8507 results about "Enzyme" patented technology

The present disclosure relates to materials and methods for spatially analyzing nucleic acids that have been fragmented with a transposase enzyme, alone or in combination with other types of analytes.

The present disclosure relates to materials and methods for spatially analyzing nucleic acids that have been fragmented with a transposase enzyme, alone or in combination with other types of analytes.

The present disclosure relates to materials and methods for spatially analyzing nucleic acids that have been fragmented with a transposase enzyme, alone or in combination with other types of analytes.

The present disclosure relates to materials and methods for spatially analyzing nucleic acids that have been fragmented with a transposase enzyme, alone or in combination with other types of analytes.

The present disclosure relates to materials and methods for spatially analyzing nucleic acids that have been fragmented with a transposase enzyme, alone or in combination with other types of analytes.

The present disclosure relates to materials and methods for spatially analyzing nucleic acids that have been fragmented with a transposase enzyme, alone or in combination with other types of analytes.

The present disclosure relates to materials and methods for spatially analyzing nucleic acids that have been fragmented with a transposase enzyme, alone or in combination with other types of analytes.

Methods and analyte sensors including at least a first working electrode having a first active area thereon, and performing a dip coating operation to deposit a bilayer membrane upon the first working electrode and the first active area. The bilayer may include an inner layer having a first membrane polymer and an outer layer having a second membrane polymer, the first membrane polymer and the second membrane polymer differing from one another. The dip coating operation may comprise one or more first dips in a first membrane formulation to form the inner layer of the bilayer membrane and one or more second dips in a second membrane formulation to form the outer layer of the bilayer membrane upon the inner layer.

Multiple enzymes may be present in one or more active areas of an electrochemical analyte sensor for detecting one or more different analytes. In particular, an analyte sensor may comprise a sensor tail configured for insertion into a tissue and one or more working electrodes having a glucose-responsive active area and an ethanol-responsive active area to detect glucose and ethanol in vivo.

The present disclosure relates to materials and methods for spatially analyzing nucleic acids that have been fragmented with a transposase enzyme, alone or in combination with other types of analytes.

The invention belongs to the technical field of bioengineering, and discloses a recombinant yarrowia lipolytica capable of efficiently and rapidly producing sclareol, which is characterized in that optimized sclareol synthase related coding genes ScLPPS, ScSCS, GGPPS1, GGPPS2, ERG20 and tHMGR coding genes are inserted into a yarrowia lipolytica genome; inserting optimized pentadienol utilization pathway coding genes such as IPK, THIM and IDI coding genes; and inserting an optimized ATP: citrate lyase coding gene ACLY to obtain the ATP: citrate lyase coding gene ACLY. The recombinant yarrowia lipolytica is based on knockout of a yarrowia lipolytica strain responsible for coding a non-homologous recombination gene ku70, so that the homologous recombination capability of the yarrowia lipolytica strain is enhanced, gene integration is realized through the homologous recombination function of the yarrowia lipolytica, the genetic stability of the introduced gene can be greatly improved, and the recombinant yarrowia lipolytica strain has a good application prospect. The construction method of the recombinant yarrowia lipolytica strain is efficient and simple to operate.

The invention provides a method for improving synthesis of nucleic acid under a non-diagnostic constant-temperature condition, a kit and application, and relates to the technical field of bioengineering. Provided are: a nucleic acid having an F2 region capable of annealing with F2c in an F region on the same chain at the 3'end, and an R2c region capable of annealing with R2 in an R region on the same chain at the 5 'end; taking the nucleic acid as a template, taking the 3'end of the F1c region annealed with the F1 region as a synthesis starting point, and synthesizing a complementary nucleic acid chain of the nucleic acid; and performing complementary chain synthesis through a polymerase catalytic chain displacement type synthesis reaction so as to replace the complementary nucleic acid chain synthesized in the previous step. The method has the main advantage that a new combination of short-chain primers can be obtained through simple transformation to trigger rapid amplification of gene fragments by applying a single-enzyme constant-temperature system to short-chain nucleic acid fragments.

