380 results about "Ammonium sulfate precipitation" patented technology

The invention discloses a method for high-effectively quickly purifying and preparing fragment antibody from animal cell scale culture. Said method adopts the processes of dilatant column bed adsorption, anion chromatography concentration and hydrophobic chromatography purification, and includes the following steps: using dilatant column bed cation chromatography to adsorb antibody IgG from large-scale cell culture, changing pH value of buffer system to make elution, making collected elution peak be directly andergone the process of anion chromatography, using pepsinum to directly enzyme-out the fully-concentrated and purified antibody IgG to obtain fragment antibody, making hydrophobic chromatography and purification so as to obtain final fragment antibody whose purity is up to above 97%, and its whole purification process can be completed within 12 hr. Said invented method can be substituted for traditional ammonium sulfate precipitation, concentration method, and has the characteristics of high efficiency, rapid speed, high purity and stable operation and result, etc.

The invention discloses a method for separating high purity phycocyanin from spirulina, comprising the following steps: adopting 0.05-0.5M of phosphate buffer to elute a spirulina powder raw material by an elution method, enabling the phycocyanin to be effused from spirulina cells, adding soluble chitosan to phycocyanin crude extract, controlling the mass concentration of chitosan in the phycocyanin solution to be 0.1-1%, and collecting supernate by centrifuging; adding activated carbon to the supernate, centrifuging and collecting supernate, adopting ammonium sulfate precipitation with the mass concentration of 30-60% to precipitate the phycocyanin in the solution, adopting ion-exchange chromatography, and performing gradient elution by phosphate buffer containing sodium chloride to obtain the high purity phycocyanin solution. The invention has the advantages of low cost, short elapsed time, convenient and rapid operation, low energy consumption, and easy massive preparation, is an efficient method for massively preparing phycocyanin with high purity and high activity and is favor of scaled development and utilization of the phycocyanin in our country.

The invention belongs to the technical field of medicament, and particularly relates to preparation and application of phycocyanin extract. The phycocyanin extract is prepared by the following steps of: crushing spirulina cell walls of spirulina powder to obtain a crude extract by a repeated freeze-thawing method at the temperature of between 20 DEG C below zero and 4 DEG C; depositing the crude extract to obtain a crude protein extract by using 50 to 60 mass percent of ammonium sulfate; and then extracting and purifying the crude protein extract by adopting double aqueous phases, and removing salt by using excess infiltration to obtain phycocyanin extract solution. The phycocyanin extract mainly comprises phycocyanin, and the aqueous solution of the phycocyanin extract has the absorbance characteristics that A620 / 280 is slightly equal to 3.0+ / -0.2 and A620 / 650 is slightly equal to 2.2+ / -0.1. The phycocyanin extract is obtained by mainly combining the processes of ammonium sulfate deposition, double aqueous phase extraction, dialysis and salt removal and the like; and the phycocyanin extract administrated to S180 sarcoma mice with different dosages shows good tumor inhibiting effect and has no obvious toxicity on the spleen and the thymus gland.

The invention discloses a submerged fermentation phellinus linteus mycelium glycoprotein, a usage and a separation extraction preparation method thereof, the complex is the complex of heteropolysaccharide and protein, wherein, the content of the heteropolysaccharide is 15 to 20 percent, and the heteropolysaccharide is composed of three monosaccharides of glucose monosaccharide, xylose and mannose; the content of the protein is 80 to 85 percent, and the protein is composed of 18 amino acids of aspartic acid, glutamic acid, arginine and so on; and the weight-average molecular weight is 20 to 40KD. The glycoprotein complex uses bran extract liquid as a main ingredient for preparing a culture medium, the phellinus linteus mycelium is produced by the submerged fermentation of the phellinus linteus bacterial strain liquid, the homogenization, the cold-water extraction, the centrifugalization, the collection of supernatant liquid, the precipitation of ammonium sulfate, the dialysis and the DEAE-Sepharose Fast Flow column chromatography are carried out for system separating and purifying the glycoprotein complex. The anti-bacterial glycoprotein is used for preparing the anti-bacterial dugs which have inhibitory effects on escherichia coil and staphylococcus aureus. At the same time, the glycoprotein complex can be used for the separation and the purification of the glycoprotein from the mycelium obtained by the submerged fermentation of various medical and edible fungi.

