544 results about "Enzyme protein" patented technology

The invention provides a functional oil microencapsulation and a manufacturing method thereof and relates to a microencapsulation, and the functional oil microencapsulation has the advantages of high stability and immobilized proteinaceous pellicle. The oil microencapsulation is prepared from the following raw materials: functional oil, plant oil, an antioxidant, aqueous-phase main-wall materials, aqueous-phase auxiliary-wall materials, bio-enzyme protein and water. The plant oil serving as a carrier is mixed with the functional oil, then the antioxidant is added, and then the materials are heated and dissolved in water bath, thus obtaining an oil-phase core material solution; the aqueous-phase main-wall materials and the aqueous-phase auxiliary-wall materials are added into deionized water, and then the materials are heated and dissolved in water bath, thus obtaining an aqueous-phase wall material solution; the oil-phase core material solution is added in the aqueous-phase wall material solution, the mixture is sheared and emulsified by a shearing machine, and then is homogenized by a homogenizer, thus obtaining a nanometer-level solution with uniform oil drop diameter; and bio-enzyme is added into the nanometer-level solution, the mixture is placed in hot-water bath for heating and stirring reaction, temperature is raised after the reaction so as to enable the bio-enzyme to be inactivated, and drying is carried out, thus obtaining the functional oil microencapsulation.

Disclosed are a fusion protein comprising enzyme N-acetylgalactosamine-6-sulfate sulfatase and a short peptide consisting of 4-15 acidic amino acids attached to the enzyme on its N-terminal side, a pharmaceutical composition containing the fusion protein, and a method for treatment of type A Morquio disease using the fusion protein. Compared with the native enzyme protein, the fusion protein exhibits higher transferability to bone tissues and improved, higher stability in the blood.

The invention provides an enzyme electrode usable as a highly sensitive sensor, a high-output bio fuel cell or an electrochemical reaction device of a high reaction efficiency. The enzyme electrode includes an electroconductive base member and an enzyme, wherein the enzyme is formed by an associated protein in which two or more different enzyme proteins are associated.

The invention relates generally to isolated leucoanthocyanidin reductase LAR polypeptides of the Reductase-Epimerase-Dehydrogenase (RED) protein family, and nucleic acid molecules encoding same and their use in regulating the biosynthesis and accumulation of proanthocyanidins in plants. The invention is further directed to isolated nucleic acid molecules of plants, which encode leucoanthocyanidin reductases of the RED protein family. The isolated polypeptides and nucleic acid molecules of the present invention are useful for modifying the pasture quality of legumes, and, in particular, for producing bloat-safe forage crops, or crops having enhanced nutritional value, enhanced disease resistance or pest resistance, or enhanced malting qualities.

The invention relates to a diluent being capable of maintaining the high stability of enzyme marker solution and also relates to a method for preparing the diluent and applications thereof. The diluent comprises buffer solution, protein matrix solution, amino acid solution, sugar solution, a specific additive and a preservative. The diluent is a novel enzyme marker solution developed based on the characteristics of antibody protein and enzyme protein. With the adoption of the dilution, the stability of enzyme markers in storage and application can be effectively maintained. Tests show that the enzyme marker diluent can be stored at temperature of 37 DEG C for more than one week to fully meet the requirements for the quality of in-vitro diagnostic reagent kits. The diluent can be directly applied to the preparation of the in-vitro diagnostic reagent kits and meet the increasingly growing market demand for the in-vitro diagnostic reagent kits.

The present invention provides porcine Forssman synthetase (FSM synthase) (Globoside α-N-acetylgalactosaminyltransferase) protein, cDNA, and genomic DNA sequence. Furthermore, the present invention includes porcine animals, tissue and organs as well as cells and cell lines derived from such animals, tissue and organs, which lack expression of functional FSM synthetase. Such animals, tissues, organs and cells can be used in research and in medical therapy, including in xenotransplantation. In addition, methods are provided to prepare organs, tissues, and cells lacking the porcine FSM synthetase gene for use in xenotransplantation.

