712 results about "Complex protein" patented technology

The instant invention provides an economical flow-through method for determining amount of target proteins in a sample. An antibody preparation (whether polyclonal or monoclonal, or any equivalent specific binding agent) is used to capture and thus enrich a specific monitor peptide (a specific peptide fragment of a protein to be quantitated in a proteolytic digest of a complex protein sample) and an internal standard peptide (the same chemical structure but including stable isotope labels). Upon elution into a suitable mass spectrometer, the natural (sample derived) and internal standard (isotope labeled) peptides are quantitated, and their measured abundance ratio used to calculate the abundance of the monitor peptide, and its parent protein, in the initial sample

The instant invention provides an economical flow-through method for determining amount of target proteins in a sample. An antibody preparation (whether polyclonal or monoclonal, or any equivalent specific binding agent) is used to capture and thus enrich a specific monitor peptide (a specific peptide fragment of a protein to be quantitated in a proteolytic digest of a complex protein sample) and an internal standard peptide (the same chemical structure but including stable isotope labels). Upon elution into a suitable mass spectrometer, the natural (sample derived) and internal standard (isotope labeled) peptides are quantitated, and their measured abundance ratio used to calculate the abundance of the monitor peptide, and its parent protein, in the initial sample.

The invention discloses a special method to extract collagen from fish peel and bone with complex prolease, which comprises the following steps: choosing salmon, AnKang fish peel and processing waste of fish product as raw material; unfreezing; washing; scaling off; degreasing; grinding; enzymolyzing; separating; de-oiling; ultra filtering; condensing; spray-drying; getting the product. This invention possesses simple craft and short production circle.

The invention discloses small molecule polypeptide Ca-chelate of fishbone, which belongs to the field of functional food or additive. The method for preparing the small molecule polypeptide Ca-chelate of the fishbone comprises the following steps: boiling the fishbone at high temperature and high pressure; pulping to obtain the fishbone paste; utilizing the compound protease to execute enzymolysis; centrifuging to obtain the liquid supernatant and the sediment; filtering the liquid supernatant to obtain the small molecule polypeptide liquid of the fishbone; executing the composite acid acidification through citric acid and lactic acid for the sediment so as to obtain the fishbone calcium liquid; mixing the small molecule polypeptide liquid of the fishbone with the fishbone calcium liquid; executing the chelation at 40-50 degrees centigrade while the pH value is 7.5-8.5; and drying to obtain the small molecule polypeptide Ca-chelate of the fishbone. The invention has high Ca-chelate rate which reaches over 90%, and has high biological absorption rate of the calcium which reaches over 70%. The invention has no side effects, has good flavor, can be directly absorbed from intestinal mucosa after being eaten, solves the problems that the solubility of the traditional calcium source is bad, the absorption rate is low and the side effect is large, and solves the problem of producing calculi. In addition, the invention has a simple preparation method and low cost.

Provided are previously uncharacterised markers of cancers, for example colorectal cancers, and uses of these as diagnostic and prognostic markers of cancers, and in particular colorectal cancers. The markers are SEQ ID NO: 1—hnRNP-K; SEQ ID NO:2—HMG-1; SEQ ID NO:3—proteasome subunit alpha type 1; SEQ ID NO:4—bifunctional purine biosynthesis protein; SEQ ID NO:5—ST11; SEQ ID NO:6—annex in IV; SEQ ID NO:7—60 kDa heat shock protein; SEQ ID NO:8—T complex protein 1 beta subunit; SEQ ID NO:9—T complex protein 1 epsilon subunit; SEQ ID NO: 10—mortalin; and SEQ ID NO: 11—TER-ATPase. The invention further provides related methods and materials for the use of the markers in therapeutic intervention in colorectal and other cancers e.g. to specifically target neoplastic cells without causing significant toxicity in healthy tissues, and to provide methods for the evaluation of the ability of candidate therapeutic compounds to modulate the biological activity of cancerous cells from the colon, rectum and other tissues.

