34 results about "Immobilized cellulase" patented technology

Belonging to the technical field of nano materials, the invention relates to a multi-arm magnetic graphene oxide composite microsphere, a preparation method and application thereof. The method includes: subjecting GO@Fe3O4 and 4arm-PEG-NH2 to covalent binding to prepare GO@Fe3O4@4arm-PEG-NH2, then dissolving a certain amount of GO@Fe3O4@4arm-PEG-NH2 in a citrate buffer solution, then adding glutaraldehyde into the solution, and carrying out activating reaction for certain period of time; and washing the obtained product with the citrate buffer solution, and then conducting vacuum drying to obtain the material needed by the invention. The material obtained by the invention has superparamagnetism, the carry capacity of the composite microsphere applied to immobilized cellulase is increased to 575mg/g, and the obtained immobilized enzyme has the advantages of high temperature resistance, strong storage capacity and high reusability.

The invention discloses a preparation method of immobilized cellulase, relating to an enzyme immobilization method. The method comprises the following steps of: firstly, mixing chitosan with the degree of deacetylation of 90% with an acetic acid solution, then injecting the mixed solution into a 2% sodium tripolyphosphate solution, filtering and washing to obtain neutral spherical chitosan-polyethylene glycol carriers, then activating the carriers by a glutaraldehyde solution, carrying out immobilization reaction of the carriers and cellulase, and filtering and washing to obtain immobilized cellulase. The immobilized cellulase prepared by the method has the advantages of high mechanical strength, good enzyme stability, high enzyme activity, strong binding force between the enzyme and the carriers and high rate of immobilization.

The invention provides a method for improving enzymatic activity and utilization rate of immobilized enzymes in an alternating magnetic filed three-phase fluidized bed. The method can effectively improve the enzymatic activity and the utilization rate of magnetic immobilized cellulolytic enzymes by applying an alternating magnetic field to a magnetic field three-phase fluidized bed and regulating and controlling various parameters of the alternating magnetic field, such as magnetic field strength, magnetic field action time, gas velocity, fluid velocity and the like. The magnetic immobilized cellulolytic enzymes used by the method of the invention have excellent stability and can be repeatedly used for 9 times after being recycled, and the recovery ratio of the enzymatic activity is over 42 percent. The method can improve the enzymatic activity and the utilization rate of magnetic immobilized cellulolytic enzymes so as to effectively reduce cost and simultaneously can improve the degradation rate of macromolecular substances and provides the basis for realizing continuous production.

The invention discloses an immobilized cellulose and a method for hydrolyzing plant material thereby for extracting active ingredients. The carrier of the immobilized cellulose is N-succinyl-chitosan, and the cellulose is selected from Trichoderma viride, aspergillus niger or fusarium cellulose; in the process of using the immobilized cellulose for hydrolyzing plant material, the immobilized cellulose can be dissolved in the solution or separated out in gel form by controlling the pH value of the solution, thus not only leading the immobilized cellulose to fully contact with the plant material and further improving the hydrolysis efficiency of the immobilized cellulose, but also faciliating the separation and recycling of the immobilized cellulose.

Cellulase immobilization and cellulose hydrolysis methods are provided. According to the cellulase immobilization method, sodium alginate is adopted as a carrier, a principle that an ion exchange reaction occurs between oxygen atoms in OH- and COOH- in the sodium alginate and Ca<2+> is utilized to form gel, polyethylene glycol is adopted as a pore-forming agent and a hydrophilic agent, glutaraldehyde is adopted as a crosslinking agent, and a polyelectrolyte membrane formed between chitosan and the sodium alginate-polyethylene glycol immobilized enzyme is utilized to improve stability of the immobilized enzyme. The immobilized cellulase is used for cellulose hydrolysis, and repeated utilization of the cellulase can be achieved. Through addition of the polyethylene glycol and the chitosan, the enzyme activity immobilization rate and enzyme activity of the immobilized cellulase are improved, the Michaelis constant is reduced, pore density of a carrier surface is increased, and the reducing sugar content of an enzyme hydrolysis reaction, the enzyme hydrolysis yield and the TS degradation rate are increased. The preparing and application methods are simple to operate, immobilization conditions are mild, investment is low, and the methods are environmentally friendly.

