32results about How to "Immobilization method is simple" patented technology

A process for producing biological diesel oil by fixed enzyme method is carried out by taking sodium alginate, kaoline, glutaraldehyde, gelatin and lactose as co-fixer, fixing the lipase, piling the spherical fixed enzyme in filled bed columnar reactor naturally, alcoholyzing animal and plant oil fatty with biological diesel oil as solvent and producing biological diesel oil. It is cheap, has less consumption and by product and better quality. It can be used for continuous production.

The invention discloses the preparation method of the ultra fine fiber membrane of an affinity acrylonitrile base copolymer. The acrylonitrile base copolymer is dissolved in solvent to form solution and then electrostatic spinning is conducted to prepare the fiber membrane. The mass percentage of the acrylonitrile base copolymer in the solution is two to eight percent; the spinning voltage of the electrostatic spinning is six to twenty five kv; the flow velocity of spinneret solution is 0.5 to 2.0 ml per hour with the deposition distance ranging from eight to twenty centimeters. The acrylonitrile base copolymer is acrylonitrile / allylamine copolymer, acrylonitrile / methyl allylamine copolymer, acrylonitrile / acrylamide copolymer or acrylonitrile / methacrylamide copolymer with the viscosity-average molecular weight being eighty to three hundred thousand; the solvent is one or various kinds which are mixed with random percentages of dimethyl sulfoxide, dimethyl formyl ammonia or dimethyl acetyl ammonia. After the glycan of the fixed shell of a chemical bond at the surface of the membrane, the membrane is used for separating and purify protein with simple operation, good chemical stability and low cost.

The invention provides a radioactive wastewater treatment method. The radioactive wastewater treatment method provided by the invention comprises the following steps: adding active calcium silicate to radioactive wastewater with the pH value of greater than or equal to 2, and then performing solid-liquid separation to obtain purified water and radioactive calcium silicate, wherein the grain diameter of the active calcium silicate is smaller than 50 [mu]m, the bore diameter is 2-50 nm, and the specific surface area is 400-700 m<2> / g. According to the radioactive wastewater treatment method provided by the invention, the active calcium silicate is taken as a treating agent, the active calcium silicate is not easy to be influenced by the water quality and pH value of wastewater when the radioactive wastewater is treated, and radioactive elements and other metal ions in the radioactive wastewater can be effectively removed under a smaller additive amount; and moreover, an immobilization method for the radioactive calcium silicate formed after treatment is simple, and the immobilization effect is good.

The invention discloses an immobilized recombinant penicillin G acylase and an application thereof to preparation of (S)-2-aryl-amino acid and cephalosporin antibiotics. The immobilized recombinant penicillin G acylase disclosed by the invention is capable of converting an S-type substrate into an S-type product within shorter time, high in substrate tolerance and strict in S selectivity for the substrate and structural analogues thereof; due to the addition of cobalt ions as well as a protective agent with phenylacetic acid and glycerin as enzyme active centers, the immobilized recombinant penicillin G acylase disclosed by the invention is prevented from being successfully subjected to multipoint covalent immobilization on the surface of an epoxy resin carrier under the condition that enzyme molecules are seriously inactivated in an immobilization process, and the immobilization method is simple and feasible, cheap in raw materials and suitable for large-scale operation; the immobilized recombinant penicillin G acylase can be used for synthesizing cephalosporin antibiotics and splitting the prepared (S)-2-aryl-amino acid, and can be continuously used for more than 50 batches before the activity of the immobilized enzyme is not remarkably reduced.

The invention discloses an L-lactic acid fermentation method by using polyvinyl alcohol to solidify rhizopus oryzae. The method comprises the following steps: heating polyvinyl alcohol and sodium alginate in a boiling water bath to be jointly dissolved; mixing with a rhizopus oryzae spore suspension; using an injector to drip the mixed solution to a saturated boric acid solution containing calcium chloride while using a magnetic stirrer to continuously stir so that particles are spherical and good immobilized particles can be hardened at a low temperature; and then, treating by a monobasic sodium phosphate solution and washing with distilled water after solidified cells are propagated and grow so as to be used for the batch fermentation and the repetitive batch fermentation of the L-lactic acid. The improved L-lactic acid fermentation method by using the polyvinyl alcohol to solidify the rhizopus oryzae overcomes the defects of spherical carriers and poor permeability of the prior polyvinyl alcohol and has the advantages of great mechanism strength, large acid production amount, high acid production speed and good repetitive use effect.

