107 results about "Zymogen" patented technology

The invention provides a genetically engineered bacterium and a construction method of the genetically engineered bacterium, and the genetically engineered bacterium is characterized in that a green fluorescent protein sfGFP is used for mediating protein glutaminase, PG (protein glutaminase), PGF (protein glutaminase), PGF, PGF, PGF, PGF, PGF, PGF, PGF, PGF, PGF, PGF, PGF, PGF, PGF, PGF, PGF, PGF and PGF; eC 3.5. 1.44) is efficiently secreted and expressed in engineering bacteria, and belongs to the technical field of biological engineering. The method comprises the following steps: an obtained sfGFP gene is derived from an NCBI database (GenBank numbering: CP035486.1), an obtained PG zymogen gene PP (Propeptide-Protein glutaminase) is derived from a Chryseobacterium prion prgA gene (GenBank numbering: AB046594.1) in the NCBI database, a recombinant plasmid pHT01 / sfGFP-PP is introduced into bacillus subtilis WB800N to obtain an engineering bacterium, extracellular secretion expression of PP is realized, the sfGFP-PP is activated by trypsin to obtain PG, and the PG is subjected to enzyme activation to obtain the recombinant bacillus subtilis. The expression quantity and the secretion efficiency of the PG in the bacillus subtilis are greatly improved. The recombinant PG produced by the method can generate 23.5 U / mL enzymatic activity under activation of trypsin, can be used for improving protein functional characteristics, and has relatively high industrial production and application values.

The application belongs to the technical field of genetic engineering, and discloses a kind of saccharomyces cerevisiae for producing rare ginsenoside by seaweed biomass and a construction method and application thereof.The saccharomyces cerevisiae has the following characteristics: overexpression of agarase, neojuncanohydrolase, hydroxymethylglutaryl coenzyme A reductase, isopentenyl diphosphate delta isomerase, dammaradienol synthase, protopanaxadiol synthase, cytochrome P450 reductase, protopanaxatriol synthase and glycosyltransferase.The application combines enzymolysis of seaweed biomass with fermentation of rare ginsenoside, which not only endows wild-type yeast with the ability to degrade seaweed biomass that it originally does not have, but also effectively improves the yield of squalene and downstream terpenes (rare ginsenoside Rh1) by overexpression of tHMG1 and IDI1.The saccharomyces cerevisiae can effectively utilize seaweed biomass to obtain squalene and rare ginsenoside Rh1, and has the characteristics of simplicity, economy and ecological friendliness, can convert cheap biomass into high-value products, and provides a way for the development of marine bioeconomy.

The present invention includes a method of treating a subject having relapsing-remitting multiple sclerosis (RRMS) that is undergoing relapse, comprising: determining a level of expression for two or more biomarkers selected from urokinase plasminogen activator (uPA), kallikrein-8 (hK8), kallikrein-11 (hK11), or desmoglein-3 (DSG3) in a biological sample of a subject when compared to the same type of sample from a subject or a population of subjects that do not have RRMS; diagnosing that the subject in undergoing relapse if the level of expression of the two or more biomarkers has decreased in the subject; and administering a therapeutically effective amount of treatment for RRMS to the subject.

The present disclosure relates to enzymes, compositions and methods for performing conditional homologous recombination of a targeted DNA molecule or genome by using modified proteins comprising nucleic acid DNA binding proteins with protease-conditional recombinase (hereinafter “ProRec”) activity. Namely a new cre recombinase pro enzyme, compositions / kit and sensors comprising the cre recombinase pro enzyme of the present disclosure. Namely, a cre recombinase pro-enzyme, for identifying and quantifying proteolytic activity in a disease or an infection, comprising intein mediated circularization, a flip-excision cassette comprising an output reporter and / or effector protein; a linker sequence cleavable by a protease.

Provided herein are compositions, systems, kits, and methods for treating a patient with a thromboembolism by administering nanoconjugates comprising nanoparticles encapsulating and / or conjugated to: i) tissue-type plasminogen activator (tPA), and ii) at least one antioxidant enzyme selected from the group consisting of: superoxide dismutase, glutathione peroxidase, glutathione reductase, and a catalase.

