191 results about "Immobilizine" patented technology

To provide an organic substance-immobilized structure employing a novel immobilizing technique and a manufacturing method thereof using the novel immobilizing technique, where, when an organic substance, particularly a biological substance is immobilized on the surface of a substrate, the organic substance, particularly the biological substance can be stably immobilized on the surface of the substrate through orientation of the organic substance, particularly the biological substance suitable for exerting physiological functions thereof. At least part of the substrate's surface is constructed of one or more substrates containing aluminum oxide. The immobilization of the organic substance to the surface of the substrate is carried out by binding at least part of the binding domain to the surface of the substrate through a binding domain containing a peptide having an affinity to aluminum oxide and composed of at least one or more amino acids, which is coupled with the organic substance.

Provided herein are processes for the production of biodiesel. In particular, provided is an esterification process in which an alcohol reacts with free fatty acids in a lipid material comprising free fatty acids and glycerides in the presence of an immobilized zirconium(IV) metal salt to form fatty acid alkyl esters. Also provided is combination process in which an esterification reaction converts the free fatty acids in a lipid material to fatty acid alkyl esters and a transesterification reaction converts the glycerides in the material to fatty acid alkyl esters.

The invention relates to the field of biotechnology in the pharmaceutical industry, and discloses the preparation of an immobilized whole-cell catalyst and its application in the synthesis of a Puley intermediate (R)-2-hydroxy-4-phenyl-butyric acid. The recombinant cells containing the D-type lactate dehydrogenase gene and the formate dehydrogenase gene are established and fixed to prepare the immobilized whole cell catalyst. Using this catalyst to synthesize the intermediate (R)-2-hydroxy-4-phenyl-butyric acid of lisinopril, the reaction conditions are mild, the action is specific, and there are no by-products. It is used in medicine and food industries. value.

Embodiments of the present disclosure provide bisbiotin ligands and related conjugates and methods. The bisbiotin ligands, combined with streptavidin, can be used in the separation, labelling, targeting, and immobilization of biomolecules.

The invention provides an immobilized metal porphyrin enzyme catalyst and a preparation method thereof. The catalyst can oxidatively catalyze hydroxymethyl furfural (HMF) to generate a furandicarboxylic acid (FDCA) on presence of a selective oxidant.

The invention provides immobilized beta-glucosaccharase and a preparation method and application thereof. The method comprises the following steps of (1) macroporous resin pre-treatment; (2) macroporous adsorption resin NKA-9 polyetherimide surface modification; (3) macroporous resin absorption beta-glucosaccharase; (4) glutaric dialdehyde cross linking reaction modification. The metal ion Zn<2+> is used as an activating agent and a heat stabilizer of the immobilized beta-glucosaccharase for using macroporous adsorption resin for the first time. Through the participation of Zn<2+>, the catalysis efficiency of the beta-glucosaccharase is improved by 20 percent or higher; the catalysis efficiency of the immobilized enzyme NKA-9II is improved by 92 percent or higher. Relative to the initial enzyme activity, through the participation of Zn<2+>, after the incubation for 7h under the condition of 85 DEG C, the enzyme activity of the immobilized enzyme NKA-9II is maintained at a value being 80 percent or higher; after the incubation for 7h under the condition of 90 DEG C, the enzyme activity of the immobilized enzyme NKA-9II is maintained at a value being 54 percent or higher.

The present invention provides a method for transesterifying an ester, comprising combining the ester, a C1-C3 alcohol, and a heterogeneous catalyst of formula (I) or formula (II): wherein R′, R″ and R′″ are each H, (C1-C8)alkyl, (C6-C9)aryl, or (alk)3Si, wherein each alk is (C1-C4)alkyl; L is an organic linking moiety and X is a solid support material, and the salts thereof under conditions wherein the catalyst catalyzes the formation of the (C1-C3) ester of the acid portion of the ester and the corresponding free alcohol of the ester.

A chromatography media such as silica controlled pore glass or agarose containing an affinity ligand such as 2-Aminobenzimidazole (ABI) or aminomethylbenzimidazole (AMBI). The ligand is present in density of from about 30 to about 80 μmole / ml.

The invention relates to the process for the preparation of N,N-disubstituted imidazolium salts, N-heterocyclic carbene ligands and ruthenium catalysts containing N-heterocyclic carbene ligands, i.e. compounds of the general formulae (I) and, (II), compounds of the general formulae (III) and (IV) and compounds of the general formulae (V) and (VI), immobilised on inorganic oxide supports. The invention furthermore relates to the use of the immobilised compounds of the general formulae (I-IV) in organic, organometallic or transition metal-catalysed synthesis and to the use of the compounds of the general formulae (V) and (VI) as catalysts in organic and organometallic synthesis, in particular for C—C coupling reactions, such as olefin metathesis.

