72 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.

The invention relates to the technical field of biological fermentation, in particular to a recombinant yeast cell and application thereof in production of thaumatin. The recombinant yeast cell contains nucleic acids corresponding to thaumatin, PDI and MSN4 genes, and can functionally express the thaumatin, PDI and MSN4 genes. The invention provides the recombinant yeast cell for co-expressing the chaperonin PDI and the transcription factor MSN4 to improve the secretory expression of the thaumatin, and accidentally, under the synergistic effect of the chaperonin PDI and the transcription factor MSN4, the secretory expression quantity of the exogenous thaumatin is remarkably increased by 3.25 times.

The invention discloses a method for improving the heterologous soluble expression efficiency of cytochrome P450, which is characterized in that plasmids containing coding genes of fungus-derived cytochrome P450 truncated protein and plasmids containing molecular chaperones are jointly transferred into host bacteria to obtain recombinant genetically engineered bacteria for improving the heterologous soluble expression efficiency of cytochrome P450. Through host screening, molecular chaperonin and cytochrome P450 gene modification, the soluble expression efficiency of cytochrome P450 in Escherichia coli is improved, and the problems of poor protein solubility and low expression efficiency when CYP450 is expressed in Escherichia coli at present are solved.

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.

Disclosed herein are compounds and methods of use thereof effective for the treatment of neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS), a disease that affects nerve cells in the brain and spinal cord, eventually causing loss of muscle strength. The compounds of the disclosure include, cutamesine, a synthetic sigma receptor agonist selective for the 81 receptor, and also a chaperone protein of the central nervous system that plays a key role in the modulation of calcium ions and apoptosis, as well as a sapogenin, such as smilagenin, a non-peptide neurotrophic factor that aids in the reversal of free radical neurotoxicity.

The invention relates to the field of biotechnology and medicine, and discloses a polypeptide targeting molecular chaperonin TRIC / CCT and application thereof. The sequence of the polypeptide is consistent with or truncated from the C-terminal sequence of a CCT subunit, and the polypeptide can be competitively combined with CRL4-DCAF12 ubiquitin ligase, so that the ubiquitination of the CCT subunit is reduced, and the formation of a TRIC / CCT compound is inhibited; the polypeptide inhibits the formation of a TRIC / CCT compound and blocks the activation of key metastasis promoting signal channels such as YAP, STAT3 and mTOR, thereby inhibiting the metastasis of lung cancer. According to systematic in-vivo and in-vitro experiments, the compound has excellent performance in the aspect of inhibiting lung cancer cell migration and metastasis, has the characteristics of clear target spot, clear action mechanism and the like, and provides a new treatment strategy for developing a new generation of lung cancer metastasis resisting medicines.

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.

The invention discloses an RNA chaperonin, a coding gene, a high-stress-resistance recombinant bacterium for expressing the RNA chaperonin and a preparation method of the high-stress-resistance recombinant bacterium, and relates to the field of bioengineering. Wherein the RNA chaperonin PyCsp has the activity of binding RNA. A recombinant plasmid for expressing an RNA molecular chaperone PyCsp is constructed and transferred into a target bacterium to prepare a recombinant bacterium, namely the recombinant bacterium with high stress resistance, and the stress resistance is low-temperature tolerance, high-pressure tolerance or high-temperature tolerance. According to the invention, a heat-resistant RNA partner is excavated from hyperthermophilic and barophilic microorganisms (the optimum growth temperature is 95 DEG C), and the heat-resistant RNA partner, as a stress-resistant element, has important application value in the aspect of improving the stress resistance of the microorganisms to environmental stress conditions such as high temperature, low temperature, high hydrostatic pressure and the like; in bioengineering application scenarios involving high temperature and high pressure, the strain has unique advantages in the aspect of improving the stress resistance of strains used for fermentation.

The present invention discloses a Thermobifida fusca cutinase mutant and a method for its soluble expression, belonging to the technical field of enzyme engineering. In the present invention, the mutant D204C / E253C and the periplasmic protein disulfide isomerase DsbC are co-expressed in the Escherichia coli mutant strain E. coli Origami B(DE3). However, during the expression process of the obtained recombinant strain E. coli Origami B(DE3) / pSCDsbC-D204C / E253C, it is prone to misfolding to form a large number of inclusion bodies, and the soluble expression ratio is extremely low. The present invention further realizes the efficient soluble expression of the cutinase mutant D204C / E253C through co-expression with the molecular chaperone protein DsbC in the Escherichia coli mutant strain E. coli Origami B(DE3), and has certain industrial application prospects.