The invention discloses a sea cucumber polypeptide functional food and a preparation method thereof which not only remarkably improves the content of polypeptide, but also has no chemical residues; the contents of salt and arsenic are low; the food is healthier and safer to eat; simultaneously the food is simply operated, easily controlled, is effective and saves energies. The key technical scheme includes: selecting a compound protease of Protamex and needing not to adjust the pH value of materials; carrying out processes of desalting and arsenic removing on an enzymolysis liquid; more than 80 percent of the molecular weight of the product is between 100 to 6000Dalt; wherein, the small polypeptide of 100 to 2100Dalt is more than 70 percent; the product components and the content weight percentages are as follows: 50 to 60 percent of polypeptide, 5 to 10 percent of free amino acids, 2.5 to 7.5 percent of mucoitin as well as containing the inherent nutrition components of a plurality of minerals and vitamins of the sea cucumber. The product has the effects of resisting knub, reducing blood pressure, preventing cardio-cerebrovascular diseases, resisting fatigue, delaying senescence, improving the immunity. The food can be used as a healthy food to eat and can also be used as a food and a medicine additive.

The invention provides a theoretical design and technical design of cell-free protein synthesis for DNA replication, transcription and translation coupling, a preparation, a kit and a preparation method. Specifically, with the application of the in-vitro cell-free synthesis system provided by the invention, complex protein can be synthesized, and moreover, DNA and mRNA can be synthesized; and effective, high-throughput and quite convenient protein synthesis can be completed with the use of a DNA template by a minute quantity (nanogram-microgram).

The invention discloses a method and process for extracting animal micro-molecule polypeptides and amino acids by utilizing a complex enzyme. The method comprises the following steps: (1) crushing animal tissues so that the animal tissues can pass through a sieve of 80-200 meshes, and adding water to prepare a suspension with the concentration of 8-30 percent; (2) raising the temperature to be 35-40 DEG C, regulating the pH value to be 8.0-9.0 by using sodium carbonate, adding a complex enzyme I accounting for 0.5-2.0 percent of the animal tissues, performing ultrasonic enzymolysis at 14-16 kHz for 1-2 hours so that the proteins between cells are subjected to full enzymolysis, regulating the pH value to be 6.5-7.5, adding a complex enzyme II accounting for 0.5-3.0 percent of the animal tissues, fully and uniformly stirring, controlling the temperature of the water bath to be 37-45 DEG C, and performing ultrasonic enzymolysis at 15-17 kHz for 1-6 hours; (3) raising the temperature of the enzymolysis solution to be 90-100 DEG C to inactivate the enzyme for 20-30 minutes after the enzymolysis is finished; (4) filtering the solution subjected to enzymolysis through a 0.22mu m microfiltration membrane; and (5) concentrating the filtrate to the relative density of 1:1.0-1:1.2 to prepare the concentrated solution, wherein the complex enzyme II consists of Alcalase, Flavourzyme, Protamex, neutral protease and papain. According to the method, the enzymolysis efficiency can be improved, and the production cost can be reduced.

A system and method for performing rapid, automated and high peak capacity separations of complex protein mixtures through the combination of fluidic and electrical elements on an integrated circuit, utilizing planar thin-film micromachining for both fluidic and electrical components.

The invention provides a protein synthesis system for in-vitro protein synthesis, a kit and a preparation method for a protein through in-vitro synthesis. Specifically, the in-vitro cell-free expression system provided by the invention is capable of extremely efficiently synthesizing proteins and synthesizing complex proteins. Moreover, due to the in-vitro cell-free expression system disclosed bythe invention, the relative light unit value of the activity of the synthesized luciferase is higher than that of the conventional commercial system (such as a rabbit reticulocyte in-vitro expressionsystem) by at least one order of magnitude (more than or equal to 10 times or higher).