This invention discloses a fusion protein targeting the urokinase-type plasminogen activator receptor and its application in the field of anti-aging. The fusion protein is composed of a hepatocyte growth factor active domain and a humanized anti-uPAR single-domain antibody linked by a flexible linker peptide, and its specific amino acid sequence is SEQ ID NO:3. This fusion protein can specifically bind to uPAR, which is highly expressed on the surface of senescent cells, with intramolar affinity, achieving targeted delivery of the hepatocyte growth factor active domain. In vitro experiments have demonstrated that this protein can significantly reverse the aging phenotype of human fibroblasts, effectively reduce aging-related β-galactosidase activity, p16INK4a protein expression, and interleukin-6 secretion, and promote cell proliferation. In a rapidly aging mouse model, this protein can systematically improve age-related physiological functional decline. The fusion protein of this invention achieves specific targeting and efficient treatment of senescent cells, providing a new solution for the development of anti-aging drugs.

To provide methods of identifying new substrates for proteases and methods of using these substrates in a variety of therapeutic, diagnostic and prophylactic indications.SOLUTION: The invention relates generally to: polypeptides that include a cleavable moiety that is a substrate for at least one protease selected from matriptase and u-plasminogen activator (uPA); activatable antibodies and other large molecules that include the cleavable moiety that is a substrate for at least one protease selected from matriptase and u-plasminogen activator; and methods of making these polypeptides that include the cleavable moiety that is a substrate for at least one protease selected from matriptase and u-plasminogen activator, and methods of use in a variety of therapeutic, diagnostic and prophylactic indications.SELECTED DRAWING: Figure 4A

The present application relates to a method for promoting the degradation of pathological TDP-43 protein, comprising administering to a subject a therapeutically effective amount of one or more compounds selected from components of the plasminogen activation pathway, compounds that can activate plasminogen directly or indirectly by activating upstream components of the plasminogen activation pathway, compounds that mimic the activity of plasminogen or plasmin, compounds that can upregulate the expression of plasminogen or plasminogen activators, plasminogen analogs, plasmin analogs, tPA or uPA analogs, and antagonists of fibrinolysis inhibitors. The present application also relates to agents that promote the degradation of pathological TDP-43 protein and uses thereof.

Disclosed herein, are peptides that bind urokinase-type plasminogen activator receptors. The peptides may comprise amino acid sequences of IPPWEAPK (SEQ ID NO: 1), DLAQCQTPTQAAPPTPVSPR (SEQ ID NO: 2), or LHVPLMPAQPAPPK (SEQ ID NO: 3), or retro-inverso amino acid sequences of the above enumerated sequences. Also described herein, are methods of administering compounds comprising peptides that bind urokinase-type plasminogen activator receptors to subjects for the treatment of ovarian cancer and improving wound closure.

The current invention relates to a polypeptide comprising an immunoglobulin single variable domain and an urokinase-type plasminogen activator (uPA) domain wherein the immunoglobulin single variable domain is able to bind to fibrin via a fibrin-binding domain.

The invention discloses a polypeptide asparagine ligase VdiPAL1 and an application of the polypeptide asparagine ligase VdiPAL1. The polypeptide asparagine ligase VdiPAL1 is newly found from viola plants, 12 mg / L zymogen protein is obtained through expression of an escherichia coli prokaryotic system, and compared with homologous enzymes VyPAL2 and VdiPAL1 which are widely applied at present, the polypeptide asparagine ligase VdiPAL1 has the advantages of high expression, high catalytic efficiency and substrate specificity and has good development and application potential. The invention also shows the value of the VdiPAL1 for guiding homologous enzyme modification, and particularly discloses the mutant protein VyOPT of the polypeptide asparagine ligase VyPAL2 and a coding cDNA (complementary deoxyribonucleic acid) sequence of the mutant protein VyOPT. The invention also discloses a mutant VyOPT, which comprises five mutations obtained by modification guided by the VdiPAL1. Compared with the expression quantity of the natural VyPAL2, which is about 0.1 mg / L, the expression quantity of the VyOPT reaches 12 mg / L.

