77results about How to "High immobilization efficiency" patented technology

The invention relates to a preparation method of a petroleum degrading bacteria solid inoculant. The preparation method comprises the following steps: A, screening and domestication of the petroleum degrading bacteria; B, preparation of a seed culture solution; C, fermentation of the solid inoculant; and D, drying, pulverization, measurement and packaging of composted products. The petroleum degrading bacteria solid inoculant has the beneficial effect of improving the petroleum dissolution effect by taking a nonionic surfactantpolysorbate-80 (tween 80) as a solubilizer. The preparation method has the advantages that solid fermentation raw materials are easy to get and the process is relatively simple; and the solid inoculant contains a large amount of carbon and nutrient elements so as to provide more proper matrix for the growth of the bacteria, has strong affinity for the microorganisms, has immobilization efficiency, is capable of improving the competitiveness and degrading efficiency of the added microorganisms and indigenous microorganisms, and is convenient to transport and operate agriculturally and suitable for the large-scale in-situ biological remediation of the petroleum-polluted soil.

The invention discloses co-immobilization glucose oxidase/catalase microspheres and application thereof in preparation of gluconic acid (salt) for catalytic oxidation of glucose. The co-immobilization glucose oxidase/the catalase microspheres can effectively promote covalent linkage of carriers and enzyme molecules, immobilization efficiency is improved greatly, service life of immobilization glucose oxidase (GOD)/catalase (CAT) is prolonged remarkably, the co-immobilization glucose oxidase/the catalase microspheres can be used repeatedly, are low in production cost, and facilitate sustainable development. Gluconic acid (salt) prepared by the co-immobilization glucose oxidase/the catalase microspheres is high in yield, reaction condition is temperate, a device is simple and easy to obtain, and the production method is environment-friendly.

This invention relates to a plastic substrate for a microarray chip, which is characterized in that an aminoalkylsilane with an aldehyde group derived from glutaraldehyde and introduced onto an amino group of the aminoalkylsilane exists on a surface of the plastic substrate, and also to a process for its production and a method of its use. Substrates according to the present invention have no variations in the immobilized amount of DNA fragments upon immobilization of DNA fragments, permit reproducible hybridization with high efficiency, and are excellent in storability.

The invention provides a method for preparing immobilized lipase by utilizing an amphipathy chitosan microsphere carrier, and relates to a method for preparing immobilized lipase. The method comprises the following steps: taking micron-order chitosan microsphere resin prepared by an inverse suspension crosslinking method as an immobilization carrier matrix, performing C2-bit benzene formolatiom on the chitosan microsphere matrix, leading in hydrophobic interaction groups, then using epoxy chloropropane to activate C6-bit hydroxyl, and coupling tetraethylenepentamine to prepare chitosan microspheres with hydrophobic phenyl and hydrophilic polyamine molecules, and taking the chitosan microspheres as the covalent immobilization carrier of lipase after coupling glutaraldehyde. According to the invention, the carrier is adopted to immobilize the antarctic candida lipase B, and the hydrophobic groups and hydrophilic arm molecules are led in, so that the immobilization efficiency of chitosan microspheres to the lipase is improved. When the quality of added lipase of the dry carrier per gram is 40 mg, and the antarctic candida lipase is immobilized for 3 h at room temperature under the condition that the pH is 7.5, the obtained immobilized lipase has the activity recovery rate of 197%, and has the activity of 481.6 U/g.

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

The invention discloses an immobilized porcine pancreatic lipase carrier, which is a macroporous bead crosslinked polymer containing succinimidyl ester groups and has a structure represented by the formula I. The carrier forms a high polymer with acrylic acid as a reactive monomer and N,N'-methylenebisacrylamide as a crosslinking agent, and in the meantime, incorporates the succinimidyl ester active group so that the time required for immobilizing the porcine pancreatic lipase is short, namely only 1 hour is required, and the immobilizing efficient is high and stability of activity retention of the immobilized enzyme is excellent; the apparent activity of the immobilized enzyme is high and can reach 750U/g and above. The invention also discloses a preparation method and application of the immobilized porcine pancreatic lipase carrier.