The invention discloses an immobilization method of a proteinase molecule by using a nano-material, namely a carbon nano-tube. The method comprises the following steps: (1) dispersing the carbon nano-tube to form a uniform solution; (2) acidifying the carbon nano-tube, wherein the carbon nano-tube obtained in the step (1) is acidified with mixed acid of concentrated nitric acid and concentrated sulfuric acid at a volume ratio of 1:2-1:5; and (3) functionalizing the carbon nano-tube, wherein a -NH2 functional group is functionally connected to the wall of the carbon nano-tube by utilizing NHS (N-hydroxy-succinamide) and EDC (1-ethyl-3-(3-dimethylaminopropyl) carbodiimide), and reaction is carried out with -NH2 of enzyme to immobilize the enzyme on the tube wall. The preparation process has convenient and quick procedure, stable and firm bonding and high efficiency. The carbon nano-tube for bio-catalysis or transformation in energy, environmental and biochemical pharmacy and other fields has low price, is easy for scale utilization, and is a nano-enzyme catalysis means with practical application prospect.

The invention relates to a method for catalytic synthesis of alpha-arbutin by using dissociative or immobilized candida lipase. The method comprises the process steps of selection of a reaction system, preparation of immobilized lipase, optimization of reaction conditions, extraction of a product, etc. According to the invention, hydroquinone and sugar mixed according to a mol ratio of 1: 1 to 1: 5 are added into a reaction medium, and lipase is used as a catalyst for a reaction; the reaction lasts for 10 to 120 h at a temperature of 20 to 60 DEG C, and a conversion rate of alpha-arbutin is as high as more than 95%; and the usage amount of lipase is 0.01 to 5 times of the weight of sugar. Alpha-arbutin is synthesized under the catalysis of dissociative or immobilized lipase in the method for the first time, and the method has the advantages of mild reaction conditions, repeated usability of lipase, a short production period, a simple extraction process for the product, etc., thereby enabling production cost to be substantially reduced.

The invention discloses immobilization hydrolase as well as a preparation method and application thereof. The immobilization hydrolase is prepared in a way that hydrolase is adsorbed and crosslinked on a magnetic chitosan composite microsphere by virtue of glutaraldehyde, and the magnetic chitosan composite microsphere is formed by crosslinking Fe3O4 nano particles with chitosan by virtue of glutaraldehyde. The preparation method of the immobilization hydrolase comprises the following steps: (1) adding Fe3O4 nano particles into a chitosan solution, and then adding a glutaraldehyde solution, so that a magnetic chitosan composite microsphere is obtained; (2), adding the magnetic chitosan composite microsphere into the glutaraldehyde solution for performing reaction, and adding the obtained carrier product into a hydrolase solution, so that the immobilization hydrolase is obtained. The immobilization hydrolase has strong stability on heat, strong acid, strong alkali, high ion strength, organic solvent and the like, is hardly inactivated and can be recycled. The immobilization hydrolase can be applied to an enhanced sludge hydrolysis process and can be used for solving the problems that an enzyme is used up in the enhanced sludge hydrolysis process and the enzyme can not be recycled.

The invention provides a preparation method of immobilized lipase. The preparation method is characterized by using amino-functionalized magnetic Fe3O4@C nanoparticles as carriers and immobilizing lipase on the surfaces of the nanoparticles through covalent binding. The method is specifically divided into two methods. The first method is characterized by firstly binding an aldehyde group at one terminal of glutaraldehyde with the amino groups on the surfaces of the amino-functionalized magnetic Fe3O4@C nanoparticles by utilizing nucleophilic reaction and then binding an aldehyde group at the other terminal of glutaraldehyde immobilized on the surfaces of the magnetic Fe3O4@C nanoparticles with the amino groups of lipase. The second method is characterized by converting the amino groups on the surfaces of the magnetic Fe3O4@C nanoparticles to carboxy groups by utilizing succinic anhydride and immobilizing lipase on the surfaces of the Fe3O4@C nanoparticles by utilizing amidation after activation with a mixture of N,N-dicyclohexylcarbodiimide and N-hydroxysuccinimide in a molar ratio of 1:(0.5-2). The preparation method has the beneficial effects that the preparation method is simple and mild; the lipase immobilization amount is large; the immobilized lipase has high enzyme activity and enzyme recovery and good stability, can be simply and conveniently recycled, effectively reduces the real cost of production and has wide application prospect.

