97results 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 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 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 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 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.

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 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.

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.

This invention belongs to the field of microbial inoculant technology and discloses a corn stalk decomposing agent product and its preparation method. The decomposing agent is composed of microbial inoculants, enzyme preparations, nutrient adjuvants, and a modified carrier in specific weight proportions. The microbial inoculants are a compound of Bacillus subtilis, Trichoderma viride, and Aspergillus niger; the enzyme preparation is a complex of cellulase, xylanase, and laccase; the nutrient adjuvant is a mixture of urea, sucrose, and potassium dihydrogen phosphate; and the carrier is an alkali-modified complex of wheat bran and straw powder. The preparation method includes preparing each microbial inoculant separately, alkali-modifying the carrier, and mixing all components. This invention also discloses the application method of this decomposing agent in corn stalk decomposition. This product achieves rapid, efficient, and thorough decomposition of corn stalks through the synergistic effect of multiple components, and the preparation process is stable, the product has a long shelf life, and it is suitable for large-scale production and field application.

The present invention specifically relates to an enzyme combination comprising a glucosidase enzyme, a secoisolariciresinol dehydrogenase enzyme and a glucose dehydrogenase enzyme, which are capable of carrying out reactions to convert secoisolariciresinol diglucoside (SDG) into matairesinol with a high yield. The invention also relates to the uses of the above combination, and to methods for producing matairesinol.

The invention relates to the field of bio-enzyme catalysis, and discloses a bio-enzyme catalysis process for reducing agent modification, which comprises the following steps: S1, enzyme solution preparation: mixing a bio-enzyme preparation with a buffer solution with the pH value of 5-8 to prepare an enzyme solution with the enzyme concentration of 0.5-5.0 g / L; the biological enzyme preparation is a compound enzyme composed of one or more of cellulase, hemicellulase and lignin degrading enzyme; s2, enzymolysis modification: mixing a carbonaceous reducing agent raw material with the enzyme solution prepared in the step S1 according to a mass ratio of 1: 5-1: 20, carrying out an enzymolysis reaction at 40-60 DEG C, and meanwhile, applying ultrasonic waves to carry out auxiliary treatment; the power of the ultrasonic waves is 100-500 W, and the frequency of the ultrasonic waves is 20-40 kHz And S3, solid-liquid separation: after the reaction is finished, separating the modified reducing agent from the enzymatic hydrolysate to obtain the solid modified reducing agent. Compared with the prior art, the method has the advantages that a biological enzyme catalysis process for modifying the reducing agent is provided, the energy consumption is low, the environment is protected, the cost is low, and the performance of the reducing agent can be remarkably improved.

The present invention relates to a nano-vesicle reactor and a method for producing the same. Specifically, the present invention provides a composition for producing a nano-vesicle reactor, characterized by comprising a first vesicle containing a first receptor on its surface and a second vesicle containing a second receptor on its surface, wherein the first receptor and the second receptor are bound with each other through a ligand system so that the first vesicle and the second vesicle are fused to form the nano-vesicle reactor ; and a method for producing the above nano-vesicle reactor. The nano-vesicle reactor of the present invention can be utilized in production of energy inside a cell through cellular enzymatic reaction, a drug delivered system, and a diagnostic system.

The invention discloses an organic selenium composition, a preparation method and application of the organic selenium composition in preparation of antitumor drugs, and belongs to the technical field of enzyme-containing preparations, the preparation method comprises five steps of carrier activation, immobilization of double enzymes, immobilization of a third enzyme, in-situ synthesis and separation and purification; aminated mesoporous silica nanoparticles are used as a carrier, selenocysteine beta-lyase, UDP-glucosyltransferase and glucose dehydrogenase are immobilized through glutaraldehyde crosslinking, an immobilized three-enzyme system is constructed, and organic selenium is synthesized in situ by catalyzing substrates such as jerusalem artichoke polysaccharide and selenocysteine in a nitrogen atmosphere. And separating and purifying to obtain the organic selenium composition. The problems of easy enzyme inactivation, large coenzyme dosage, poor product stability and the like in the prior art are solved, and the prepared organic selenium composition is high in selenium content, concentrated in molecular weight, excellent in bioavailability and low in toxicity, can be used as an active component for preparing antitumor drugs, and has a good application prospect.