The invention relates to an enzyme immobilization method based on a hyperbranched polymer, which belongs to the technical field of bioengineering. The method comprises: in a synthetic hyperbranched polymer solution with a large number of diazonium salt groups on the periphery, or In the dilute solution of the hyperbranched polymer solution diluted with water or organic solvent N,N-dimethylformamide less than 100 times, the carrier is soaked for more than half an hour; then the carrier is taken out, washed with water, and then immersed in In the enzyme-containing buffer solution, adsorb at 4°C for more than 4 hours to achieve saturated adsorption; this method can greatly simplify the operation steps of immobilized enzymes, without the need for pre-modification of materials and enzymes, and has high universality; it can be used On a variety of enzymes and carriers, and has no obvious effect on the catalytic activity of the enzyme.

The invention discloses an immobilized glutamate decarboxylase and a preparation method thereof. The immobilized glutamate decarboxylase is prepared according to the method comprising the following steps of: 1) adding acetic acid-acetate buffer solution into lactic acid bacteria to obtain bacterial suspension containing 300-500cfu bacteria per milliliter, performing ultrasonic crushing on the bacterial suspension under the condition of an ice-water bath to obtain crude enzyme solution of the glutamate decarboxylase; 2) using cotton flannel fabric as a carrier, and sequentially performing pretreatment with sodium hydroxide, pyridine and para-toluenesulfonyl chloride; and 3) arranging the cotton flannel fabric after the pretreatment in the crude enzyme solution of the glutamate decarboxylase for soaking, adding glutaraldehyde into the solution, and realizing immobilization of the glutamate decarboxylase. The immobilization method of the glutamate decarboxylase is simple, the conditions are required to be mild and easy to implement, and the obtained immobilized glutamate decarboxylase has the advantages of high activity, large loading amount, long catalytic reaction service life and the like.

The invention discloses a preparation method and an application of an immobilized lactobacillus. The preparation method includes the steps: S1 lactobacillus concentrated solution preparation: centrifugally concentrating lactobacillus suspension after enrichment culture to obtain lactobacillus concentrated solution; S2 lactobacillus immobilization: 1) swelling sericin protein by water to obtain sericin protein solution, uniformly mixing the sericin protein solution and the lactobacillus concentrated solution to obtain liquid A; 2) dripping a cross-linking agent into the liquid A acquired in thestep 1) to obtain sericin protein hydrogel containing lactobacilli; 3) dissolving cellulose powder in water to obtain swelling cellulose solution, adding the swelled cellulose solution into the sericin protein hydrogel acquired in the step 2), uniformly mixing mixture, and crosslinking the mixture; 4) slowly dripping calcium chloride solution into crosslinked solution acquired in the step 3) through a constant-flow pump, filtering mixture, washing the mixture by deionized water to obtain the immobilized lactobacillus, and storing the immobilized lactobacillus in a refrigerator with the temperature of 4 DEG C. The immobilized lactobacillus is used for preparing gamma-aminobutyric acid in a fermented manner.

The invention discloses a method for preparing a series of neoagarool igosaccharode with single degree of polymerization by utilizing an immobilized enzyme. The method comprises the following steps: (1) acquiring agarase powder from vibrio natriegens HJPHYXJ-1; (2) preparing an immobilized carrier; (3) preparing the immobilized enzyme; (4) preparing a series of neoagarool igosaccharode. Accordingto the method, an immobilized agarase is prepared by adopting an immobilized enzyme technology and is applied in the preparation of the neoagarool igosaccharode; through simple magnetic separation, reutilization of the agarase can be realized, and aftertreatment steps are simplified, therefore, the production cost of the neoagarool igosaccharode is greatly lowered; additionally, the immobilized carrier used in the method is simple to prepare, and an immobilization method is simple and effective, so that the theoretical and technical supports can be provided for future industrial large-scale production of the neoagarool igosaccharode.

The invention provides an enzyme membrane reactor. The enzyme membrane reactor comprises a microfiltration membrane and an immobilized enzyme, wherein the immobilized enzyme comprises a horseradish peroxidase which is fixed to the microfiltration membrane through an embedding medium. The invention also provides a method of using the enzyme membrane reactor to treat wastewater. The enzyme membranereactor and method have the advantages that the enzyme membrane reactor has high reaction activity and fast reaction speed and can basically complete reaction in ten minutes; reaction conditions are mild, and the reactor operates at a normal temperature and a normal pressure; an enzyme immobilization method is simple, and the production cost is low; an enzyme membrane has good stability, an enzymeis unlikely to run off, the membrane is unlikely to be blocked, and the service life is long; the enzyme membrane reactor can treat various pollutants such as phenols and azo, and has a wide application range; only a small amount of hydrogen peroxide is added, and the treatment cost is low; and no poisonous and harmful chemical agents are added, and thus, the secondary pollution of environments is avoided.

