136 results about "Proteinase C" patented technology

The invention discloses a preparing method and fertilizer product of agricultural amino acid, which is characterized by the following: separating microbe bacteria 37-1 to produce secretory proteinase from wild pig gastrointestinal; seeding the bacteria in the waste protein; emzymlyzing solid for 5d (30-50 deg.c); making free amino acid content at 12% as raw material of fertilizer.

The invention relates to a preparation method of a hydrolyzed protein mixture and a protein peptide powder feed additive, wherein a multienzyme complex enzyme method is used for hydrolyzing vegetable protein to prepare the hydrolyzed protein mixture and protein peptide powder. The technology of the method is simple, and a non-starch polysaccharide enzyme and a proteinase are compounded so as to be capable of improving the hydrolyzation rate and degree of protein as well as the utilization rate of feed and reducing the production cost. Products prepared by the preparation method can be used as the feed additive to be capable of significantly improving the laying rate of egg-laying poultry, improving the weight increment of meat poultry, reducing the feed-meat rate and improving the production performance of table poultry as well as reducing the incidence rate of diarrhea in young animals and the feed-meat rate of piggy; and the preparation method is applied to aquatic livestock to be capable of improving the weight increment of fish and reducing the bait coefficient and content viscosity in the intestinal tract.

The present invention relates to thermostable proteinases from thermophilic Bacillus species and their uses in the preparation of nucleic acid samples. The enzymes of the invention are stable and active at 65-80 ° C., but are readily autolysed or denatured above 90° C.

The invention discloses an engineered Saccharomyces cerevisiae producing heat-stability recombinant trypsin, and its application, and belongs to the genetic engineering field. A trypsin gene obtained through in-vitro amplification is fused with a leading short peptide YVEF, and is wholly connected to a Pichia pastoris GS115 chromosome to construct engineered Saccharomyces cerevisiae efficiently secreting and expressing recombinant trypsin, and the recombinant trypsin having an improved stability is obtained after purification, and the heat stabilities of the recombinant trypsin at 40DEG C, 50DEG C and 60DEG C are 1.77, 2.6 and 31 times wild trypsin respectively, so the problem of the low stability of trypsin is solved. The production of trypsin through applying the engineered Saccharomyces cerevisiae has the advantages of high output, simple technology, intelligible heredity and application background of engineered Saccharomyces cerevisiae, and convenient industrial application.

The invention discloses an oral liquid containing soybean peptides and fungi polysaccharides and a preparation method thereof, and belongs to the technical field of non-alcoholic beverages, wherein the soybean peptides contain oligopeptides, short peptides and polypeptides mixtures. The oral liquid comprises the following raw materials in percentage by weight: soybean peptides containing oligopeptides, short peptides and polypeptides mixtures 2-12%, lentinan or ganoderma polysaccharide 0.03-0.1%, citric acid 0.1-0.15%, malic acid 0.1-0.15%, xylitol 5-10%, brown sugar 5-7%, honey 1-3%, CMC 0.2%, various kinds of vitamins, and the balance of water. The soybean protein powder which has been completely deodorized and one of akaline proteinase and flavor proteinase are treated at 55 DEG C and at the pH value of 9.0, and then centrifuged at 3000rpm. The supernatant is subjected to spray drying, so as to obtain the soybean polypeptide powder, and the soybean polypeptide powder is mixed with different kinds of fungi polysaccharides and the other raw materials such as citric acid, so as to prepare the oral liquid. The oral liquid has very good physiological and nutritive function, and the oral liquid and the preparation method develop a new way for deep processing of soybeans.

Assays are provided for rapid detection, with high specificity of the pathogenic form of prion protein responsible for neurodegenerative diseases affecting humans and animals, such as transmissible spongiform encephalopathy in bovine, sheep, and cats. Also provided are assays for testing animal feedstock, such as animal feed, for the presence or concentration of pathogenic prion protein. Results are available in from about 0.5 to about 20 minutes and preferably within from about 5 to about 10 minutes. The assays employ proteinase-K to remove normal prion protein from a biological sample, so that the sample may be analyzed by immunochromatography to determine the presence and concentration of pathogenic prion protein. Because the proteinase-K is immobilized on a solid support for in situ removal of interfering components, the present invention obviates the need for subsequent extraction of the desired analyte. All aspects of the present invention are suitable for quantifying the minimal detectable amount of pathogenic prion protein in a biological sample. Moreover, the simplicity of sample preparation makes the present invention suitable for use in the field.

