1170 results about "PROTEASE M" patented technology

The invention provides compositions and methods for releasing and for isolating nucleic acids from biological samples, preferably from whole tissue, using cationic surfactants and proteases. The surfactant-protease combinations, when used with whole tissue, macerate the tissue, lyse individual cells, release nucleic acids, and inactivate nucleases. Kits for isolating nucleic acids from biological samples, particularly from whole tissue, are also provided.

Compositions and methods for controlling plant pests are disclosed. In particular, novel nucleic acid sequences encoding modified Cry3A toxins having increased toxicity to corn rootworm are provided. By inserting a protease recognition site, such as cathepsin G, that is recognized by a gut protease of a target insect in at least one position of a Cry3A toxin a modified Cry3A toxin having significantly greater toxicity, particularly to western and northern corn rootworm is designed. Further, a method of making the modified Cry3A toxins and methods of using the modified cry3A nucleic acid sequences, for example in microorganisms to control insects or in transgenic plants to confer protection from insect damage, and a method of using the modified Cry3A toxins, and compositions and formulations comprising the modified Cry3A toxins, for example applying the modified Cry3A toxins or compositions or formulations to insect-infested areas, or to prophylactically treat insect-susceptible areas or plants to confer protection against the insect pests are disclosed.

The invention provides a method of determining activity of a protease. The method can include the steps of (a) providing a protease substrate including a protein moiety attached to a nucleic acid moiety and a ligand moiety; (b) contacting the protease substrate with a protease under conditions wherein the protease catalyzes cleavage of the protein moiety, thereby producing a proteolytic product wherein the nucleic acid moiety is separated from at least a portion of the protein moiety and the ligand moiety; (c) contacting the proteolytic product with a receptor under conditions wherein the ligand moiety binds to the receptor to form a complex; (d) separating the complex from the nucleic acid moiety, thereby forming a separation product including the nucleic acid moiety; (e) contacting the separation product with a probe nucleic acid under conditions wherein the nucleic acid moiety hybridizes to a complementary sequence of the probe; and (f) detecting hybridization of the separation product to the probe, thereby determining activity of the protease.

Catalytic monoclonal antibodies (abzymes) with selective protease activity in the pathologies characterized by the presence of plaques and fibrillar aggregates with protein component; methods for the preparation thereof and the use thereof as medicaments in the treatment of pathologies such as Alzheimer's disease, amyloidosis, atherosclerosis, prions diseases.

Disclosed are useful constructs and methods for the expression of proteins using primary translation products that are processed within a recombinant host cell. Constructs comprising a single open reading frame (sORF) are described for protein expression including expression of multiple polypeptides. A primary translation product (a pro-protein or a polyprotein) contains polypeptides such as inteins or hedgehog family auto-processing domains, or variants thereof, inserted in frame between multiple protein subunits of interest. The primary product can also contain cleavage sequences such as other proteolytic cleavage or protease recognition sites, or signal peptides which contain recognition sequences for signal peptidases, separating at least two of the multiple protein subunits. The sequences of the inserted auto-processing polypeptides or cleavage sites can be manipulated to enhance the efficiency of expression of the separate multiple protein subunits. Also disclosed are independent aspects of conducting efficient expression, secretion, and / or multimeric assembly of proteins such as immunoglobulins. Where the polyprotein contains immunoglobulin heavy and light chain segments or fragments capable of antigen recognition, in an embodiment a selectable stoichiometric ratio is at least two copies of a light chain segment per heavy chain segment, with the result that the production of properly folded and assembled functional antibody is made. Modified signal peptides, including such from immunoglobulin light chains, are described.

Fish derived serine proteinases including trypsins and chymotrypsin derived from cod such as Atlantic cod is used for treating and / or preventing a variety of diseases and disorders such as inflammatory diseases, infectious diseases caused by viruses, bacteria and fungal species and diseases where a receptor binding mechanism is involved in the pathogenesis. Pharmaceutical and cosmetic compositions comprising the proteinases are described.