A process for producing biological diesel oil by fixed enzyme method is carried out by taking sodium alginate, kaoline, glutaraldehyde, gelatin and lactose as co-fixer, fixing the lipase, piling the spherical fixed enzyme in filled bed columnar reactor naturally, alcoholyzing animal and plant oil fatty with biological diesel oil as solvent and producing biological diesel oil. It is cheap, has less consumption and by product and better quality. It can be used for continuous production.

Technology for fixing fungi and bacterium with vermiculite as carrier is carried out by modifying while moistening for vermiculite as carrier material, steam sterilizing at high-temperature and high-pressure, inoculating mixed bacterium, and multiplication culturing to obtain the final product. It's simple, porous and efficient, has better growth environment, degradation and resistance to poisonous, larger specific area and competition ability.

The invention provides a production method of inosinic acid. The method comprises the step of adopting adenosine deaminase fixed in anion resin to catalyze the generation of the inosinic acid from adenylic acid. According to the method provided by the invention, the adenylic acid is taken as a substrate, the supply of raw materials is sufficient, the requirements for equipment are low, the separation and extraction are convenient, the semi-continuous production of the inosinic acid can be realized, the substrate is convenient and easy to obtain, the transformation is thorough, and the cost is relatively low.

The invention discloses an ampere type biosensor electrode and a method for making the same. Dimethylformamide is taken as solvent, acrylonitrile copolymer and an electric mediator are taken as raw materials, an electrochemical electrode is taken as an accepting electrode, a composite fiber membrane of the acrylonitrile copolymer and the electric mediator is prepared on the surface of the electrode through electrostatic spinning, then a covalent method is used to fix xidoreductase on the surface of fibers to form an activated layer of the fibers on the surface of the electrode. The activated layer of the fibers has large specific surface area, high porosity rate and a through-hole structure; at the same time, the electric mediator is provided, so the electrode has higher response strength, linear detection range and stability. The mediator electrochemical enzyme electrode has the characteristics of simple preparation process, repeated use, large surface area of an effective enzyme membrane, strong anti-jamming ability and so on, and is suitable for batch production.

The invention relates to a cell immobilization technology, in particular to an immobilization bacteria production method used for repairing soil contaminated by polycyclic aromatic hydrocarbon. The carrier material is 0.25 to 1.00mm of red brick powder, fly-ash, vermiculite, peat soil, diatomite or coke powder, which are sterilized and added with bacteria liquid to do multiplication culture, so as to make the required immobilized particles. The immobilized biological product made according to the prescription of the invention has good repairing effect on the soil contaminated by the polycyclic aromatic hydrocarbon and is a reliable, novel and environment-friendly biological product.

An immobilized organism co-metabolism and decolouring process for the dyeing sewage includes such steps as adding nutritive liquid to non-sterilized water to obtain bacterial suspension, adding it to an up-flow fixed bed reactor filled with attapulgite, and adding carbon source and nitrogen source substances to the sewage to be treated. Its advantage is high average decolouring rate up to 80%.

The invention relates to the cell immobilization technology, specifically speaking, it's about the preparing method of a solidified granule which can be used in the repair of soil which is polluted by polycyclic aromatic hydrocarbon. The main carrier is corncob. Mix the fresh, dry and pulverized corncob (the diameter is about 1.5-2cm) with cotton kernel pastry, wheat bran, bean cake powder, rice bran and / or wheat bran, of which the corncob accounts for 70-85%, the accessories account for 15-30%; regulate the water content to 40-55wt% and pH value to 6.3-6.5; after autoclaving, inoculate the foundation stock full of fungal spore and enrichment cultivate to get the solidified granule. The advantage of the invention includes: low cost, high solidifying efficiency, simple operating method and suitable to repair the soil in large scale.

The invention provides a production method of inosinic acid. The method comprises the step of adopting adenosine deaminase fixed in anion resin to catalyze the generation of the inosinic acid from adenylic acid. According to the method provided by the invention, the adenylic acid is taken as a substrate, the supply of raw materials is sufficient, the requirements for equipment are low, the separation and extraction are convenient, the semi-continuous production of the inosinic acid can be realized, the substrate is convenient and easy to obtain, the transformation is thorough, and the cost is relatively low.