The invention provides a penicillin G acylase mutant for synthesis and an application thereof in the preparation of amoxicillin. Penicillin G acylase of Achromobacter xylosoxidans origin is mutated by computer aided design in connection with semi-rational design of site-saturation mutagenesis technique and enzyme engineering modification of orthogenesis, thus acquiring the penicillin G acylase mutant lower in hydrolytic activity, better in synthetic activity, higher in synthesis-hydrolysis ratio (S / H), higher in acid resistance and better in stability, and amoxicillin can be catalytically synthesized more effectively and quickly. Immobilized enzyme hydrolytic activity of the mutant SPGA-4 obtained is decreased by 8.7 times, synthetic activity is increased by 5.6 times, the S / H ratio is increased by 8 times, the activity remains at 79% for 60 min under the condition of pH 2.0, amoxicillin is catalytically synthesized by a solid method at 10 DEG C or 20 DEG C, substrates 6-APA and D-HPM are directly charged in a solid form without dissolving, reaction pH need not be controlled, substrate conversion rate is higher than 99%, continuous use is available in more than 300 batches, and good operational stability is given.

The invention discloses a yeast which has the stereoselective lipase activity and the application of the yeast in the preparation of an S-type betaxolol hydrochloride by using a biological separation method. The method selects one yeast with the stereoselective lipase activity by screening from the soil, utilizes an immobilized cell which is obtained by immobilizing the wet bacteria or sodium alginate-activated carbon-polyethylenimine as an enzyme preparation, carries out an enantiomer separation to a substrate which contains acyl group and prepares the S-type betaxolol hydrochloride. The conversion method is simple, the cost is low and the stereoselectivity is better. The usage of the bacterial strain can carry out the chiral separation of Beta-receptor blocker of betaxolol etc., ephedrine hydrochloride, epinephrine, levodropropizine, salbutamol sulfate, captopril, zofenopril and other compounds which contain hydroxy group or acyl group at the chiral center, thereby having important application value for promoting the development process of the chiral drugs of China.

The invention discloses a method for producing L-2-aminobutyric acid by double immobilized multi-enzyme systems. The method provided by the invention comprises the following steps that 1, threonine dehydrogenase and leucine dehydrogenase are fixed on reversely-dissoluble pH-sensitive polymer carriers so that a co-immobilized multi-enzyme system is obtained; and 2, alcohol oxidase, formaldehyde dehydrogenase and formate dehydrogenlyase are fixed on reversely-dissoluble pH-sensitive polymer carriers so that a co-immobilized coenzyme regeneration system is obtained. The method for producing L-2-aminobutyric acid by the double immobilized multi-enzyme systems utilizes dissolution reversibility of co-immobilized enzymes, realizes effective separation of the co-immobilized enzymes and products, improves accessibility between the co-immobilized enzymes and reactants, improves recovery and utilization rates of threonine dehydrogenase, leucine dehydrogenase, alcohol oxidase, formaldehyde dehydrogenase and formate dehydrogenlyase, improves coenzyme regeneration efficiency, reduces follow-up separation purification processes, simplifies a process flow and reduces a production cost.

The invention discloses immobilized cyclodextrin glucoside transferase and a preparation method and application thereof. The preparation method comprises the following steps of: firstly, adding a molecular sieve into enzyme liquor; secondly, adding glutaraldehyde to crosslink mixed liquor; and finally embedding with sodium alginate and calcium chloride to obtain the immobilized cyclodextrin glucoside transferase. The immobilized cyclodextrin glucoside transferase prepared by the method has higher thermal stability and storage stability, can be immobilized by crude enzyme liquor and greatly reduces immobilization cost. The immobilized enzyme can be used for producing 2-O-glucosyl ascorbic acid, the conversion rate is up to 0.22 g*L<-1>*h<-1>, products after conversion are easy to extract, and the immobilized enzyme is also easy to recycle and can be reused.

The invention provides a degumming method of hydrated degummed soybean oil through immobilized phospholipase A2. Influences of various degumming conditions such as degumming temperature, degumming initial pH, additive amount of the immobilized phospholipase A2 and deguming reaction time on soybean oil degumming effects are researched, and a response surface analytical method is adopted to optimize the researched factors. According to the method, the activity and stability of an enzyme in a reaction system are improved, the oil and fat refining cost is greatly reduced; additionally, the immobilized phospholipase A2 is adopted to degum the soybean oil, the phosphorus content of the finally obtained degummed soybean oil through an enzymic method can be reduced to 9.83 ppm, the activity of the immobilized enzyme can be maintained above 54% after reused for seven times, and significance is provided for the oil and fat refining.