The invention provides an extraction method of total DNA of metagenome of fish enteric microorganisms and a kit. The extraction method comprises the following steps: (1) fish sample sampling and fixing treatment of microorganisms on the surface of the fish; (2) acquisition of enteric microorganisms and collection of thalli; (3) embedding and cracking of cells; (4) extraction of DNA; and (5) DNA preservation. The kit comprises the following main components: a solution A which is a mixed solution of Tris and EDTA; a solution B which is a mixed solution of Tris, EDTA, NaCL, PVP, guanidinium isothiocyanate, Triton-X100, PMSF, and beta-mercaptoethanol; a solution C which is a mixed solution of Tris, EDTA, NaCl and DTT; a solution D which is a mixed solution of proteinaseK, a lysozyme, Tris, NaCl and SDS; a solution E which is a mixed solution of Tris saturated phenol and chloroform; F which is isopropanol; and G which is an ethanol aqueous solution. The DNA fragment obtained by the invention is high in purity, and the content of DNA of a host and the microorganisms on the surface of the fish is small.

The present invention discloses the tumor invasion and metastasis resisting function and application of venin cysteine proteinase inhibitor, and belongs to the field of biomedicine. Through designing and synthesizing sv-Cystatin cDNA according to 99 amino acid sequences of Chinese cobra venin Cystatin protein, cloning to pPICZ alphaA vector and transforming Pichia yeast, stable and high expression engineering bacterium GS115-sv-Cystatin is screened out. Through further inducing expression and purification, the extracorporeal bioactivity experiment shows that the recombinant sv-Cystatin protein has the functions of inhibiting the activity of papain and inhibiting the tumor cell invasion and metastasis obviously, and so do the intracorporeal experiment. The present invention also constitutes pcDNA-sv-Cystatin eukaryotic expression vector. The present invention has huge application foreground in preventing and treating tumor.

The invention relates to a Bacillus licheniformis engineering bacterium capable of efficiently secreting nattokinase, and belongs to the technical fields of enzyme engineering and microbes. The method adopts a molecular biology technology to screen and replace signal peptides with important effects in an exogenous protein transhipment process, and the signal peptides are replaced by signal peptides from levanase SacC in Bacillus subtillis 168 from signal peptides from extracellular serine proteinase Vpr in Bacillus licheniformis WX-02 on the basis of nattokinase producing Bacillus licheniformis engineering bacterium BL10 (pP43SNT) preserved in a laboratory in advance in order to afresh construct the nattokinase producing Bacillus licheniformis engineering bacterium BL10 (pP43SacCNK). The bacterium disclosed in the invention can substantially improve the nattokinase secreting level under liquid fermentation conditions, and the maximum enzyme activity can reach 33.83 FU / mL. The bacterium is preserved in China Center for Type Culture Collection on June 16, 2014 with the preservation number of CCTCC NO: M2014253.

The invention relates to a deoxyribonucleic acid (DNA) extraction technology, in particular to a method for simply, conveniently and quickly extracting trace total DNA of single roes and fries. The method comprises the following steps of: fixing and preserving collected roes or fries; washing to remove stationary liquid, adding DNA extraction buffer solution and lysis buffer solution, shearing the roes or the fries, adding proteinase K and ribonucleic acid (RNA) enzyme, incubating and digesting; and precipitating protein by using high-concentration NaCl solution, adding isopropanol to precipitate DNA, washing a precipitate by using 70 percent ethanol to remove salt ions, and adding ultrapure water or tris-ethylenediaminetetraacetic acid (TE) solution and dissolving after the ethanol is volatilized. The method solves the problem of the limitation of the application the conventional total DNA extraction method to samples with the low content of genomic DNA, such as roes or fries and thelike.

