399 results about "Zymogen" patented technology

Disclosed are methods of producing eukaryotic disulfide bond-containing polypeptides in bacterial hosts, and compositions resulting therefrom. Co-expression of a eukaryotic foldase and a disulfide bond-containing polypeptide in a bacterial host cell is demonstrated. In particular embodiments, the methods have been used to produce mammalian pancreatic trypsin inhibitor and tissue plasminogen activator (tPA) in soluble, biologically-active forms, which are isolatable from the bacterial periplasm. Also disclosed are expression systems, recombinant vectors, and transformed host cells.

Disclosed are methods and compositions for producing heterologous disulfide bond containing polypeptides in bacterial cells. In preferred embodiments the methods involve co-expression of a prokaryotic disulfide isomerase, such as DsbC or DsbG and a gene encoding a recombinant eukaryotic polypeptide. Exemplary polypeptides disclosed include tissue plasminogen activator.

The invention discloses a micro plasminogen mutant with a function of inhibiting platelet aggregation, which has an amino acid sequence of KLYDYCKGDWPCAAPSFDCGKPQVEPKKCPGRVVGGCVAHPHSWPWQVSLRTRFGMHFCGGTLISPEWVLTAAHCLEKSPRPSSYKVILGAHQEVNLEPHVQEIEVSRLFLEPTRKDIALLKLSSPAVITDKVIPACLPSPNYVVADRTECFITGWGETQGTFGAGLLKEAQLPVIENKVCNRYEFLNGRVQSTELCAGHLAGGTDSCQGDSGGPLVCFEKDKYILQGVTSWGLGCARPNKPGVYVRVSRFVTWIEGVMRNN. A preparation method of the micro plasminogen mutant comprises the following steps of: replacing a D-V-P-Q sequence at 7 to 10 positions of a micro plasminogen sequence with a K-G-D-W-P sequence; forming a convex Loop structure on the replaced sequence; and placing a K-G-D sequence at the top end of the Loop structure. Meanwhile, the invention provides application of the micro plasminogen mutant to preparation of a medicament for preventing and treating thrombotic diseases and ophthalmic diseases.

The invention relates to a microbial foliage fertilizer, and a preparation method and application thereof. Every liter of microbial foliage fertilizer is prepared by mixing the following raw materials: 0.3-0.5L of microbial zymogen liquid, 0.2-0.3L of trace element solution, 0.05-0.15kg of K2SO4, 0.05-0.15kg of NH3H2PO4, 0.1-0.25kg of urea, 0.05-0.15kg of yeast fermentation powder slag, 3-4g of diethylaminoethanol hexanoate and the balance of water. Multiple microbes with the functions of nitrogen fixation, phosphorus decomposition, potassium decomposition and disinsection are added into the foliage fertilizer, thereby being beneficial to enhancing the crop yield, quality and disease / insect resistance.

The invention discloses a recombinant microorganism for preparing dharma diene and protopanoxadiol and a construction method of the recombinant microorganism. The construction method of the recombinant bacteria comprises a step of adding dharma diene synthase, protopanoxadiol synthase and nicotinamide adenine dinucleotide phosphate - cytochrome P450 reductase encoding gene into saccharomyces cerevisiae to obtain recombinant bacteria I. According to the recombinant microorganism for preparing the dharma diene and the protopanoxadiol and the construction method of the recombinant microorganism, by means of homologous recombination, the dharma diene synthase, the protopanoxadiol synthase and the nicotinamide adenine dinucleotide phosphate - the cytochrome P450 reductase encoding gene are all added into the saccharomyces cerevisiae to obtain the initial recombinant bacteria, and the effect that the initial recombinant bacteria can produce trace amount of the dharma diene and trace amount of the protopanoxadiol is discovered; tHMG1 activity of the initial recombinant bacteria is further improved, and therefore intermediate recombinant bacteria are obtained, and by means of the intermediate recombinant bacteria, the yield of the dharma diene and the yield of the protopanoxadiol are significantly increased; the activity of one or two or three of ERG1, ERG9 and ERG20 are improved on the basis of the intermediate recombinant bacteria, and the effect that the recombinant bacteria which can be used to increase the yield of the dharma diene and the yield of the protopanoxadiol are constructed is also discovered. By means of the recombinant microorganism for preparing the dharma diene and the protopanoxadiol and the construction method of the recombinant microorganism, the foundation is laid for artificially synthesizing the dammar diene and the protopanoxadiol.