The invention relates to a process method of oxidized graphene directional immobilization glucose oxidase. The process method comprises the following steps that 1) oxidized graphene is dissolved in distilled water, and oxidized graphene turbid liquid with the concentration being 1mg/mL is obtained through ultrasonic mixing; N-hydroxy sulfo-succinimide and 1-ethide-3-(3-dimethyl aminopropyl) carbodiimide are added into the turbid liquid, the washing and the separation are carried out after shaking, and esterified oxidized graphene is obtained; and 2) activated concanavalin A is added into the uniform esterified oxidized graphene, the centrifugation is carried out after reaction, and sediments are fully washed; and then, glucose oxidase solution is added, the full washing is carried out after reaction, and the directional immobilization glucose oxidase is obtained after the vacuum freeze-drying for 24 hours. The method for preparing the immobilized enzyme has the advantages that the immobilization efficiency is high, and the defects that the orientation of enzyme activity sites is inconsistent, or the enzyme activity sites are covered, and the like are avoided, so the activity of the immobilized enzyme is improved, the enzyme activity loss is low, and the enzyme activity can reach 150 to 195U/mg.

The invention provides a purification coupled fixation method of myrosinase. By means of concanavalin combined with myrosinase, concanavalin is loaded on a base, myrosinase is linked to the concanavalin, and therefore, the myrosinase is specifically fixed to the base; purification and fixation of the myrosinase are combined together such that difficulty in purifying the myrosinase is greatly reduced, and fixing efficiency is improved; the concanavalin adsorbs glycosyl on the surface of the myrosinase so as to adsorb the myrosinase but not adsorb ESP protein, fewer nitriles are generated, the yield of isothiocyanates is increased, and conversion rate of glucosinolates into isothiocyanates is greatly increased.

The invention relates to a recombinant beta-glucosidase and an expression and purification method and the immobilization application thereof, and belongs to the technical field of biological enzyme engineering. The recombinant beta-glucosidase is recombinant beta-glucosidase Glu-linker-ELP-GB (GLEGB) obtained by linking binary labels ELP and GB with beta-glucosidase (Glu) through a universal linking peptide linker, wherein the sequence of GB polypeptide is linked to the 3' end of an ELP gene. By virtue of a reversible phase change property of an ELP label on recombinant enzyme, separation andpurification of the enzyme can be achieved; the recombinant enzyme containing the ELP label can be precipitated only by adding a certain amount of (HN4)2SO4 for incubation for a period of time; by virtue of a temperature response behavior of the ELP label, one-step rapid separation and purification of a target enzyme molecule is achieved; the enzyme purifying effect of the expression and purification method is superior to that of an ammonium sulfate precipitation method, and the purification cost of the expression and purification method is much lower than that of a chromatography method.

The invention discloses a method for synchronously purifying and immobilizing linoleate isomerase. The method comprises the steps of: treating attapulgite by using hydrochloric acid, adjusting the pH value by using solution of sodium hydroxide, stirring uniformly, filtering, washing, and drying to obtain acid modified attapulgite; mixing the acid modified attapulgite and solution of magnesium sulfate, stirring, filtering, and drying to obtain attapulgite treated by the magnesium sulfate; mixing the attapulgite treated by the magnesium sulfate, gamma-(methacryloyloxy)propyltrimethoxysilane, water and dimethylbenzene and stirring, filtering, washing, and drying to obtain modified attapulgite; mixing enzyme fluid of linoleate isomerase produced from Lactobacillus delbrueckii subsp bulgaricus and the modified attapulgite, stirring, filtering, and drying to obtain the purified and immobilized linoleate isomerase. The method for synchronously purifying and immobilizing the linoleate isomerase is simple and convenient, simple in steps, low in cost, good in purification effect, and high in immobilization efficiency; the activity recovery rate of enzyme reaches over 60 percent during the purification; and the immobilized enzyme has better pH adaptability, thermal stability and operation stability.