The invention relates to a method for producing a recombinant functional plasminogen in micro-organisms, and to a method for identifying plasminogen activators. The nucleic acid sequence coding for the functional part of the plasminogen is fused with a nucleic acid molecule coding for at least one signal peptide. The nucleic acid molecule coding for the plasminogen and the nucleic acid molecule coding for the signal peptide are combined with codons for interfaces of proteases which ensure the separation of the signal peptide. The recombinant plasminogen or the corresponding plasmine is suitable for treating wounds which are slow to heal or not healing, by application of the enzyme in an appropriate formulation.

The invention relates to targeted post translational modifi-cation of metallo-beta-lactamase by truncation and inser-tion of a dipeptide at the amino terminal end to reduce amino terminal heterogeneity in a recombinant DNA pro-duction system. A protein K-T-E-ΔBL is expressed, and modified by host proteases to E-ΔBL. Appropriate nucleotide molecules, vectors and hosts are also de-scribed. E-ΔBL is useful in a pharmaceutical composition for treating antibiotic induced adverse effects in the intes-tine of patients treated with beta-lactam antibiotics.

The invention relates to a method for enzymatically obtaining protein hydrolysates for human consumption, animal feed and cosmetics. The process involves the use of a proteolytic composition derived from fish, such as Cod (Gadus morhua), to obtain hydrolysates which have a non-bitter taste and retain the flavor and aroma of the protein-containing material which is hydrolyzed: e.g. when hydrolyzing protein-containing material from marine organisms or parts thereof, such as fish, shrimp, lobster or other seafood according to the invention, a protein hydrolysate is produced that has a characteristic natural flavor of the organism Also provided are food products comprising the hydrolysates of the invention, such as soup, sauce, cheese, HVP, meat extract and flavoring agent, broth, paté, mousse, frying dough, orly dough, and pastries.

Provided are methods and compositions useful in detecting protease activity in a sample, as well as methods of identifying agents that modulate protease activity. The methods and compositions provide a modified luciferase polynucleotide sequence and a luciferase polypeptide containing protease recognition sequences, wherein cleavage of the recognition sequence by a protease inhibits luciferase activity. Further provided are methods and compositions for detecting and modulating caspase activity and apoptosis.

The invention provides functional soy peptide fermented milk and a preparation method thereof and relates to a manufacturing method of fermented dairy products in the field of food processing. The method is carried out according to the following steps: (1) the preparation of fermented base stock; (2) the preparation of mother cultures; (3) synchronous enzymolysis and fermentation; (4) blending and homogenizing; (5) ultrahigh pressure sterilization and filling; by adopting the steps, a finished product is obtained. The method adopts protease and lactic acid bacteria synchronous-enzymolysis fermentation technology, and ultrasonic waves are adopted to process slurry-residue mixture which is ground, so as to increase the dissolving-out rate of the protein and prepare acidity soy-bean milk by ultrahigh pressure and non-heat processing. The product taste is coordinated without enzymolysis odour; the enzymolysis fermentation time can shorten to 2-4h, and the production cost is low. The ultrahigh pressure processing can remarkably improve the product taste; the method can keep and produce new functional peptide, the lactic acid bacteria is inhibited to ferment continuously, and a certain micro-organism viable counts are kept; the shelf life of the acidity soy-bean milk is prolonged, so as to prevent the acid milk from being whey-separated owning to continuous increasing of acidity; inaddition, the product can be stored and marketed at normal temperature.

Gene expression profiling and hierarchial clustering analysis readily distinguish normal ovarian epithelial cells from primary ovarian serous papillary carcinomas. Laminin, tumor-associated calcium signal transducer 1 and 2 (TROP-1 / Ep-CAM; TROP-2), claudin 3, claudin 4, ladinin 1, S100A2, SERPIN2 (PAI-2), CD24, lipocalin 2, osteopontin, kallikrein 6 (protease M), kallikrein 10, matriptase and stratifin were found among the most highly overexpressed genes in ovarian serous papillary carcinomas, whereas transforming growth factor beta receptor III, platelet-derived growth factor receptor alpha, SEMACAP3, ras homolog gene family, member I (ARHI), thrombospondin 2 and disabled-2 / differentially expressed in ovarian carcinoma 2 (Dab2 / DOC2) were significantly down-regulated. Therapeutic strategy targeting TROP-1 / Ep-CAM by monoclonal chimeric / humanized antibodies may be beneficial in patients harboring chemotherapy-resistant ovarian serous papillary carcinomas. Claudin-3 and claudin-4 being receptors for Clostridium Perfringens enterotoxin, this toxin may be used as a novel therapeutic agent to treat ovarian serous papillary tumors.