The invention provides a novel oil extraction method which combines enzymolysis with a solvent and aims at overcoming the defects that lixiviation time of a lixiviation method is relatively long, the dosage of enzyme of an aqueous enzymatic method is high, and the enzyme cannot be used repeatedly. The novel oil extraction method comprises the steps that cellulase and alkali protease are immobilized under an optimum immobilization condition, and an enzyme membrane is taken out to be wound on the surface of a cylinder, so as to obtain an immobilized cellulase roll and an immobilized alkali protease roll; then the moisture of germ flakes is adjusted by adjusting the humidity of the air under a vacuum condition, and the optimum relative humidity is 75%; the immobilized cellulase roll is used to roll the germ flakes continuously for 30min to allow the cellulase to perform the enzymolysis on plant cell walls; and then the immobilized alkali protease roll is used to roll the germ flakes continuously for 30min for separating protein from oil. After the enzymolysis of the germ flakes is finished, the solvent is used to extract the oil, the extraction time is only required to be 35min, and the residual oil reaches below 0.5%. In addition, an immobilized enzyme column can act on the material germ continuously for more than 24h, and the enzymic activity is still kept at 80%.

The invention provides a method for improving enzymatic activity and utilization rate of immobilized enzymes in a pulsed magnetic filed three-phase fluidized bed. The method can effectively improve the enzymatic activity and the utilization rate of magnetic immobilized cellulolytic enzymes by applying a pulsed magnetic field to a magnetic field three-phase fluidized bed and regulating and controlling various parameters of the pulsed magnetic field, such as magnetic field strength, magnetic field action time, gas velocity, fluid velocity and the like. The magnetic immobilized cellulolytic enzymes used by the method of the invention have excellent stability and can be repeatedly used for 9 times after being recycled, and the recovery ratio of the enzymatic activity is over 42 percent. The method can improve the enzymatic activity and the utilization rate of magnetic immobilized cellulolytic enzymes so as to effectively reduce cost and simultaneously can improve the degradation rate of macromolecular substances and provides the basis for realizing continuous production.

The invention belongs to the technical field of immobilized enzymes, relates to a soluble-insoluble UCST type PMAAc vector as well as an immobilized enzyme and an application thereof, and in particular to a preparation method of the soluble-insoluble high critical solution temperature (UCST) type sodium polymethylacrylate (PMAAc) vector, an application of the immobilized enzyme and an applicationof an immobilized enzyme degraded cellulose material thereof. The preparation method comprises the following steps: dissolving acrylic acid and ammonium persulfate through de-ionized water; slowly dropping methacrylamide which is dissolved by de-ionized water; uniformly shaking an obtained solution, filling nitrogen until bubbling and transferring an obtained solution into a single-mouth flask; implementing vacuumizing in nitrogen protection; conducting a reaction under magnetic stirring; implementing centrifuging after conducting cooling; discarding supernatant liquid; and heating up a resuspended material via de-ionized water, so that the finished product (the PMAAc vector) is obtained. The vector provided by the invention is applicable to immobilized cellulase, and the vector is used for degrading cellulose materials; the immobilized enzyme prepared by the invention is excellent in storage stability and reuse effect; and efficiency of hydrolyzing and catalyzing the cellulose materials is also obviously improved.

The invention relates to the field of biochemical engineering, and discloses a preparation method of nano immobilized cellulase, which comprises the following steps: adding a HCl solution into ethyl orthosilicate while stirring, and hydrolyzing to obtain a transparent solution; cooling the transparent solution, recovering ethanol, and controlling the ethanol volume percent of the transparent solution to be less than 1%; adding a glucose solution into the transparent solution while stirring, then adding a cellulase citric acid-sodium citrate buffer solution, finally regulating the pH value to approximate neutral, transferring to 4-10 DEG C, and gelatinizing; and grinding the wet gel obtained after gelatinizing, washing with water to remove the glucose, performing vacuum filtration on the mixed solution after water washing to obtain a white gel, drying the gel, grinding, and screening for later use. Experiment shows that the enzyme activity is almost kept unchanged after the nano immobilized cellulase prepared by the method is repeatedly used multiple times; and the method can be used for large-scale production of cellulosic ethanol.