The invention belongs to the technical field of edible fungus processing, and particularly relates to a method for extracting soluble nitrogen-containing compounds from edible fungi by combining screw extrusion with enzymolysis. The method includes the steps: (a) treating raw materials; (b) performing screw extrusion pretreatment; (c) performing enzymolysis; and (d) performing centrifugal treatment to obtain supernatant liquor of the soluble nitrogen-containing compounds and edible fungus residues. By combining modified wall breaking of screw extrusion with further proteinase enzymolysis, the advantages of modified wall breaking of screw extrusion and enzymolysis are combined, delicious substances in thallus of the edible fungi are effectively released, the extraction rate of the soluble nitrogen-containing compounds of mushrooms is increased, and the method is green and environment-friendly.

The invention discloses a new Pseudomonas corrugate strain (P94 GMCC No.1895), which is characterized by the following: manufacturing relative HCN to prevent bacteria, proteinase and phospholipase and IAA; inhibiting partial plant pathogenic fungus (Botrytis cinerea, Ceratocystis fimbriata, Monilinia laxa, Magnaporthe grisea, Pythium aphanidermatum and Phytophthora capsici) and partial pathogenic bacteria (Pseudomonas syringae, Acidovorax avenae and Ralstonia solanacearum); accelerating the growth of tomato and cucumber to improve production by 22%-28%.

The invention relates to a method for improving the silkworm chrysalis protein functional characteristics using an ultrasonic wave coordinated with enzymolysis technology. The method uses ultrasonic wave coordinated with neutral proteinase to improve the functional characteristics of silkworm chrysalis protein. The preparation method comprises the following steps: preparing a suspension liquid from silkworm chrysalis protein using distilled water, regulating the pH, putting the suspension liquid together with the neutral proteinase into a reaction tank, inserting an ultrasonic probe for ultrasonic wave coordinated enzymatic hydrolysis, and spray drying the reaction mixed solution to obtain a powder modified silkworm chrysalis protein product. The method has the advantages that the production time is short, the operation is convenient, the function characteristics of solubleness, frothiness, emulsibility and the like of the silkworm chrysalis protein can be obviously increased, and the obtained product meets functional food requirements.

The present invention relates to comprehensive development and application of oyster, and belongs to the field of food biotechnology. Fresh oyster material is steamed to leach out glycogen, centrifugated to obtain supernatant and oyster dregs as precipitate; and the supernatant is processed with alcohol solution of 20-95% concentration and centrifugated to obtain precipitate and the precipitate is dried to obtain glycogen product. The oyster dregs are hydrolyzed at pH 3.0-10 and 30-80 deg.c with proteinase and centrifugated to obtain enzymolyzed liquid, and the enzymolyzed liquid is spray dried to obtain oyster peptide powder. The present invention has high comprehensive utilization and may be used in industrial production.

There is a demand for improved turbidimetric immunoassays for human Cystatin C in biological samples, especially in human clinical samples of body fluids. The present invention provides a turbidimetric immunoassay method and reagent set enabling measurement of human Cystatin C by turbidimetric methods, resulting in a surprisingly stronger and faster turbidimetric signal than in the present state of the art. The increased and faster signal is accomplished by the use of new reagents and compositions, and enables shorter assay times and kinetic reading with a stronger signal, improving overall assay speed and quality. Improved robustness to lipid interference and improved linearity is achieved.

The invention relates to a plant proteinase extraction method and obtained plant proteinase. The method comprises the following steps: mixing slurry or a crushed material of plants with water in a solid-liquid ratio of 1:(5-20), then carrying out water extraction, and collecting an extracting solution; filtering the obtained extracting solution by virtue of a plate-and-frame filter press and a ceramic membrane sequentially, wherein the pore size of filter cloth of the plate-and-frame filter press is 100-150 meshes, and the pore size of the ceramic membrane is 50-200nm; carrying out primary ultrafiltration treatment on a filtrate by using an ultrafiltration membrane with the molecular weight cutoff of 30-100KDa, collecting the filtrate, carrying out secondary ultrafiltration treatment on the filtrate by using an ultrafiltration membrane with the molecular weight cutoff of 5-15KDa, and collecting trapped fluid; and concentrating the trapped fluid to obtain a concentrated solution, and carrying out freeze drying on the concentrated solution, so that the plant proteinase is obtained. The method provided by the invention has the advantages of simple technology, no pollution to the environment, the feasibility of large-scale industrial production and high proteinase activity and purity. The plant proteinase has the advantages of good activity, high purity and no solvent residue and meets usage requirements of the food, daily chemical and medicine industries.