The preparation method of high-purity fungous polysaccharide uses powdered carposoma or deep-fermented mycelium as raw material and includes the following steps: water extraction, then using NaOh solution to extract residue, respectively using ammonium sulfate to precipitate water extract and alkaline extract neutralized by hydrochloric acid, centrifuging to remove precipitate, using ultrafiltering membrane separation technique to desalt supernatant and concentrating, freeze-drying concentrate or spray-drying the concentrate to obtain the fungous polysaccharide whose polysaccharide purity is up to above 99%, and said fungous polysaccharide can be made into capsule or tablet. The above-mentioned concentrate can be directly used as raw material for making water preparation or injection preparation.

The invention discloses a method for extracting high-purity beta-glucosan from oat bran. According to the method, oat bran powder is sequentially treated by adopting the following steps: (1) carrying out ethyl alcohol reflux and enzyme deactivation; (2) stirring and extracting in hot water; (3) centrifuging and collecting supernatant; (4) carrying out enzymolysis and isoelectric precipitation; (5) carrying out alcohol precipitation; (6) centrifuging, collecting and precipitating; (7) carrying out ammonium sulfate precipitation; (8) dialyzing and removing impurities; and (9) carrying out freeze drying, so that the oat beta-glucosan is obtained. The high-purity oat beta-glucosan is obtained by adopting a mild water extraction method through extraction and treatment steps such as enzymolysis, isoelectric precipitation, alcohol precipitation, ammonium sulfate precipitation and dialysis. The high-purity oat beta-glucosan has a certain antioxidation effect on oil and can be added to yoghourt so as to improve the texture characteristics and nutritional ingredients of the yoghourt.

The present present invention relates to a method of simultaneously extracting phycoerythrin and agar from seaweeds such as asparagus, etc in seaweeds biological chemical field. The concrete operation process is that cells of the asparagus frond are crushed. A phycobiliprotein crude extract is obtained from water soluble extracts after ammoniym sulfate precipitation. The crude extract is extracted by Phenyl-Sepharose expansion columns and phycoerythrin products are obtained. Q-Sepharose iron exchange columns are used to purify the phycoerythrin products to obtain purified phycoerythrin products. Then, the left asparagus residues after the phycoerythrin extract are used to extract the agar. The present invention can simultaneously obtain the phycoerythrin and the agar from the asparagus. And the purified phycoerythrin purity is larger than 2.954, and the yield is 0.097mg per gram of fresh algae. The yield of the obtained agar is 2.31percent (the algae is wetly weighted), which is equivalent to the yield of an agar direct extraction method. Therefore, the utilization ratio of the asparagus can be greatly improved by adopting the method.

The invention discloses a preparation method of high-purity phycocyanin, and is characterized in that the preparation method comprises the following steps: (1) taking a fresh spirulina powder, fully mixing with a phosphate buffer solution evenly, repeatedly freezing and thawing for 7-10 times to break and remove cell walls, centrifuging to remove spirulina mud, and thus obtaining a supernatant; (2) adopting a two-step precipitation method with a 20%-30% ammonium sulfate and a 40%-60% ammonium sulfate to obtain a phycocyanin crude extract; (3) after dialyzing the crude extract, loading the sample onto a weak anion exchange column DEAE Sepharose FF, carrying out gradient elution after ion exchange, collecting an outflow component with A620 / A280 of more than 3; and (4) dialyzing the collected sample, then loading the sample onto a strong anion exchange column SOURCE30Q, carrying out gradient elution after ion exchange, collecting an outflow component with A620 / A280 of more than 4, again carrying out one-time ammonium sulfate precipitation concentration, and thus obtaining the high-purity phycocyanin having the purity of more than 4.5. The extraction purification method is simplified in process, wide in source of the raw material spirulina, simple in required equipment, and high in purity of the product, and has quite high application value.

