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1745 results about "Enzyme method" patented technology

Process of knocking out Wnt3a gene and verification method thereof

The invention discloses a process of knocking out Wnt3a gene and a verification method thereof. The knockout and verification of Wnt3a gene are finished through the following steps: establishment of a Cas9 lentiviral vector for Wnt3a gene, culture and passage of HepG2 cell, lentivirus infection and screening of target cell, verification of gene knockout efficiency through a mispairing enzyme method, cell protein analysis and cell proliferation detection by a CCK-8 method. The invention has the following advantages: the Wnt3a gene is knocked out by establishing a Cas9 double-vector lentivirus system for the first time; Crispr/Cas9 is a technology for accurately editing specific site of the genome of any species, and the cell-level single gene or multiple genes can be knocked out by the technology; compared with other gene editing technologies, the method has the advantages that the targeting accuracy is higher; only if the RNA target sequence is completely matched with the genome sequence, can the Cas9 cut the DNA and realize simultaneous knockout of multiple sites of the target gene; and moreover, the experimental period of vector establishment is short, the time and the cost are remarkably saved, and species limit is avoided.
Owner:AFFILIATED HOSPITAL OF NANTONG UNIV

Method for extracting polysaccharide from cordyceps militaris medium

The invention discloses a method for extracting polysaccharide from a cordyceps militaris medium. The cordyceps militaris is crushed and mixed with water, is extracted in an enzyme liquor and an alkali liquor under 30 DEG C to 80 DEG C, and an obtained solution is centrifugalized. An insoluble substance is extracted repeatedly, and supernatants are combined. Amylase enzymolysis is performed on the obtained product under 50 DEG C to 75 DEG C. After centrifugation, a supernatant is condensed to be with a sugar content of 1.5 percent to 3.0 percent. The supernatant is added with ethanol and alcohol precipitation is performed on the obtained product under 0 DEG C to 4 DEG C, a product is obtained by drying the obtained product after centrifugation. After deproteinization and dialysis, crude cordyceps polysaccharide is obtained. A single enzyme method, a double enzyme method, a compound enzyme method and a supersonic wave enzyme method can be adopted, and protease, trypsin, etc. can be adopted. The method optimizes the technology route for extracting the cordyceps polysaccharide, enhances an obtaining rate and an extraction efficiency of the cordyceps polysaccharide. Extraction, separation and purification of the cordyceps polysaccharide are carried out effectively. The method provides a solution approach for the utilization of agricultural wastes, and provides a technology base for further exploitation of the cordyceps polysaccharide.
Owner:DALIAN POLYTECHNIC UNIVERSITY

Technique for extracting grease from oil-tea camellia seed by enzyme method

InactiveCN101235399AReduce emulsificationReduce foaming and other problemsFermentationCamellia oleiferaDry weight
The invention relates to an enzymatical extraction process of oil in tea seed oil kernel, which comprises the following steps: firstly, utilizing dried tea seed oil kernel to be raw material, immersing in water in high temperature after disintegrating, stirring in uniform speed under 70-100 DEG C, secondly, enzyme hydrolyzing, adding acid protease which is 0.02-1% dry weight of disintegration samples or mixed enzyme of acid protease/cellulose in 45-55 DEG C to stir in uniform speed, and enzymatic hydrolysis for 2-4 hours, thirdly, separation emulsion breaking processing, directly separating out primary free boiled oil after enzymolysis liquid is centrifuged, getting secondary free boiled oil through centrifuging after missible oil demulsifies, combining the primary free boiled oil and the secondary free boiled oil, and getting the total boiled oil. The invention has the advantages that first, the invention reduces the problem of emulsification, foaming and the like which are faced when a grinding method is utilized to destroy cell structures, second, the invention prevents various enzymes in cells from degrading oil, which has high oil extraction rate to a further disintegrated function to tea seed oil kernel cell, third, the invention increases the treatment to missible oil, which increases free boiled oil yield through demulsifying, centrifuging, stewing to layer and the other modes.
Owner:江西春源绿色食品有限公司

Preparation method of instant oat flour

InactiveCN104642910AModerate tastePreserve soluble nutrientsFood preparationCelluloseAdditive ingredient
The invention discloses a preparation method of instant oat flour and relates to a preparation method of oat flour. The preparation method comprises the following steps: baking, cleaning, soaking, steaming and cooling oats, serving as raw materials, sequentially; glue milling the oats after pretreatment to prepare oat pulp, carrying out enzymatic hydrolysis, heating, boiling, and killing enzyme; and cooling an enzymolysis liquid, glue milling, homogenizing, drying, sterilizing and packaging a finished product. According to the preparation method disclosed by the invention, with the oats as the raw materials, by virtue of pretreatment, enzymatic hydrolysis technological treatment and spray drying, the instant oat flour which can keep multiple nutritional components and fragrance of oat is prepared; the prepared instant oat flour is moderate in taste, has self flavor of oat, keeps the soluble nutritional components in the oats, is also capable of providing extra plant proteins, fats and soluble dietary fibers and can be directly used for preparing an oat beverage; as macromolecular substances including starch, cellulose and proteins in the oats are hydrolyzed by virtue of an enzyme method, the viscosity of the starch of the oats is lowered, and the problems of water separating, gelation, precipitation and the like caused when the oats are applied to beverages are solved.
Owner:厦门北大泰普食品科技有限公司

Synchronous aqueous enzymatic ultrasonic extraction for walnut oil and walnut protein peptide

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.
Owner:KUNMING UNIV OF SCI & TECH
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