The present invention provides a gene modification method to realize its soluble and active expression: take the lipase gene with polyamine tags at the N-terminus and C-terminus as the target gene, and add coding A nucleotide sequence of m histidines, m arginines or m lysines, m=2-10, and at the same time add n histidines and n sperms to the C-terminus of the lipase parent gene sequence amino acid or n lysine nucleotide sequence, n=2-10, and primers are designed for the target gene; the N-terminal and C-terminal lipase genes with polyamine tags are obtained by PCR. After the gene is expressed, a soluble lipase with a polyamine tag is produced, which can induce the formation and immobilization of biomimetic nano-silicon. The invention can obtain the immobilized enzyme with high activity and immobilization efficiency under mild conditions and in a short time; the operation is simple, the immobilization conditions are easy to control, and the cost is low.

The invention relates to a preparation method of immobilized water purifying agent, which comprises the following steps: mixing immobilized carrier and composite microbial agent according to a volumeratio of 1: (6-7), stirring and culturing at 28-30 DEG C for 36-48h. The composite microbial agent comprises of acinetobacter, candida mycoderma bacteria and bacillus subtilis. The water purifying agent can obviously reduce the COD value of the eutrophic water body and inhibit the large-scale growth of the water hyacinth, thereby having popularization value and wide application prospect.

The invention discloses an immobilized glutamate decarboxylase and a preparation method thereof. The immobilized glutamate decarboxylase is prepared according to the method comprising the following steps of: 1) adding acetic acid-acetate buffer solution into lactic acid bacteria to obtain bacterial suspension containing 300-500cfu bacteria per milliliter, performing ultrasonic crushing on the bacterial suspension under the condition of an ice-water bath to obtain crude enzyme solution of the glutamate decarboxylase; 2) using cotton flannel fabric as a carrier, and sequentially performing pretreatment with sodium hydroxide, pyridine and para-toluenesulfonyl chloride; and 3) arranging the cotton flannel fabric after the pretreatment in the crude enzyme solution of the glutamate decarboxylase for soaking, adding glutaraldehyde into the solution, and realizing immobilization of the glutamate decarboxylase. The immobilization method of the glutamate decarboxylase is simple, the conditions are required to be mild and easy to implement, and the obtained immobilized glutamate decarboxylase has the advantages of high activity, large loading amount, long catalytic reaction service life and the like.

The invention discloses a method for immobilizing cells containing glucose isomerase. The method is to add wet bacteria obtained through fermentation and culture of recombinant genetically engineered bacteria containing glucose isomerase genes into a buffer to prepare a bacterial suspension solution; add the carrier to the bacterial suspension, stir and mix, then add polyethylenimine, then add glutaraldehyde for cross-linking, stir and cross-link at 0-30°C for 1-2 hours, filter, and wash the filter cake with distilled water Extrude into long strips with an axial extruder, air-dry at room temperature, and pulverize into granules to obtain immobilized glucose isomerase cells containing glucose isomerase; the immobilization method provided by the present invention has the advantages of low cost of immobilized materials, The operation is simple, the mechanical strength is high, and it is relatively stable under high temperature conditions. The immobilized glucose isomerase-producing recombinant E. coli whole cells prepared are used to catalyze the production of D-glucose from D-glucose under high-temperature conditions. At 85°C, the conversion The yield is as high as 54%, and after repeated use for 10 batches, it still has more than 90% of the enzyme activity, and has a good industrial application prospect.

The invention discloses a preparation method and an application of an immobilized lactobacillus. The preparation method includes the steps: S1 lactobacillus concentrated solution preparation: centrifugally concentrating lactobacillus suspension after enrichment culture to obtain lactobacillus concentrated solution; S2 lactobacillus immobilization: 1) swelling sericin protein by water to obtain sericin protein solution, uniformly mixing the sericin protein solution and the lactobacillus concentrated solution to obtain liquid A; 2) dripping a cross-linking agent into the liquid A acquired in thestep 1) to obtain sericin protein hydrogel containing lactobacilli; 3) dissolving cellulose powder in water to obtain swelling cellulose solution, adding the swelled cellulose solution into the sericin protein hydrogel acquired in the step 2), uniformly mixing mixture, and crosslinking the mixture; 4) slowly dripping calcium chloride solution into crosslinked solution acquired in the step 3) through a constant-flow pump, filtering mixture, washing the mixture by deionized water to obtain the immobilized lactobacillus, and storing the immobilized lactobacillus in a refrigerator with the temperature of 4 DEG C. The immobilized lactobacillus is used for preparing gamma-aminobutyric acid in a fermented manner.