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%.

The present invention discloses a method for improving the acid tolerance of E. coli by integrating the molecular chaperone CbpA of extreme acidophilic bacteria, belonging to the fields of biochemistry and molecular biology. The present invention provides a molecular chaperone protein CbpA from Acidithiobacillus thermophilum acidophilus and its encoding gene; the molecular chaperone protein CbpA can improve the acid stress resistance of Escherichia coli, and through gene editing technology, the genome of E. coli BL21 is modified to obtain recombinant Escherichia coli. By detecting the growth status and organic acid tolerance of wild-type BL21, defective BL21-ΔcbpA, and recombinant strain BL21-ΔcbpA / AccbpA strains under acidic conditions, its acid tolerance was confirmed. The present invention lays a foundation for later improving the acid tolerance performance of engineering strains through macromolecular repair means such as molecular chaperone proteins, so as to improve the robustness of industrial microorganisms.

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.

The invention disclosure provides compositions that inhibit amyloid protein aggregation and / or dissociate the already formed amyloid protein aggregates, both in vitro and in vivo. According to the invention disclosure, the provided compositions are useful for the treatment of neurodegenerative diseases - amyloidoses. The compositions provided herein have the advantage of using one or more chaperones of human endoplasmic reticulum (ER), selected from calnexin (CNX), BiP / GRP78, ERp57, and calreticulin (CRT). The compositions may comprise combinations of several chaperones. This description also provides method of use of such compositions, as well as pharmaceutical compositions, wherein the compositions comprise one or more of said chaperones and a pharmaceutically acceptable carrier.

Disclosed herein methods of treating proteinopathies involving proteins that are targeted by karyopherin beta 1's (Kpnβ1) chaperoning function. Therefore the methods of the present application are for increasing Kpnβ1's chaperoning function and comprise administering to a subject in need thereof, an effective amount of one ormore compounds of Formula I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, XV, and XVI. The methods can be used to, for example, to treat proteinopathies of TDP- 43, SOD1 or a-synuclein, or fragments and / or isoforms thereof, and / or proteinopathies other proteins that arc targeted by Kpnβ1's chaperoning function, and therefore to treat subjects with neurodegenerative disorders. Formula (I)

The invention discloses a preparation method and application of a bivalent feline calicivirus VLP vaccine, an insect cell endogenous molecular chaperone protein HSP60 is creatively utilized to construct a recombinant plasmid co-expressing HSP60 and FCV VP1, and the expression quantity of VP1 is successfully improved, the expression quantity of GII VP1 is improved by 9.2 times, and the expression quantity of GI VP1 is improved by 17 times. By screening assembly buffer solutions with different salt ion concentrations, pH values and components, the assembly conditions of GI VLP and GII VLP are optimized, and the yield and yield of VLP are improved. Animal model evaluation shows that the bivalent VLP vaccine has an immune synergistic effect, the immune effect of the bivalent VLP vaccine is remarkably superior to that of a monovalent VLP vaccine and a commercially available inactivated vaccine, the same immune effect of a high dose can be achieved by a low dose of the bivalent VLP vaccine, the antigen dose demand is reduced by 4-8 times, meanwhile, protection on two FCV types is provided, and a new direction is provided for research and development of FCV vaccines.

The application discloses a plant coding gene and protein with improved cadmium resistance and promoted biomass and a transgenic strain. The application carries out RNA-seq sequencing on switchgrass under cadmium stress treatment, screens a differential expression gene with the highest up-regulation fold, and genome annotation shows that the gene codes a member of a heat shock protein 70 family (Hsp70), a chaperone BiP (Binding Protein) protein, and the gene is named as PvBiP2. An overexpression vector of the gene is constructed and is transferred into a cadmium-sensitive yeast mutant strain (ycf1), and the growth condition of the transgenic yeast on a cadmium-containing culture medium is obviously better than that of a control group. Overexpression of the PvBiP2 gene in switchgrass is found to significantly improve the cadmium resistance of the transgenic switchgrass and promote the biomass of the switchgrass.

