131results about "Multi-enzyme systems" patented technology

The invention provides a preparation method and application of a bacterium-enzyme-magnetic nano-cluster composite detoxicating agent, the detoxicating agent takes a Fe3O4-HAP nano-cluster as a magnetic core, the surface of the detoxicating agent is subjected to functional modification, a composite flora composed of bacillus amyloliquefaciens, lactobacillus plantarum and saccharomyces cerevisiae is loaded to form sclerotia, a chitosan shell layer is constructed on the outermost layer, and the bacterium-enzyme-magnetic nano-cluster composite detoxicating agent is prepared. And a compound enzyme consisting of aflatoxin Bdegrading enzyme, zearalenone hydrolase and deoxynivalenol invertase is immobilized. The carrier can efficiently and synchronously degrade aflatoxin, vomitoxin and zearalenone, and can realize rapid separation and recovery through an external magnetic field.

The invention belongs to the technical field of soil organic matter improvers, and discloses a soil organic matter improver based on a compound microbial agent and a preparation method thereof, and the soil organic matter improver comprises the following components: a compound enzyme, a modified chitosan microsphere complex, a modified polyelectrolyte, an inhibitor L-methionine sulfoximine, a substrate, an auxiliary factor and the compound microbial agent. A compound enzyme system is used as a core, soil ammonium ions are circularly and efficiently assimilated through glutamine synthetase / glutamate synthetase, and the soil ammonium ions are converted into organic nitrogen; and energy guarantee is provided by virtue of polyphosphate kinase. The modified chitosan microsphere complex is used for protecting and slowly releasing enzyme and a substrate, and controlled release of the inhibitor L-methionine sulfoximine is realized through the modified polyelectrolyte, so that carbon-nitrogen balance of soil is regulated and controlled. The system has a synergistic effect with a compound microbial agent, the problem of soil organic matter reduction caused by excessive nitrogen application is effectively solved, meanwhile, the crop root biomass is remarkably increased, and the soil organic matter content is increased.

The invention discloses a method for preparing artificial bear gall powder through double enzymes, the artificial bear gall powder and application of the method, and belongs to the technical field of biology. The method comprises the following steps: respectively immobilizing two enzymes, and filling the two immobilized enzymes into a column or a reactor according to a ratio. The method can reasonably and effectively control the proportion of the amount of two enzymes participating in the reaction, so that the enzymatic reaction is effectively controlled, and the stability and consistency of the batch-to-batch conversion reaction are ensured. And after the enzymatic reaction is finished, the immobilized enzyme can be recycled in manners of centrifugation, filtration, magnetic separation and the like and is fixed according to the enzymatic reaction requirement, so that the immobilized enzyme can be recycled for multiple times, the use amount of the enzyme is saved, and the use efficiency of the enzyme is improved. The method provided by the invention is strong in operability, good in repeatability and easy in linear amplification, the ratio of TUDCA to TCDCA in the obtained product can reach 1.33 or above, and the method is suitable for industrial production of artificial bear gall powder.

The invention relates to the technical field of red algae polysaccharide extraction, and provides a preparation technology for preparing instant red algae polysaccharide through a compound enzyme method, and the preparation technology comprises the following steps: S100, washing a red algae raw material, and sequentially carrying out mechanical crushing treatment and freeze drying treatment to obtain red algae powder; s200, sequentially performing enzymolysis treatment, centrifugal treatment and filtration treatment on the red algae powder to obtain supernatant liquid; and S300, sequentially carrying out concentration treatment, purification treatment, spray drying treatment and fluidization granulation treatment on the supernatant to obtain the red algae polysaccharide. The red algae raw material is efficiently separated and purified by adopting the compound enzyme, so that the efficient degradation of the red algae cell wall and the efficient extraction and preparation of the red algae polysaccharide are realized, a new technical way is provided for the efficient preparation and wide application of the red algae polysaccharide, and the method has important industrial application value and market prospect.