The invention provides a method for enzymatically modifying stevioside mother liquor sugar with immobilized biological enzyme, belonging to the technical field of food additives. Bitter taste of the stevioside is always an important factor limiting the practical application of the stevioside, and, currently, a main method for solving the problem is to extract rebaudioside-A from the stevioside or perform enzymatic conversion to change the taste of the stevioside. In the method, the immobilized biological enzyme is adopted for enzymatically modifying the stevioside, and the method comprises the following steps of: immobilizing cyclodextrin glucanotransferase by adopting an embedding-crosslinking method; screening influencing factors in the process of enzymatic modification; recycling the biological enzyme after the enzymatic modification is completed; and spray-drying the enzymatically modified products. By the method, the prepared product has the enzymatic modification rate of more than 90 percent, the adverse aftertaste of the stevioside is effectively improved, the biological enzyme can be recycled, and the production cost is effectively reduced.

The invention discloses a preparation method of a polyphenol oxidase immobilizing carrier and a method for immobilizing polyphenol oxidase by using the same. The method comprises the following steps: inserting a chitosan molecular chain into a montmorillonite lamella by adopting a solution intercalation method so as to prepare a chitosan/montmorillonite intercalation compound; performing in-situ reduction on chlorauric acid by using chemical structures with multiple hydroxyl groups and multiple amino groups of chitosan as a stabilizing agent and a reducing agent so as to realize one-step green synthesis of nano-gold particles; and preparing a chitosan-nano-gold/montmorillonite intercalation compound, and taking the intercalation compound as a carrier to immobilize polyphenol oxidase. The intercalation compound is an immobilizing carrier for polyphenol oxidase, and a prepared immobilized enzyme has the advantages of high immobilizing efficiency, high enzymatic activity, good stabilizer and the like.

The invention discloses a preparation method of an immobilized microbial agent from a modified wheat bran material. The preparation method comprises the following steps of S1, preparing a bacterial solution for later use; S2, carrier preparation, specifically, gradually carrying out amylohydrolysis and proteolytic modification on the wheat bran to prepare a carrier for later use, wherein the hydrolysis sequence is irrespective; and S3, microbial agent preparation, specifically, uniformly mixing the bacterial solution obtained in the step S1 and the carrier obtained in the step S2 according toa ratio of 1mL: (0.5-1) g, and drying to constant weight. Common wheat bran is modified and used as the carrier of the microbial agent, and the immobilization efficiency is high; due to the fact thatthe specific surface area of the modified wheat bran is greatly increased, a rich micropore growth environment is created for microorganisms, the adsorption amount and the containing amount of the microorganisms are increased, the modified wheat bran can be used as a novel carrier, and the viable count in the microbial agent prepared by adopting the carrier far exceeds the executive standard.

The invention discloses a genetically engineered bacterium for efficiently immobilizing enterococcus faecium glutamate decarboxylase and an immobilization method. The invention provides the genetically engineered bacterium, namely Escherichia coli (DH5alpha-LNSF1), for efficiently immobilizing enterococcus faecium GAD. The engineered bacterium is preserved in the Guangdong Microbial Culture Collection Center on September 18th, 2018, and the preservation number is GDMCC No.60445. The genetically engineered bacterium can be used for efficiently producing enterococcus faecium GAD immobilized enzyme. According to the method, a modified chitin affinity adsorption CBM-GAD fusion enzyme crude enzyme solution is used for obtaining the enterococcus faecium GAD immobilized enzyme. Modified chitin and CBM-GAD fusion enzyme which are used in the method are high in affinity and firm in combination, and the obtained GAD immobilized enzyme is high in activity and stability. The problems that GAD is difficult in immobilization, short in half-life period and high in cost are solved. In addition, the immobilization method is rapid and high in efficiency, the cost is greatly reduced, and the method has wide application prospects in the aspect of industrial production of efficient production of enterococcus faecium GAD immobilized enzyme.