The invention provides a nano enzyme as well as a preparation method and application thereof. The preparation method of the nano enzyme comprises the following steps: adding a stabilizer into a preheated solvent, and carrying out ultrasonic treatment; then adding ferric salt and cobalt salt, and adding alkali to adjust the pH value of the solution; then transferring the solution into a reaction kettle for reaction; and after the reaction, washing and separating to obtain the product. The obtained nano-enzyme is appropriate and uniform in particle size and good in dispersity, the enzyme activity is as high as 9.56 U / mg, and the application of the nano-enzyme in immunochromatography detection and plant canker prevention and treatment is widened. When the immunochromatography detection card prepared from the nano enzyme is used for detecting human hemoglobin, the detection limit can reach 0.05 ng / mL and is at least 10 times lower than that of a commercially available kit. When the nano-enzyme suspension is used for preventing and treating the kiwifruit canker, the nano-enzyme suspension with the concentration larger than or equal to 0.1 mg / mL is used, and the plant canker can be well controlled.

The invention provides a preparation method of a polygonatum sibiricum fermentation product, which comprises the following steps: A) pretreating a polygonatum sibiricum raw material, and mixing with water to obtain polygonatum sibiricum slurry; b) heating the polygonatum sibiricum slurry, mixing the heated polygonatum sibiricum slurry with a nitrogen source and a phosphorus source, and performing primary sterilization to obtain sterilized polygonatum sibiricum slurry; c) inoculating the sterilized rhizoma polygonati slurry with saccharomycetes, and fermenting to obtain fermentation liquor; d) adding a carbon source into the fermentation liquor, performing secondary sterilization, inoculating lactic acid bacteria, performing secondary fermentation, and performing third sterilization to obtain secondary fermentation liquor; and E) adding a compound enzyme into the secondary fermentation liquid for enzymolysis, then sterilizing for four times, and centrifuging to obtain the polygonatum sibiricum fermentation product. 3000 Da), a small molecule polypeptide (lt; the polygonatum kingianum fermentation product prepared by the method disclosed by the invention has the advantages that the conversion of polygonatum kingianum saponin can be promoted, the contents of diosgenin and trilliin are increased and calibrated, and the polygonatum kingianum fermentation product has the effects of improving the reaction capability and the exercise capability.

Methods and analyte sensors including at least a first working electrode having a first active area thereon, and performing a dip coating operation to deposit a bilayer membrane upon the first working electrode and the first active area. The bilayer may include an inner layer having a first membrane polymer and an outer layer having a second membrane polymer, the first membrane polymer and the second membrane polymer differing from one another. The dip coating operation may comprise one or more first dips in a first membrane formulation to form the inner layer of the bilayer membrane and one or more second dips in a second membrane formulation to form the outer layer of the bilayer membrane upon the inner layer.

Methods and analyte sensors including at least a first working electrode having a first active area thereon, and performing a dip coating operation to deposit a bilayer membrane upon the first working electrode and the first active area. The bilayer may include an inner layer having a first membrane polymer and an outer layer having a second membrane polymer, the first membrane polymer and the second membrane polymer differing from one another. The dip coating operation may comprise one or more first dips in a first membrane formulation to form the inner layer of the bilayer membrane and one or more second dips in a second membrane formulation to form the outer layer of the bilayer membrane upon the inner layer.

The invention relates to a cascade gating activated CRISPR / Cas12a system which is used for synchronous detection of miRNA155 and FEN1, the system comprises recognition assembly and enzyme digestion of two DNA hairpins, and Cas12a is activated by an enzyme digestion product to generate a fluorescence signal. The DNA hairpin 1 comprises a recognition part of miRNA155, a hidden foothold of the DNA hairpin 2 and a part of Cas12a activation chain, and the DNA hairpin 2 comprises a part combined with the hidden foothold of the DNA hairpin 1 and a part of Cas12a activation chain. In the application, two DNA hairpins are combined and assembled under the catalysis of miRNA155 to be called as an FEN1 enzyme cutting substrate, a CRISPR / Cas12a system can be activated in a cascade gating manner, and signals are generated only when the two targets exist, so that the specificity synchronous detection of the two targets is completed.