The invention relates to a microneedle patch, a preparation method thereof and a blood glucose colorimetric detection method, and belongs to the technical field of biomedicine. The microneedle patch comprises a needle tip layer and a backing layer, the needle tip layer is a methacrylated natural biopolymer matrix in which immobilized enzyme is uniformly distributed, and the backing layer is a methacrylated natural biopolymer matrix in which a chromogenic substrate is uniformly distributed, the immobilized enzyme is composed of a zinc-based imidazoline metal organic framework MAF-7 carrier and a natural enzyme loaded in the framework. The immobilized enzyme with double-enzyme activity is loaded into the needle tip, the spatial distance of the cascade catalytic reaction is limited in the metal organic framework, the mass transfer efficiency is high, the reaction is rapid, the natural enzyme is encapsulated in the metal organic framework MAF-7, and the stability of the enzyme is enhanced; according to the method, color development is carried out by firstly extracting the interstitial fluid and then dropwise adding the color development substrate, the color development effect is obvious, and the time is shorter.

The invention discloses a functional microbial agent for purifying breeding sewage and a preparation method of the functional microbial agent, and belongs to the technical field of functional microbial materials. The microbial agent is composed of a core functional unit and a porous carrier according to a mass ratio of 3: 1 to 5: 1, the core functional unit comprises a symbiotic engineering flora and a compound enzyme system in a volume ratio of 7: 1, and the symbiotic engineering flora is compounded by sphingomonas aeruginosa, paenibacillus splendidus and acinetobacter hemolyticus according to a ratio of 1: 1: 1; the porous carrier is a low-cost porous carbon material prepared on the basis of an animal waste blood / iron complex. Through systematic cooperation of a specific functional flora, a compound enzyme system and a special porous carrier, synergism of adsorption protection and biodegradation is achieved, the stress resistance and purification efficiency of the microbial agent in an antibiotic and heavy metal combined pollution environment are remarkably improved, and the industrial problem that an aquaculture sewage biological treatment system is prone to collapse is particularly solved; meanwhile, waste is treated by waste, and the method is low in cost and suitable for large-scale application.

The invention discloses a Pickering IL emulsion microreactor constructed on the basis of temperature-sensitive double-enzyme Janus spheres and application of the Pickering IL emulsion microreactor to enhanced catalysis of conversion of CO2 into formic acid. A Janus emulsion template method is combined with a photocuring technology to synthesize a nano-scale dual-functional temperature-sensitive Janus sphere, one half of the nano-scale dual-functional temperature-sensitive Janus sphere is a hydrophilic hydrocarbon polymer, the other half of the nano-scale dual-functional temperature-sensitive Janus sphere is a hydrophobic fluorocarbon polymer, and rapid inversion of a Pickering emulsion can be realized by utilizing temperature sensitivity. The method comprises the following steps: preparing a Janus particle hydrophilic / hydrophobic hemisphere, selectively immobilizing formate dehydrogenase FDH and carbonic anhydrase CA on the Janus particle hydrophilic / hydrophobic hemisphere to obtain a temperature-sensitive FDH-Janus-CA composite sphere, introducing a fluorine-functionalized ionic liquid to regulate and control the internal microenvironment of a microreactor, and constructing a Pickering IL emulsion microreactor. The CO2 absorption capacity is increased by 47 times compared with that of a pure water system, the formic acid yield reaches 27.5 mM and is higher than the maximum value 22.8 mM of current enzyme electrocatalysis, the storage and catalysis stability is good, and an efficient and sustainable scheme is provided for CO2 recycling.