The invention provides a multifunction active agent of fermentation beds. Chinese herbal medicament components are added in the active agent to kill ova and increase lean meat yield. The aim of the invention is realized by comprising the compositions in portion: 2 portions of zymogen and the following Chinese herbal medicaments: 0.1 to 0.5 portion of folium cortex eucommiae, 0.1 to 0.5 portion of glossy privet fruit, 0.5 to 2 portions of hawthorn, 0.5 to 2 portions of astragalus root and 0.1 to 0.8 portion of Chinese cinnamon. The multifunction active agent is characterized in that the active agent ensures that livestock and poultry excretions undergo fermentation in the fermentation beds to provide abundant organic fertilizer for plants and that the temperature generated by fermentation creates favorable growth conditions for livestock, poultry and plants; harmless treatments such as quick fermentation, decomposition, adsorption, disinfestation and ovum killing of the livestock and poultry excretions are carried out through beneficial strain and the Chinese herbal medicaments; meanwhile, the added Chinese herbal medicaments can promote the multiplication of beneficial bacteria in pig intestines so as to accelerate growth; and lean meat ratio can be increased by approximately 15 percent.

The affinity chromatographic process for producing high purity chymotrypsin includes the following technological steps: freezing fresh ox or pig pancreas as the material, crushing and extracting protein, stepped salting out and crystallizing zymogen, enzymolyzing to obtain coarse product, affinity chromatographic separating and purifying, ultrafiltering and concentrating sterilizing, and vacuum freeze drying to obtain product. Compared with available technology, the present invention has the advantages of suitability for large scale produce, high efficiency, specificity, low cost and high product quality.

The invention discloses a recombinant bacillus subtilis and a method for producing transglutaminase by utilizing the recombinant bacillus substilis. The recombinant bacillus subtilis is obtained by the following steps of: (1) by virtue of fusion PCR (polymerase chain reaction), connecting a P43 promoter and a signal peptide gene with a subtilisin-like protease gene, and converting a fusion PCR amplification product into bacillus subtilis WB800 to obtain bacillus subtilis B.subtilis WB800S; and (2) carrying out PCR amplification on streptoverticillium mobaraense transglutaminase protogene, andthen converting amplification product into the bacillus subtilis B.subtilis WB800S to obtain bacillus subtilis B.subtilis WB800S / proMTG. The bacillus subtilis B.subtilis WB800S / proMTG is fermented and purified to obtain soluble transglutaminase. By adopting the method disclosed by the invention, the transglutaminase with biological activity can be directly obtained in supernatant of fermentation liquor, fussy processes of breaking cells and activating proenzyme after purification are eliminated, and a fermentation production process is simplified.

The invention discloses one kind of bread zymogens agents for plants straw especially fit to fermentation of dried and yellow straw contained plant lactobacillus, microzyme, trichoderma reesei, bacillus subtilis, which is preparation by mixing bacterial liquid in proportion after third class general cultivation in solid test tube, liquid triangle bottle and liquid malt wort for all bacterial, and state fermentation cultivation to grow up. All bacterial in this invention are cultivated separately, mixed till the stage of producing bacterial in liquid, which is more suitable to different cultivation conditions of different bacterial and enables every bacterium with the strongest activity.