The present invention relates to an inhibitor of heat shock protein 90, and discloses a bivalent inhibitor based on heat shock protein 90, a preparation method, and an application thereof. The compound has 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 linking group of PEG and alkane, or a rigid linking group having an aryl, heteroalkyl, or heteroaryl group. The bivalent inhibitor can effectively inhibit the activity of the molecular chaperone protein HSP90, hinder the folding and modification of substrate proteins, degrade substrate proteins through the ubiquitin-proteasome pathway, induce non-native dimerization of HSP90, and interfere with protein-protein interactions associated with HSP90. At the same time, it also reduces the heat shock effect caused by HSP90 inhibition, and exhibits potent activity in degrading substrate proteins.

Disclosed herein are fusion proteins comprising a truncated thrombostasin protein having at least 85% sequence homology to a thrombostasin protein, wherein the thrombostasin protein has a carboxy terminal deletion; and a fusion partner protein that is a non-thrombostasin protein. Further disclosed are vaccine compositions thrombostasin proteins having a comprising a carboxy terminal deletion, and methods for inhibiting a response to a thrombostasin protein in a host in need thereof, comprising the disclosed fusion proteins or vaccine compositions. Further disclosed are methods for the preparation of a fusion protein composition.

The present disclosure provides a cell-free method for synthesizing an Invasion Plasmid Antigen B (IpaB) antigen associated with a Shigella bacterium comprising exogenous addition of the purified chaperone protein IpgC to the cell-free synthesis mixture. The disclosure further provides IpaB antigen mutants comprising non-natural amino acids incorporated during cell-free synthesis, enabling covalent conjugation to a Shigella O-antigen polysaccharide. Further provided are Ipa B antigens and conjugates thereof, as well as immunogenic compositions prepared with the synthesized IpaB antigens and conjugates thereof and methods of use.

The invention belongs to the technical field of bioengineering, and discloses an engineering strain applied to biosynthesis of 16 alpha-hydroxyprednisolone, and the engineering strain comprises a gene for coding cytochrome P450 enzyme and a redox chaperonin gene thereof. The cytochrome P450 enzyme gene, the redox chaperonin gene and the combination of the cytochrome P450 enzyme gene and the redox chaperonin gene are screened through an overexpression technology. Compared with a wild strain, the initial production rate of a target product of the obtained excellent engineering strain is improved by 12.17%-161.96%, the maximum yield is improved by 0.37%-38.08%, meanwhile, the fermentation period is shortened, the production intensity is improved by 82.33%-147.64%, and theoretical guidance and application cases are provided for preparation of 16 alpha-hydroxyprednisolone.

The application discloses a kind of recombinant engineering bacteria for co-expression BIP production antitumor ginseng peptide, the engineering bacteria is preserved in China general microbiological bacterial strain preservation center, and the preservation number is CGMCC NO:29804, and the preservation date is January 29, 2024.The engineering bacteria of the application is by genetic means, with pichia pastoris GS115 as host bacteria, simultaneously express the ginseng polypeptide gene with strong anti-colon cancer activity and the molecular chaperone protein BIP that can help protein correct folding and secretion, realize stable and efficient production of recombinant antitumor ginseng peptide F0630.

The invention discloses a bacillus amyloliquefaciens genetically engineered bacterium with high antibiotic resistance as well as a construction method and application thereof, the construction of the engineered bacterium comprises the following steps: bacillus amyloliquefaciens is taken as a starting strain, an RNA (Ribonucleic Acid) molecular chaperonin hfq coding gene is knocked out, and the nucleotide sequence of the RNA molecular chaperonin hfq is shown as SEQ ID NO.1. The RNA molecular chaperonin hfq is obtained through cloning, the tolerance of engineering bacteria to erythromycin and ampicillin is increased by 2.44 times and 3.79 times by knocking out the gene or reducing the expression of the gene, and the enhancement of the antibiotic resistance of bacillus is crucial to the threat of drug-resistant bacteria, the improvement of environmental adaptability and the development of biotechnology application. By strengthening the resistance gene, drug-resistant pathogens can be efficiently inhibited, the antibacterial curative effect can be improved, the metabolic activity can be maintained in a complex antibiotic environment, and the pollution repair and industrial fermentation efficiency can be guaranteed.