The invention belongs to the technical field of bioengineering, and particularly relates to collagen tripeptide as well as a preparation method and application thereof. The method comprises the following steps: carrying out depolymerization treatment on a raw material containing collagen to construct a magnetic immobilized enzyme; by utilizing the magnetic immobilized enzyme, raw materials subjected to depolymerization treatment are subjected to enzymolysis at a certain temperature, pH and reaction time, and with the assistance of a magnetic field, rapid separation and cyclic utilization of the immobilized enzyme can be realized. And finally, carrying out enzyme deactivation, solid-liquid separation and ultrafiltration to obtain the collagen tripeptide with GPH content of more than or equal to 59wt.%, and compared with the existing collagen tripeptide, the collagen tripeptide has higher GPH content. The preparation method is simple in process, easy and convenient to operate, low in cost, capable of achieving mass production and suitable for industrialization, and has remarkable practicability and economic benefits.

The invention provides a leavening agent and a preparation method thereof, and belongs to the technical field of leavening agents. According to the scheme, an efficient fermentation system is constructed through the synergistic effect of bacillus amyloliquefaciens, bacillus licheniformis, lactobacillus plantarum and saccharomyces cerevisiae, in the initial stage, the bacillus amyloliquefaciens secretes amylase and phytase degradation substrates, and conditions are created for the saccharomyces cerevisiae to metabolize monosaccharide to generate flavor substances and vitamins; in the middle stage, bacillus licheniformis decomposes protein to release amino acid, lactobacillus plantarum regulates pH through lactic acid metabolism to inhibit infectious microbes, and a multi-layer encapsulated compound enzyme preparation is introduced to synergistically degrade a complex substrate; in the later stage, fructooligosaccharide in the prebiotics-buffer stabilizer promotes proliferation of probiotics, mannitol and chitosan protect the activity of thalli and the function of metabolic enzymes, and the fermentation environment is stabilized; according to the whole system, the functionality, the environmental adaptability and the storage stability of the leavening agent are improved through flora synergistic metabolism, dynamic fermentation condition control and the action of the multifunctional additive, and the diversified requirements in industrial production are met.

The present invention relates to enzymatic reactor cells and related methods of use, e.g., to produce a compound or product by using an enzymatic reactor cell, wherein the enzymatic reactor cell includes a surface, a linker, and one or more enzymes.

The invention belongs to the technical field of xylulose production, and discloses a method for catalyzing hemicellulose to generate xylulose by an immobilized enzyme cascade reactor. According to the method, hemicellulose is used as a raw material, aminated macroporous resin Namberlyst-15 is used as a carrier, xylanase and xylose isomerase are subjected to cross-linking adsorption to obtain an immobilized double-enzyme Namberlyst-15-Xyn-XI cascade reactor, and the xylulose is produced by the immobilized double-enzyme Namberlyst-15-Xyn-XI cascade reactor. According to the method, efficient immobilization and recycling of xylanase and xylose isomerase are achieved, the catalytic efficiency and selectivity are improved, the problems of high raw material cost and poor enzyme stability in the xylulose production process are solved, technical reserve is provided for the xylulose production process, and meanwhile technical support is provided for construction of a high-stability enzyme cascade reactor.

The invention discloses a method for catalytically synthesizing adenosine triphosphate by using microcrystalline cellulose carrier purification immobilized enzyme, which comprises the following steps: preparing bacterial suspension by using escherichia coli wet thalli and Tris-HCL buffer solution, crushing, centrifuging, and taking supernate to obtain crude enzyme; adding a microcrystalline cellulose carrier and the wet thalli into the crude enzyme liquid, and washing the immobilized enzyme after immobilization to obtain the immobilized enzyme; the method comprises the following steps: adding immobilized enzyme adenosine kinase and immobilized polyphosphate kinase into a substrate system with the volume of 1L according to a weight ratio, and starting a catalytic reaction under the conditions that the pH is 5.5-7.5 and the temperature is 20-40 DEG C; after catalysis is finished, the immobilized enzyme adenosine kinase and the immobilized polyphosphate kinase are subjected to solid-liquid separation with reaction liquid through a Buchner funnel, the immobilized enzyme continues to be subjected to catalytic reaction, and the method for preparing the immobilized enzyme is simple in mode, low in cost, easy to industrially produce, high in stability and capable of being repeatedly used for many times, and products are easy to separate.

