55results about How to "High enzyme loading" patented technology

A multilayer enzyme immobilization process is provided comprising adsorbing a polyethyleneimine (PEI) solution in a fibrous matrix, and adding an enzyme to the fibrous matrix, which comprises a plurality of fibrils. The process further comprises forming at least two layers of PEI-enzyme aggregates on the fibrils, and cross-linking the multilayer PEI-enzyme aggregates. The process can further comprise washing the fibrils containing the cross-linked PEI-enzyme aggregates with distilled water and acetic acid buffer subsequent to cross-linking. However, the PEI-containing matrix is not washed prior to the addition of enzyme. The enzyme can be β-galactosidase and the fibrous matrix can be cotton cloth. The multilayer immobilized enzyme can be employed in a biocatalyst reactor for production of galacto-oligosaccharides from lactose and the hydrolysis of lactose to glucose and galactose.

The invention belongs to the field of biological catalysis, and particularly relates to a novel immobilized enzyme preparation, and a preparation method and application thereof. The preparation method of the novel immobilized enzyme preparation comprises the following steps: combining nano magnetic chitosan microspheres and a metal-organic framework material having so many properties such as multiple pores, large specific area, multiple metal sites and the like through graft copolymerization to obtain a nano magnetic chitosan-metal organic framework material having a core-shell structure with a porous network structure; and mixing the nano magnetic chitosan-metal organic framework material used as a carrier and an enzyme solution, and immobilizing at 0-25 DEG C for 1-12 hours to obtain the novel immobilized enzyme preparation. The method can immobilize enzymes such as lipase, proteinase, peroxidase and the like, and has the advantages of low cost, high immobilization efficiency, favorable enzyme stability, high enzyme loading amount and high enzyme recovery rate; no bifunctional reagent is used in the immobilization process; and the method is simple to operate, low in cost and mild in reaction.

The invention provides a transaminase immobilization method. The method comprises the following steps: oxidizing a hydroxyl group on polyvinyl alcohol sponge, adding transaminase to convert a carbonyl group into an amino group, flocculating, and carrying out cross-linking immobilization by using polyaldehyde to obtain immobilized transaminase. The immobilization method has the advantages of high enzyme activity unit and enzyme purification.

The invention relates to a grease catalyzing and separating biphasic enzyme-membrane reactor and a method for preparing and using hydrolyzed edible glyceride. The method comprises the steps of filling polyphosphazene superfine fiber complex film into a mould with an inner and an outer cylinders to form a roll membrane device; sequentially injecting activating agent solution and lipase solution into the membrane device, and causing the lipase to be chemically fixed on the surface of the superfine fiber film to obtain an enzyme-membrane reactor; and injecting edible glyceride and phosphate buffer respectively into the inner side and the outer side of the superfine fiber film to conduct hydrolysis reaction on the surface of the superfine fiber surface. The reactor can be used for catalyzing the hydrolysis of the glyceride, and for producing diglyceride or monoglyceride edible oil and extracting functional lipid by treating lard or fish oil. The glyceride hydrolyzate accounts for more than 85%. The grease catalyzing and separating biphasic enzyme-membrane reactor is designed by combining efficient biocatalysis and complex film separation, and has the advantages of repeatable use, realized continuous efficient production, reduced cost, saved resource and good development prospect.

The invention relates to a preparation method for polyacrylonitrile nanofiber membrane immobilized enzyme. The preparation method comprises the following steps of dissolving a polyacrylonitrile polymer into N,N-dimethylformamide and preparing a polyacrylonitrile nanofiber membrane through electrostatic spinning; then soaking the polyacrylonitrile nanofiber membrane into an absolute alcohol solution, aerating hydrogen chloride gas for 10-30 minutes, and fixing enzyme into the nanofiber membrane through chemical bonding. According to the preparation method, high voidage and high mechanical strength of the polyacrylonitrile nanofiber membrane are utilized for preparation of the nanofiber membrane immobilized enzyme which has high enzyme content, is easy to recycle from a reaction system and can be used repeatedly. The immobilized zymophore can be used for increasing the use ratio of enzyme and lowering the production cost, and has the advantage of simple preparation processes, and the like.

The invention discloses a preparation method of immobilized lipase used for grease hydrolysis, comprising the following steps of: dissolving cellulose in a dimethyl acetamide/acetone mixed solution to prepare a uniform solution which is rested for removing bubbles to obtain a spinning solution; then preparing cellulose superfine fibrous membrane by electrostatic spinning; placing the cellulose superfine fibrous membrane in the ethanol solution of potassium hydroxide for hydrolysis to generate hydroxyl group which is activated after being washed for a plurality of times by de-ionized water; and then placing the activated cellulose superfine fibrous membrane in an enzyme solution, and obtaining the immobilized lipase of the cellulose superfine fibrous membrane after reaction for a plurality of hours. The prepared immobilized lipase can be used for grease hydrolysis. The method adopts the cellulose superfine fibrous membrane and has the advantages of simple and fast technology, good repeatability, low cost and safe operation and being environment-friendly and suitable for large-scale industrialized production, furthermore, the prepared immobilized lipase has large enzyme-loading capacity, and catalytic efficiency for grease hydrolysis is high.