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 discloses fishbone bioactive polypeptide calcium powder and a preparation method, belonging to the field of functional food or food additives. The preparation method comprises the following steps: freezing fishbone, coarse grinding, fine grinding, adding water, stirring, adding a compound protease to hydrolyze, centrifugating, and respectively collecting the supernatant and the precipitate; ultrafiltering the supernatant by a filter membrane, concentrating the filtrate, and freeze-drying to obtain the fishbone protein polypeptide powder; washing the precipitate, drying and pulverizing; activating by using citric acid, and drying to obtain the soluble calcium; and mixing the fishbone protein polypeptide powder and the soluble calcium to obtain the fishbone bioactive polypeptide calcium powder. The fishbone bioactive polypeptide calcium powder enables the biological absorptivity of calcium to reach above 65%, and meanwhile, maintains the functional characteristics of polypeptide. Hydrolysis is carried out by using the food-grade protease, and thus, the fishbone bioactive polypeptide calcium powder has the advantages of high safety and no side effect. The method of the invention has the advantages of simple and feasible realization, and low cost. The fishbone bioactive polypeptide calcium powder can be used as a food additive or directly eaten as nutritional food.

The invention provides flavor soup-stock seasoning which comprises the following raw materials: pork, pig bone clear soup, lard, chicken, chicken bone clear soup, chicken oil, water, L-cysteine hydrochloride monohydrate, DL-methionine, VC, glucose, a yeast extract, table salt, aginomoto, soybean peptides, gingers, onions, modified starch, edible essence, compound protease, flavourzyme, light soy sauce, hydrolyzed vegetable protein powder, CMC, white sugar, I+G and potassium sorbate. The meat raw materials and the soup are subjected to Maillard reaction step by step to obtain the flavor soup-stock seasoning. According to the soup-stock seasoning, the optimum formula of the flavor soup-stock seasoning is determined; the soup-stock flavor is enhanced through pork enzymolysis, chicken enzymolysis and Maillard thermal reaction; through combination of the soybean peptides, the pig bone clear soup and the chicken bone clear soup and with the aid of spice and the edible essence, the prepared seasoning looks like milk-white thick paste, has a strong bone soup flavor and tastes delicious and mellow; a dish can be delicious with a little seasoning; and the seasoning is convenient to use.

The invention provides egg white powder with high foamability and a preparation method thereof and relates to reconstruction of whey protein structure and functional properties, belonging to the technical field of biological processing of foodstuffs. According to the invention, on the basis of preliminary work, enzymatic hydrolysis of lipase in advance and cooperative enzymatic hydrolysis of composite protease are utilized for treatment of egg white; total usage amount of lipase and composite protease is less than usage amount of individually used lipase or protease, and however, foamability and foam stability of egg white powder obtained by combined utilization of lipase and composite protease are higher than those of egg white powder obtained by individual utilization of lipase or composite protease; the egg white powder obtained in the invention can meet demands for high-grade products on the market, and the advantages of a simple process and high cost performance are achieved in the invention. According to the invention, the ratio of active usage amount of Aspergillus oryzae protease, papain and trypsin is determined to be 1:1:1; the usage amount and other technological parameters cooperatively allow egg white powder with high foamability to be obtained; egg white powder with high foamability provided in the invention enables the additional output value of eggs to be increased, lays a technical foundation for development and industrial production of special-purpose egg white powder products and increases economic benefits for enterprises.

The invention provides yak bone protein peptide with DPP-IV inhibitory activity and a preparation method thereof. The yak bone protein peptide provided by the invention is optimally prepared from the enzymolysis products by conducting multiple-step enzymolysis by multilevel compound protease on the yak bone protein prepared from yak bone, and the compound protease contains alkali protease, neutral proteinase, trypsin and flavourzyme. The yak bone protein peptide provided by the invention has excellent DPP-IV inhibition and antioxidation functions, and can be used as a DPP-IV inhibitor and antioxidation raw material to be applied to special foods and nutritional foods.