The invention discloses a mutant human plasminogen kringle5 gene mK5 and an amplification primer of the mK5 gene, the mutant human plasminogen kringle5 gene modified by adding glutathione-S-transferase before the gene mK5, and a method for preparing protein mK5 recombinant protein of two gene codes, glutathioneS transferase (GST)-mK5 fusion protein and two proteins, wherein the mK5 gene and the GST-mK 5 gene can be applied to preparing medicaments for treating angiogenesis diseases. By the invention, the number of exogenous amino acid in the recombinant K5 protein molecules obtained by a gene engineering method is remarkably reduced, the K5 bioactivity of the obtained mK5 is improved, while the mK5 activity of the GST-mK5 fusion protein is maintained, the stability and water solubility of the protein are improved, the purification steps of an expressed product are simplified, and the purity of the product is improved.

The invention relates to a soilless seedling culturing substrate for resisting diseases and root nematode of plants and a production method thereof. In the invention, organic wastes such as straws are taken as raw materials, inoculated with zymogen according to certain proportion, subjected to moisture content regulation, evenly mixed and stacked for primary fermentation; when the temperature of a pile reaches 60 DEG C, the pile is turned; the pile is turned and oxygen and moisture content are supplemented every seven to ten days afterwards; after the pile is turned for three times, when the temperature is lower than 35 DEG C, the pile is inoculated with a bio-antagonist for secondary fermentation, turned every three to four days and kept at a temperature of between 35 and 40 DEG C; and after 15 to 20 days, the seedling culturing substrate containing the 10 bio-antagonist is obtained. The seedling culturing substrate has chemiotaxis for interfering pathogenicbacteria and nematode, can reduce infestation chance of the pathogenicbacteria and nematode on the root surface, and has double effects of resisting phytophthora disease and root nematode disease through the secondary fermentation and microorganism activation technology.

The invention discloses an efficient microbial soil remediation agent, belonging to a propagation culture technique of microbes. The efficient microbial soil remediation agent is prepared by mixing the following components in percentage by volume: 10-25% of photosynthetic bacteria zymogen solution, 25-35% of Bacillus subtilis zymogen solution, 20-35% of microzyme zymogen solution and 20-40% of actinomycetes zymogen solution. The four strains are respectively subjected two secondary amplification culture under the conditions of proper temperature, ventilation, culture medium, pH value and culture time; and the culture process is carried out in a fermentation tank without foreign microbes. By using the modern biotechnology, single microbe strains are subjected to high-concentration pure culture and then compounded to produce the efficient microbial soil remediation agent.

Method and device, including a uniquely operative applicator, and pharmaceutical compositions, for the controlled, non-surgical removal and retrieval of cells from a variety of skin lesions and tissue surfaces are disclosed. A synergistic effect of proteolytic digestion of the intracellular matrix and "stripping" flow is achieved by treating a defined area with a controlled, continuous stream of proteolytic enzyme solution, causing gentle but effective tissue erosion. Isolated cells from the skin lesion and / or tissue surface may be collected from the protease solution stream for histological analysis and / or cell culture, affording a method of "enzymatic biopsy". The protease solution may be supplemented with anesthetics, coagulants, anticoagulants and antibiotics to decrease the discomfort, erythema, bleeding, risk of infection and scarring traditionally associated with surgical treatment of skin lesions. Delivery of precise levels of catalytic activity is ensured by controlled activation of stable, inactivated enzyme stock solutions and powders shortly prior to application.

An aspartic enzyme having a high homology with a cathepsin D precursor, which is a protein having the N-terminal amino acid sequence LVRIPLHKFT (SEQ ID NO: 1) and showing a molecular weight of about 45 kDa in non-reductive SDS electrophoresis and can degrade plasma proteins, typically plasminogen, to produce plasma protein fragments having an inhibitory activity to metastasis and growth of cancer; the plasma protein fragments having an inhibitory activity to metastasis and growth of cancer which is prepared via the degradation with the above enzyme; a process for preparing the protein fragments which comprises degrading plasma proteins with the above enzyme; and a medicament for treating and preventing metastasis and growth of cancer which comprises as a major ingredient the above enzyme or the plasma protein fragments.

