35results about How to "Reduce the amount of enzyme" patented technology

The invention discloses a method for preparing gelatin with protease degradation ossein. The method is characterized in that derosination and demineralization ossein is mainly adopted as the raw material, and acidic or neutral protease degradation ossein is used for preparing gelatin. Compared with the traditional technology, the method has the advantage that long liming period and complicated neutralization washing process are omitted, so that the production period of gelatin is shortened, and a great amount of water resource and manpower cost are saved; and compared with the new technology of preparing gelatin with skeleton grains, and the content of foreign protein and inorganic acid in gelatin is greatly reduced, so that freezing strength and viscosity of the product are improved.

The invention relates to a gene encoding of firefly luciferase, its preparation method and application, which comprises the following steps: 1. introducing mutation sites of mutant enzymes; 2. identifying recombinant plasmids; 3. sequencing to obtain polymerase chain reaction (PCR) fragment with 1060 mutation sites; 4. obtaining escherichia by using restriction endonucleasecoli purifying after recovering and purifying through gel; 5. identifying recombinant plasmids to obtain a separating protein gene; 6. culturing escherichia coli, 7. converting recombinant plasmids into BL21 competent cell to obtain bacterial strain capable of expressing mutant enzymes, 8. culturing the obtained BL21 expression bacterial strain; 9. centrifuging bacteria liquid of protein, centrifuging to obtain supernatant through ultrasonic fragmentation; 10. determining to obtain firefly luciferase with high heating stability and high enzyme activity. The invention relates to application of firefly luciferase gene on foodstuff, medicine, thalline detection. The luminous intensity of the mutant enzyme is about 15 to 20 times than that of wild type, and the mutant enzyme has good heat stability than that of the wild type.

The invention relates to a method for producing grape skin health-care beverage and belongs to the technical field of beverage processing. The method comprises the following steps of: selecting and cleaning fresh grape skin serving as a raw material; adding 0.02 percent of isoVc sodium and 0.15 percent of citric acid to protect color; crushing and pulping the grape skin to prepare grape skin pulp; promoting composite enzymolysis ultrasonically; filtering by using a filtering material with 100 to 350 meshes or performing centrifugal separation at 3,000 to 5,000 r/min; mixing; performing vacuumdegassing under vacuum degree of 0.09 MPa for 20 to 25 minutes; homogenizing under 30 to 40 MPa; after performing encapsulation, performing sterilization at the temperature of between 135 and 150 DEGC for 35 to 60 seconds; cooling; performing labeling; spraying codes; and preparing the finished product after the product is inspected to be qualified. In the method, the enzymolysis reaction time is shortened by 50 percent and the enzyme adding amount is reduced by 20 percent, so that enzymolysis efficiency is improved and production cost is reduced. The processed health-care beverage has rich nutrients and good quality and is suitable for people of all ages to drink. The quality guarantee period of the beverage is more than 12 months. By the method, the grape skin is utilized at high value; waste is changed into treasure; and obvious economic, social and ecological benefits are achieved.

The invention relates to a method for synchronously extracting soybean oil, soybean milk powder and soybean dietary fiber by an aqueous enzymatic method, which belongs to the technical field of plant grease processing. The method comprises the following steps: 1) dehulling soybeans and crushing the soybeans, extruding and puffing the material to obtain a puffed product; 2) adding water into an enzymatic hydrolysis reaction vessel, heating steam and adding the puffed product to obtain a mixed liquor, performing alkaline extraction on the mixed liquor, adding alkali protease for performing enzymatic hydrolysis, using a horizontal centrifuge after enzymatic hydrolysis is carried out for performing centrifugation to obtain an emulsified oil, a hydrolysate and residues; 3) using a disk centrifuge for performing centrifugation on the emulsified oil to obtain the soybean oil and a water phase, adding the water phase into the hydrolysate for concentration, and drying the material to obtain the soybean milk powder; and 4) adding water for mixing the water and the residues for homogenizing and drying a mixture to obtain the soybean dietary fiber. The method has the advantages of simple process, less enzyme usage amount, short reaction time, and low cost. The prepared high-quality soybean oil can obtain the high-nutrition partly-defatted soybean milk powder and high-oil high-protein soybean dietary fiber, and is suitable for industrial continuous production.

The invention relates to water-soluble poly amino benzenes high temperature resisting conductive material and method for preparation, wherein the material is prepared from p-phenylene diamine 54-216 parts, monomer propenoic acid 35-71 parts, horseradish peroxidase 0.05-3 parts as raw material, p-phenylene diamine and propenoic acid as monomers, deionized water as solvent, horse radish peroxidase HRP as catalyst, H2O2 as oxidation agent through copolymerization, acetone deposition and vacuum drying 48 hours at 50 deg C.