Bacteria which co-express protease inhibitors and protease sensitive therapeutic agents, which are surface displayed, secreted and / or released and result in their localized production and maintenance within a target tissue and inactivation outside of the target tissue, thereby increasing therapeutic activity and reducing the systemic toxicity. The bacteria may be attenuated, non-pathogenic, low pathogenic or a probiotic. Protease sensitivity may be further accomplished by engineering protease degradation sites within the therapeutic agents, further enhancing the inactivation outside of the target tissue while retaining activity within the target tissue through co-expression of a protease inhibitor. Novel chimeric proteins secreted by bacteria, including chimeric toxins targeted to neoplastic cells, tumor matrix cells and cells of the immune system, and combination therapies of these protease inhibitor:chimeric toxin-expressing bacteria together with small-molecule and biologic agents are also described. Non-conjugative bacteria limiting exchange of genetic material, and antibody resistant bacteria are also provided.

The invention discloses a method for preparing collagen peptide from fish scales, which is characterized by comprising the following steps of: 1, taking the fish scales and immersing hydrochloric acid to remove calcium; washing to be neutral, drying and crushing; 2, adding the calcium-removed, dried and crushed fish scales obtained by the step 1 into an enzyme reactor; adding water and mixed protease to extract the collagen peptide; after the extraction, carrying out enzyme deactivation and centrifuging to obtain an enzymolysis solution, wherein the mixed protease is a mixture of neutral protease, papain and flavored protease; and 3, carrying out film concentration on the enzymolysis solution obtained by the step 2; and freezing and drying a concentrated solution to obtain fish scale collagen peptide powder. According to the fish scale collagen peptide obtained by the invention, the neutral protease, the papain and the flavored protease are used for extracting the fish scale collagen peptide by an enzymic method at one step, so that not only can the enzymolysis time (9 hours)be reduced, but also the extracting rate (72.72%) of the collagen peptide is improved, and solves the special flavor problem of bitter taste of the collagen peptide and the like.

Based on a principle that is different to that of a conventional enzymatic method, the present invention provides a novel method for assaying a glycated protein by a simple procedure, within a short period of time, and with high accuracy, and a reagent kit for assaying used in the method. The method for assaying a glycated protein in a sample is realized by treating a glycated protein-containing sample with protease to liberate a glycated peptide, preferably an α-glycated peptide, particularly preferably an α-glycated dipeptide, from a glycated protein, allowing an oxidase to react with the liberated glycated peptide, and determining the produced hydrogen peroxide.

The present invention uses co-expression of protease inhibitors and protease sensitive therapeutic agents that results in their localized production within the target tissue and inactivation outside of the target tissue, thereby increasing therapeutic activity and reducing the systemic toxicity. Inactivation is also accomplished by engineering protease degradation sites within the therapeutic construct for proteases, preferably those that are under-expressed within the target tissue yet present in non-target tissues within the body, resulting in therapeutic activity within the target tissue and inactivation outside of the target tissue. Novel chimeric proteins secreted by bacteria are also described. The chimeric proteins include chimeric toxins targeted to neoplastic cells and cells of the immune system. Novel combination therapies of these protease inhibitor:chimeric toxin-expressing bacteria together with small-molecule and biologic agents are also described. Non-conjugative bacteria capable of delivering phage / phagemids expression cassettes for DNA and RNA-based therapeutics are also described.

The invention relates to an enzyme method ultrasonic extraction method of walnut oil and walnut protein, which belongs to the food and the functional food field. Water is added into walnut kernel or walnut powder to be grinded into paste, protease or compound protease are added into to be performed hydrolization, and simultaneously ultrasonic processing is performed, then the walnut kernel or walnut powder is agitated to perform enzymatic extraction, and then centrifugal separation oil phase, protein peptide oil water phase and residual solid phase are performed; walnut oil is acquired by refining the obtained oil phase, which meets the requirements of green foods; the protein peptide aqueous solution can be directly used to produce degreased walnut protein nutrient milk, or to prepare walnut antioxidation peptide after performed nanofiltration, which is used in health products, food additives, cosmetics or daily chemical articles, or to produce nutrient condensed milk after being performed low temperature concentrating, or to produce walnut protein nutrition powder after performed spray drying; the solid phase residue is prepared into diet fiber food after being dried and grinded into powder; walnut nutrient protein peptide products can be obtained through performing vacuum concentration and spray drying to walnut protein peptide extracting solution containing nutrient content.