The invention discloses a bacterial laccase mutant protein, which is characterized in that the mutant protein amino acid sequence is obtained by deletion mutation of the 323rd glycine residue to the 332rd glycine residue in the bacterial laccase amino acid sequence shown as SEQ ID No.1. Through a genetic engineering reconstruction method, a stability improved bacterial laccase protein coding gene, its expression plasmid and engineered bacteria can be obtained, and after large-scale fermentation and induced expression of the engineered bacteria, the stability improved bacterial laccase protein can be obtained. According to the invention, the marine uncultured microorganism source bacterial laccase Lac15 is taken as the foundation, and by means of genetic engineering reconstruction, mutant gene can be obtained. At the same time, a recombinant escherichia coli is employed to conduct high-density culture for high-efficiency expression of the bacterial laccase mutant protein. According to the invention, the stability and yield of the bacterial laccase are greatly improved.

The present invention relates to novel fluorescent dyes, novel fluorogenic and fluorescent reporter molecules and new enzyme assay processes that can be used to detect the activity of caspases and other enzymes involved in apoptosis in whole cells, cell lines and tissue samples derived from any living organism or organ. The reporter molecules and assay processes can be used in drug screening procedures to identify compounds which act as inhibitors or inducers of the caspase cascade in whole cells or tissues. The reagents and assays described herein are also useful for determining the chemosensitivity of human cancer cells to treatment with chemotherapeutic drugs. The present invention also relates to novel fluorogenic and fluorescent reporter molecules and new enzyme assay processes that can be used to detect the activity of type 2 methionine aminopeptidase, dipeptidyl peptidase IV, calpain, aminopeptidase, HIV protease, adenovirus protease, HSV-1 protease, HCMV protease and HCV protease.

The present invention provides methods and compositions for modulating the phosphorylation of DARPP-32 in a serotonergic receptor intracellular signaling pathway. The invention provides methods and compositions for modulating the activities of DARPP-32, casein kinase 1 (CK1), cyclin-dependent kinase 5 (Cdk5), AMPA receptors, protein phosphatase-1 (PP-1), protein phosphatase 2C (PP2C), protein phosphatase 2B (PP2B) and / or protein phosphatase 2A (PP2A) in cells or tissues. The invention provides methods of treating serotonergic intracellular signaling pathway disorders, e.g., depression. The invention provides methods of treating dopamine-related disorders. The invention provides methods of identifying agents that modulate the activities of serotonergic receptor intracellular signaling molecules, DARPP-32, casein kinase 1, cyclin-dependent kinase 5, AMPA receptors, protein phosphatase-1, protein phosphatase 2C, protein phosphatase 2B and / or protein phosphatase 2A, for use in such treatments. The invention also provides methods of modulating phosphorylation-dependent activation of AMPA receptors for use in such treatments.

The invention relates to a host cell and a method for the host cell applied in efficient secretion expression of a recombinant protein, which belongs to the filed of microbiological engineering and fermentation engineering. The invention provides a bacillus licheniformis host cell CBB3008 which is preserved in the China Center for Type Culture Collection with the preserving number of CCTCC NO: M 208236. The invention transforms an expression plasmid of an industrial enzyme which is obtained by a gene cloning technology into the host cell to synthesize an industrial enzyme preparation in the host cell efficiently, and then secretes the synthesized enzyme protein into a culture medium efficiently through a protein secretion system of the host cell, thus the invention can guide efficient secretion production of the industrial enzyme preparation. Therefore, the invention is helpful to reduce the fermentative production cost of the industrial enzyme preparation, simplify the fermentative production process and reduce the environmental pressure of the fermentation industry. A method for host cell screening and genetic improvement of the invention can also be used for other types of host cells, in particular for the breeding of the host cells of bacillus subtilis, bacillus megaterium, bacillus pumilus, bacillus starch solution and the like.