The invention belongs to the technical field of food additives, and provides a method for preparing blue-green algae pigment. The method comprises the steps of performing enzymolysis on blue-green algae, and performing high-pressure homogenization. The method for preparing the blue-green algae pigment specifically comprises the following steps of: performing enzymolysis on blue-green algae cells by using an enzyme preparation, crushing the blue-green algae cell walls, circulating blue-green algae cytochylema which is obtained after enzymolysis treatment in a high-pressure homogenization mode, and finally centrifuging, and performing ammonium sulfate precipitation to obtain the blue-green algae pigment. By the method, edible, high-purity, and high-valeur natural cyanine which also keeps the primary nutrition of protein of blue-green algae pigment can be obtained.

The present invention provides a preparation method of Grifola frondosa protein peptide-selenium chelate, comprising: extracting the protein in Grifola frondosa frondosa by alkali-dissolving acid precipitation method or ammonium sulfate precipitation method; using alkaline protease, neutral protease or compound Grifola frondosa protein enzymatic hydrolyzate is prepared by protease after restriction enzymolysis, and the enzyme is inactivated; the selenium in the inorganic substance sodium selenite is used to chelate the grifola frondosa protein peptide-selenium Chelates. The invention can significantly improve the extraction rate of grifola frondosa protein by alkali-dissolving and acid-precipitating method; the degradation degree of protein can be effectively controlled by controlling the enzymatic hydrolysis time; the preparation process of polypeptide-selenium chelate is simple; the mineral element-peptide prepared by the invention Chelates have a unique chelation system and transport mechanism, are easily absorbed, safe and non-toxic, low in price, and can supplement amino acids and mineral ions at the same time, and will definitely become the first choice for mineral element supplements; the present invention is the application of Grifola frondosa Provided a new way of thinking.

The invention discloses lactobacillus casei bacteriocin and use thereof in feed. The lactobacillus casei provided by the invention is separated from Tibet plateau traditional fermented yak yogurt, and the produced bacteriocin is obtained by ammonium sulfate precipitation, Sephadex G-100 gel filtration and reversed-phase high-performance liquid chromatography (RP-HPLC). The bacteriocin has thermal and acid stability, can be degraded by protease and has a wide antibacterial spectrum. When the bacteriocin is added into feed, and the growth of bacteria can be inhibited. The bacteriocin has a bright prospect in use as a feed additive.

The invention discloses a biological fresh-keeping agent for hairtail. The biological fresh-keeping agent contains the following components in parts by weight: 5-8 parts of tea polyphenol, 2-4 parts of allicin, 3-5 parts of chitosan, 3-5 parts of vitamin E, 0.4-0.5 part of vegetable acid, 2-4 parts of bioenzyme, 2-4 parts of an active peptide ferrous chelate and 100-120 parts of distilled water. Active peptide is prepared through fermentation of actinomycetes and is purified through centrifugation and ammonium sulfate precipitation, and then the active peptide ferrous chelate is prepared. The biological fresh-keeping agent has the beneficial effects that the biological fresh-keeping agent has excellent effects of killing bacteria, resisting oxidation, inhibiting enzymatic activity and the like, the fresh-keeping limit of the hairtail can be prolonged from several days to 40-60 days, so that the fresh-keeping agent is very good; and by utilizing the active peptide ferrous chelate capable of scavenging free radicals, the fresh-keeping agent has extremely strong oxidation resistance, so that the biological fresh-keeping agent is safe, non-toxic and efficient, the original quality of the hairtail is well maintained, and the shelf life of the hairtail can be remarkably prolonged.

The invention is an protein exciton, able to induce the disease resistance of plant system, separated from phytophthora and the preparing method. It uses the deposition of ammonium sulfate to obtain a coarse protein exciton, and then makes purification by ion-exchange chromatography and hydrophobe-interaction churomatography to obtain a 90kDa protein excition. The excition can be applied to induce system broad-spectrum disease resistance of tobacco and little Chinese cabbage. The use concentration is 0.1-10nM. The effect of prevention and treatment is above 80%.

