving the expression / functional response of olfactory receptors are described herein. Genetically modified cells and methods can generally modulate the levels of chaperone proteins that help target receptors to the cell membrane. Genetically modified cells and methods can also provide a platform for use in studying the function of a broader group of chemosensory receptors, including taste receptors and TRP channels.

Provided herein are methods and host cells for producing a polypeptide containing two chains, such as an antibody, half-antibody, antibody fragment, or one-armed antibody. The methods and host cells allow for two-chain polypeptide production using expression of polynucleotides encoding the polypeptide chains from extra-chromosomal polynucleotide(s), and expression of one or more chaperone protein(s) (e.g., peptidyl-prolyl isomerases and / or protein disulfide oxidoreductases) from the host cell chromosome using non-native combination(s) of promoters and translational units encoding a chaperone protein.

This invention belongs to the field of nanobiomedicine technology, specifically relating to a mitochondrial-targeting copper-based nanoparticle (Qu@Cu-SS31) and its application in the preparation of drugs for treating myocardial infarction. The nanoparticles consist of quercetin (Que) and copper ions (Cu²⁺). + The nanoparticles, consisting of a self-assembly of the mitochondrial-targeting peptide SS31, exhibit a uniform, near-spherical morphology with an average particle size of 17.2 ± 1.6 nm. These nanoparticles specifically target cardiomyocyte mitochondria, responsively releasing Cu²⁺. + Furthermore, Qu@Cu-SS31 restores cardiomyocyte copper homeostasis by downregulating the copper metabolism negative regulator COMMD1, the copper uptake protein CTR1, and the copper chaperone protein CCS. Based on this, Qu@Cu-SS31 exhibits remarkable SOD-like and CAT-like enzyme activities, efficiently scavenging various reactive oxygen species / reactive nitrogen species and significantly reducing oxidative stress. Simultaneously, it inhibits NLRP3 inflammasome activation and Caspase-1-mediated pyroptosis, regulates the Bcl-2 / Bax balance to inhibit apoptosis, and promotes vascular endothelial growth factor expression and angiogenesis.

The invention discloses a method for improving the expression level of bacillus subtilis recombinant protein based on low PAM (Polyacrylamide)-dependent CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) regulation, and belongs to the field of gene engineering. According to the invention, the CRISPRa / i system VcaQCas-RpoZ capable of mediating gene expression activation and inhibition regulation in bacillus subtilis at the same time is obtained by adding an activation module RpoZ. After the system is integrated and expressed into a bacillus subtilis genome, transcription of chaperonin PrsA and penicillin binding protein PonA is activated, meanwhile, the transcriptional level of a regulatory factor MurR is inhibited, and the expression level of recombinant protein is remarkably improved. The enzyme activity of the ultrahigh-temperature amylase is improved to 2.46 times of that of a control strain. The bacillus subtilis provided by the invention provides a new gene expression regulation strategy as a foreign protein expression host, and also provides reference for site selection of gene regulation.

The invention belongs to the technical field of enzyme engineering, and particularly relates to chaperonin assisted folding-based high-activity hyaluronidase and a preparation method thereof, and the preparation method comprises the following steps: S1, constructing a recombinant vector pET32 (a)-TrxA-6H-DDDDK-Hylp1 containing a Hylp1 fusion protein coding sequence; s2, transforming the recombinant vector into Escherichia coli host cells, screening positive engineering bacteria, and performing low-temperature induced expression to obtain expression bacteria; s3, crushing the expression thalli to obtain a cell lysis supernatant; s4, taking the cell lysis supernatant, carrying out nickel column affinity chromatography, carrying out desalination treatment on the fusion protein by adopting pretreated sephadex G-25 chromatography, and collecting a fusion protein solution; s5, performing enterokinase site-specific cleavage treatment on the fusion protein solution to obtain an enzyme-digested product; s6, carrying out nickel column affinity chromatography on the enzyme-digested product again, and collecting a flow-through liquid containing the target protein; and S7, carrying out anion exchange chromatography on the flow-through liquid to obtain the high-purity recombinant hyaluronidase Hylp1.

The application discloses an anti-tumor ginseng peptide producing heterologous recombinant Pichia pastoris engineering bacteria, the engineering bacteria are preserved in the China General Microbiological Culture Collection Center, the preservation number is CGMCC NO:29803, and the preservation date is January 29, 2024. The engineering bacteria are integrated with a FKEHGY polypeptide gene segment with anti-colon cancer activity derived from fresh ginseng leaves by a gene means, the FKEHGY polypeptide gene segment is enhanced in activity by 30 times of repetition, and a molecular chaperone protein HAC1 is expressed simultaneously, the homologous characteristics of HAC1 and Atf / CREB1 are fully utilized, protein correct folding and secretion are helped, and the engineering bacteria have the characteristics of stably and efficiently producing recombinant anti-tumor ginseng peptide F0630.