The invention provides a preparation method for an immobilized cellulase of an enteric coating material, which comprises the following steps: 1) evenly dispersing the enteric coating material Eudragit L-100 into the acetic acid-sodium acetate buffer solution with a pH value of 5.5; 2) adding in the cellulase solution and crosslinking agent solved into the acetic acid-sodium acetate buffer solution with the pH value of 5.5 in sequence, and agitating and stewing for 3 to 6 hours under room temperature; 3) using 3M acetic acid to adjust the pH into 3.8, and centrifuging the solution for 10 min at the speed of 5000rpm; 4) then precipitating and dissolving the enteric coating material into the acetic acid-sodium acetate buffer solution with the pH value of 5.8, thus obtaining the immobilized cellulase. By changing the pH value of the solution, the immobilized cellulase can conveniently complete the homogeneous enzymolysis and heterogeneous recovery of the immobilized enzyme, and is able to ensure that the immobilized enzyme conducts high efficiency enzymolysis of the insoluble cellulose material in homogeneous state, and the by-product lignin after the enzymolysis can be quickly separated. Therefore, the immobilized cellulase of an enteric coating material has the advantages of capacity for repeated use, minor change in the enzymatic activity retention rate, simple and convenient preparation process, much lower costs of using the cellulase, and good popularization and application values.

The invention discloses a preparation method of sustained-release immobilized cellulase for feeding, which belongs to the field of cellulase preparation. The method comprises the following steps: taking gelatin, mixing the gelatin with deionized water, a glucose aqueous solution and a polyethylene glycol aqueous solution, and mixing the obtained mixture with glycerin, a glacial acetic acid solution and a crosslinking agent so as to obtain a sustained-release coating material for standby application; and taking and mixing mannitol, tryptone, yeast and agar, adjusting the pH value of the obtained mixture so as to obtain a clostridium perfringens culture medium, inoculating the clostridium perfringens culture medium, inoculating clostridium perfringens in a rapeseed bran cake so as to obtain a rapeseed bran cake-glucose solution, after the clostridium perfringens are subjected to sealed culture, opening the culture medium for feeding air, evaporating clostridium perfringens containing rapeseed bran cake particles until the particles are dry, mixing the dry particles with the standby sustained-release coating material, pressing the obtained mixture in a die, and drying the obtained product, so that the sustained-release immobilized cellulase is obtained. The method disclosed by the invention has the beneficial effects that the preparation cost is reduced by 18% in comparison with traditional methods, the production efficiency is high, the enzyme activity of the cellulase is increased by 42%, and the enzymatic activity is greater than 9250 IU/g.

The invention discloses a preparation method of modified nano-hydroxyapatite carried immobilized cellulase, and relates to the field of preparation of bio-enzyme preparations. Specifically, the method comprises the following steps: firstly, preparing modified nano-hydroxyapatite; then, preparing a cellulase water solution which is 1-2% in mass concentration, mixing and stirring a buffer solution which is constituted by the cellulase water solution as well as salicylic acid and sodium salicylate, regulating pH value of the solution to 3-5, adding the prepared modified nano-hydroxyapatite, and implementing stirring for 1-4h; and finally, adding a glutaraldehyde water solution which is 5% in mass concentration, conducting stirring at room temperature for 1-2h, then implementing filtering, washing an obtained solid by virtue of de-ionized water for 2-3 times, and implementing drying, so that the immobilized cellulase is obtained. The immobilized cellulase prepared by the invention is good in stability, excellent in reuse performance, safe and non-toxic, and low in preparation cost.

