38 results about "Coupling enzyme" patented technology

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.

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 and S-adenosylhomocysteine hydrolase enzyme activity.

The invention discloses a method for preparing pharmaceutical dextran and levulose by bi-enzyme immobilization coupled multistage membrane separation, which comprises the following steps: co-immobilizing dextran sucrase and dextranase in sodium alginate microspheres to obtain bi-enzyme-immobilized spheres; adding the bi-enzyme-immobilized spheres into a sucrose solution used as a substrate to form an immobilized bi-enzyme / sucrose reaction system; and after the reaction product is separated by a multistage membrane system composed of hyperfiltration membranes and a nanofiltration membranes, directly obtaining the pharmaceutical dextran in the hyperfiltration membrane separation stage, and obtaining the high-purity levulose solution in the last nanofiltration membrane separation stage, wherein the enzymes and macromolecular dextran contained in the trapped solution of the first-stage hyperfiltration membrane return to the immobilized bi-enzyme / sucrose reaction system to participate in the reaction again, thereby implementing cyclic utilization of the enzymes. The invention has the advantages of lower cost, environment friendliness, high safety and high efficiency, is simple to operate, implements oriented adjustable production of high-quality pharmaceutical dextran with target molecular weight, and obtains the high-purity high-added-value levulose byproduct.

Aldehyde and ketone reductase crude enzyme system catalytic synthesis of recombinant bacteria ( S )‑N,N‑Dimethyl‑3‑hydroxy‑3‑(2‑thiophene)‑1‑propylamine (( S )‑DHTP), which belongs to the technical field of biocatalytic asymmetric transformation. The present invention is to add auxiliary substrate and initial amount of coenzyme NADP in recombinant bacterial crude enzyme system + Promote the regeneration cycle of the coenzyme NADPH in the system, the reaction substrate is N,N-dimethyl-3-ketone-3-(2-thiophene)-1-propanamine (DTKP), and the recombinant bacteria express the aldehyde and ketone reductase gene of E. coli BL21(DE3)(pETCPAR4), aldehyde and ketone reductase gene cpar4 from Candida parapsilosis ( Candida parapsilosis ) CCTCC NO: M 203011, the gene encodes aldehyde and ketone reductase CPAR4, which catalyzes the asymmetric reduction of DKTP to ( S )‑DHTP. The invention utilizes the cell-free system to catalyze the reaction without additionally adding coupling enzymes required for coenzyme regeneration in the reaction system, improves the efficiency of the direct action of the enzyme and the substrate, shortens the reaction time, and obtains better conversion effects.

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.

A sterilization indicator system and method of determining the efficacy of a sterilization treatment using the system. The system comprises a bottle comprising a first compartment containing one or more microbial spores; a second compartment containing a growth medium comprising disaccharides, oligosaccharides or polysaccharides, wherein the bottle Contains no monosaccharides; enzymes capable of acting on said monosaccharides to produce reaction products and electron transfer, said enzymes being disposed on two or more electrodes adapted to carry the electrical signal of the electrode pair positioned to contact the contents of the merged first and second compartments during incubation; and a device connected or connectable to the electrodes and adapted to The electrical signal generated by electron transfer when the enzyme acts on monosaccharides is detected and measured.

The invention discloses a device and method for gradient concentration by a coupling enzyme-membrane method using residual heat for mechanical supercharging. The device comprises an ultra filtration membrane filtering unit, a reverse osmosis membrane filtering unit and a mechanical supercharging concentrating unit, wherein the ultra filtration membrane filtering unit comprises a balancing barrel, a first enzymatic hydrolysis tank and an original ultra filtration membrane which communicate each other through a pipeline. The device disclosed by the invention can concentrate raw material soup under low energy consumption level by level so as to obtain a concentrated solution of different concentrations; the first enzymatic hydrolysis tank is additionally arranged in the ultra filtration membrane filtering unit, so that protein in the raw material soup can be separated through membranes with different bore diameters so as to obtain soup in a target molecular weight segment, and the characteristic attributes of products are improved; mechanical supercharging concentration is performed, so that the material soup with a high concentration is obtained. Through the device and the application method disclosed by the invention, the target concentrated solution with different concentrations and different molecular weights can be obtained, the original flavor of materials is well maintained, a great number of energy resources are saved, and the method and the method are particularly suitable for concentration of bone soup.

The invention discloses a high-efficiency polypeptide-polypeptide coupling system and method based on a disordered protein coupling enzyme. By designing and splitting the protein domain CnaB2 and optimizing it, a three-component system is obtained: spy tag (SpyTag), BD tag (BDTag) and spy stapler enzyme (SpyStapler), so that SpyStapler can catalyze the coupling between SpyTag and BDTag Linkage reaction, and prepare different fusion proteins containing SpyTag and BDTag two tags and carry out coupling or cyclization reaction. Based on the polypeptide-polypeptide coupling system of the present invention, an active protein with high purity and stable function can be obtained by means of gene coding; the spy stapler enzyme coupling spy label and BD label have high reaction efficiency, and some of the staples are stapled after the reaction The mechanical enzyme falls off, and the coupling part of the obtained cyclization product has a smaller molecular weight. The protein tag of the present invention is applicable to various functional proteins and can be fused and expressed at any position of the protein.

The present invention relates to genetically modified enzymes obtained by rational design of the active site binding pocket of the prototypic enzyme 4-hydroxyphenylacetate 3-hydroxylase (4HPA3H) for hydroxylating a 4-hydroxyphenyl compound to yield a 3,4-dihydroxyphenyl compound and to biotechnological methods including in vivo and in vitro methods using said enzymes or catalytically active fragments thereof. Further provided is a method either using a suitable oxidase or hydroxylase further enabling the subsequent site specific methylation of the 3,4-dihydroxyphenyl compound in a coupled enzymatic reaction by providing a suitable O-methyltransferase. Finally, compositions obtainable by the aforementioned methods are disclosed.