38 results about "Coupling enzyme" patented technology

Isolated and/or recombinant enzymes that include surface binding domains, surfaces with active enzymes bound to them and methods of coupling enzymes to surfaces are provided. Enzymes can include large and/or multiple surface coupling domains for surface coupling.

The invention provides a human proinsulin chemiluminescence immunoassay kit and a preparation method thereof. The kit provided by the invention comprises an anti-proinsulin reagent 1, an anti-proinsulin reagent 2, a magnetic separation reagent, a proinsulin calibrator, a chemiluminescence substrate, and a washing liquid. The invention also provides a preparation method of the kit. The preparation method comprises the following steps: coupling antibody and biotin, preparing the anti-proinsulin reagent 1, coupling enzyme and antibody, preparing the anti-proinsulin reagent 2, preparing the magnetic separation reagent, preparing the proinsulin correction sample, and preparing the chemiluminescence substrate. Compared to the proinsulin kit existing in the present market, the kit provided by the invention has the characteristics of high detection sensitivity, good repeatability, and low cost.

The invention relates to a method for extracting, separating and purifying four main anthocyanins from the blackcurrant residue. The object is to provide a simple, economic, green, eco-friendly, and high-yield method for extracting, separating and purifying four main high-purity anthocyanins from the blackcurrant residue. According to the method, the blackcurrant waste residue produced from industrial production is taken as a raw material, and homogenized pulverized coupling enzyme hydrolysis technology, negative-pressure cavitation reinforced extraction technology, macroporous adsorption resin concentration technology, medium-pressure quick-flash reverse chromatographic separation technology and low-temperature crystallization and recrystallization technology are adopted to obtain four anthocyanin monomer ingredients including delphinidin 3-O-glucoside, delphinidin 3-O-rutinoside, cyanidin 3-O-glucoside, and cyanidin 3-O-rutinoside and having the purity reach 95%. The method is abundant and available in raw materials, short in time consumption, less in solvent consumption, high in purity of obtained anthocyanins, low in production cost, and suitable for massive industrial production.

The invention discloses an ELISA (enzyme linked immunosorbent assay) kit for porcine circovirus type 2 (PVC2) antibody detection. The kit includes an antibody detection board, a sample diluting solution, an enzyme-labeled antigen, a washing solution, a substrate color developing solution, a stop solution, a positive serum and a negative serum. The Cap protein coated with antibody detection board of the kit is PCV2 eukaryotic expression nucleocapsid Cap protein, and the non-specific reactions are greatly reduced by using the Cap protein coupled enzyme-labeled antigen, so that the antibody detection accuracy is improved and the kit is more favorable for large-scale popularization and use.

A sterilization indicator system and method of using the system to determine efficacy of a sterilization process. The system includes a vial having a first compartment containing spores of one or more species of microorganism; a second compartment containing a growth medium with a disaccharide, an oligosaccharide or a polysaccharide in which the vial is free of monosaccharide; an enzyme, capable of acting upon the monosaccharide to yield reaction products and electron transfer, disposed on two or more electrodes adapted to carry an electrical signal resulting from the electron transfer, the pair of electrodes positioned to contact the combined contents of the first compartment and the second compartment during incubation; and an apparatus linked or linkable to the electrodes and adapted to detect and measure the electrical signal resulting from electron transfer when the enzyme acts upon the monosaccharide.

The invention relates to an enzymatic activity test method using combining integration and initial velocity method, which is characterized in that the classical initial velocity is measured when the concentration of initial substrate is 70% higher than that of the preset substrate, and the classical initial velocity is still within linearity range; when the substrate consumption proportion in 80% record period is higher than the lower limit required in integration, the fitting reaction curve of integral velocity equation is used to determine the maximum reaction velocity using reaction time as independent variable, then the maximum reaction velocity is calculated as the initial velocity with preset substrate concentration is 93%; meanwhile, the slope of classical initial velocity and computation initial velocity to the enzyme amount reaction curve; when the substrate consumption proportion of the coupling enzyme reaction system in 80% record period is higher than the lower limit required in integration, the fitting enzyme reaction curve is used to determine the initial velocity using corresponding numerical integral velocity equation, otherwise the classical initial velocity is directly measured; total time for the optimization monitoring reaction guarantees the combination of the two methods.

The invention discloses a method catalytically synthesizing (S)-N, N-dimethyl-3-hydroxy-(2-thiofuran)-1-propylamine((S)-DHTP) by aldehyde ketone reductase recombinant strain crude enzyme system, and belongs to the technical field of biological catalysis asymmetric conversion. An auxiliary substrate and original bulk coenzyme NADP<+> are added in the recombinant strain crude enzyme system to accelerate regeneration cycle of coenzyme NADPH in the system; the reaction substrate is N, N-dimethyl-3-ketone-(2-thiophene)-1-propylamine (DTKP); the recombinant strain is E. coliBL21(DE3)(pETCPAR4); aldehyde ketone reducing enzyme gene cpar4 comes from Candida parapsilosis; the gene coded aldehyde ketone reducing enzyme gene CPAR4 catalytically and asymmetrically reduces the DKTP into (S)-DHTP. The method utilizes the cell-free system to conduct catalytic reaction; coupling enzyme required by regeneration of the coenzyme is not extra added in the reaction system; direct acting efficiency of the enzyme and the substrate is improved; reaction time is shortened; the conversion effect is relatively good.