The present invention provides methods for modifying a donor organ, e.g., liver, or a portion thereof. The methods comprise contacting the donor liver with lipid nanoparticles comprising RNA. Compositions of lipid nanoparticles comprising RNA are also provided. The disclosed LNPs may comprise an siRNA, e.g., targeting tissue plasminogen activator (tPA) or plasminogen activator inhibitor 1 (PAI-1). The LNPs may comprise an mRNA encoding a protein, e.g., Nuclear Factor-erythroid factor 2-related factor 2 (NRF2) or thrombomodulin.

The invention relates to the technical field of membrane separation, and discloses a high-protein milk concentration process based on a membrane filtration technology, the process adopts an electric response membrane grafted with an electric response polymer brush on the surface, and an electric control zymogen modified by genetic engineering is added into milk feed liquid to be treated. The zymogen is modified with an electric field responsive anchoring group and an electric signal activated ferrocene protecting group. In the filtering stage, a first electric field is applied to stretch the polymer brush, so that anti-pollution filtering is realized; and in the cleaning stage, switching to a second electric field to enable the polymer brush to shrink and drive the zymogen to be anchored on the surface of the membrane, and applying an activation pulse to trigger the activity of the zymogen so as to degrade pollutants in situ. On the premise of not interrupting production, not adding chemical cleaning agents and not damaging target product protein, milk concentration is efficiently and continuously carried out, and long-term stable high permeation flux is maintained.

The present invention relates to double stranded ribonucleic acid (dsRNA) compositions targeting the plasminogen (PLG) gene, as well as methods of inhibiting PLG expression, and methods of using such dsRNA compositions to treat subjects that would benefit from reduced PLG expression, such as subjects having a PLG-related disease, disorder or condition.

The invention discloses a PAI-1 protein targeting peptide compound (called P peptide for short) with a thrombus dissolving effect and application thereof, and belongs to the field of biological medicine. The PAI-1 targeted thrombolytic polypeptide compound comprises P1, P2, P3 and P4, and is characterized in that the compound is derived from a core structure 'X-Asp-Glu-Leu-Cys-X-Tyr-Tyr-X-X-Cys-X' (cyclization is performed through a disulfide bond between Cys, and X represents any type of amino acid), and the compound can be specifically bound with a key binding site of PAI-1. The polypeptide can significantly reduce the inhibitory effect of PAI-1 on plasminogen activators (tPA, uPA), enhance the activity of a fibrinolytic system, and generate in-vivo and in-vitro thrombolysis effects. The polypeptide compounds P1, P2, P3 and P4 not only have a thrombolysis effect, but also can reduce the expression level of pro-inflammatory cytokines (IL-1beta, TNF-alpha, IL-6 and the like), which shows that the polypeptide compounds also have an anti-inflammatory effect. In conclusion, the polypeptide compound can be used for preparing a PAI-1 antagonist, a thrombolytic agent and an anti-inflammatory agent related to thrombus.

The disclosure provides urokinase plasminogen activator receptor (uPAR) binding polypeptides (e.g., uPAR antigen binding proteins), and fusion proteins comprising said uPAR binding polypeptides and a cytokine or variant thereof. The disclosure further provides pharmaceutical compositions comprising the uPAR binding polypeptides and fusion proteins of the disclosure and methods of treating cancer with the same. The disclosure further provides engineered IL-12 p35 and p40 polypeptides.

This invention relates to the field of biological detection, and more particularly to the preparation method and application of the t-PAI·C detection kit. The invention provides a detection reagent comprising: antibody-coated magnetic microparticles and enzyme-labeled antibodies; the antibodies include: tissue plasminogen activator-plasminogen activator inhibitor-1 complex antibody. In the detection reagent provided by this invention, when the sample contains the tissue plasminogen activator-plasminogen activator inhibitor-1 complex, the antibody coated with the magnetic microparticles and the enzyme-labeled antibody undergo an immune reaction with the complex, forming a sandwich-like immune complex consisting of the antibody coated with the magnetic microparticles, the complex, and the enzyme-labeled antibody; the enzyme on this complex acts on a luminescent substrate and emits light, and the concentration of the complex in the sample is calculated by detecting the luminescence signal.