The invention relates to a method for improving the extraction rate of crude oil by using industrial waste water and industrial waste gas. The method comprises the following steps: injecting the industrial waste water containing carbohydrates and the industrial waste gas containing carbon dioxide into an oil layer, carrying out fermentation reaction in the oil layer environment, and generating metabolites, such as a biosurfactant, CO2, H2 and the like. The method comprises the following steps: A, preprocessing the industrial waste water to remove larger particulate matters and adjust pH to be equal between 6.0 to 9.0; B. making the proportion of the preprocessed industrial waste water to an activating agent to a zymogen liquid be 100,000:10 to 100:1 to 5, and synchronously injecting the industrial waste water, the industrial waste gas and oil well injecting water into stratum by the industrial waste water delivery capacity of 5 to 100m / h and the industrial waste gas delivery capacity of 0.5 to 10 x 10Nm / h at a pressure of less than the oil failure pressure; and C, continuously injecting the industrial waste water, the industrial waste gas and the oil well injecting water into the stratum at any proportion for a long time, carrying out the microorganism fermentation reaction in the stratum, and generating the metabolites which are favorable for improving the extraction rate of the stratum crude oil. The method can be widely used in the petroleum exploitation process.

The invention provides a method for quickly aging and tenderizing beef, which comprises the following steps: during the beef chilling aging process, cattle pancreas is prepared into homogenate; Ca2+ is utilized to activate various zymogens in the pancreas; active pancreas injection solution is prepared and directly injected into the beef, so that trypsin in the beef can decompose muscle protein; and simultaneously, by the action of ultrasonic wave, the original structure of muscle is damaged, the tenderness is improved, the aging purpose can be achieved within three to five days, and the natural aging time can be shortened by about 50%. In the aspect of keeping quality, vacuum packaging and radiation sterilization are combined, so that the storage period of the cut beef can be prolonged for 3-5 times; in the aspect of the quality of the beef, the cut beef prepared by the method has organoleptic, physicochemical and other indexes which can meet the requirements of 'GB2707 Hygienic Standard for Fresh (Frozen) Meat of Livestock'; and the cutting force is 4.2-5.3kg, and people can obtain the best mouthfeel. The method maintains the specific flavor and nutrition of the beef, and has the characteristics of quick aging speed, long shelf life, energy saving effect and high production efficiency.

The invention belongs to the field of subsidiary agricultural products, in particular to an organic fertilizer prepared by using dregs of WangLoKAT herb residue and a preparation method thereof. Organic content of the organic fertilizer is more than 70 wt percent, and the total content of NPK is more than 4.0 wt percent. The preparation method comprises the following steps: cutting the dregs of WangLoKAT herb residue in short, adding proper water according to the requirement, adding zymogen agent, uniformly stirring, bedding a vent plate in a fermentation plate, piling the WangLoKAT herb residue, inserting a vent pipe into a middle upper part of the dreg pile and covering with jute cloth or a venting plastic film, piling and fermenting, turning the pile, supplementing proper water, airing, grinding into fine powder when the water content is less than 20 wt percent to obtain the organic fertilizer. The product meets the standard of commercial organic fertilizer.

Disclosed is a method for treating pig manures to produce an organic fertilizer, which inoculates horse manures in pig manures and purifies and amplifies high temperature fiber bacteria thereof to prepare a zymogen agent through two times of high temperature fermentation, then composts are inoculated in a solar energy isolation greenhouse to produce the organic fertilizer. The method has advantages of fast compost starting, low cost of nitrogen protecting and deodorization agent, broad sources and good effects; and the whole process for producing the organic fertilizer is conducted in the solar energy isolation greenhouse, thereby effectively preventing secondary pollutions such as pig manures and diffusion of nutrients and realizing the clean treatment to the pig manures and production to the organic fertilizer. The method is suitable for the southwest area and the like in China abound with rain and moisture and with rather low temperature in winter, and has advantages of low investment, low energy consumption, low cost, simple operation, strong practicability and easy promotion.