The invention discloses a forage liquid phytase preparation with high enzyme activity retention rate. The forage liquid phytase preparation is prepared by adding polyethylene glycol 6000, trehalose, sodium benzoate and pure water to original phytase enzyme liquid; and adjusting a pH value to 4.5-5.5, wherein hydroxide radicals in the polyethylene glycol 6000 are bonded with carboxyls, aminos, and the like on the surfaces of an enzyme protein molecule and an activity center thereof by covalent bonds, hydrogen bonds and Van der Waals force so that the activity center of the enzyme protein molecule and the whole enzyme protein molecule are protected; the trehalose (stabilizing agent) improves the permeation pressure of enzyme solution to reduce the stretching degree of the enzyme protein molecule, and the enzyme protein molecule is relatively in a dewatering state to shrink, thereby improving the stability; and the sodium benzoate (preservative) takes the effect of improving the biological stability of liquid enzymes and the pH range is optimally selected from 4.5 to 5.5 . The invention is used for the field of fodder additives.

The invention discloses a composite stabilizing agent for preserving a liquid cellulase, which can preserve the vitality of the cellulase for a long time under the liquid condition. The composite stabilizing agent is prepared from the raw materials including sucrose, sodium chloride, potassium sorbate and sodium hyposulfite by blending and stirring. The weight percentages of the raw materials in the liquid cellulase are as follows: 30 to 50 percent of the sucrose, 5 to15 percent of the sodium chloride, 0.01 to 0.03 percent of the potassium sorbate and 0.01 to 0.02 percent of the sodium hyposulfite. The composite stabilizing agent uses the sucrose and the sodium chloride so that cellulase protein molecules keep the stable structures easily in water molecules and are not easy to denaturalize and lose the activity, uses the potassium sorbate to control the colony count so as to meet the requirements on food conditions conveniently, and uses the sodium hyposulfite for preventing enzyme protein from being oxidized to lose the activity. The four raw materials of the composite stabilizing agent is easily available, the food grade is relatively high so that the cellulase activity cannot be reduced within two years at room temperature, the cost for building a refrigerated storage for keeping the cellulase activity is saved, and the preservation and transportation of the liquid cellulase are convenient.

The invention discloses biological enzyme protein feed additives which comprises microbe live bacteria cells, active biological enzymes, amino acid, starch, fibers and water. The microbe live bacteria are Candida utilis bacteria, lactobacillus acidophilus and Bacillus licheniformis. The active biological enzymes are protease, amylase, beta-glucanase, xylanase, pectase, glucoamylase and fat acid enzyme, and amount of the active biological enzymes is 20000-24000 mu / g. The additives are full of microbe live bacterial and active biological enzymes for growth, metabolism, digestion, absorption, immunoregulation of livestock and poultry, intestinal tract flora balance capability, intestine mucous membrane epithelial cells restoration capability, intestinal villus growth capability, vitamin and trace element supply capability of the livestock and poultry are maintained. Absorption and utilization rate of the feed additives is high. According to the feed additives, drain amount of nitrogen, phosphor and calcium in night soil of the livestock and poultry is reduced, qualities of livestock, poultry and aquatic products are raised, and environmental pollution is reduced. Offensive odor of the livestock and poultry night soil is weakened and culture environment is purified.

The invention relates to a preparation method of a levan-containing yoghourt, and the method comprises the following steps of: (1) preparing a fermented culture solution by implementing activation and enzyme production fermentation to a levan strain; (2) centrifugally collecting the supernatant of the fermented culture solution or the thallus; (3) adding ammonium sulfate powder into the supernatant of the fermented culture solution, centrifugating and collecting albumen precipitation; or crushing and centrifugating the thallus by using ultrasonic waves, adding the ammonium sulfate powder into the supernatant, centrifugating and collecting the albumen precipitation; and purifying to prepare pure enzyme protein levansucrase; and (4) adding sugar and the pure enzyme protein levansucrase into raw milk, then vaccinating with lactobacillus, and preparing the levan-containing yoghourt after fermentation. According to the preparation method of the levan-containing yoghourt, levan is produced from microorganism levan sucrase when yoghourt is fermented, so that a novel yoghourt containing functional polysaccharide is directly acquired without complex polysaccharide purification processes. The preparation method of the levan-containing yoghourt has wide prospect of application.