The invention relates to the technical field of enzyme engineering, in particular to an aminopeptidase mutant, a preparation method of the aminopeptidase mutant and application of the aminopeptidase mutant to preparation of L-carnosine. The aminopeptidase mutant is obtained by mutating wild type aminopeptidase and can be used for preparing the L-carnosine through reverse hydrolysis of beta-alanine methyl ester and L-histidine. The L-carnosine produced by using the aminopeptidase mutant disclosed by the invention has the following advantages: 1, the conversion rate is greater than or equal to 95%, the conversion time is short, and relatively cheap alanine methyl ester can be used as a raw material to catalytically prepare the L-carnosine; 2, the production process is simple, the reaction conditions are mild, and the green chemistry concept is met;

The preparation method comprises the following steps: mixing a tannic acid solution and a tetravalent cerium salt solution, adjusting the pH value to 9-11, and reacting to obtain the cerium tannate nano-enzyme, the molar ratio of the tannic acid to the tetravalent cerium salt is (1-4): 1; the hydration dynamic diameter of the cerium tannate nano-enzyme is 20 to 80 nm. The TA-Ce nano-enzyme prepared by the preparation method disclosed by the invention is smaller in particle size, can keep stable and good dispersibility in a physiological environment, and has good biocompatibility, low hemolysis rate and small systemic toxicity; meanwhile, the compound has good kidney targeting property, can be effectively enriched into the injured kidney, eliminates various ROS (reactive oxygen species) and relieves oxidative stress by utilizing unique redox cycle between Ce < 3 + > and Ce < 4 + > states, and has a good effect in treating acute kidney injury induced by rhabdomyolysis and / or cis-platinum. The research lays a foundation for the metal polyphenol nano material in the treatment of acute kidney injury.

The invention discloses a nano-enzyme as well as a preparation method and application thereof, and relates to the technical field of nano-enzymes. According to the preparation method, natural polyphenol, a cerium source and polyethylene glycol are mixed to prepare the novel polyphenol nano-enzyme, the preparation method is simple and easy to implement, the preparation cost is low, the prepared nano-enzyme has good biocompatibility, and a new way is provided for effective treatment of inflammatory diseases.

The invention belongs to the field of molecular biology and immunology, and particularly relates to an anti-CD3 nano antibody capable of activating the activity of T cells as well as a preparation method and application of the anti-CD3 nano antibody. The invention provides a preparation method of the anti-CD3 nano antibody. The preparation method comprises the following steps: constructing a CD3 plasmid and an overexpression cell line, and immunizing alpaca to obtain peripheral blood mononuclear cells (PBMC) of the alpaca; extracting RNA (Ribonucleic Acid) and carrying out reverse transcription to obtain cDNA; carrying out PCR amplification on the antibody gene segment and cloning the antibody gene segment to a phage display vector; and electrically transforming the vector into SS320 competent cells to obtain the nano antibody library. After two rounds of panning, detecting and screening an antibody clone capable of being combined with the CD3 protein through ELISA (Enzyme-Linked Immunosorbent Assay); an antibody sequence is selected through sequencing, and an antibody protein is expressed by using a mammalian cell high-throughput expression system; and then detecting and screening the antibody which is only specifically combined with the CD3. The method disclosed by the invention is simple, convenient and rapid to operate, and can be used for preparing active antibodies for treating potential tumors and proliferative diseases.