The invention relates to a micromolecule walnut peptide and a preparation method thereof. The preparation method sequentially comprises the following steps of protein extraction, twice enzymolysis, filtering, purification, concentration and drying. In the twice enzymolysis process, alkaline protease with the mass being 1-3% that of the walnut protein is added into a walnut protein extracting solution for once enzymolysis; enzyme deactivation is performed on a once enzymolysis solution after the once enzymolysis is finished; one or more kinds of proteinase including the flavored proteinase, the neutral proteinase and the papain with the mass being 1-3% that of the walnut protein are added after the once enzymolysis solution obtained after enzyme deactivation is performed is cooled for secondary enzymolysis. According to the preparation method, the twice enzymolysis method is adopted, due to the specificity of the structure of the walnut protein structure, sufficient enzymolysis cannot be performed on the walnut protein through enzymolysis of one proteinase, and the twice enzymolysis method is adopted for ensuring that the enzymolysis is brought into play on the most suitable condition of each proteinase, and therefore the obtained walnut peptide content is high.

The invention discloses a fungal alpha-amylase-containing beer complex enzyme and a preparation method thereof, belonging to the field of enzyme preparation processing. The high-activity fungal alpha-amylase and other food grade enzyme preparations, plant extracts, antioxidants, protective agents, activating agents and the like can be scientifically compounded by taking concentrated maltase and concentrated malt juice powder as main raw materials; the prepared beer complex enzyme is complete in proteases, high in enzyme activity, difficult to deactivate, mellow in malt aroma, and capable of providing abundant nitrogen source for malt juice, wherein the activity of fungal alpha-amylase in fermenting liquor during the preparation of fungal alpha-amylase is 17000-21000 U / mL. The complex enzyme can be stored for 12 months under the conditions of 0DEG C and 40DEG C, the single enzyme activity loss in the complex enzyme are respectively 0.50% and 0.72%, the enzyme deactivation caused by environment change, and inappropriate operation methods during packaging, storage, transportation, use and the like can be effectively prevented, especially the enzyme deactivation caused by high temperature can be prevented.

The present invention provides compounds that efficiently and specifically inhibit lethal factor (LF) protease activity of anthrax toxin.

The invention relates to technology for fermenting and decomposing protein feed with protease in two steps by using probiotics, which is characterized in that the protein is fermented and decomposed with protease of aerobic fermentation matter in the anaerobic condition, soybean dregs, cotton dregs, vegetable dregs meal, wheat bran and other vegetable protein raw materials are sterilized at high temperature and then are aerobically fermented for 24-36h, feather meal and other animal protein raw materials are aerobically fermented for 20-26h, the inoculating temperature is controlled at 60-80 DEG C, the fermented materials are not dried and are directly mixed with fresh raw materials, corn steep liquor and other nutrient solution are added or not added; lactic acid bacillus are inoculated to the mixture, and water is supplemented to the mixture until the water content of the mixture reaches 40-60 percent, the mixture is compacted and sealed to be anaerobically fermented for 1-3d above 20 DEG C or 2-5d below 20 DEG C, and the mixture is dried after being fermented and decomposed with protease.

The invention relates to a method for biosynthesis preparation of human glucagon-like peptide-1 (GLP-1) and an analogue thereof. With adopting of a gene engineering technology, a recombinant escherichia coli expressed GLP-1 fusion protein is constructed, and a protein enzyme digestion site is designed in the fusion protein; a fusion gene has a gene sequence with a form of A-B-C structure, wherein A is a chaperonin gene, B is a nucleotide sequence encoding a connection peptide containing the enzyme digestion site, and C is a gene encoding the GLP-1 or the analogue thereof. After recombinant engineering bacteria is subjected to induced expression, the fusion protein is purified and subjected to enzyme digestion, and then the GLP-1 and the analogue thereof are obtained and are detected to have biological activity. The preparation method of the GLP-1 and the analogue thereof provided by the invention is simple and quick, the production conditions are mild, the product is convenient to separate and extract, the process is simple, and the industrialization prospect is good.