A method comprising the steps of: (a) assessing an expression level of at least one protein, selected from Table 1, 2 or 3 in a sample from a subject, where in the at least one protein may be selected from the list comprising: Beta-Ala-His dipeptidase, Apolipoprotein L1, Methanethiol oxidase, Vitamin K-dependent protein S, von Willebrand factor, Plasminogen, Selenoprotein P, Protein disulfide-isomerase A6 and Inter-alpha-trypsin inhibitor heavy chain H3, and (b) using the expression level to determine whether the subject has endometriosis.

The application discloses an engineered exosome targeting human urokinase-type plasminogen activator receptor uPAR and a preparation method and application thereof. Firstly, a high-affinity anti-human uPAR single-domain antibody VHH is prepared; then, a human umbilical cord mesenchymal stem cell engineering cell line expressing a VHH-PTGFRN fusion protein is constructed, and the engineered exosome is prepared and purified by combining three-dimensional hypoxic culture, chemical induction and size-exclusion chromatography technology. The engineered exosome comprises four comparative samples, wherein the core sample Exo^(VHH / miR) can simultaneously display the anti-uPAR VHH on the membrane surface to realize targeted delivery and enrich miR-193b-3p in the chamber to realize gene silencing, and the synergistic effect of the two can efficiently remove the uPAR high-expression senescent cells and inhibit the senescence-associated secretory phenotype SASP. The engineered exosome has the advantages of standardized preparation process, high yield and high purity, strong targeting, excellent curative effect and good safety, and can be used for preparing a treatment preparation for anti-aging and aging-related diseases, and has important clinical application value.

The invention belongs to the technical field of medicines, and particularly relates to an analogue of a cysteine zymogen activator (a PAC-1 analogue containing a carvone structure) and application of the analogue in cancer treatment. The invention relates to an analogue of a cysteaspase-3 activator, which is a PAC-1 analogue containing a carvone structure as shown in a formula I. The PAC-1 analogue disclosed by the invention is obtained by retaining structures such as piperazine ring and hydrazide, introducing a monoterpenoid carvone and carrying out structural modification on benzene rings on two sides, and a novel procaspase-3 activator is developed. The derivatives have stronger physiological activity. Pharmacological tests prove that the PAC-1 analogue can inhibit proliferation of various tumor cells and has good anti-cancer activity, and the preparation method of the PAC-1 analogue is simple, feasible and easy to operate.

The invention belongs to the technical field of enzyme engineering and molecular biology, and particularly relates to serine peptidase, a nucleic acid molecule and application of the serine peptidase. According to the invention, three kinds of serine peptidase SpBcA, SpBcB and SpBcC are identified in B. cardium for the first time, and the structures and functions of the three kinds of serine peptidase SpBcA, SpBcB and SpBcC are analyzed. SpBcA and SpBcB have good serine peptidase activity, can tolerate high temperature (the optimum enzyme reaction temperature is 60 DEG C), and have good activity under an alkaline condition. It is further found that SpBcA is generated in the form of zymogen and is subjected to self-splitting. An inhibitory propeptide is arranged at the N end and can be used as a molecular chaperone to inhibit the activity of the molecular chaperone, and a novel activity mechanism is shown. In addition, the SpBcA serine peptidase can degrade various substrates such as casein, fibrinogen, antibacterial peptide LL-37, gelatin and the like. Therefore, as a novel serine peptidase, the serine peptidase has potential industrial and medical application values.

Provided are modified u-PA polypeptides and fusion proteins containing the modified u-PA polypeptides. The u-PA polypeptides are modified to have altered activity and / or specificity so that they cleave a complement protein, such as complement protein C3, to thereby inhibit complement activation. The modified u-PA polypeptides and fusion proteins that inhibit complement activation can be used for treatment of diseases and conditions that are mediated by complement activation, or in which complement activation plays a role. These disorders include ischemic and reperfusion disorders, including myocardial infarction and stroke, sepsis, autoimmune diseases, diabetic retinopathies, age-related macular degeneration, transplanted organ rejection, inflammatory diseases and diseases with an inflammatory component.