The invention discloses an isothermal nucleic acid amplification reaction reagent which comprises Tris buffer solutions, potassium acetate, magnesium acetate, dithiothreitol, polyethylene glycol, adenosine triphosphate (ATP), deoxy-ribonucleoside triphosphate (dNTPs), phosphocreatine, creatine kinase, a primer group, single-strand binding (SSD) protein, colon bacillus helicase RecQ protein, UvsY protein and DNA polymerase. The invention further discloses an isothermal nucleic acid amplification method which includes steps of extracting DNA or performing inverse transcription after DNA extracting, and adding the isothermal nucleic acid amplification reaction reagent and reacting the mixture at a temperature in a range between 25 DEG C and 45 DEG C for 10 minutes to 60 minutes to complete the nucleic acid amplification. Compared with traditional polymerase chain reaction (PCR) technologies and according to the technology, nearly no professional instrument is needed, the operation is simple, technical requirements for operators are low, and the reaction time only needs about half an hour.

The invention relates to a method for reducing the viral and microbial content of biological extracts which contain solids. It is provided that the method comprises the steps(a) provision of the biological extract which contains solids, comprising a biologically active substance, selected from enzymes, proteins and peptides, or a mixture of such substances; and(b) subjecting the biological extract provided in step (a) to a high-pressure treatment;wherein the biological activity of the biological extract which contains solids after the high-pressure treatment is at least 50% of the biological activity of the biological extract which contains solids before the high-pressure treatment.

The invention discloses a mutant zymoprotein of D-amino acid oxidase and preparation method of the mutant zymoprotein. The preparation method comprises the following steps: according to homologous sequence comparison result, respectively performing mutation on 115, 119 and 286 sites of D-amino acid oxidase in primary Arthrobacterprotophormiae (DSM15035) through site-specific mutagenesis technology to construct three-site mutation expression vector Pet-e115A / N119D / T286A; and carrying out prokaryotic expression and purification to obtain zymoprotein. An apparent secondary speed constant Kcat / Km of the obtained mutant protein to substrate D-Met is 1.39*10 to the power of 5s-1.M-1, is 5.03 times of wild type DAAO (D amino acid oxidase), and is 55.96 times of pKDAAO protein of pig kidney source. The detection efficiency of partial D-amino acid can be improved, and the good application value is realized.

The present invention comprises methods and compositions for the reduction of oxalate in humans, and methods for the purification and isolation of recombinant oxalate reducing enzyme proteins. The invention provides methods and compositions for the delivery of oxalate-reducing enzymes in particle compositions. The compositions of the present invention are suitable in methods of treatment or prevention of oxalate-related conditions.

The invention relates generally to isolated leucoanthocyanidin reductase LAR polypeptides of the Reductase-Epimerase-Dehydrogenase (RED) protein family, and nucleic acid molecules encoding same and their use in regulating the biosynthesis and accumulation of proanthocyanidins in plants. The invention is further directed to isolated nucleic acid molecules of plants which encode leucoanthocyanidin reductases of the RED protein family. The isolated polypeptides and nucleic acid molecules of the present invention are useful for modifying the pasture quality of legumes, and, in particular, for producing bloat-safe forage crops, or crops having enhanced nutritional value, enhanced disease resistance or pest resistance, or enhanced malting qualities.