The present invention belongs to the field of biopharmaceutical technology, and specifically relates to a method for electrically promoting the synthesis of NADPH by immobilized enzyme catalysis. The present invention adopts a three-electrode system to + The buffer solution is used as the reaction solution, and the immobilized enzyme electrode is used as the working electrode to perform an electrocatalytic enzyme reduction reaction to obtain the NADPH; the immobilized enzyme electrode includes a semiconductor substrate, a porous conductive filler coating disposed on the surface of the semiconductor substrate, and a ferredoxin-NADP+ reductase loaded in the pore structure of the porous conductive filler coating. The present invention adopts a three-electrode system. Under an electric environment, NADP + As a substrate, NADPH is synthesized by catalysis of an immobilized enzyme electrode loaded with ferredoxin-NADP+ reductase (FNR). The method provided by the invention has the characteristics of high yield, high purity, simple synthesis route, low cost, and environmental friendliness.

The invention belongs to the technical field of biological extraction, and particularly relates to a preparation method of sturgeon protein peptide for enhancing immunity. Soluble salts of Cu < 2 + >, Zn < 2 + > and Ti < 4 + > are added into an ethanol-water mixed solution, the pH is adjusted by ammonia water to form a precipitate, and a hydrothermal reaction is carried out. A microbial carrier is soaked or coated in sol, a uniform gel layer is formed on the surface of the carrier through a sol-gel method, and the carrier is converted into a Cu-Zn-Ti three-way catalyst layer after being dried and roasted. And then, adding a microbial carrier loaded with a catalyst and a compound enzyme into the minced sturgeon, carrying out enzymolysis reaction under the conditions that the temperature is 25-50 DEG C, the pH is 6.8-7.2 and the speed is 100-200 rpm, and carrying out membrane separation and drying on a product to obtain the sturgeon protein peptide. According to the method, the Cu-Zn-Ti three-way catalyst is combined with the microbial carrier, so that the enzymolysis efficiency and the uniformity of the molecular weight of the product are improved, the active ingredients are effectively protected, and the production cost is reduced.

The application discloses a multi-enzyme immobilized enzyme for glucose-6-phosphate synthesis, which is obtained by immobilizing three enzymes, starch debranching enzyme mutant AcDBE_E131R, maltodextrin phosphorylase mutant AtMalP_N188D and glucose phosphomutase mutant TkPGM_F360P, on a resin. The application also discloses a preparation method and application of the multi-enzyme immobilized enzyme. The application screens enzyme mutants with good thermal stability and high catalytic activity by site-directed mutagenesis of starch debranching enzymes, maltodextrin phosphorylases and glucose phosphomutases from different sources, and prepares the multi-enzyme immobilized enzyme AcDBE_E131R+AtMalP_N188D+TkPGM_F360P. The multi-enzyme immobilized enzyme is used as a catalyst, maltodextrin is used as a substrate, and glucose-6-phosphate is efficiently synthesized by one-pot method, and the conversion rate reaches 71.2%. After 50 times of reuse, the relative enzyme activity of the multi-enzyme immobilized enzyme still remains 85.1%.

The invention provides fruit anthocyanin as well as an extraction method and application thereof. The extraction method comprises the following steps: S1, raw material pretreatment: selecting fresh fruits, removing impurities, cleaning, draining surface moisture, performing crushing treatment, then performing vacuum drying, and finally performing crushing to obtain fruit powder; s2, extracting three-layer microspheres: preparing an extraction substrate solution, adding the three-layer microspheres, and stirring for 8-12 minutes at the temperature of 40-45 DEG C and the rotating speed of 100-150r / min; the rotating speed is kept at 100-150 r / min, the power is set to be 300 W, the temperature is 42-45 DEG C, working is conducted for 4-6 min, pause is conducted for 1-2 min, and intermittent ultrasonic extraction is conducted for 3-4 h; after the extraction is finished, centrifuging an extracting solution, and collecting supernate to obtain a crude extracting solution; and S3, obtaining a final product of the fruit anthocyanin: filtering, adsorbing, eluting and purifying, concentrating and drying the crude extract to obtain the fruit anthocyanin. A targeted attachment-step conversion-enrichment protection and release linkage system is formed among the three-layer structures of the three-layer microspheres, and 1 + 1 + 1gt is realized; and 3, due to the synergistic effect, the activity and the quality stability of the anthocyanin are guaranteed while the extraction rate of the anthocyanin in the fruits is increased.