To provide compositions and methods for modulating the plasmin system for treating and / or preventing various pathological conditions.SOLUTION: The present invention relates to antigen binding proteins comprising an antigen binding domain that binds to plasminogen, wherein the antigen binding protein reduces the activation of plasminogen, pharmaceutical compositions comprising the same, and methods and uses thereof.SELECTED DRAWING: None

This invention belongs to the field of biomedical technology, specifically relating to a recombinant human plasminogen mutant, its preparation method, and its application. A recombinant human plasminogen mutant, the amino acid sequence of which is shown in SEQ ID NO.3. The recombinant human plasminogen mutant of this invention, compared to wild-type mPLG, can be directly expressed solublely in the supernatant of *E. coli* fermentation broth. Soluble expression allows for direct chromatography of the fermentation broth to obtain the target protein, which has significant advantages over inclusion body renaturation treatment in terms of process simplification, activity loss, time cost, and production cost. Furthermore, the activity of the mutant protein was tested using the fibrin plate method and kit method. The results showed that the titer of the target protein was approximately 8 UI / mg, which is similar to the titer of mPLG obtained by renaturation before mutation and PLG obtained from human blood. In summary, compared to the wild type, the mutant strain exhibits significantly improved solubility while maintaining high protein activity.

A product, composition, suspension and / or method to dissolve vaso-occlusive clots in vivo in a sickle diseased subject utilizing Target-specific Thrombolytic Urokinase Spheres (TTUS). The TTUS can target vaso-occlusive clots in vivo and then lyse the obstructive blood clots with high efficiency can be intravenously administered to a subject in need thereof. The TTUS are configured to dissolve vaso-occlusive clots by converting plasminogen in blood of the subject into plasmin molecules that dissolves the clot. The nanospheres can encapsulate an enzymatically active bioreactive agent, such as urokinase. Since the active enzyme is targeting and binding essentially only to obstructive clots, there is minimal risk of side-effects to the subject. The nanospheres are less than one micron in diameter, resulting in a size much smaller than red blood cells.

The invention relates to bacillus subtilis for high-yield protein glutaminase and application of the bacillus subtilis. Enzyme activity detection shows that extracellular proteases AprE, Bpr and Epr can activate zymogen of the protein glutaminase, so that the effect of improving the enzyme activity of the protein glutaminase is achieved; the knockout of the extracellular protease WprA can improve the enzymatic activity of the protein glutaminase, and based on the experiments, it is found that the bacillus subtilis BS1-PG for high yield of the protein glutaminase is constructed, the enzyme activity is 18.05 U / mL after fermentation in a shake flask for 96 hours, the enzyme activity reaches 45.58 U / mL after fermentation in a 5L fermentation tank for 60 hours, and the enzyme activity is remarkably improved compared with that of an original strain. On the basis, protease related genes lytC, spoIVB, amyE, malA, srfAC and mdxL in a bacillus subtilis genome are knocked out, and the enzyme activity is further improved to 65.68 U / mL in a 5L fermentation tank, so that the production cost of the protein glutaminase is reduced, and large-scale production and application of the protein glutaminase in the field of food industry are promoted.

The utility model relates to the technical field of endoscope instrument cleaning, in particular to a portable instrument pretreatment device which comprises a reagent tube, a reagent tube cover is connected to the reagent tube in a sealed mode, an insertion round hole is formed in the reagent tube cover in a penetrating mode, and the insertion round hole is used for the front end of an endoscope instrument to enter the reagent tube. The inside of the reagent tube is hermetically connected with two diaphragms, the inside of the reagent tube is divided into a chamber I and a chamber II by the diaphragms, the chamber I is used for storing pure water, and the chamber II is used for storing a multi-enzyme stock solution. According to the multi-enzyme cleaning device, the multi-enzyme stock solution and the pure water are stored in the reagent tube at intervals, and after the front end of an endoscope instrument enters the reagent tube, the multi-enzyme stock solution and the pure water are mixed to form the multi-enzyme cleaning solution, so that the multi-enzyme cleaning solution can be prepared and used in the laparoscopic surgery, and the multi-enzyme cleaning device has the beneficial effects of guaranteeing the pretreatment effect, improving the working efficiency and prolonging the service life.