The invention relates to the technical field of nano material application. The invention provides a ruthenium-doped Prussian blue nano-enzyme and a preparation method thereof. The ruthenium-doped Prussian blue nano-enzyme is obtained by reacting metal ruthenium with Prussian blue nano-particles at normal temperature. The invention further provides a nano-enzyme library containing the ruthenium-doped Prussian blue nano-enzyme and a construction method of the nano-enzyme library. The invention also provides the SNP-PB nano-enzyme obtained by loading sodium nitroprusside on the ruthenium-doped Prussian blue nano-enzyme. The ruthenium-doped Prussian blue nano-enzyme and the SNP-PB nano-enzyme both have good antibacterial and anti-inflammatory performance, after NO gas release capacity is given, the problem of resistance of a biological membrane is better solved, then oxidative stress resistance and periodontal bacterial infection resistance are better exerted, and a new treatment method is provided for periodontal diseases.

The invention discloses a multi-layer controllable degradation coating composition as well as a preparation method and application thereof, and belongs to the field of biodegradable high polymer materials and coating compositions. The coating composition comprises an outer protective layer and an inner functional layer, and the inner functional layer is arranged between a substrate and the outer protective layer and comprises a plurality of microcapsules. The key innovation lies in that the microcapsule encapsulates an enzyme capable of catalytically degrading the outer protective layer and is configured to be released after a predetermined delay time. The pH buffer system is integrated in the inner functional layer, and an optimal catalytic microenvironment is created for released enzyme, so that the outer protective layer is actively and quickly degraded, and programmable time-controlled removal is realized. The problem that the degradation rate of an existing biodegradable material is passive and uncontrollable is solved, and the coating has a remarkable application value in the field of seed coating or fertilizer controlled release by endowing the coating with a unique two-stage degradation characteristic.

The invention provides recombinant escherichia coli with high yield of ergothioneine and a method for preparing ergothioneine, the recombinant escherichia coli co-expresses ergothioneine anabolic enzymes Egt1 and Egt2, the methyltransferase structural domain of the Egt1 is derived from trichoderma reesei, mycobacterium smegmatis EgtD or rhizopus nigricans, and the methyltransferase structural domain of the Egt2 is derived from the trichoderma reesei, the mycobacterium smegmatis EgtD or the rhizopus nigricans. The histidine sulfoxide synthase structural domain is derived from trichoderma reesei, schizosaccharomyces pombe or rhizopus nigricans, and the Egt2 is derived from claviceps purpurea, neurospora crassa or aspergillus niger. After the recombinant escherichia coli obtained by screening is cultured for 68 hours in a 5L fermentation tank, the yield of ergothioneine can reach 5g / L or above; wherein after recombinant escherichia coli co-expressing Egt1 enzyme (the structural domain of methyltransferase is derived from mycobacterium smegmatis EgtD, and the structural domain of histidine sulfoxide synthase is derived from trichoderma reesei) and Egt2 enzyme (derived from neurospora crassa) are cultured in a 5L fermentation tank for 68 hours, the yield of ergothioneine can reach 8.7 g / L, and the production intensity reaches 127.9 mg / (L.h).

The invention discloses a method for large-scale production of rare ginsenoside Rk3. According to the invention, wild type beta-glucosidase derived from Sulfobus acidocaldarius is modified, so that the beta-glucosidase has efficient enzymatic activity of catalyzing the conversion of ginsenoside Rg1 into ginsenoside Rh1, and meanwhile, the heat-resistant and acid-resistant characteristics of the enzyme are retained, so that the ginsenoside Rg1 can be converted into ginsenoside Rk3 by adopting a one-pot method. The technology for large-scale production of ginsenoside Rk3 is obtained through optimization, and a foundation is laid for large-scale application of ginsenoside Rk3 in the market.