The invention discloses a method for preparing (R)-phenylglycol from SD-AS sequence coupled (R)-carbonyl reductase and glucose dehydrogenase and belongs to the technical field of biological catalytic asymmetric transformation. The invention provides recombinant escherichia coli E. coli RIL/pET-R-SD-AS-G. The recombinant escherichia coli E. coli RIL/pET-R-SD-AS-G is preserved in the China center for type culture collection (CCTCC) and has a preservation number of CCTCC NO: M2015170. R-carbonyl reductase and glucose dehydrogenase are coupled by a coexpression way, and the coupled enzyme, a cultured recombinant strain and 2-hydroxyacetophenone as a substrate undergo a catalytic asymmetric transformation reaction under optimized biotransformation reaction conditions to produce (R)-phenylglycol with optical purity of 100% and a yield of 99.9%. The method utilizes a SD-AS sequence high efficiency coupling double-enzyme technology, solves the problem of limitation of chiral catalytic reaction coenzyme regeneration cycle and provides an effective approach for high efficiency bio-preparation of the (R)-phenylglycol.

The present invention relates to a composition comprising a coupled enzyme system for the rapid and efficient production of hydrogen peroxide by the coupling of a first enzyme system capable of hydrogen peroxide generation, to a second enzyme system which utilizes the non hydrogen peroxide product of the first enzyme system, and optionally is capable of generating further hydrogen peroxide.

A method for producing chitosan oligosaccharides coupled with enzymatic degradation of chitosan and membrane separation is characterized in that the production process is as follows: (1) preparing chitosan oligosaccharides by enzymatic degradation of chitosan; (2) adopting hollow fiber or flat ultra- Combine the filter with the degradation reaction to separate the active oligosaccharides in situ; (3) filter the ultrafiltrate with a nanofilter to separate high-concentration active chitosan oligosaccharides. The invention realizes the integration of reaction and separation, and prevents secondary degradation of active oligosaccharides. Since the process realizes continuous operation, the degrading enzyme can be used continuously, the yield is improved, the cost is reduced, and it is beneficial to realize industrialization.

An economical and expedient method is disclosed for the preparation of alpha-hydroxy-carboxylic acids or salts thereof in very high enantiomeric purity which comprises oxidizing a corresponding alpha-amino-carboxylic acid or salt thereof using an amino acid deaminase followed by reducing the corresponding alpha-keto-carboxylic acid or salt produced using a D- or L-lactate dehydrogenase in the combination with an electron donor and an enzyme/substrate system for recycling the electron donor. The resulting alpha-hydroxy-carboxylic acids, hydrates, and salts thereof are valuable components and intermediates in the preparation of chiral compounds, especially pharmaceuticals. This invention also relates to the use of alpha-amino-carboxylic acids, hydrates, and salts thereof and a coupled enzyme system in the production of alpha-hydroxy-carboxylic acids, hydrates, and salts thereof.

The present invention relates generally to enzyme assays and more particularly to a rapid, sensitive, reliable and robust homogeneous assay for acetyl CoA carboxylase activity comprising a coupled enzyme assay in a scintillation proximity format suitable for high-throughput screening. In one aspect, the assay couples ACC and FAS and the product, palmitic acid is then detected by scintillation counting. The invention also relates to the identification of modulators of ACC activity.

Bertrand(1982) hydrolytic generates trophicardyl from trophicardyl acid by nucleotidase, and gains H2O2 through the reaction of the enzyme1 and enzyme2. Use of H2O2 from Trinder's reaction to combine hydrogen provider3,5-double CL-2-hydroxide benzene sulphur acid and 4- to create a amaranth in the situation of the catalase. By watching the velocity of increase of the degree of the amaranth absorbing light at 510 nms to gain 5' nucleotidase activity. The invention uses the aniline compound ADOS, ADPS, ALPS, TODB, TOOS and TOPS as hydrogen provider for Trinder's reaction and increases the reaction ability. The invention also involves a reagent box used for implementing the method mentioned above.

The present invention relates to coupled enzyme assays. In particular, the present invention provides a coupled fluorescent assay for detection of S-adenosylmethionine (AdoMet)-dependent methyltransferase activity.

The invention discloses a quantitative determination method of cardiac myosin binding protein C. According to the method, firstly, antibody coupling super paramagnetic micro particles and samples to be tested take specific combination reaction to obtain a first reactant, wherein the antibody coupling super paramagnetic micro particles are products obtained by coupling super paramagnetic micro particles and anti-cMyBP-C monoclonal antibodies A; then, the first reactant is taken to perform specific combination reaction with enzyme-labeled articles to obtain a second reactant, wherein the enzyme-labeled articles are products obtained by coupling enzymes and anti-cMyBP-C monoclonal antibodies B; finally, chemiluminescence substrates and the second reactant are taken to take enzymatic reaction;luminous signals are determined; the concentration value of cMyBP-C is obtained. The technology mode of the invention is a sandwich method. Compared with a cMyBP-C ELISA kit, the detection method andthe prepared kit have the advantages that the detection sensitivity and the linear range can be greatly improved. The technology evaluation on the prepared kit shows that the analysis sensitivity is0.05 ng/ml; the detection range is 0.1 to 50 ng/ml; high correlation and coincidence rate to clinic AMI patients are high.

The invention discloses a high-specificity coupling enzyme YND mutant of 3', 5'-adenosine diphosphate based on molecular docking, the gene sequence of the YND mutant is SEQ ID NO.2, SEQ ID NO.3 or SEQ ID NO.4, and the amino acid sequence of the YND mutant is SEQ ID NO.6, SEQ ID NO.7 or SEQ ID NO.8. The invention designs and modifies a coupling enzyme with strong PAP specificity by taking yeast 3 ', 5'-adenosine diphosphate enzyme with a known crystal structure as a starting point. The modified enzyme is subjected to inducible expression by an escherichia coli expression system and stored in a storage solution, which can maintain activity for 3 months at-80 DEG C, modified enzyme is used by being diluted with a corresponding buffer system, the use is convenient and the reaction system can be replaced, and the problem of limitation of a system buffer substance on ST activity detection is solved.