An efficient method of xylanase immobilization and improving reutilization effect thereof, the immobilization method is: blending Eudragit L-100 and xylanase by a certain proportion, agitating continuously 1.5-3 hours at 25 DEG C to obtain xylanase. After reaction of the xylanase and substrate, adding coagulator, regulating pH value from 3.8 to 4.2 by acetic acid, quiescence 25-40 minutes, centrifugating 30 minutes in condition of 5000xg to finish recovery of immobilized enzyme. The advantages of the invention used to industrialized enzyme method for producing xyloolgosaccharides are: efficient immobilized efficiency for immobilizing enzyme on the carriers; the immobilized enzyme can homogeneous react with substrate to improve enzyme efficiency; using method of improving reutilization of the immobilized enzyme can improve reutilization efficiency of immobilized xylanase using reversible solution material Eudragit L-100remarkbly as carrier.

The invention relates to a chemical modified fiber type immobilization carrier as well as a preparation method and an application thereof. The invention provides a method for preparing the chemical modified fiber type immobilization carrier. The method comprises the steps as follows: 1), the fiber type immobilization carrier is soaked in a polyethyleneimine solution, so that the carrier modified by polyethyleneimine is obtained, wherein, polyethyleneimine molecules are formed in the surface of the carrier modified by the polyethyleneimine; 2), the carrier modified by the polyethyleneimine is soaked in a glutaraldehyde solution, so that amino groups and aldehyde groups of glutaraldehyde in the polyethyleneimine molecules have reactions, and the chemical modified fiber type immobilization carrier is obtained. According to an embodiment of the chemical modified fiber type immobilization carrier, the chemical modified fiber type immobilization carrier can be taken as a cell immobilization carrier and can be filled to form an immobilization fiber bed, and is applied to single-batch fermentation and consecutive batch fermentation of biobutanol.

The invention belongs to the field of immobilized enzymes and in particular relates to an immobilized enzyme of a core-shell structure as well as a preparation method and application of the immobilized enzyme. The preparation method comprises the following steps: preparing an enzyme solution from zymoprotein, adding a modifier into the enzyme solution, uniformly mixing, and reacting for 2-10 hoursto obtain a surface double bond modified zymoprotein; and adding a monomer in an amount of 1-10 times that of the mass of the solution into the double bond modified enzyme solution, performing vortexblending uniformly, removing dissolved oxygen, sequentially adding a cross-linking agent, an initiator and a catalyst, reacting the solution at room temperature for 1-5 hours, dialyzing and purifying, thereby obtaining the immobilized enzyme of the core-shell structure. Compared with the traditional immobilized enzyme method, the method disclosed by the invention has the advantages that the enzyme is immobilized on a small scale, and the enzyme immobilization efficiency is improved, so that all the enzyme molecules are embedded and fixed by the finally formed 'shell' structure. Meanwhile, theprepared immobilized enzyme system has excellent properties such as high enzyme activity retaining rate and environmental stability, wide application field and the like.

The invention relates to immobilized lysine decarboxylase, a preparation thereof, a 1, 5-pentane diamine preparation method and a product. The invention specifically relates to immobilized lysine decarboxylase comprising a diazo group-modified polymer carrier and lysine decarboxylase, and a method for preparing immobilized lysine decarboxylase by the diazo group-modified polymer carrier; a methodfor preparing 1, 5-pentane diamine by the immobilized lysine decarboxylase, and the prepared 1, 5-pentane diamine thereof. The immobilization method has high enzyme immobilization efficiency and goodstability in use, improves the use efficiency of enzyme, solves the problem of poor use stability of free lysine decarboxylase (cells) in traditional processes, greatly reduces the use cost of the carrier and the biological production cost of 1, 5-pentanediamine, simplifies the separation step of 1, 5-pentane diamine solution and enzyme, increases the degree of automation of the production of 1, 5-pentane diamine, and promotes the process of industrialization of biological production of 1, 5-pentanediamine.