The invention discloses a method and a system for adjusting a tea leaf fixation process and a medium, and aims to realize cooperative regulation and control of enzyme activity and aroma substances in a fixation process. The method comprises the following steps: collecting multi-modal sensing information in a fixation process, and carrying out feature extraction on the multi-modal sensing information to obtain multi-dimensional features; dynamically constructing an LSTM prediction model based on the multi-dimensional features, and outputting an enzyme activity prediction value sequence; the enzyme activity predicted value sequence is input into a fuzzy PID controller for reinforcement learning optimization, a temperature-rotating speed adjusting instruction is generated, and adjusting parameters of the controller are dynamically updated through a reward function of the chlorophyll retention rate and the energy consumption ratio; according to the temperature-rotating speed adjusting instruction, adjusting the hot air temperature of each temperature area of the enzyme deactivation chamber, the rotating speed of the enzyme deactivation cylinder and the axial segmentation wind speed; and collecting fixation end point judgment information, and terminating fixation when a preset condition is met. According to the invention, the problems of unstable quality and overhigh energy consumption caused by parameter solidification in the traditional enzyme deactivation process are solved, and dynamic optimization and accurate control of the enzyme deactivation process are realized.

The invention belongs to the technical field of medical examination, and particularly relates to preparation and application of a polymerase-labeled secondary antibody. The preparation method comprises the following steps: activating enzyme by divinyl sulfone (DVS), polymerizing with hyperbranched aminated polyethylene glycol to obtain polymer enzyme, activating the polymer enzyme again, and incubating with a second antibody to obtain a polymer-labeled second antibody. According to the method, the main defects of the secondary antibody in the current market are overcome, the enzyme label density on the antibody is improved, and the number of enzyme molecules labeled on the secondary antibody in unit volume is greatly increased; the volume of the polymer enzyme is reduced; the cell penetrability of the enzyme-labeled secondary antibody is improved, and the sensitivity and the stability are higher.

The invention discloses a formate dehydrogenase mutant and application thereof in catalytic synthesis of D-mannitol, and belongs to the technical field of bioengineering. According to the formate dehydrogenase mutant provided by the invention, the 18th-site glutamic acid of wild-type formate dehydrogenase with an amino acid sequence shown as SEQ ID NO.2 is mutated into proline, the 57th-site asparagine is mutated into glutamic acid, the 70th-site histidine is mutated into tryptophan, the 235th-site serine is mutated into threonine, and the 316th-site valine is mutated into threonine, so that the formate dehydrogenase mutant is high in enzyme activity and catalytic activity; the coenzyme circulation efficiency during the production of D-mannitol can be obviously improved, so that the yield of D-mannitol can be effectively improved. When the genetically engineered bacterium co-expressed by the formate dehydrogenase mutant and the mannitol dehydrogenase mutant is used for catalyzing 150 g / L fructose to produce D-mannitol, the yield of D-mannitol within 15 h can reach 140 g / L or above.

The present invention belongs to the field of biopharmaceutical technology, and specifically relates to a method for preparing GLP-1 or its analogs. The present invention provides a leader peptide, and a fusion protein comprising the leader peptide. The leader peptide or fusion protein is used to prepare a semaglutide precursor polypeptide, and the results show that the yield of the prokaryotic expression fusion protein can reach 26g / L, and the purity of the precursor polypeptide obtained after enzyme cleavage is higher than 95%. The precursor polypeptide is further side-chain modified and connected with a dipeptide to efficiently prepare semaglutide.

The invention belongs to the field of nano-drug preparations, and discloses a multifunctional nano-enzyme platform and a preparation method and application thereof, and the multifunctional nano-enzyme platform is mainly composed of a nano-enzyme, a carrier material and a targeting substance. The synthesis of the nano-enzyme, the loading of the nano-enzyme on a carrier material and the modification of a targeting substance on the surface of the nano-enzyme are carried out through methods such as stirring synthesis, coprecipitation, strong acid oxidation, chemical reduction, covalent binding, a solvothermal method and physical adsorption. The targeting substance on the surface of the nano-enzyme platform can be accurately and efficiently positioned to an inflammation part, the meta-acid microenvironment of the inflammation part enables the targeting substance and the carrier material to be subjected to structural depolymerization, so that various nano-enzymes are released, and the nano-enzymes not only realize self-cascade and synergistic catalysis enhancement effects of the enzymes at the inflammation part, but also can be used for preparing the nano-enzyme. And a substrate self-circulation catalysis system is also formed, so that the effect of efficiently relieving and treating various inflammation-related diseases is achieved.