The invention discloses recombinant saccharomyces cerevisiae for producing dammarenediol and protopanoxadiol using xylose and a construction method. The construction method comprises the steps of replacing a promoter of a saccharomyces cerevisiae xylulokinase gene XKS1 with a promoter PFBA1 by virtue of a homologous recombination method, introducing xylose reductase XYL1 and a xylitol dehydrogenase XYL2 expression cassette, increasing the activities of transketolase TKL1 and transaldolase TAL1 so as to obtain recombinant bacteria 1, introducing a farnesyl-diphosphate farnesyltransferase gene ERG9, a squalene monooxygenase gene ERG1 and a dammarenediol synthase gene DS into the recombinant bacteria 1 so as to obtain recombinant bacteria 2, and introducing a nicotinamide adenine dinucleotide-hydroxymethylglutaryl coenzyme A reductase gene NADH-HMGr, farnesyl diphosphatesynthase ERG20 and a protopanoxadiol synthase-cytochrome P450 reductase fusion protein gene PPDS-ATR1 into the recombinant bacteria 2, so as to obtain recombinant bacteria 3. According to the recombinant saccharomyces cerevisiae, dammarenediol and protopanoxadiol can be artificially synthesized by virtue of xylose.

The invention provides a silage additive special for paper mulberries and an application of the silage additive. The additive comprises zymogen and complex enzymes, wherein the zymogen consists of lactobacillus plantarum (lactobacillus plantarum) plantarum ) ACCC11016 and enterococcus faecalis (enterococcus faecalis) faecalis ) CICC20440 according to the ratio of the viable count of the lactobacillus plantarum to that of the enterococcus faecalis being (1-10) to (0-1), and the complex enzymes consist of cellulase, pectinase and glucose oxidase. According to the silage additive, through synergistic effects of lactic acid bacteria, an enzyme preparation and soluble carbohydrate, the retention period of a fresh paper mulberry coarse feed is prolonged, besides, various organic acids and various nutrient substances are produced, the palatability and the digestion and utilization rate of paper mulberry branches and leaves are increased, the content of neutral washing fibers and the content of acidic washing fibers are reduced, increase of the food consumption of livestock and poultry is facilitated, and the silage additive can be used for producing an environmental-friendly biologic feed.

A functional mutant of human plasminogen disclosed in the invention, respectively, is hPLG-delta K: human plasminogen protein Pro<544>-Asn<791> polypeptide; Pro<559> in RGD-hPLG-delta K: hPLG-delta K is mutated to Asp<559>; and Gly<560> in RHP-hPLG-delta K: hPLG-delta K polypeptide is mutated to His<560>. The invention also discloses a preparation method for the functional mutant. The product is obtained by using a plasmid containing a full-length cDNA sequence of human plasminogen as a template to carry out a PCR to construct a plasmid and using Pichia Pastoris expression. The functional mutant has a dual-function of fibrinolysis and inhibiting platelet aggregation or inhibiting fibrin monomer polymerization.

The invention provides a fusion protein formed by the combination of urokinase-type plasminogen activator a chain and melittin, the preparation method thereof, and the application of the fusion protein in tumor treatment.

The technological process of preparing zymogen with waste beer yeast liquid includes the following steps: heating waste beer yeast liquid to 50-55 deg.c and maintaining at the temperature for certain period while controlling pH 5.5-6.5; adding salt, potassium dihydrogen phosphate and magnesium chloride to promote cell wall breaking and autolysis while maintaining the temperature and stirring; adding enzyme preparation before maintaining for 25-30 hr to dissolve cell completely and to make amino acid, nucleotide and other nutrients dissolved into solution; and further centrifugation, concentration and disinfection to obtain zymogen. Owing to the addition of helicase and neutral proteinase as enzyme preparation and the strict control in reaction temperature and pH value, the zymogen prepared is delicious and mellow and may be used as seasoning. In addition, the production process is short and high in yield.

The present invention relates to the use of molecules capable of specifically binding a urokinase plasminogen activator receptor (uPAR) as diagnostic reagents for the detection of metastases in vivo. Such metastases can include, but are not limited to, micrometastases.