The invention relates to a preparation method of a block polymer immobilized enzyme. The method comprises the following steps of dissolving polymethyl methacrylate-ethyl acrylate block polymer into N,N-dimethylformamide and tetrahydrofuran which are in a certain mass ratio; preparing polymethyl methacrylate-ethyl acrylate nano fiber film through an electrostatic spinning method; with the polymethyl methacrylate- ethyl acrylate nano fiber film as an immobilized carrier of the enzyme, and fixing the enzyme onto the nano fiber film through a method for ammonificating polyethylenimine and activating glutaraldehyde. By utilizing the high specific surface area and high void ratio of the nano fiber film, the nano fiber film immobilized enzyme with high enzyme carrying capacity and high catalyzing efficiency can be obtained. The immobilized enzyme has the advantages that the immobilized enzyme is easily recovered from a reaction system and can be reutilized for multiple times, the utilization rate of the enzyme can be increased, the production cost can be reduced, and the preparation process is simple.

The invention provides a method of preparing linoleate isomerase micro-capsules on the basis of a polyelectrolyte layer-by-layer self-assembly technology and can solve a problem that free linoleate isomerase is poor in tolerance, is poor in stability and cannot be recycled in industrial production. The method of the invention includes the following steps: with an amorphous calcium carbonate crystal, which is obtained through co-precipitation of calcium carbonate and the linoleate isomerase, as a template and with polyallylamine hydrochloride and sodium polystyrenesulfonate as coating materials, preparing the linoleate isomerase micro-capsules through the polyelectrolyte layer-by-layer self-assembly technology. The micro-capsule is high in activity, excellent in stability and long in half life.

The invention discloses an enzyme biofuel cell for generating electricity from trehalose produced by Escherichia coli in sewage. The invention successfully prepares a two-chamber enzyme biofuel cell capable of generating electricity by using trehalose from sewage by immobilizing engineering Escherichia coli containing trehalose hydrolase TreA and glucose oxidase GOx after whole-cell permeability treatment on an anode carbon felt and immobilizing bilirubin oxidase on a cathode carbon felt. The fuel cell assembled by the invention has a maximum output power of up to 16.2. M u.W/cm2, and the output current density of the fuel cell can still be maintained at 72% of the initial maximum output value after 60 hours of continuous power generation under the condition that the raw material is sufficient. The method for preparing the fuel cell of the invention is simple and low in cost, and the reaction is carried out at room temperature, and the prepare enzyme electrode has good catalytic performance and good application prospect.

The invention discloses a method for producing high fructose syrup by immobilized glucose isomerase. The method comprises the following steps: immobilizing glucose isomerase in an ultra-filtration membrane with an asymmetric structure in a filtering mode to prepare a membrane bioreactor of the immobilized glucose isomerase; performing reaction on the glucose solution in batches or continuously by utilizing the membrane bioreactor to produce the high fructose syrup. According to the method, an enzyme catalysis process and a membrane separating process are integrated, so that the production process is simplified, and the activity and the stability of an enzyme are improved; meanwhile, an enzyme membrane reactor can be used repeatedly, solvent cleaning and batchwise feeding are avoided, the development direction of green chemistry is met, continuous production is realized, the production efficiency is improved, and reference is provided for industrial production of fructose.

The invention provides a poly(methacrylic acid-itaconic acid) modified membrane, as well as a preparation method and application thereof. The preparation method comprises the following steps: mixing poly(methacrylic acid-itaconic acid)(IA-MAA), polyvinylpyrrolidone (PVP) and N,N-dimethyl formamide to obtain a casting solution; reacting the casting solution in a sealed condition at 55-65 DEG C for40-50 hours, defoaming, and preparing a flat membrane by adopting a tape casting method; soaking the flat membrane with the casting solution in a gel bath, and curing to form a membrane. The poly(methacrylic acid-itaconic acid) modified membrane prepared by a conversion method by using immobilized carrier as a phase, lipase is efficiently immobilized by modifying chitosan and activating glutaraldehyde. The method is simple in operation, the material has stable performance, the selectivity of transesterification can be greatly improved. The poly(methacrylic acid-itaconic acid) modified membranehas excellent industrial application prospect.

The invention relates to a preparation method of a laccase/multihole nanometer titanium dioxide modified electrode. The preparation method comprises the steps of material preparation, preparation for combine use of hydrothermal method and calcination method of a multihole nanometer titanium dioxide carrier, adsorption and immobilization of laccase on the carrier and electrode modification. Immobilized laccase is uniformly scattered in Nafion dilute solution so as to modify a glassy carbon electrode which is used as a work electrode, so that a three-electrode system is established, and the three-electrode system is connected with an electrochemistry work station so as to detect the electrochemistry response of the three-electrode system on water pollutant hydroquinone through cyclic voltammetry. Compared with the prior art, the method disclosed by the invention has the advantages that the cost is low, the preparation is simple, the enzyme activity of the immobilized laccase is high, the sensibility is high, and the like.

The invention provides a method for immobilizing lipase by block polymer micelle. The method comprises the following steps: dithiodipropionic acid is activated by an activator to activate a carboxyl group and reacted with hyaluronic acid to prepare an intermediate, the intermediate is activated by the activator to react with polylactic acid-glycolic acid to obtain a block polymer, and the block polymer is dissolved in deionized water to prepare a block polymer micelle solution; the lipase powder is dissolved in a phosphate buffered saline solution of pH being 8.0, and the material is stirred and centrifuged, a supernatant is transferred to a volumetric flask, and a volume is metered with a buffer solution of pH being 8.0 to obtain enzymatically synthesized cyhalofoprin lipase; the polymermicelle solution is mixed with the enzymatically synthesized cyhalofoprin lipase, a mixture is filtered, deionized water is added for dissolving, and washed with the buffer solution of pH being 8.0 toremove excess lipase, a volume is metered to the 1L of a phosphate buffer solution of the pH being 8.0, and lipase immobilized by block polymer micelle can be obtained after freeze drying. The process of the invention is simple.