The invention discloses an immobilized glutamine transaminase preparation method. The method includes that natural macromolecular compounds such as sephadex, agarose gel, carrageenan and gelatin are adopted for enzyme immobilization, an immobilized embedded substance is immersed into a liquid mixture of alcohols, carbohydrates, an antioxidant and the like, and accordingly stability of an immobilized enzyme preparation is greatly improved. The immobilized glutamine transaminase preparation method has advantages that preservation time of glutamine transaminase at a low temperature, the normal temperature and a high temperature is greatly prolonged, and stability of glutamine transaminase is remarkably improved.

The invention discloses a multi-enzyme system with immobilized polyethylenimine and metal coordination and a method for preparing the multi-enzyme system, and belongs to the technical field of immobilized enzymes. The method includes immobilizing glycerol dehydrogenase, coenzyme oxidase and glycerol dehydrogenase-coenzyme oxidase fusion enzymes by the aid of coordination of polyethylenimine and metal ions; forming netted polyethylenimine frameworks by polyethylenimine molecules under coordination effects of imine and the metal ions in immobilizing procedures; forming coordination bonds by the metal ions coordinated on the polyethylenimine, C-end histidine tags of the glycerol dehydrogenase and the coenzyme oxidase and C-end histidine tags of the glycerol dehydrogenase-coenzyme oxidase fusion enzymes so as to acquire the immobilized multi-enzyme system. Coordination crosslinking effects can be realized by the metal ions in the immobilizing procedures, and the catalysis efficiency of the multi-enzyme system can be improved. The multi-enzyme system and the method have the advantages that the multi-enzyme system is high in multi-enzyme coupling efficiency, preparation conditions are mild, and processes are simple and feasible; the immobilized enzymes are high in immobilizing rate and activity recovery rate and good in reuse stability, the temperature stability can be obviously improved, and the like.

The invention discloses a method for immobilizing D-psicose3-epimerase through polydopamine-magnetic Fe3O4 nanoparticles, and belongs to the technical field of food processing. The method includes thesteps of preparing the D-psicose3-epimerase and polydopamine-magnetic Fe3O4 nanoparticles, and immobilizing the D-psicose3-epimerase through the polydopamine-magnetic Fe3O4 nanoparticles. The polydopamine-magnetic Fe3O4 nanoparticles have strong magnetic responsibility, can be effectively controlled by magnetic force under the effect of an external magnetic field, can enhance the immobilizing effect through high-activity phenolic hydroxyl groups and other functional groups on the surface of polydopamine, and improves the stability of a carrier and the capacity of an enzyme, and the enzyme activity of the immobilized enzyme can reach 450.45 U per gram of carrier or above.

The invention relates to a phosphatidase A1 immobilization method. The method comprises the following steps: 1, allowing a carrier selected from macroporous adsorption resin or ion exchange resin to contact with phospholipase A1 in order to immobilize the phospholipase A1 on the carrier; and 2, drying the phospholipase A1 immobilized carrier obtained in step 1 in order to obtain immobilized phospholipase A1. The invention also provides immobilized phospholipase A1 prepared through the method, and an application of the immobilized phospholipase A1. The immobilized phospholipase A1 has the functions of esterification and acid hydrolysis, can be repeatedly used in the oil industry, and has the advantages of high reaction efficiency low cost and potential industrial application.

The invention relates to novel immobilisable imidazolium salts of the general formulae (I) and (II) which contain a group R carrying SiR′n(OR′)3−n on one of the two nitrogen atoms and a sterically demanding cyclic hydrocarbon radical on the other of the two nitrogen atoms of the heterocyclic ring. The invention furthermore relates to the use of the compounds of the general formulae (I) and (II) as starting materials for immobilisable catalyst ligands, catalyst precursors and catalysts, and to the use as solvents in organic, organometallic and transition metal-catalysed synthesis and as catalysts in organic synthesis.