The invention relates to a process of continuous production of casein bioactive peptide by enzymolysis and filtering membrane concentration which comprises, using caseinum as raw material and at least one prolease action, obtaining biologically active polypeptides having multiple functions in multi-stage enzyme membrane reactor combined from enzymolysis tank and hyperfiltration, nano filter membrane of dissimilar entrapment molecular weight.

The problem of the present invention is to provide a preventive and / or therapeutic agent for diabetes and / or complications of diabetes based on the novel mode of action. The protease-inhibiting compound according to the present invention is a compound represented by the general formula (I) [wherein all the symbols have the same meanings as described in the specification], its salt or solvate, or a prodrug thereof, is useful as a preventive and / or therapeutic agent for diabetes and / or complications of diabetes.

The invention relates to the technical field of bioactive peptides, in particular to a casein-derived antioxidant peptide mixture. Casein is hydrolyzed with protease, various pre-products of the casein antioxidant peptide are obtained, then separation, desalination and concentration are performed through ion exchange column chromatography, and the casein antioxidant peptide rich in basic amino acid is obtained. The casein antioxidant peptide has higher cell antioxidant capacity and low-density lipoprotein oxidation inhibition capacity and can still keep good antioxidant capacity after being digested and absorbed by gastrointestinal tracts, so that the casein antioxidant peptide rich in basic amino acid can be applied to fields of food and healthcare products and has very high social and economic benefit.

The invention belongs to the field of medicine, and relates to D-configuration polypeptide with high stability and capable of realizing mediated targeting of acetylcholine receptor high-expression cells and crossing corresponding barrier membranes and a nano drug delivery system thereof as well as an application in in-vivo and in-vitro brain targeting and in treatment of brain diseases and the like. Test results indicate that DCDX and the acetylcholine receptor are combined with IC50 to obtain 84.5nM which is stable in serum and tolerates hydrolysis of protease; the model drug carried by DCDX is specifically taken in by the positive cells expressing the acetylcholine receptor and has an ability of crossing the barrier formed by the kind of cells; and the nano drug delivery system made of a DCDX-modified polymer carrier material can deliver the entrapped model drug to the target tissue while the drug effect is remarkably improved. The D-configuration polypeptide DCDX provided by the invention can mediate active targeting of the drug or nano drug delivery system and has a good application prospect in the diagnosis and treatment of multiple diseases.

The present invention relates to thermostable proteases having an amino acid sequence which homologous to the amino acid sequence of proteases derived from Nocardiopsis, and the production thereof by wild-type and recombinant host cells including transgenic plants and non-human transgenic animals. The proteases are effective in animal feed, in particular fish feed, and detergents. The proteases are capable of degrading the soybean Bowman-Birk inhibitor, and other antinutritional factors such as soybean agglutinin and the Kunitz trypsin inhibitor, as well as the isolated soy storage proteins glycinin and beta-conglycinin. Characteristic structural features of relevance for the thermostability of these proteases of peptidase family S2A or S1E are disclosed.

The invention discloses a muting sensitive lactalbumin hydrolysate and a preparation method thereof. The method mainly comprises the following steps: dissolving condensed whey protein in the water, then carrying out denaturation under high temperature, carrying out hydrolysis by using the protamex composed of alkali protease, papain and flavourzyme, treating the lactalbumin hydrating solution with desalinization and drying to obtain the lactalbumin hydrolysate of the invention. The lactalbumin hydrolysate of the invention has high suppression ratio of Beta-lactoglobulin and Alpha- lactalbumin, can reduce or eliminate cow milk allergic reaction; furthermore, the lactalbumin hydrolysate can also produce other active materials, thus also having the functions of promoting immunity, being easy to digest and absorb and reducing blood pressure; and the lactalbumin hydrolysate has high security and no side effect, bitter taste or other bad flavors.