The invention relates production method of soybean peptide, using the degreasing soya bean waste / soybean powder as raw material, comprising the following steps: 1 solid fermenting: mixing the water with raw material in proportion by weight, inoculating zymocyte; 2 liquid enzymolysis: mixing the solid culture with water in proportion by weight, adding composite proteinase, carrying out enzymolysis; 3 extinguishing bacterium: catabolite extinguishing bacterium; 4 filter liquor: separating soybean peptide solution from enzymolysis liquid; 5 decolourization; 6 refining; 7 sterilization, and getting the product. The advantages are following: absorbing the advantages of liquid invoice method and liquid enzymolysis method, overcoming the defects of them, reducing the cost, decreasing the content of added enzyme, getting the high hydrolysis rate, and extending the soybean peptide application.

The invention discloses a black coarse grain composite drink with black rice, black sesame, black beans and black peanuts as the main raw materials and a production method thereof. The four kinds of black coarse grains are compounded. During preparation, the method of roasting is adopted for increasing the aroma and removing the beany flavor and the colloid compounding technology, colloid milling, high-pressure homogenization and other ultrafine comminution technology are adopted to fully break and levigate the material granules, so that the functional factors contained in the granules completely enter into the drink. The prepared black coarse grain composite protein drink further has nutritional value because of being rich in melanin substances, proteins, linoleic acid and trace elements. Meanwhile, the drink has natural color and exquisite mouthfeel, is fragrant, sweet, smooth, stable and free of precipitates and lamination, has long shelf life, can be suitable for various consumer population in all seasons and can be used as the thirst-quenching drink for replenishing moisture and the nutritional drink at breakfast.

The invention discloses a method for preparing lactalbumin peptides, comprising the following steps: (1) adding compound protease into lactalbumin solution to carry out enzymatic hydrolysis; (2) inactivating the compound protease; (3) depositing the enzyme solution and selecting supernatant fluid to carry out centrifugation; (4) carrying out decoloring processing on the centrifugalized supernatant fluid and then carrying out desalting processing; (5) carrying out nanofiltration on the desalted enzyme solution; (6)concentrating; (7) embedding the concentrated product into wall material; (8) spray drying, then the product is obtained. By applying compound protease in the invention, the lactalbumin is zymohydrolyzed with effectively improving enzyme hydrolysis degree, prominently improving usage factor of the lactalbumin, high product yield, good distribution range of molecular weight of the lactalbumin peptides, high peptide content and good product quality. By detecting, the content (counted by dry basis) of the lactalbumin peptides prepared by the method in the invention is up to 75.15 percent, the molecular weight distribution range of the peptide smaller than 2258 dalton can reach 98.83 percent.

The invention discloses a preparation process of high molecular weight silk fibroin freeze-dried powder. The silk fibroin freeze-dried powder can be rapidly and completely dissolved in water, and is applicable to various fields of biomedicine clinic. The preparation process, from controlling an interaction between molecules of the silk fibroin, changes pH value, ionic strength and concentration of a silk fibroin solution, and implements high pressure and high temperature sterilization to induce transformation of the silk fibroin from complex protein structures such as single molecule, polymer and nanoparticle to stable nanoparticle which is about 50-300nm, so as to inhibit interaction as well as coagulation and cross-linking between the molecules of the silk fibroin; therefore, the freeze-dried powder can be prepared through a basic freeze-drying procedure. The silk fibroin, which has been sterilized at high temperature before being freeze-dried, can be directly applied to clinic treatment without further disinfection treatment, thus greatly simplifying operation steps and saving cost.

The invention discloses a unique extracting method of maize albumen powder from maize, which comprises the following steps: using composite proteinase to enzymolyze zein; separating; hyperfiltering; condensing; spraying; drying; obtaining the white powder shaped product. The invention simplifies the technique and shortens the manufacturing period with good taste and small molecular weight of hybrid peptide, which can be clinical nutrition, hygienic food, sports food and cosmetics.