The invention discloses a method for preparing immobilized cellulase. Cellulase serving as a raw material is immobilized by using UiO-66MOFs material as a carrier. The method comprises the following steps: (1) dissolving 50-100mg of cellulase in ultrapure water, adding 50-100mg of a UiO-66MOFs material into the mixture, performing oscillation in a thermostatic water-bath at 20-40 DEG C for 8-20 hours to obtain an oscillation product; (2) centrifuging the oscillation product in the step (1) to obtain centrifugal supernate and a centrifugal product; and (3) drying the centrifugal product in thestep (2) to obtain the immobilized cellulase. The method has the advantages of high enzyme loading and high activity recovery rate. The immobilized cellulase can be used for effectively degrading cellulose macromolecules in reconstituted tobaccos when being applied to raw material pre-treatment of reconstituted tobaccos, and main aroma components are added to reduce nicotine and improve the aromaand harmony of the reconstituted tobaccos.

The invention discloses an enzymatic method for removing seed coats of walnut kernels in combination with infrared drying. The walnut kernels are added into immobilized cellulase for enzymatic hydrolysis and then filtered for use; the walnut kernels are added into a protease solution, the pH is adjusted, heat preservation and enzymatic hydrolysis are conducted at 55-65 DEG C, and then the solutionis filtered for use; the walnut kernels are fished out for ultrasonic rinsing, the epidermis is filtered, and the water is wiped out; then the walnut kernels are placed in a drying box for drying at50-65 DEG C until the moisture content is equal to or lower than 8%; finally, the walnut kernels are subjected to winnowing to obtain peeled walnut kernels. According to the provided method for removing the seed coats of the walnut kernels, the cellulase is prepared into an immobilized enzyme agent to react with the protease first and the walnut kernels later, so that mutual inhibition of the twoenzymes due to different reaction conditions is avoided, the hydrolysis efficiency of the enzymes is improved to the maximum degree, and the peeling time is shortened; by immobilizing the cellulase, the activity duration of the cellulase is prolonged, and the half-life period of the activity of the enzymes is prolonged.

The invention relates to a method for preparing immobilized cellulase. As free enzyme is mixed and dissolved in a reaction system and is difficult to not only separate and recover but also use repeatedly or continuously, free cellulase is immobilized. Cellulose acetate is immobilized on polypropylene composite films with different pore diameters, an effect of immobilizing the cellulose acetate to the composite films is observed through a scanning electron microscope, a polypropylene composite film with an optimal pore diameter is selected out, and the selected optimal composite film is immobilized on a roll. Conditions for immobilizing the free cellulase are that the glutaraldehyde concentration is selected to be 0.45-0.55%, the immobilization temperature is 48-49 DEG C, the pH (potential of hydrogen) value for immobilization is selected to be 3.9-4.0, the crosslinking time is 3-4 hours, the adsorption time is 3.0-3.1 hours, and the cellulase concentration is 1.0-2.0%. The activity of the free cellulase is measured to be 5.103 U/cm<2>, the enzyme activity still keeps about 80% after the roll for immobilizing the cellulase continuously presses soybean embryo slices for 24 hours, and the activity of the obtained immobilized cellulase is 4.732 U/cm<2>.

The invention provides a process for transforming geniposide under an enzymatic action of polyurethane foam immobilized cellulase. By using unit perforated polyurethane foam as a carrier of the immobilized cellulase, the process for transforming the geniposide under the enzymatic action of the polyurethane foam immobilized cellulase is obtained, so that the cellulase is fixed in a pore unit of thepolyurethane foam in the embedded form; by optimizing a foaming material and the usage of the foaming material, the opening percentage and aperture size are controlled to control the contact area between the cellulase and a substrate; and therefore, the cellulase repeated use frequency and use efficiency are enhanced in the case of guaranteeing a certain geniposide transformation rate. It is shown by an experimental result that the geniposide cumulative transformation rate of the polyurethane foam immobilized cellulase is 413.3% of equivalent-quality free cellulase after the polyurethane foamimmobilized cellulase is used for seven times in circulation; and moreover, selectable ranges of an enzyme and an glucoside suitable for the process are very large; according to the differences between the glucosides needing to be transformed, different invertases are selected to be immobilized. In addition, the process for transforming the geniposide under the enzymatic action of the polyurethane foam immobilized cellulase, provided by the invention, has the advantages that adopted instrument equipment is simple, the operation is simple and convenient, the preparation cost is low and the industrialization is easy to expand.