Enzyme formulations should stay active and devoid of all possible inactivation processes during transport and long-term storage. The stabilization of enzymatic activity, especially in presence of critical functional components for specialized application, has been a long-standing obstacle when stored over an extended period of time. One such obstacle is the stability of enzyme in presence of weak acids in formulation, which is often desired to efficiently break down biopolymer filtercake embedded in a carbonate matrix, in the open-hole section of a horizontal well, for hydrocarbon production. The invention comprises methods and compositions to enhance the long-term storage stability of an enzyme formulation consisting of an in-situ acid precursor system, where in-situ generated precursors are mainly ester compounds that dissociate by slow hydrolysis to generate organic acid, shifting the pH of the formulation outside the pKa of the enzyme protein. The methods and compositions include the addition of high concentrations of metal halide salts, a pH control additive, and a water activity reducing agent. The methods and compositions of the invention provide a high degree of stability for enzyme formulation at long-term storage conditions.

The invention provides and discloses an optimized nucleotide sequence of alkaline pectinase pel168s and a high-level expression method thereof. According to the method, a pel168 gene sequence (wherein the gene is Bacillus subtilis 168 the accession number of which in a GenBank is AL009126) is optimized by DNA works software, restriction enzyme cutting sites SalI and PmeI are shielded, a restriction enzyme cutting site EcoRI is added at the 3' end of a primer, and a restriction enzyme cutting site NotI is introduced at the 5' end of the primer. After the procedures of PCR (Polymerase Chain Reaction) amplification, connection transformation and sequencing verification, the optimized gene sequence of the alkaline pectinase pel168s is obtained. Recombinant plasmid pel168s-9k is constructed according to the sequence, and then is transformed into pichia yeast GS115, thereby obtaining a positive recombinant strain GS115 / pel168s-9k. According to the invention, when alkaline pectinase is produced by adopting the optimized nucleotide sequence of the alkaline pectinase pel168s and utilizing the pichia yeast, the target protein expression index is high, the purge process is simple, the production cost of the alkaline pectinase is reduced greatly, and the utilization rate of an enterprise on the alkaline pectinase is enhanced.

The invention discloses immobilized crosslinked enzyme aggregates (CLEAs) and a preparation method thereof. The crosslinked enzyme aggregates are fixed on a porous microsphere; the preparation method thereof comprises: dispersing detached enzyme needing to be immobilized into the porous microsphere; carrying out depositing and crosslinking on zymoprotein in the porous microsphere in sequence, and finally rinsing the porous microsphere, thus preparing the immobilized crosslinked enzyme aggregates. The method of the immobilized enzyme can be used for immobilizing the enzyme of various types or sources; the immobilized method is simple and easy to be carried out, is easy to be operated, does not need special devices and has low production cost; the prepared crosslinked enzyme aggregates have high stability and strong capacity for resisting trypsinase, has a certain physical form and a stable structure, can be recycled and repeatedly used and can be used for continuous industrial production.

The invention discloses a method for rapidly dissolving chitosan and preparing chitosan with high-concentration substrate enzymatic method; a high-speed shearing dispersion machine is adopted to rapidly dissolve and disperse the chitosan to form high-concentration substrate solution, and the adding part is utilized to degrade hydrolytic enzyme of the chitosan to lead the viscosity of the solution to be diluted, and the concentration of the chitosan substrate in a reaction system is improved to 10-20 percent. After enzyme reaction is carried out, chitosan hydrolytic enzyme protein and chitosan macromolecules in product solution are removed by an ultrafiltration method, permeate is concentrated for desalinating through nanofiltration, and the solid content of the chitosan in the product can reach 20-35 percent through vacuum concentration process; the method can effectively solve the problems that the dissolution time of the chitosan is long, the produced solution concentration is low, and energy consumption is high owning to low substrate concentration, the energy consumption level of subsequent processing such as separation purification, concentration and drying processes is greatly reduced, and the production cost is lowered.