Belonging to the immobilized enzyme field, the invention discloses a preparation method for an immobilized 3-phenoxy benzoic acid degrading enzyme. The method includes the steps of: (1) centrifuging a Sphingomonas sp. SC-1 fermentation liquid, collecting thalli, mixing the thalli with a Tris-HCl buffer solution evenly, then conducting ultrasonic crushing under ice water bath, performing centrifugation to remove intracellular enzyme, adding the Tris-HCl buffer solution to cell debris deposits to a volume before crushing to make an enzyme solution; and (2) mixing the enzyme solution with a sodium alginate solution in certain proportion, dripping the mixed solution in a calcium chloride solution uniformly to perform embedding immobilization. The immobilized 3-phenoxy benzoic acid degrading enzyme prepared by the invention has high enzyme activity under a wide temperature range (4DEG C-40DEG C) and a wide pH range (5.0-9.0), and has strong continuous reaction performance. The preparation method is simple, and the production cost is low.

The invention belongs to the field of immobilized enzyme and discloses a method for double solidification of enzyme by using in-situ free radical polymerization technology and carbon nanometer material. According to the method, firstly, that carbon nano material is dispersed in a phosphate buffer solution to obtain a carbon nano tube dispersion; enzyme nano-capsule solution was prepared by in situradical polymerization to modify and protect enzyme molecules; then adding enzyme nano-capsule solution into the carbon nano-material dispersion, uniformly mixing and standing and, finally, the mixedsolution was separated and washed to obtain a precipitate which was called double immobilized enzyme. Compared with the traditional immobilized enzyme, the double immobilized enzyme system is based on nanocapsules. Compared with the traditional immobilized enzyme, the surface of the enzyme molecule is modified with a polymer layer, which greatly improves the stability of the enzyme molecule. Thedouble-immobilized enzyme obtained by the invention retains high enzyme activity and has higher environmental stability and reusability than free enzyme and traditional immobilized enzyme.

The invention provides a preparation method for synthesizing a polyphenol compound by using an enzyme immobilization technology. The preparation method comprises the following steps: mixing metal ionsor metal clusters with an organic ligand and tyrosinase, and immobilizing the tyrosinase to obtain a tyrosinase-metal organic framework compound; and using the tyrosinase-metal organic framework compound as a catalyst and a monophenol compound as a reactant so as to synthesize the polyphenol compound. A metal organic framework is formed by coordination of the metal ions or the metal clusters andthe organic ligand under the induction of the tyrosinase. Meanwhile, the metal organic framework forms a tyrosinase protective shell, so that the tyrosinase is immobilized, and the obtained immobilized tyrosinase has relatively high catalytic performance. Monophenol compounds are catalyzed by the immobilized tyrosinase to synthesize an ortho-hydroxylated polyphenol compound, and a reducing agent is added to inhibit the polyphenol compound from being further oxidized, so that the yield of the polyphenol compound is increased.

The invention discloses a mussel protein antihypertensive peptide with an antihypertensive activity as well as a preparation method and an application thereof. The amino acid sequence of the antihypertensive peptide is Val-Ser-Trp-Pro-Cys-Arg-Trp (VSWPCRW), and the molecular weight measured by ESI-MS is 935.07 Da. The active peptide Val-Ser-Trp-Pro-Cys-Arg-Trp (VSWPCRW) is obtained through shelling and meat taking, enzymolysis, ultrafiltration, agarose magnetic microsphere immobilized ACE adsorption and reversed-phase high-performance liquid chromatography (RP-HPLC) purification. The active peptide prepared by the preparation method disclosed by the invention is remarkable in angiotensin converting enzyme (ACE) inhibitory activity, and capable of being used for hypertension treatment-related medicines.

The invention discloses an application of acetone / isopropanol circulation in preparation of ursodeoxycholic acid (UDCA) by an enzyme method, and provides an environment-friendly, economic and efficient new idea for preparation of UDCA by a two-step enzyme method. The method comprises the following steps of 1, reducing acetone into isopropanol by using ketoreductase, and converting NADPH / NADH intoNADP+ / NAD+ to provide NADP+ / NAD+ for catalyzing chenodeoxycholic acid (CDCA) to generate 7KLCA by using 7alpha steroid dehydrogenase (7alpha HSDH); in the second step, isopropanol being oxidized intoacetone through isopropanol dehydrogenase, NADP+ / NAD+ being converted into NADPH / NADH, and NADPH / NADH being provided for catalyzing 7KLCA to generate UDCA through 7beta steroid dehydrogenase (7beta HSDH). According to the two-step enzyme method, immobilized enzyme is utilized, the process is simple, second-step reaction can be carried out through simple treatment after first-step reaction is finished, and intermediate purification is not needed; the addition amount of NADP+ / NAD+ in the reaction is small, and acetone and isopropanol can be recycled, so production cost and the environmental protection pressure are greatly reduced.