The invention relates to a method for ultrasonically improving the activity of immobilized cellulase, comprising the following steps of: (1) preparation of the immobilized cellulase: accurately weighing and dissolving chitosan and 2% by volume of acetic acid water solutions into transparent colloidal solution, adding 5% by weight of NaOH water solution to a certain amount of the transparent colloidal solution to obtain a flocculent substance, and then adding 5% by volume of glutaraldehyde water solution to obtain crosslinked chitosan; and adding cellulase to the crosslinked chitosan, washing the mixture 3-4 times in water in the second day to remove free enzyme, and then freeze-drying to obtain the immobilized cellulase; and (2) ultrasonic preprocessing of the immobilized cellulase: processing the immobilized cellulase by adopting an ultrasonic generator for 5-30 min, wherein the power of the ultrasonic generator is 10-80 W, and the frequency is 18-29 KHz. The method greatly improves the activity of the immobilized cellulase by combining an enzymologic bioengineering technique with an ultrasonic technique.

The invention discloses a method for reducing the chromaticity of waste liquid in litsea cubeba essential oil, and the method mainly comprises the following steps: 1, adding immobilized cellulase andimmobilized glucanase into litsea cubeba essential oil extraction waste liquid, and performing stirring, wherein the amount of the immobilized cellulase is 1-40% of the weight of the litsea cubeba essential oil extraction waste liquid, the amount of the immobilized glucanase is 1-40% of the weight of the litsea cubeba essential oil extraction waste liquid, and the stirring time is 1-120 minutes; and 2, adding immobilized glucose oxidase into the mixed solution treated in the step 1, performing stirring for 1-120 minutes, and conducting standing for 30-1040 minutes, wherein the amount of the immobilized glucose oxidase is 1-40% of the weight of the mixed solution. The method has simple production and processing steps, is convenient to operate, can effectively reduce the chromaticity of thewaste liquid in the litsea cubeba essential oil, the processing cost is greatly reduced, and the product quality is stable and reliable.

The invention provides immobilized cellulase as well as a preparation method and application thereof, and belongs to the technical field of papermaking. According to the method, cellulose nanofiber liquid and cellulase liquid are mixed to obtain the immobilized cellulase. The immobilized cellulase provided by the invention can maintain the strength of paper sheets while improving the slurry treatment efficiency of the biological enzyme.

The invention discloses a preparation process of a high-corrosion-inhibition cleaning agent used for a motor vehicle water circulation system. Gelatin is dissolved in redistilled water, a ferrous chloride solution and a hydrogen peroxide solution are added, stirring is carried out, a sodium hydroxide solution is added dropwise, and reacting is carried out for several hours in a boiling water bathto obtain nano-scale magnetic gelatin particles; cellulase is added, oscillating is carried out for several hours at the room temperature, the redistilled water is used for washing till no enzyme protein is washed out, so that the magnetic immobilized cellulase is obtained; then a certain amount of plant straw is taken and is subjected to crushing, then the quantitative water and magnetic immobilized cellulase are added, standing is carried out for several hours after stirring, and pressure filtration is carried out to remove residues to obtain a straw extracting solution; and the straw extracting solution, an organic cosolvent, a surfactant and a defoaming agent are added into a reaction kettle to be stirred and mixed to obtain the cleaning agent. According to the process, a crop straw extract is used as a main raw material, so that the source is rich, the process operation is simple, and the cost is low; and one-time and quick clearing under the conditions of multiple types of material and multiple types of dirt can be achieved, and the environment is not polluted.