The invention relates to a three-dimensional composite material of the titanium carbide, and a preparation method thereof, and an application in constructing a thrombin aptasensor. A substrate of thethree-dimensional composite material TiO2 / Ti3C2TX is formed by the self-growth of TiO2 nanorods on the surface of Ti3C2TX by one-step hydrothermal method; meanwhile, the three-dimensional composite material M NPs / TiO2 / Ti3C2TX is obtained by reducing noble metal nanoparticles (M NPs) on the surface of the three-dimensional material. The material has a highest current intensity, due to the fact thata variety of enhancements are achieved in one step. The three-dimensional structure of the composite material may provide a particularly large accessible surface area, which facilitates the anchoringof the M NPs. The introductions of transition metal carbonitrides (MXenes) and metal nanoparticles can improve the efficiency of the charge separation, and accelerate the electron transfer rate. A sensitive unlabeled aptamer is successfully established through the combination of M NPs and aptamer chains, and is used for determining the enzyme protein. According to the three-dimensional compositematerial of the titanium carbide, and the preparation method thereof, and the application in constructing thrombin aptasensor, the aptasensor has a good electrochemical performance, a wide linear range, and a relatively low detection limit, thereby indicating that the M NPs / TiO2 / Ti3C2TX will be promising for the use as electrode materials in electrochemical biosensors.

The present invention relates to an enzyme having novel N-acetylglucosaminyltransferase (OMGnT) activity, a polynucleotide encoding the enzyme, a recombinant polynucleotide comprising the polynucleotide, a host cell comprising the recombinant polynucleotide, a method for producing an enzyme protein having OMGnT activity by culturing the host cell in a medium, and a method for modifying a sugar chain structure with the enzyme and a carbohydrate having a sugar chain structure modified by the method. This enzyme enables the production of complex carbohydrates, which could not be formed with conventional glycosyltransferases.

The invention provides an analytical method of correlation of xylanase heat resistance and an N-terminal disulfide bond, and belongs to the technical field of microbial genetic engineering. A result that the N-terminal disulfide bond is one of factors which affect the xylanase heat resistance is proved by adopting a scheme of combining three kinds of bioinformatics methods including the homologous comparison, the homologous modeling and the molecular dynamics simulation of a primary structure of the zymoprotein, and the result is also proved by combining an experimental means of site-specific mutagenesis and by analyzing a heat resistance mechanism of the EvXyn11TS. The research result establishes a solid foundation for the heat resistance transformation of the 11 family normal temperature high specific activity xylanase which has a similar primary structure with the EvXyn11TS.

The invention belongs to the technical fields of molecular biology and genetic engineering, and provides an encoding sequence for encoding cDNA of Jatropha curcas farnesyl pyrophosphate synthase. The farnesyl pyrophosphate synthase catalyzes isopentenyl pyrophosphate and dimethylallyl pyrophosphate to carry out continuous condensation reaction to form farnesyl pyrophosphate, is key enzyme for metabolic pathway of Jatropha curcas isoprene, and contributes to reducing the content of phorbol esters and precursor thereof in the Jatropha curcas. Due to the use of the Jatropha curcas protein, substances interacting with the Jatropha curcas protein, or receptors, inhibitors or antagonists, and the like can be screened by various conventional screening methods. The encoding sequence for encoding the cDNA of the Jatropha curcas farnesyl pyrophosphate synthase has great application prospects.

The invention relates to beta-galactosidase and application thereof, in particular to a beta-galactosidase with a cellulose adsorption zone and application thereof in the production of galactooligosaccharide, and belongs to the technical field of enzyme protein engineering. The beta-galactosidase gene with the cellulose adsorption zone has a nucleotide sequence shown as SEQ ID No.1; and the beta-galactosidase coded by the gene has an amino acid sequence shown as SEQ ID No.2. The related GOS production method has the advantages of low cost, high yield, simple and reasonable steps, and the like, and has great industrialized application prospect.