The invention provides a method for rapidly extracting DNA from whole blood. The method comprises the following steps: S1, centrifuging a blood sample at a high speed, removing a supernatant, keeping precipitate, adding a lysate, uniformly mixing till a clear state, keeping for 40-60min in a water bath with the temperature of 50-60 DEG C, cooling, performing suction extraction by using an organic solvent, centrifuging, and taking a supernatant; S2, adding isopropanol and NaAc into the supernatant, uniformly mixing, collecting the precipitate, cleaning the precipitate and airing; S3, adding a TEN buffer solution and ribonuclease into the precipitate, uniformly mixing, incubating for 30min, then adding a proteinase K containing TEN buffer solution, uniformly mixing, performing water bath for 30min, cooling, centrifuging, taking a supernatant, and purifying to obtain the pure DNA. By the method, the sample treating amount is large; most of red blood cells in the supernatant are removed through high-speed centrifugation first and then blood cells (white blood cells) in the precipitation are lysed by using the self-prepared whole cell lysate by one step so as to rapidly obtain a large number of DNAs applicable to three generations of sequencing from blood.

The invention discloses a total DNA (Deoxyribonucleic Acid) extraction method of bottom mud containing high content of humus at the river mouth. The method comprises the following steps: washing bottom mud by stroke-physiological saline solution to remove part of impurities such humus, and meanwhile, concentrating microorganisms; then, multigelating in liquid nitrogen and a water bath at 65 DEG C by means of CTAB (Cetyltrimethyl Ammonium Bromide), proteinase K and lysozyme for cell disruption, and then adding SDS (Sodium Dodecyl Sulfonate) and insulating for 2 hours in the water bath at 65 DEG C to combine protein; adding isovolumetric phenol / chloroform / isoamylol (25:24:1) to remove protein; adding isovolumetric isopropanol to set DNA, washing precipitate with 70% ethanol, and dissolving DNA by TE (pH8.0) after airing; adding 3 times of absolute ethyl alcohol in volume to re-set DNA, washing the precipitate with 70% ethanol, dissolving the precipitated DNA by TE after airing, and storing at minus 20 DEG C for later use. The method provided by the invention is simple to operate and cheap in price. The total DNA of bottom mud extracted by the method provided by the invention has the advantages of high purity and integral biodiversity, and can be used for molecular biology study and microbial diversity analysis.

The invention discloses an efficient and economical soil microbial DNA extraction method. The method comprises the following steps: step 1, obtaining a microbial suspension; weighing 1 g of soil, adding 2 ml of suspension Buffer C1, 1 g of glass beads having the particle diameter of 0.09-0.12 mm, 500 muL of humic acid adsorbent Buffer C2 and 10 muL of proteinase K (10 mg / mL), mixing and shaking; step 2, obtaining microbial DNA; step 3, obtaining crude DNA; placing the material obtained in the last step in a water bath of 60 DEG C while stirring, then centrifuging at a high speed of 4 DEG C, and collecting the first supernatant; and step 4, obtaining purified DNA. The method is based on the requirements of soil-contaminated microbial detection, can extract DNA containing no inhibiting factor from complex environmental samples such as soil and feces, and is high in extraction speed and accurate in analysis result.