The invention relates to the production of a compound fertilizer, in particular to the production of a microorganism organic-inorganic compound fertilizer, belonging to the organic fertilizer production field. The invention consists of pretreated chicken manure, maize straw power, dry coal slurry, microorganism zymogen inoculum, carbamide, lemery, fused calcium magnesium phosphorus potash fertilizer, humic acid and attapulgite soil. The invention has the advantage of having available for raising the content of organic matter in fertilizer through adding proper amount of coal slurry into the materials of the microorganism organic-inorganic compound fertilizer, which can be proved by actual production. Replacing such phosphate fertilizer as superphosphate with fused calcium magnesium phosphorus potash fertilizer in the materials of the microorganism organic-inorganic compound fertilizer does not influence the effect of phosphate fertilizer, can raise fertility and promote crops to grow owing to the added trace elements necessary for crop growth.

The present invention is directed to peptide sequences that were identified from combinatorial libraries and could serve as substrates of plague plasminogen activator (Pla). Another aspect of the present invention is drawn to peptides derived from the substrates for Pla as a result of chemical modifications leading to specific inactivation of the proteolytic activity of Pla. Additionally, the present invention is directed to the use of the substrates identified herein in the detection of bacteria expressing omptin family of proteases which includes Y. pestis. Furthermore, the present invention is also directed to the use of the inhibitors identified herein in the prevention and treatment of infection caused by these bacteria.

The invention is about a method to purify the recombined human urokinase by the chromatography. The invention relates to recovery the gene engineering production from the mammal cell culture using the Streamline-SP, Sephacryl S-200, Sepharose Fast Flow and DEAE-Sepharose Fast Flow method. The recombined human urokinase can meet the SFDA standard and the percent recovery is above 70%, the purity is above 99% by using the method.

The invention discloses a raw material formulation for vanilla-seed fermented milk, which includes raw milk, lactalbumin powder, a milk-fat product, a sweet substance, a thickening agent, zymogen and vallina-seed jam with viscosity not being higher than 10cm / (20 DEG C, 60s); and the invention discloses a semiquantitative analysis method for addition of the jam, which comprises steps of 1) covering vanilla-seed fermented milk uniformly on an entire filter-paper surface; 2) drying filter-paper surface with the vanilla-seed fermented milk for 15 to 40 minutes; and 3) comparing with a series standard sample. The invention also discloses a preparation method of the vanilla-seed fermented milk, which comprises the steps of: 1, mixing the fermented milk and the vanilla-seed jam to be canned; and 2, implementing the analysis according to the semiquantitative analysis method and fetching the vanilla-seed fermented milk reaching the target addition of the jam. The invention also discloses the vanilla-seed fermented milk prepared through the preparation method. The vanilla-seed fermented milk combines the light fermented flavor of the fermented milk and the fragrance of the vanilla and has rich color, the addition of the jam can be semiquantitatively monitored, the quality of different product batches is stable, and the market potential is remarkable.

The invention discloses a cigarette perfume solvent and a preparation method thereof. The preparation method of the cigarette perfume solvent comprises the steps of mixing water-solubility solvent and tobacco fragments or tobacco smalls, heating the mixture for inverse-flow extraction, filtering the extracted liquid, adding organic acid and sugar to the extracted liquid, adding zymogen strain at 35 DEG C, fermenting the reaction product for 5-15 days in a sealing mode under the temperature of 80-85 DEG C, filtering and removing deposition, stirring under normal atmosphere and heating to 80-85 DEG C for distilling, and obtaining the distilment which is the needed cigarette perfume solvent. Compared with edible alcohol and general grain brewing wine, the cigarette perfume solvent has obvious ferment smell characteristic, better mouthfeel and special ferment perfume, enables the perfume of cigarette to be cooperative, sweet-scented, mellow and exquisite, remarkably improves the aftertaste and has remarkable function for slowing down stimulation. The cigarette perfume solvent can dissolve multiple essences and flavors through adjusting the alcohol degree according to requirement, can be used as solvent for essences and flavors, and can be independently used for adding flavors, thereby having favorable application foreground.