An oral spray for preventing and treating the decayed tooth is prepared from vitelline antibody IgY, alcohol, wetting agent, surfactant, essence, antiseptic, pigment and deionized water through extracting IgY by water diluting method, PEG deposition method, membrane filtering method, or ammonium sulfate depositing method, and preparing oral spray.

A process for preparing the acetylcholinesterase used to detect the content of residual agricultural chemical in fruit and vegetable includes such steps as extracting the coarse liquid extract from animal blood, purifying by ammonium sulfate deposition method, removing ammonium sulfate by dialysis method, concentrating and drying. Its advantages are widely available raw material, low cost and high purity and sensitivity of product.

This invention discloses a method to purify rhIL-12, belonging to the protein purification technology, which technical points are that the cell culture supernatant of rhIL-12 is conducted filtering, cation or anion exchange chromatography, precipitation of ammonium sulfate, anion or cation exchange chromatography and molecular sieve chromatography, and during the precipitation of ammonium sulfate, pH is far away from the isoelectric point of rhIL-12. This method uses ammonium sulfate precipitation to remove most of hybridproteins, making the future separation simple and effective.

the invention discloses a lactic acid bacteria bile salt hydrolase and manufacturing method and specific manufacturing strain, which comprises the following steps: 1) fermenting Lactobacillus casei (KTx CGMCCNo..1739); gathering bacteria; 2) suspending bacterial through phosphoric buffer; grinding in the icy bath through supersonic wave; collecting the rough enzyme liquid; sedimenting protein through ammonia sulfate; centrifuging to obtain the product.

The invention relates to a separation and purification method for a recombinant hepatitis B core antigen. The method includes the steps of thermal denaturation and clarification; ammonium sulfate precipitation; ultrafiltration and concentration, washing filtering and liquid exchange; depolymerization; first-step molecular sieve chromatography; ultrafiltration and concentration, washing filtering and liquid exchange; repolymerization; second-step chromatography. By means of the method, the high-purity, low-host-residue and high-stability recombinant hepatitis B core antigen with a uniform granular structure can be obtained, that is to say, the method has the advantages that the antigen purity is high, the residue of host protein and host nucleic acid is low, antigen particles are uniform, and industrial enlargement is easy.

The present invention relates to a method for preparing high-purity laver phycoerythrin by one-step chromatography, belonging to the preparation technology of the functional ingredients of halobios. Laver is swelled in PBS (1mM, pH6.8) buffer solution with a volume fifty times larger than the volume of the laver and smashed, then 800 watts of ultrasonic waves carry out cell disruption for 800 seconds to produce crude extract, which is precipitated by ammonium sulphate with 45 percent of saturation, microfiltrated via 0.1Mu m of film, ultrafiltered via a 10kDa film, goes through reversed-phase precipitation by ammonium sulphate with 20 percent of saturation and is precipitated and crystallized by ammonium sulphate with 65 percent of saturation under the temperature of 4 DEG C, and one week later, the crude extract passes through a DEAE cellulose-52 anion-exchange chromatography column, is gradiently eluted by PBS (1mM, pH6.8) buffer solutions with different concentrations of NaC1, separated and purified to produce the laver phycoerythrin. The purity (A562nm / A280nm) reaches 5.24, which accords with the reagent-grade requirement.

The invention discloses acidproof medium-temperature alpha-amylase and a preparation method thereof. The preparation method comprises the following steps of: A1, amplifying a precursor alpha-amylase gene; A2, performing site-directed mutagenesis on the precursor alpha-amylase; A3, constructing a mutant alpha-amylase expression vector; and A4, transforming bacillus subtilis by using the expression vector. A recombinant strain obtained by the method can be used for industrialized production of an acidproof alpha-amylase mutant. The required acidproof alpha-amylase mutant can be obtained by the following steps of: culturing recombinant cells in a liquid culture medium with selection pressure without induced expression, precipitating supernatant by using ammonium sulfate, dialyzing a precipitate, desalting, adding an allyl dextran S300 gel column, and eluting by using eluent.