The invention belongs to the field of enzyme engineering, and particularly relates to a method for purifying and immobilizing gamma-lactamase and splitting (+/-) gamma-lactam by one step. Gamma-lactam hydrolase is immobilized on a Ni-NTA agarose affinity column, and (+) gamma-lactam is hydrolyzed on the column so as to split the (+/-) gamma-lactam. Plasmids of heat-resistant gamma-lactam hydrolase genes with sulfolobus solfataricus are transferred into colon bacillus to construct gene engineering bacteria capable of expressing the gamma-lactam hydrolase with histidine marks at an N end and a C end or gene engineering bacteria capable of expressing the gamma-lactam hydrolase with histidine marks at only one end of the N end and the C end. And then, the enzyme with the histidine marks is further purified and immobilized on the Ni-NTA agarose affinity column, and a (+/-) gamma-lactam substrate is continuously converted on the column under the conditions that the pH value is 5 to 10 and the temperature is between 30 and 100 DEG C; the conversion rate can be kept at more than 80 percent; and the optical purity of a product is more than 99 percent.

The invention discloses thermal stability and trehalose yield improved MTSase mutant and belongs to the technical field of enzyme engineering and protein engineering. The maltooligosyl trehalose synthase mutant disclosed by the invention is prepared by mutating one amino acid site of maltooligosyl trehalose synthase. Compared with parent maltooligosyl trehalose synthase, the prepared maltooligosyltrehalose synthase has higher thermal stability, a lower enzyme adding amount and a higher trehalose producing conversion rate. Compared with a wild type, a half-life period of the maltooligosyl trehalose synthase mutant disclosed by the invention at 50 DEG C is increased by 12 hours, and the half-life period of the maltooligosyl trehalose synthase mutant disclosed by the invention is 1.3 times of the half-life period of the wild type.

The invention relates to firefly luciferase and an encoding gene and an obtaining method thereof. The defect that the original firefly luciferase in North America is poor in heat stability and cannot be used at a high temperature is overcome; and the original firefly luciferase in North America is mutated, so that mutative enzyme of which the heat stability is obviously improved is obtained; and the application value is improved. An amino acid sequence is SEQ ID NO.1; and a protein deoxyribonucleic acid (DNA) sequence in encoding is SEQ ID NO.2. An encoding gene and a preparation method of firefly luciferase mutant in North America are provided. By the method, a recombinant expression vector containing a wild firefly luciferase gene is built; and the protein is purified by using an affinity chromatography method. The method is simple and feasible, simple to operate, and low in cost.

The invention discloses thermal stability improved maltooligosyl trehalose synthase mutant and belongs to the technical field of enzyme engineering and protein engineering. The maltooligosyl trehalosesynthase mutant disclosed by the invention is prepared by mutating one amino acid site of maltooligosyl trehalose synthase. Compared with parent maltooligosyl trehalose synthase, the maltooligosyl trehalose synthase mutant has higher stability. Compared with a wild type, a half-life period of the maltooligosyl trehalose synthase mutant disclosed by the invention at 50 DEG C is increased by 41 hours; the half-life period of the maltooligosyl trehalose synthase mutant disclosed by the invention is 2 times of the half-life period of the wild type; namely, compared with the wild type, the stability of the maltooligosyl trehalose synthase mutant disclosed by the invention is improved by twice.

The invention relates to the technical field of processing of vegetable protein, in particular to a vegetable protein peptide and a preparation method thereof. The preparation method of the vegetableprotein peptide comprises the following steps of (a) performing enzymolysis for 0.5 to 4 h on a mixture of vegetable protein and water under the vacuum condition of 50 to 55 DEG C under the effect ofan alkaline protease so as to obtain a mixed material; and (b) after the mixed material is subjected to enzymolysis for 1 to 2 h under the conditions of 50 to 55 DEG C and normal pressure under the effect of an enzyme A, performing enzymolysis for 0.5 to 2 h under the effect of a flavourzyme, wherein the enzyme A is selected from a neutral protease, a papain and a trypsin. The vegetable protein peptide and the preparation method provided by the invention have the advantages that the yield of the vegetable protein peptide can be greatly improved; the protein content and the polypeptide contentin the vegetable protein peptide are improved; the product quality is improved; the enzymolysis time can be reduced; the preparation period is shortened; pH regulation is not needed; the production cost is reduced; the production process is simplified; and the vegetable protein peptide and the preparation method can be used for mass production.

The invention provides a method for preparing a high-glucan cell wall product. The method adopts a flow path shown as a figure 1 in the specification. The cell wall product prepared with the method has a stable glucan content of 40% or above, which is greatly higher than that of the existing product in the market; in addition, a raw material used in the invention is a byproduct obtained during yeast cell extraction, and is low in cost; the cell wall product with high glucan content is widely applied to the fields of feed, food and the like; therefore, the method has a great economic prospect.