The invention discloses a microbial whole-cell sensor based on a riboswitch as well as a construction method and application thereof, and belongs to the technical field of biosensors. The invention provides a microbial whole-cell sensor based on a riboswitch, escherichia coli is modified by synthetic biology, so that the escherichia coli responds to mercury ions and expresses red fluorescent protein through the riboswitch, protein signals with different intensities are further generated, and the effect of detecting the mercury ions is achieved; according to the microbial whole-cell sensor based on the riboswitch, mercury ion signals can be quickly responded through the designed artificial riboswitch; through combination with transcription factor MerR, chaperonin ProQ and BL21 Star (DE3) strains, the probability of false positive results can be effectively reduced, the anti-interference capability is improved, and the detection specificity, sensitivity and accuracy of the whole-cell sensor are greatly improved.

The present application provides for compositions comprising high concentrations of acid α-glucosidase in combination with an active site-specific chaperone for the acid α-glucosidase, and methods for treating Pompe disease in a subject in need thereof, that includes a method of administering to the subject such compositions. The present application also provides methods for increasing the in vitro and in vivo stability of an acid α-glucosidase enzyme formulation.

This invention relates to the field of enzyme engineering technology, and discloses a heat-resistant chaperone protein PGI and its application in improving enzyme thermostability. Addressing the difficulties in designing linker peptides in traditional fusion protein construction and the limited applicability of existing stabilization tags, this invention provides a heat-resistant chaperone protein PGI, with the amino acid sequence shown in SEQ ID NO.1. Using the SpyTag / SpyCatcher system, the target enzyme is covalently linked to PGI in vitro, effectively improving the thermostability of the target enzyme while retaining its background catalytic efficiency. Furthermore, this invention reveals for the first time that stabilization efficiency is positively correlated with complex size, exhibiting good applicability to multimeric enzyme systems. Taking nitrile hydratase as an example, after covalent linking with PGI, it exhibits stable and sustained catalytic activity in the catalytic synthesis of acrylamide. This invention provides an effective technical strategy for modifying the thermostability of enzyme preparations.

The present invention relates to an inhibitor of heat shock protein 90. Disclosed are a heat shock protein 90-based bivalent inhibitor, and a preparation method therefor and the use thereof. The bivalent inhibitor is a compound having a structural formula as shown in formula I: A-L-B Formula I, or a pharmaceutically acceptable salt, solvate, or optical isomer thereof, wherein A and B are ATP inhibitors of heat shock protein 90, and the motif structure of L is a flexible linker group of PEG or alkanes, or a rigid linker group comprising aryl, heteroalkyl, or heteroaryl. The bivalent inhibitor can effectively inhibit the activity of molecular chaperone HSP90, hinder the folding and modification of a substrate protein, degrade the substrate protein via a ubiquitin-proteasome degradation pathway, induce non-native dimerization of HSP90, and interfere with protein-protein interactions associated with HSP90. In addition, the bivalent inhibitor reduces the heat shock response induced by HSP90 inhibition, and exhibits potent activity in degrading the substrate protein.

The present application relates to the technical field of virology, more particularly to O-type foot-and-mouth disease virus-like particle antigen and its preparation method and application. The O-type foot-and-mouth disease virus structural protein is constructed, and the recombinant expression vector comprises a Sumo-VP1 sequence, a Sumo-VP3 sequence, a Sumo-VP0 sequence and a chaperone sequence; the recombinant expression vector is transformed into a genetically engineered bacterium, and Sumo-VP1 protein, Sumo-VP3 protein, Sumo-VP0 protein and chaperone protein are expressed; the initial extraction solution is obtained by crushing; the purified extraction solution is obtained by purification; the self-assembled O-type foot-and-mouth disease virus-like particle antigen is obtained by enzyme cutting and removal of Sumo enzyme. The present application improves the soluble expression proportion and overall expression amount of the target protein in the supernatant by optimizing the construction mode and transformation strategy of the foot-and-mouth disease structural protein, and overcomes the technical bottleneck that the supernatant expression amount of the existing E. coli expression system is low and the inclusion body is prone to be formed.