The present invention relates to anticancer blood clam peptide with high anticancer activity and its preparation process and belongs to the field of medicine technology. The anticancer blood clam peptide is prepared with blood clam as one kind of marine life and through serial technological process of extraction, precipitation, chromatographic separation, desalting, etc. The preparation process includes biological extract, staged ammonium sulfate precipitation, Superdex75 gel filtering and chromatographic separation, and Sephade G-25 column desalting. The screened out peptide molecules below 3000 Dal have optimal pharmaceutical effect and economic utility. The injection with the matter of the present invention has broad spectrum anticancer curative effect and low toxic side effect.

The invention relates to a preparation method of a zearalenone biodegradation agent. The preparation method comprises the following steps: culturing through beer yeast, namely, firstly culturing in a PDA (potato dextrose agar) solid culture medium for 48hours, then taking single colony, inoculating the single colony into a PDB (potato dextrose broth) liquid culture medium for performing shaking culture for 3-5 days, adding 1-10% by weight of yeast fermentation broth by using wine lees protein feed as a carrier to prepare a crude enzyme preparation of the zearalenone biodegradation agent; extracting a zearalenone degradation enzyme of a beer yeast fermentation product, culturing through the beer yeast at the temperature of 37 DEG C for 3-5 days, taking the fermentation broth, performing freezing centrifugation under the condition of 8000r / min at the temperature of 4 DEG C for 20minutes, or separating supernatant liquid from bacterial cells by adopting a suction filtration method; and taking the supernatant liquid which is separated out, thereby obtaining an extracellular crude extraction solution, precipitating with 30-90% of ammonium sulfate, and performing the freezing centrifugation under the condition of 8000r / min at the temperature of 4 DEG C for 20minutes to obtain the zearalenone degradation enzyme which is finely extracted. The zearalenone degradation agent disclosed by the invention has the advantages that yeast can not only degrade toxins, but also adsorb the toxins.

An object of the present invention is to search and identify novel antifungal proteins capable of inhibiting the growth of plant pathogenic microorganisms including Magnaporthe grisea and Rhizoctonia solani causing two major rice diseases at relatively low concentrations, and further to clone a gene for said protein. The present invention provides an antifungal protein which can be obtained from fraction(s) precipitated by ammonium sulfate precipitation using an aqueous extract from Pleurotus cornucopiae, wherein said protein has an antifungal activity against at least rice blast, and exhibits existence of a component having a molecular weight of about 15 kDa as determined by SDS-PAGE method; a gene encoding said protein and uses thereof.

The invention discloses a method for extracting a nano-pearl organic substrate and grading by molecular weight. Nano-pearl powder is added with water for twice with stirring to ensure that the nono-pearl powder is fully extracted, a suspension is obtained through a centrifugation with high-speed rotation, impurities are filtered, and then a first pearl organic substrate extract with a molecular weight more than 5kDa and a second pearl organic substrate extract with a molecular weight less than 5kDa are separated through a centrifugal concentration separation by a centrifugal concentration filter-membrane device; further an ammonium sulfate precipitation method is used to separate pearl protein and protein which is failed to be precipitated, and Sephadex G25 gel is used for chromatography according to the molecular weights of the proteins to obtain three grades: grade A, grade B, and grade C. The method ensures that industries such as cosmetics, food and so on, can obtain pearl protein or pearl peptide with purified effective compositions for best utilization.

The invention provides a bran coat antitumor active protein, a preparation method of the protein, and applications of the protein in preparing antitumor drugs and health foods. The preparation method of the protein comprises the steps of pulverizing all natural bran coat, adding protein extracting solution, extracting at low temperature after one night, then adding precooled acetone to precipitate protein at low temperature so as to obtain bran coat protein, adding ammonium sulfate powders with different saturation levels into coarse protein extracting solution for fractional precipitation of the protein, enabling the protein precipitated by the ammonium sulfate to permeate through a Q anion exchange chromatographic column, collecting penetrating fluid, enabling the penetrating fluid to permeate through the Q anion exchange chromatographic column again, performing elution with Tris-NaCl eluant with the concentration of 0.1mol / L, and collecting eluting peaks to obtain the bran coat antitumor active protein (FMB). The protein can obviously restrain the proliferation and the metastasis of colorectal cancer cells and can be applied to preparation of the antitumor drugs and the health foods.