The invention discloses a Pseudomonas aeruginosa strain SWJSS3 and application thereof in producing proteinase. The strain is collected by General Microorganism Center of China Committee for Culture Collection of Microorganisms on June 10th, 2015; the collection address is Institute of Microbiology in Chinese Academy of Sciences, Yard 1, Beichenxi Road, Chaoyang District, Beijing City; and the collection number is CGMCC No.10973. The application method of the strain for producing proteinase comprises the following steps: adding a fermentation culture medium into a fermentation tank, wherein the liquid filling amount is 8-12%, the initial pH value is 7.0-7.5, the strain inoculum size is 0.8-1.2%, and the initial temperature in the tank is 20-40 DEG C; and fermenting under oscillating conditions for 30-80 hours to generate abundant proteinase in the tank, and separating and extracting to obtain the proteinase. The strain SWJSS3 has high proteinase production capacity; and the obtained proteinase has the characteristics of high enzyme activity and high salt tolerance. The fermentation method disclosed by the invention has the advantages of simple conditions and stable hereditary property, and is suitable for industrial production.

The invention provides a method using a composite bacterium to ferment bean dregs to manufacture protein feed. The composite bacterium is composed of trichoderma, aspergillus oryzae and saccharomyces cerevisiae. Agriculture residues or food residues of a bean dreg class are added to base materials, and composite fermentation is carried out. Products after being fermented are rich in single-cell protein, digestible energy and metabolic energy of the products are improved, the content of rough fibers is reduced, and a great amount of probiotics, abundant enzymes and bioactivator are included. Through detection to unfermented essential nutrients and fermented essential nutrients, compared with the content of bean dreg protein of unfermented bean dregs, the fact that the content of bean dreg protein of bean dregs after being fermented is increased by about 8%, amino acid nitrogen is increased by 4.9 times, the activity of proteinase containing is 1157 U / g, and the bean dregs after being fermented are ideal protein feed.

The invention relates to an immunoturbidimetry kit for detecting a cysteine proteinase inhibitor C and a preparation method thereof. An application liquid bottle, an antibody suspension liquid bottle and a standard substance bottle are placed in a box body; an application liquid is filled in the application liquid bottle and comprises the following components: 0.01-2% of surfactant, 0.01-1% of preservative, 0.1-20% of sodium chloride and 10-200 mmol / L of buffer solution; a latex suspension liquid coated with a monoclonal antibody of the cysteine proteinase inhibitor C is filled in the antibody suspension liquid bottle and comprises the following components: 0.05-1% of latex coated with the monoclonal antibody of the cysteine proteinase inhibitor C, 0.01-2% of surfactant, 0.01-1% of preservative and 10-200 mmol / L of buffer solution; and a standard substance of the cysteine proteinase inhibitor C is filled in the standard substance bottle. The purpose of large-batch quantitative detection on a biochemical instrument is achieved.

The invention discloses a method for improving the yield of cordycepin and thermoduric proteinase of cordyceps militaris (L.) link through a liquid-state fermentation method, and relates to a strain which is preserved in China Committee for Culture Collection Center for general microbiology. By utilizing the strain, through preparation of cordyceps militaris (L.) link strain by liquid fermentation and production of cordycepin by liquid strain fermentation, an experiment result proves that the content of strain deep liquid-state ventilation fermentation cordycepin reaches 4.12g / L, and the activity of the thermoduric proteinase reaches 92554U / ml. Meanwhile, the method disclosed by the invention has the advantages of simple operation, low cost, short period, and easy realization; the raw materials are rich in sources and easy to get, are low in cost and have no pollution to the environment; the prices of the used equipment and reagents are cheap; and large-scale production is convenient.

The invention provides a livestock / poultry manure-straw coupled field returning method. Cellulase, proteinase, lipase, microzyme, deodorizing bacterium and other fermentative bacteria are added into livestock / poultry manure to shorten the manure fermentation time; the livestock / poultry manure is used as a carrier for production, reproduction and application of the straw decomposing bacterium, so that the quantity of the fermented decomposing bacterium is increased by 8-20 times; and no odor emits in the manure fermentation process under the actions of the proteinase and lipase. By organically combining the livestock / poultry manure field returning and straw mechanical pulverization field returning, and the livestock / poultry manure is used as a crop straw accelerator to accelerate the rotting of the straws under the action of the mixed bacteria in the manure. Compared with the single straw mechanical pulverization field returning process, the method obviously shortens the straw rotting time, solves the problem that the emergence of seedlings of the seeds can be influenced during seed sowing since the straws can not be easily rotten and returned to the field, and is capable of obviously improving the aggregate structure in soil and enhancing the contents of organic matters, nitrogen, phosphorus and potassium in the soil. Compared with the straw pulverization-stack retting fermentation field returning process, the method is simple, simplifies the application links, is easier to operate, and saves abundant human resources and material resources.