The invention belongs to the field of food processing, and relates to a method for hydrolyzing egg-white proteins by various proteases. The method comprises the following steps of: first, heating obtained egg white to degenerate; and then, hydrolyzing egg-white proteins simultaneously or in stages by using two or more of compound protease, neutral protease, alkaline protease and flavourzyme. According to the invention, different enzymes are combined in use and compensate one another in enzyme acting sites, so that the degree of hydrolysis is improved, and the yield of egg-white protein peptide is increased.

The invention discloses a processing method of sea cucumber polypeptide, which is characterized by comprising the following steps: A. pretreatment: cleaning fresh sea cucumber, pulping and homogenizing the sea cucumber to obtain sea cucumber pulp; B. enzymolysis: adding papain and compound protease without adjusting the pH value to form a multienzyme system with the autolytic enzyme contained in the fresh sea cucumber so as to perform enzymolysis, and adding flavourzyme to continue enzymolysis until enzymolysis is finished; C. classification: commonly filtering and micro-filtering, and classifying the sea cucumber polypeptide enzymatic hydrolysate through ultrafiltration classification and gradient dilution; and D. package: drying and packaging the sea cucumber polypeptide in a moisture proof manner. According to the invention, the papain and the compound protease have a synergistic effect in the enzymolysis system to reinforce the enzymolysis effect, the whole enzymolysis process is convenient to operate and is beneficial for the mass scale production demand, and meanwhile, the separation efficiency is improved, classified sea cucumber polypeptide with high purity and quality is obtained, and no organic solvent is added in the sea cucumber polypeptide, so that the safety is high, and a guarantee is provided for developing the sea cucumber polypeptide.

The invention discloses a preparation method of egg albumen powder with a great instant property and no bitter or fishy smells, which solves the problems of bad instant property, bitter smell or fishy smell, and bad organoleptic quality of egg albumen powder in the market at present. According to the preparation method, enzymolysis treatment is performed on egg albumen protein by a compound protease (including Alcalase, neutral protease and flavourzyme) and cooperates with a two-stage enzymolysis method, the use amount of the compound protease is less than the use amount of any one protease in the compound protease during independent use, the physicochemical indexes of instant property, taste and the like of the egg albumen powder obtained via the enzymolysis are greatly improved, and unexpected technical effects are obtained; the proportioning ratio of Alcalase to neutral protease to flavourzyme in the compound protease is determined to be 1: 2: 1, and via the proportioning, the instant property and dissolution stability of the egg albumen powder are remarkably improved; and high-speed short-time superfine treatment is combined, so that the effects are better.

The invention relates to a technology for synchronously preparing walnut oil and walnut peptide by means of microwave-assisted enzymolysis, belonging to the technical field of the food or the health food. The technology comprises the following steps of: removing the coat and the core of the walnut, grinding the walnut into walnut slurry, adding the water to prepare the slurry, hydrolyzing by adding the protease or the composite protease, treating by microwave, and centrifugally separating the oil phase, the peptide water phase and the residual solid phase, wherein the obtained oil phase is the cleaning and bright walnut oil which can meet the requirement of the green food; and further physically refining the oil phase at low temperature to be taken as the raw material oil of the high-class edible oil, the health food oil, the blend oil and the like. The obtained walnut peptide water solution can be directly used for preparing the degreased walnut peptide beverage by means of the enzyme inactivating, or can be used for preparing the active polypeptide product such as the walnut antioxidant peptide by means of the ultra-filtrating for the health food, the food additive, the nutrition reinforcing agent, the cosmetic, the daily chemicals and the like, or can be used for preparing the walnut peptide powder by means of the mist spray drying at low temperature. The technology improves the extraction efficiency of the walnut oil and the walnut peptide, and is simple in equipment, short in period, safe, low in energy consumption, high in use ratio and free of three-waste pollution.