The present invention discloses a hollow ZIF-8 immobilized bienzyme and its preparation method and application, by embedding formate dehydrogenase and carbonic anhydrase in ZIF-67, then growing a layer of ZIF-8 shell on its surface, dissociating ZIF-67, and the formate dehydrogenase and carbonic anhydrase are fixed in the ZIF-8 shell in a free state, thereby maintaining a high enzyme activity to form a hollow ZIF-8 immobilized bienzyme. Utilizing 2-hydroxy-1,4-naphthoquinone coenzyme regeneration, bioelectrocatalytic CO2 reduction is achieved at the cathode. The preparation process of this method is simple to operate and easy to implement. The hollow structure is used to provide a soft microenvironment for the enzyme. Hollow ZIF-8 and carbonic anhydrase can increase the substrate CO2 adsorption capacity. Using 2-hydroxy-1,4-naphthoquinone regeneration can reduce the consumption of expensive coenzymes, and bioelectrocatalytic CO2 reduction has a higher formic acid yield.

The invention discloses a stipa brevis seed germination promoter and application, and belongs to the technical field of seed germination promoter preparation. The stipa brevis seed germination promoter is composed of the following components in parts by weight: 42-55 parts of cow dung, 28-42 parts of wheat bran, 30-45 parts of rice milk, 5-9 parts of PBS buffer, 2-3 parts of composite hydrogel, 6-10 parts of immobilized enzyme, wherein the composite hydrogel is prepared from sodium chloride, cellulose nanofiber, sodium periodate, γ-polyglutamic acid and ethylene glycol diglycidol, and the immobilized enzyme is prepared from tribenzaldehyde, acetic acid buffer, p-phenylenediamine and composite enzyme. The promoter prepared by the method can effectively improve the germination rate and plant height of stipa brevis.

To provide an immobilized enzyme composition for hexose production or an improved process for hexose production. [Solution] The present invention relates to an immobilized enzyme composition for the preparation of hexoses. Examples of hexoses include tagatose, psicose, fructose, allose, mannose, galactose, altrose, talose, sorbose, gross, idose, and inositol. The present invention also relates to an enzymatic process for preparing hexoses from sugars by contacting starch derivatives with the immobilized enzyme composition of the present invention.

The invention discloses a bio-enzyme preparation for catalytic degradation of grease as well as a preparation method and application thereof, and belongs to the technical field of bio-enzymes. The gel carrier is formed by cross-linking a chitosan-lauric acid-dopamine conjugate and Fe < 3 + > ions through covalent coupling; the bonding adaptive compound enzyme is a mixed enzyme of which the surface is grafted with a catechol group through an N-hydroxysuccinimide-dopamine ester modifier; the technical bottleneck that efficient catalysis, firm fixation, convenient recovery and long-acting stability of an industrial cleaning enzyme preparation are difficult to consider at the same time is solved, and the prepared biological enzyme preparation has the high-activity recovery rate, excellent recycling stability, intelligent separation and recovery characteristics and efficient broad-spectrum cleaning capacity; and the method has remarkable industrial application advantages and market competitiveness.