The invention discloses a recombinant beauveria brongniartii proteinase K mutant PK-M1 and a preparation method of the recombinant beauveria brongniartii proteinase K mutant PK-M1. According to the technical scheme, starting from transforming the molecular structure of proteinase K of recombinant beauveria brongniartii, the proteinase K mutant PK-M1 with the activity and stability both improved isobtained, and the amino acid sequence is as shown in SEQ ID NO:1; an expression system is optimized, by utilizing the yeast cell external secretion expression technology, the recombinant proteinase Kmutant can be fermented in large scale and efficiently expressed, and further, the yield of proteinase K is improved; and with the protein purification manner realizing efficient affinity chromatography, the yield of the proteinase K in the purifying process is improved, and thus the large-scale production of the proteinase K is realized.

The invention discloses a deegging liquid and method to strip egg in the abdominal region of hen rapidly without injuring, wherein the deegging liquid contains proteinase and sodium chloride at certain proportion. The deegging method comprises the following steps: placing the incubated hen into deegging liquid to enzymolyze; aerating continuously during enzymolytic course; stripping the whole egg from abdominal region of hen completely; finishing the deegging course within 5h; maintaining the survival rate of deegged hen and egg over 90%. The invention has first break-through of rapid and non-injured deegging of the incubating hen, which can be applied to crossbreed, do manufactured breeding, collect and manufacture fresh water shrimp egg.

The preparation process of water soluble royal jelly includes the following steps: defreezing royal jelly at room temperature; homogenizing via adding water in 1.5-3 times and stirring; enzymolyzing with proteinase at 40-45deg.c while stirring for 2-4 hr and ultrafiltering membrane or vacuum concentration at normal temperature to the amount of 1-1.5 times of royal jelly material; freeze drying; and vacuum packing. The water soluble royal jelly product has water content not more than 3 wt% and 10-HAD content not less than 4.0 wt%. The preparation process is simple and reliable, and the water soluble royal jelly product has wide dissolving temperature range, high stability, water dissolving retention rate as high as 90 % and other features.

The invention discloses small peptide chelated zinc as well as a preparation method and application thereof. The preparation method comprises the following steps: crushing coarse fishes and adding water; adjusting the pH (Potential of Hydrogen) to 5-9; heating and raising the temperature to 50 DEG C-70 DEG C; adding proteinase for enzymolysis; carrying out the enzymolysis until the hydrolysis degree is 30%; heating and carrying out enzyme inactivation and centrifuging; carrying out ultra-filtration by utilizing a 10kDa ultra-filtration membrane and taking permeate liquid; carrying out the ultra-filtration on the permeate liquid by utilizing a 1kDa ultra-filtration membrane, and taking non-permeate liquid; adding inorganic zinc salt into the non-permeate liquid, and stirring and oscillating under the conditions that the temperature is 40 DEG C-60 DEG C and the pH value is 5-7 until the chelating rate is 70%+ / -5%; centrifuging and concentrating; and freeze-drying to obtain the small peptide chelated zinc. According to the small peptide chelated zinc disclosed by the invention, the digestion and absorption of proteins and microelements of piglets are improved and the immunity of the piglets is enhanced; the small peptide chelated zinc also has good anti-oxidization and antibacterial functions and is added into feed so that the guarantee period of the feed can be effectively prolonged; and risks that the diarrhea of the piglets is caused and the immunity is reduced and the like, caused by the fact that the feed goes bad, are reduced.

The present invention provides a meat tenderer and its preparation method. The formula is formed from plant hydrolyzing proteinase, edible carbohydrate, taste regulator and special adjuvant. Said method is simple and easy to implement, the said materials are mixed uniformly to obtain the invented tenderer capable of tendering various meats, such as beef, mutton, pork, chicken meat, duck meat and fish muscle, etc. in the various cooking processes.