175 results about "Immobilizine" patented technology

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.

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 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 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 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 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 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.

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.

The invention discloses immobilized maleic acid cis-trans isomerase and a preparation method and application thereof, and belongs to the technical fields of enzymology and enzyme engineering and the field of biochemical industries. According to the immobilized maleic acid cis-trans isomerase and the preparation method and application thereof, R5 peptide fragments are connected to the N ends of the maleic acid cis-trans isomerase, R5-MaiA is crosslinked through glutaraldehyde, embedding is conducted on the crosslinked R5-MaiA by utilizing silicon reaction liquid, and immobilization of the maleic acid cis-trans isomerase is completed. The immobilized maleic acid cis-trans isomerase (R5-MI-CLEA-Si) with high stability is obtained through the method which is low in cost and easy to operate, the optimum reaction temperature of the immobilized isomerase is 55 DEG C, and the optimum reaction pH is 7.7; on the condition that the temperature is 55 DEG C, the half-life period of the immobilized isomerase is 4 h, the half-life period of the free isomerase is 0.5 h, the stability of the immobilized isomerase is eight times of that of the free isomerase, and after a catalytic reaction is repeated for eight times, the enzyme activity of the immobilized isomerase can be still retained at 75%.

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 temperature-sensitive nano gel material based on a polyquaternary phosphonium salt ionic liquid, and belongs to the field of new materials. With methanol as a solvent, azodiisobutyronitrile as an initiator and CTA1 or CTA2 as a chain transferring agent, and under the protection of N2, the temperature-sensitive nano gel material is obtained by carrying out a cross-linking reaction of a chlorinated 4-vinyl benzyl triphenyl butyl phosphonium ionic liquid. The hydrodynamic diameter of the prepared temperature-sensitive nano gel is 20 to 90 nm, the particle size distribution index is 0.112 to 0.464, and phase transformation of a macro gel and a micro gel can be achieved in a temperature range of -5 to 13 DEG C. The temperature-sensitive nano gel material has reversible temperature response, and has broad application prospects in drug release, enzyme immobilization, material separation, immunoassay, catalysis and other fields after the surface is grafted with polymerized N-isopropylacrylamide PNIPAm.

The invention discloses a magnetic immobilized lipase and an application thereof in preparation of (R)-(+)-N-acetyl-1-methyl-3-amphetamine through resolution of 1-methyl-3-amphetamine. According to the method, glutaraldehyde is replaced with carbon disulfide, dithiocarbamate is introduced to the surfaces of magnetic particles to be covalently combined with lipase. The method for preparing the magnetic immobilized lipase is simple and safe and is obviously superior to a glutaraldehyde cross-linking enzyme immobilization method. Meanwhile, the magnetic immobilized lipase is applied to an alternating magnetic field to catalyze an asymmetric acylation reaction, so that continuous preparation of (R)-(+)-N-acetyl-1-methyl-3-amphetamine is realized. The method is simple, safe, effective and easyin resolution. The enantiomer excess value of an R-configuration product obtained by resolution of 1-methyl-3-amphetamine reaches 98.5%, and the resolution effect is excellent.

The invention relates to a method for preparing an iron ion immobilization affinity chromatography monolith column, which comprises: preparing porous monolith column substrate by polymerization in capillaries with N-hydroxysuccinimide serving as a functional monomer, ethanedimethylethyl acrylate as a crosslinking agent, mixture of cyclohexanol, dodecanol and N,N-dimethylformamide as a porogen andazodiisobutyronitrile as an initiator; bonding carboxyl-containing functional groups on the surface of the monolith column substrate through surface modification for subsequent iron ion immobilization; and immobilizing iron ions by using iron ion containing solution to accomplish the preparation of the iron ion immobilization affinity chromatography monolith column. The invention has the advantages that: the preparation method is simple and convenient; and the micro structure of the obtained substrate material is uniform. Compared with other metal ion immobilization affinity chromatography monolith column, the iron ion immobilization affinity chromatography monolith column prepared by the method has high preparation repeatability, and has stable performance and high specific enriching capacity for phosphorylated peptide fragments.

The invention discloses a method for synthesizing L-tryptophan by immobilized enzyme. In the method, immobilized tryptophanase is taken as an enzyme source, L-cysteine and indole are taken as substrates, the L-tryptophan is produced through enzymatic conversion, the immobilized enzyme is removed through filtration, and the L-cysteine is obtained through further separation. In the method, the tryptophanase is immobilized through GE resin, and the optimized catalysis reaction conditions are that: the pH is 8.5, the reaction time is 3 hours, the concentration of the substrates is 10g/L of L-cysteine and 10g/L of indole, the cell concentration of the enzyme source is 20mL/L, and the concentration of phosphopyridoxal coenzyme is 0.15g/L. Under the optimum reaction conditions, the conversion rate of the cysteine substrate is 81.5 percent. Because the immobilized enzyme is adopted for direct enzymatic synthesis, the method is easy and convenient to implement, high in catalysis efficiency and low in fermentation cost, and the products are easy to separate; therefore, the method has good industrial prospect in the field of industrial production of L-tryptophan.

The invention discloses an immobilizing method of L-aspartic acid-alpha-decarboxylase, and belongs to the technical field of biological engineering, TiO2 modified by a poly amino acid is used as a carrier, the L-aspartic acid-alpha-decarboxylase is bonded onto the surface of the carrier, and the poly amino acid is gamma-polyglutamic acid or epsilon-polylysine. Immobilization studies on the L-aspartic acid-alpha-decarboxylase by respective use of the modified carrier and an unmodified carrier find that the enzyme carrier content of the modified carrier is about 8 times of that of the unmodified carrier.

The invention discloses a magnetic COFs based immobilized enzyme, a synthetic method and applications of the immobilized enzyme in preparation of biodiesel. The immobilized enzyme includes nanoparticles with core-shell structures and immobilized hydrolase; the core is Fe3O4 magnetic nanoparticles, and the shell is COFs; and the immobilized hydrolase is RML. The synthetic method includes the synthesis of the magnetic COFs and the immobilization of the enzymes; a magnetic COFs material is synthesized and then used to immobilize the RML so as to obtain the immobilized enzyme RML@Fe3O4@COF-OMe. The immobilized enzyme is good in stability; the characteristics of the RML of high catalytic activity can be reserved; and the high yield effects of the immobilized enzyme in preparation of the biodiesel can be achieved.