The invention belongs to the technical field of separation and purification of lectin, and provides a method for separating and purifying lectin from pleuratus ferulae. Immersion is performed with normal saline overnight, a supernatant is collected centrifugally, 40%-70% of ammonium sulfate precipitation is adopted, the precipitation is removed centrifugally, ammonium sulfate is removed by dialysis, freeze drying is performed, the dissolved upper-layer clear liquid passes through a DEAE-52 ion-exchange chromatographic column, an elution peak is collected, freeze drying is performed, the dissolved upper-layer clear liquid is loaded through SaphedexG-100, and the elution peak is collected, and the procoagulant activity is detected. A technological process for separating and purifying lectin from pleuratus ferulae is simpler and more convenient, the separation and purification efficiency of the lectin is higher, the production and usage costs are lower, the practicability is high, and higher popularization and application values are provided.

The invention relates to a method for extracting natural antitumor active protein and polypeptide from chickpea bean. In the method, protein and polypeptide capable of inhibiting the activity of colonic cancer cells can be obtained by the steps of extracting phosphate buffer solution, precipitating ammonium sulfate, desalting, centrifuging and the like. The method is a basic research on bioactive substances in chickpea which is a Uighur medicinal food dual-purpose plant, and provides a basis for developing related products. The method has the advantages that the extraction process is simple, and the protein and the polypeptide can not be deactivated easily. The prepared polypeptide has the advantages of high biological activity, no toxicity and good thermal stability. The method can be easy for production in a large scale, reduces the cost of the extraction process, and is an ideal method for extracting and preparing bioactive protein and polypeptide. The protein and the polypeptide prepared by the method can be used for preparing functional foods or drug additives.

A chitosan microsphere or microcapsule is prepared through proportionally mixing oil phase with surfactant, stirring, proportionally adding chitosan solution (water phase), stirring, proportionally adding ammonium sulfate solution, stirring, centrifugal separation of deposit, washing and drying.

The invention discloses an antimicrobial protein product of bacillus amyloliquefaciens NCPSJ7 and a preparation method of the antimicrobial protein product. The preparation method comprises the steps of centrifuging a bacillus amyloliquefaciens NCPSJ7 fermentation liquid, and collecting a supernatant liquid; carrying out ammonium sulfate precipitation, ultrafiltration, desalination and concentration on the fermented supernatant liquid; then, carrying out anion exchange column chromatography, ultrafiltration, desalination and concentration; next, carrying out spray drying to obtain the antimicrobial protein product. The antimicrobial protein product prepared by using the method mainly contains the two antimicrobial proteins: 24kDa and 36kDa, and the total content of the two antimicrobial proteins accounts for more than 60% of the mass of spray dried powder. The antimicrobial protein product is favorable in reconstitution properties, strong in water solubility and favorable in antimicrobial activity and has a certain industrial application prospect so as to be applied to the fields such as fruit and vegetable preservation and fresh keeping, food additives, fish medicines, veterinary medicines and the like.

The invention relates to a pseudomonas syringae, a screening method thereof and application to kill nematode. The classification name of the pseudomonas syringae is pseudomonas syringae MB03, colonies has protuberances and show milk white, the thalline is in the shape of a rod through microscopic examination and belongs to a gram-negative bacterium. The screening method comprises: acquiring frozen plant tissue, chopping and culturing in a medium, and finally picking growing single colony, and performing separation purification through streaking. The application to kill nematode means that the thalline, a fermentation supernatant, a cell disruption solution, or extracellular crude protein and / or intracellular crude protein of pseudomonas syringae MB03 subjected to ammonium sulfate precipitation is used to kill caenorhabditis elegans and meloidogyne incognita. The advantages comprise that pseudomonas syringae is low in production cost and free of pollution to environment, and has relatively killing effect on caenorhabditis elegans and meloidogyne incognita. The factor, killing nematode, in pseudomonas syringae is protein, so that pseudomonas syringae is sutiable for molecular gene operation, and has great value on microbe control resource and mechanism research.