The invention relates to a process for preparing TEMTP through turntable extraction, and belongs to the technical field of organic annular peroxides. According to the process, a rotating disc reactor is adopted to integrate a photocatalysis-enzyme catalysis-chemical catalysis triple synergistic system, and the problem of cross inactivation of a catalyst is solved through partition design and nano sieve membrane isolation. A peptide-guided magnetic MOF photocatalytic carrier is adopted as a core, a Fe3O4 magnetic core of the peptide-guided magnetic MOF photocatalytic carrier achieves magnetic field driving, a ZIF-8 frame loads a TiO2-Ag photocatalyst, PNIPAM temperature-sensitive hydrogel fixes halogenase, and the intelligent enzyme control characteristic is achieved. According to the method, TEMTP is taken as a raw material, a reaction is carried out under light source irradiation and magnetic field driving, extraction and purification are carried out through fluoroether HFE-7500 at normal pressure, a high-purity TEMTP product is finally obtained, and unification of reaction efficiency, selectivity and green production is achieved.

The present invention provides a method for producing methane by dry anaerobic fermentation, which relates to the technical field of anaerobic fermentation. In the present invention, a biochar-immobilized complex enzyme is added during the dry anaerobic fermentation process. The biochar-immobilized complex enzyme is achieved by fixing the enzyme to biochar, which can enhance the stability and resistance to protein degradation of the enzyme, extend the service life of the enzyme, and reduce the inactivation rate of the enzyme, thereby facilitating an increase in methane production. In addition, biochar has a high specific surface area and the characteristics of promoting interspecies electron transfer, which is conducive to increasing methane production. Moreover, in the biochar-immobilized complex enzyme, biochar fixes the enzyme on its surface or interior, which can provide more reactive sites, which is conducive to the enrichment of methanogens, thereby shortening the fermentation reaction cycle and facilitating an increase in the reaction rate of anaerobic fermentation. In summary, the reaction rate and methane yield of the method for producing methane by dry anaerobic fermentation provided by the present invention are both high.

The invention relates to an enzyme-loaded porous gel capable of being quickly reconstructed, a preparation method and a flow microreactor, amphiphilic nanoparticles are used as a stabilizer, bubbles are introduced into a polymer solution and polymerization crosslinking is induced to form a uniform porous gel structure with an adjustable pore diameter, the amphiphilic nanoparticles can be adsorbed on a gas-liquid interface to stabilize the bubbles, and the porous gel structure can be quickly reconstructed. Enzyme molecules can be loaded on the surface to form a porous reaction medium with catalytic activity. The obtained porous gel has good mechanical strength, controllable aperture and porosity, and can realize rapid reconstruction of a porous structure, so that the application scene of a microreactor is widened, and the effective service cycle of enzyme is remarkably prolonged when the microreactor is blocked.

The present invention relates to a process to carry out two or more enzymatic reactions in a reaction medium, wherein the process comprises providing a system comprising at least two substantially homogeneous and colloidal populations of magnetic nanoparticles (MNPs) and applying one or more external alternating magnetic field to produce the simultaneous or sequential activation of the enzymes functionalized on the surface of each population of MNPs so that the enzymatic reactions can be thermally activated.

The present invention relates to the field of bioengineering, and particularly to mutant compositions, fusion enzymes and methods for preparing collagen tripeptides. The present invention provides mutant compositions, including Mutant 1 and Mutant 2; Mutant 1 has an amino acid sequence as shown in SEQ ID NO: 1 and Mutant 2 has an amino acid sequence as shown in SEQ ID NO: 2. The preparation method provided by the present invention uses inexpensive amino acids as raw materials, and is highly efficiently converted into corresponding dipeptide or tripeptide products under the action of adenosine triphosphate (ATP) and the corresponding ligase. Introducing an ATP recycling system into the reaction system can greatly reduce its usage amount; at the same time, the amount of enzyme used in the actual production process is reduced by constructing fusion enzymes; the reaction and purification processes are further optimized by using immobilized enzyme catalysis, and the preparation scheme is very concise, with high yield, good product quality and easy to scale up.