Methods and compositions are described for analyzing complex protein mixtures, such as proteomes, using activity-based probes. In particular, probes that specifically react with and bind to the active form of one or more target proteins are employed. Labeled peptides obtained from the labeled active target proteins can be used in screening and identification procedures, and can be related to the identity, presence, amount, or activity of active members of the desired target protein class. The methods and compositions described herein can be used, for example, to provide diagnostic information concerning pathogenic states, in identifying proteins that may act as therapeutic targets, and in drug discovery.

The invention discloses a method for preparing fishskin fish-scale collagen protein. The method includes the steps of processing fishskin, processing fish scale, preparing collagen protein and the like. The fish scale processing includes the steps of preprocessing, decalcifying, neutralizing and removing immunities, washing, injecting sol, solid and liquid separation, filtering and obtaining a water-soluble collagen solution generated after fish scale is removed. The collagen protein preparing includes the steps of hydrolysis, enzymolysis, collagen protein filtering and separation, enzyme deactivation, decolorization, fishy smell elimination, drying and the like. The ash content in obtained collagen peptide is obviously lower than that of other enzymolysis methods, and no fishy smell exists. According to the method, hydrolysis is conducted on collagen through compound protease and bromelain, the yield of micromolecule collagen peptide is high, and the enzymic preparation is low in cost and free of bitter taste. By means of the method, the production cost of the collagen peptide can be lowered remarkably, the obtained collagen peptide is white and tasteless powder, the ash content is smaller than 0.5%, and peptide fragment with the molecular weight smaller than 3000 Da accounts for over 96% of total nitrogen.

The invention relates to a preparation method of a yak bone protein peptide capable of relieving fatigue and resisting oxidation and the protein peptide prepared therefrom. The preparation method is characterized in that yak bone protein liquid is pretreated by high-frequency ultrasonic wave, ultra-high pressure micro-jet and the like, and then subjected to enzymolysis step by step using various complex proteases without adding any acid or alkali, and finally separated by membrane separation, gel separation and reverse phase HPLC separation techniques. The preparation method of the yak bone protein peptide is simple. No acid or alkali is added for pH adjustment in the whole processing process, so that a product maintains good functional characteristics, and industrial production is easilyachieved.

Provided are protein microarrays, their manufacture, use, and application. Protein microarrays in accordance with the present invention are useful in a variety preoteomic analyses. Various protein arrays in accordance with the present invention may immobilize large arrays of proteins that may be useful for studying protein-protein interactions to improve understanding of disease processes, facilitating drug discovery, or for identifying potential antigens for vaccine development. The protein array elements of the invention are native or modified proteins (e.g., antibodies or fusion proteins). The protein array elements may be attached directly to a organic functionalized mirrored substrate by a binding reaction between functional groups on the substrate (e.g., amine) and protein (e.g., activated carboxylic acid). Techniques for chemical blocking of the arrays are also provided. The invention contemplates spotting of array elements onto solid planar substrates, labeling of complex protein mixtures, and the analysis of protein binding to the array. The invention also enables the enrichment or purification, and subsequent sequencing or structural analysis of proteins that are identified as differential by the array screen. Kits including protein-binding microarrays for proteomic analysis in accordance with the present invention are also provided.

A combination of “bottom up” and “top down” MS analysis of posttranslational modifications in complex proteins is described. The method comprises digestion of the protein with an enzyme that forms larger peptide fragments than trypsin (>3000 D), performing HPLC with the fragments and applying a new data acquisition strategy using on-line coupling with e.g. LTQ-FTMS, a hybrid mass spectrometer that couples a linear ion trap with a Fourier transform ion cyclotron resonance (FTICR) cell. The method is applied to analysis of posttranslational modifications of protein isoforms.