The invention discloses a method for continuously synthesizing oligo-isomalto-oligosaccharides. An aldehyde-modified chitosan magnetic carrier is used to further immobilize recombinant Pichia pastorisdisplaying alpha-glucosidase on the surface. Aldehyde groups modifying the surface of the carrier are covalently bound to glycoprotein on the surface of the recombinant Pichia pastoris, and at the same time, through the adsorption effect of chitosan on the recombinant Pichia pastoris, the effect of immobilizing a Pichia pastoris cell catalyst can be achieved. By utilizing the superparamagnetic properties of the carrier, immobilized mycelia and a reaction substrate can be rapidly separated in a magnetic field, and the recycling efficiency of the cell catalyst is improved. The immobilized mycelia have good thermal stability and good recyclability. At the same time, the invention also relates to addition of the immobilized recombinant Pichia pastoris into a fixed bed for direct continuous catalytic synthesis of IMOs, and better production efficiency of the IMOs can be achieved. The immobilization method is simple and effective, and has good operability, the reuse of the mycelia is easy to realize, and the production cost of the IMOs can be reduced.

The invention discloses an L-lactic acid fermentation method by using polyvinyl alcohol to solidify rhizopus oryzae. The method comprises the following steps: heating polyvinyl alcohol and sodium algiThe invention discloses an L-lactic acid fermentation method by using polyvinyl alcohol to solidify rhizopus oryzae. The method comprises the following steps: heating polyvinyl alcohol and sodium alginate in a boiling water bath to be jointly dissolved; mixing with a rhizopus oryzae spore suspension; using an injector to drip the mixed solution to a saturated boric acid solution containing calciumnate in a boiling water bath to be jointly dissolved; mixing with a rhizopus oryzae spore suspension; using an injector to drip the mixed solution to a saturated boric acid solution containing calcium chloride while using a magnetic stirrer to continuously stir so that particles are spherical and good immobilized particles can be hardened at a low temperature; and then, treating by a monobasic sodchloride while using a magnetic stirrer to continuously stir so that particles are spherical and good immobilized particles can be hardened at a low temperature; and then, treating by a monobasic sodium phosphate solution and washing with distilled water after solidified cells are propagated and grow so as to be used for the batch fermentation and the repetitive batch fermentation of the L-lacticium phosphate solution and washing with distilled water after solidified cells are propagated and grow so as to be used for the batch fermentation and the repetitive batch fermentation of the L-lactic acid. The improved L-lactic acid fermentation method by using the polyvinyl alcohol to solidify the rhizopus oryzae overcomes the defects of spherical carriers and poor permeability of the prior polyacid. The improved L-lactic acid fermentation method by using the polyvinyl alcohol to solidify the rhizopus oryzae overcomes the defects of spherical carriers and poor permeability of the prior polyvinyl alcohol and has the advantages of great mechanism strength, large acid production amount, high acid production speed and good repetitive use effect.vinyl alcohol and has the advantages of great mechanism strength, large acid production amount, high acid production speed and good repetitive use effect.

The invention discloses a method for preparing a Xanthoceras sorbifolia polypeptide based on a papaya seed gum immobilized enzyme. The method uses papaya seed gum as a carrier and a trivalent chromium salt as a cross-linking agent to immobilize the biological enzyme. The immobilization method is simple and has high efficiency. The immobilized enzyme is physical gel with good stability, and the immobilized enzyme is free of a costly carrier so that a cost is greatly reduced. The immobilized enzyme can realize enzymolysis of a Xanthoceras sorbifolia protein to extract the polypeptide. The method has simple processes, is easy to operate, realizes enzyme recovery and recycling and greatly reduces the waste of resources.

The invention relates to an enzyme immobilization method which is based on hyper-branched polymer. The method belongs to the biology engineering technology area. The method comprises that the carrieris immersed in the synthesized hyper-branched polymer solution with a large amount of peripheral diazo salt groups or in the diluted solution which dilutes the hyper-branched polymer solution or the organic solvent of N, N-dimethylformamide less than 100 times over 30 minutes; then the carrier is taken out from the diluted solution and washed by water; the carrier is immersed in the buffer which contains the enzyme below 4 degree for over 4 hours to reach saturation adsorption. The method can greatly simplify the operation steps of enzyme immobilization, which requires no modification in priorperiod for the material and the enzyme; the method has high universality, which can be used on a plurality of enzymes and carriers; and the method does not affect catalytic activity of the enzyme.