The present invention relates to a biocatalyst for organic synthesis, comprising a controlled porosity silica (CPS) as a support material, wherein the pore diameter is from about 20 to about 100 nm, the support material comprises an amino-functionalized surface, and one or more catalytically active enzymes immobilized on the support material. The present invention also relates to the use of such biocatalysts and a method for their production.

The invention relates to a Janus nano silicon dioxide particle with double-enzyme split-phase distribution as well as a preparation method and application of the Janus nano silicon dioxide particle. The preparation method comprises the following steps: by taking gamma-glycidyl ether oxypropyl trimethoxy silane and octyl trimethoxy silane as modifying agents, modifying nano SiO2, so as to prepare nano SiO2Janus particles with an epoxy group; then adding glucose oxidase to covalently bind with epoxy groups on the surfaces of the particles, and stirring for reaction to obtain silicon dioxide particles with glucose oxidase immobilized on the hydrophilic side; and finally, adding a CALB enzyme solution, stirring and reacting, and centrifuging, washing and drying after the reaction is completed, so as to obtain the Janus nano silicon dioxide particles with double-enzyme split-phase distribution. Epoxy groups are introduced and can react with amino groups to achieve enzyme immobilization, use of a cross-linking agent in the immobilization process is avoided, immobilization conditions are mild, and operation is easy.

Provided are a biomimetic silicon mineralized microcapsule immobilized multi-enzyme, a preparation method therefor, and a method for producing tagatose by using same. The preparation method comprises the following steps: (1) pre-mixing glucan phosphorylase, phosphoglucomutase, phosphoglucoisomerase, 6-phosphate tagatose 4-position epimerase and 6-phosphate tagatose phosphatase solutions, then adding the mixture to a calcium chloride solution, and then pouring same into a sodium carbonate solution, stirring and separating same to obtain calcium carbonate microspheres containing a multi-enzyme; (2) mixing the calcium carbonate microspheres with a polyethyleneimine solution to obtain polyethyleneimine-calcium carbonate microspheres after separation; (3) mixing the polyethyleneimine-calcium carbonate microspheres with a silicate solution to obtain biomimetic silicon mineralized-calcium carbonate microspheres after separation; and (4) mixing the biomimetic silicon mineralized-calcium carbonate microspheres with ethylenediamine tetraacetic acid for reaction to remove calcium carbonate, and separating same to obtain a biomimetic silicon mineralized microcapsule immobilized multi-enzyme

This invention relates to a carrier-free immobilized enzyme and its preparation method, wherein the carrier-free immobilized enzyme is a catalytically active inclusion body of the enzyme crosslinked with glutaraldehyde, and the enzyme's end (especially the N-terminus) is tagged with histidine-leucine-histidine-leucine-histidine-leucine.

The chemical formula of a metal organic framework 2D-NCM (Fe) in the magnetic metal-organic framework mediated multi-enzyme nano material GOx-GAA at 2D-NCM (Fe) is Fe3O6Co2Ni2C3N0.4, Gox is glucose oxidase, and GAA is glucosidase. The invention also provides a method for efficiently and quickly measuring the activity of alpha-amylase, which comprises the following steps: acquiring a to-be-measured sample solution containing alpha-amylase, and carrying out enzymolysis on the to-be-measured sample solution and a starch solution at a first preset temperature for a first preset time to obtain a first reaction solution, after the enzymolysis reaction is finished, adding a first solution containing a magnetic metal-organic framework mediated multienzyme nanomaterial GOx-GAA at 2D-NCM (Fe) and a 3, 3 ', 5, 5'-tetramethyl benzidine solution into the first reaction solution, and reacting for a second preset time to obtain a second reaction solution; measuring the absorbance of the second reaction liquid at 652 nm; and obtaining the enzymatic activity of the alpha-amylase in the to-be-detected sample solution containing the alpha-amylase according to a standard curve of association between the absorbance at 652nm and the enzymatic activity of the alpha-amylase.