The invention relates to a protein scaffold mediated multienzyme cascade system and application thereof in preparation of D-tagatose. The protein scaffold mediated multi-enzyme cascade system provided by the invention comprises fructokinase, D-tagatose-6-phosphate 4-epimerase and D-tagatose-6-phosphate phosphatase, and the protein scaffold mediated multi-enzyme cascade system comprises fructokinase, D-tagatose-6-phosphate 4-epimerase and D-tagatose-6-phosphate phosphatase, the D-tagatose-6-phosphoric acid 4-epimerase and the D-tagatose-6-phosphoric acid phosphatase are immobilized on the protein support SH3, and the D-tagatose-6-phosphoric acid 4-epimerase is immobilized on the protein support SH3. According to the system, a plurality of key enzyme spaces in a reaction path are positioned on a protein scaffold through a protein-protein interaction structural domain (SH3), so that a substrate channel and a spatial proximity effect are enhanced, and the yield of D-tagatose is increased. According to the method disclosed by the invention, the yield of the D-tagatose is 3.2-7.7 times that of the original D-tagatose by optimizing reaction conditions.

The invention relates to a method for efficiently removing penicillin G sodium in mushroom dregs by using a magnetic mushroom dreg biochar immobilized degrading enzyme. According to the method, the enzyme for degrading the penicillin G sodium is produced in an amplified mode through a 5L stirring type reactor, the working procedures comprise strain culture, shake flask first-stage culture, shake flask second-stage culture and 5L stirring type reactor fermentation, and the amplified enzyme activity reaches up to 1288.09 U / mL. Penicillin G sodium in actual fermentation mushroom dregs is removed in a 5L stirring type reactor by utilizing the mushroom dreg biochar immobilized degrading enzyme, and the removal rate of 0.1 g of immobilized enzyme on the penicillin G sodium in the fermentation mushroom dregs within 120 minutes reaches 98.72%. The invention also provides a method for extracting the residual penicillin G sodium in the mushroom dregs. The method specifically comprises three steps of extraction, purification and detection. According to the invention, a new low-cost choice is provided for efficiently repairing the pollution of the penicillin G sodium in the environment, and the treated mushroom dregs have important significance for realizing high-value utilization of hazardous wastes.

A deproteinized natural rubber latex and preparation method thereof are provided. A combined immobilization technology of a multi-enzyme system is introduced to form a multi-enzyme synergistic effect, such that high-concentration enzymes contact with proteins in natural rubber latex. Meanwhile, due to the complexity and diversity of latex proteins, the catalytic properties of different enzymes are effectively combined to realize a multi-enzyme cascade reaction, and the proteins are hydrolyzed gradually and orderly. In addition, an immobilized enzyme filter screen is introduced, and the filter screen is lifted and circulated to make rubber particles collide with high-density enzyme carrier and then complete mixing evenly to reduce damages to the rubber particles when surface proteins are removed, thereby maintaining original physical properties of the latex. The rubber latex has high stability, reusability, is easy for separation and continuous operation, and is environmentally friendly.

The invention relates to the technical field of bioengineering, in particular to a preparation method and application of an immobilized enzyme. Compared with free enzyme, the immobilized enzyme prepared from the selected specific resin can be reused and is high in stability; the immobilized enzyme is easily separated from a reaction system, so that a product purification process can be simplified, and the product yield and quality are improved. Moreover, by immobilizing a sugar nucleotide producing enzyme and four sialyltransferases, a new strategy suitable for large-scale synthesis of functional sugar chains such as sialylated sugar chains is developed, the stability of related enzymes is improved, long-term storage and reutilization of the enzymes are realized, and the production cost of the functional sugar chains is reduced.