81 results about "Lipase preparation" patented technology

A preparation technique of chitosan microspheres immobilization lipase comprises the steps that: (1) lipase from candida SP is dissolved in phosphate buffer solution by taking chitosan microspheres as an immobilization carrier and adopting glutaraldehyde cross-linking, lipase A is obtained. (2) candida SP lipase solution of 0.70-0.80mg / mL prepared by phosphate buffer solution b with a pH value of 6.5-7.5 is added with immobilization lipase A (final concentration is 0.005-0.0055g / ml) and after being stirred, ethyl (3-dimethylamino propyl) carbodiimide hydrochloride solution is added, the mixture is then stirred to be fully immobilized, chitosan microspheres immobilization lipase B is obtained after the treatment of the mixture. The chitosan microspheres immobilization lipase prepared by the preparation technology of the invention improves apoenzyme combination rate and activity recovery, has powerful immobilization lipase activity and specific activity and the immobilization lipase can be recycled for six times.

The invention discloses a strain WBRD1.11121501 producing surface active agent resistance lipase and a surface active agent resistance lipase preparation method using the strain. According to the invention, the lipase which is prepared by using the strain and the preparation method has strong stability in an anionic surface active agent SDS, nonionic surface active agents AEO-9 and Triton X-100 and a cationic surface active agent CTAB, and most of the surface active agents can enhance the reaction activity of the lipase. In addition, oxidant hydrogen peroxide can enhance the reaction activity of chryseobacterium lipase provided by the invention.

The invention discloses a preparation method of magnetic hydroxyapatite immobilized lipase. The method includes the steps of preparing a magnetic hydroxyapatite carrier so that lipase can be fixed to the carrier in the form of a chemical bond, conducting amino functional decoration on prepared magnetic hydroxyapatite, and immobilizing lipase with glutaraldehyde as cross-linking agent. The method has the advantages that magnetic nanometer particles are wrapped by hydroxyapatite, hydroxyapatite is provided with plentiful ducts, the inner walls of the ducts are provided with plentiful hydroxyl, further decoration of the carrier and immobilization of active substances are facilitated, and therefore the specific surface area of magnetic particles is greatly enlarged, the loading amount of lipase is increased, and the catalytic effect of immobilized lipase is better embodied; meanwhile, due to the good biocompatibility of hydroxyapatite, the catalytic activity of immobilized lipase is brought into better play.

The invention provides a preparation method of a magnetic micro / nano gel-coated biological charcoal immobilized lipase. The method comprises the following steps: compounding magnetic ferroferric oxide nanoparticles and a sodium oleate solution to form a nano compound, mixing the nano compound with a temperature-sensitive monomer N-isopropyl acrylamide, a crosslinking agent and an emulsifier solution, and carrying out emulsion polymerization reaction to prepare the magnetic micro / nano gel; regulating the temperature of the solution according to the phase change characteristics of the obtained micro / nano gel, so that the micro / nano gel forms a coating on the surface of the biological charcoal; mixing the biological charcoal carrier with a free lipase solution so that the lipase molecules permeates into the micro / nano gel network of the carrier surface and the catalytic active site of the lipase is activated through the oil / water interface; and carrying out glutaraldehyde crosslinking reaction to immobilize the active lipase. The prepared magnetic immobilized lipase has higher catalytic activity, and can be conveniently separated and recovered from the reaction medium by the aid of a magnet, so the application cost is low.

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 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 discloses a preparation method of modified cellulose immobilized lipase, and belongs to the field of lipase preparation. In the invention, bamboo shoots are used as raw materials, and bamboo shoot fibers are prepared by acid-soluble enzymolysis, then alkalized with sodium hydroxide, oxidatively modified by the catalysis of hydrogen peroxide and copper, and finally impregnated and solidified by vacuum freeze-drying. Thereby, the preparation method of the modified cellulose immobilized lipase is obtained. The method of the invention is unique and novel, and the preparation process is simple and feasible. It not only utilizes the spatial structure and flexibility of cellulose, but also has a large number of free hydroxyl groups. The functional group is covalently bonded, has excellent compatibility, and does not affect the activity and reaction ability of the enzyme, improves the binding rate and specific activity of the enzyme, has a huge binding capacity, and is suitable for large-scale application.

Disclosed are various enzymatic processes for the enrichments of oils with omega-3 fatty acids, and specific lipase preparations for use with these processes.

A process for the preparation of short-chain alkyl esters of fatty acids, preferably fatty acid methyl esters (biodiesel) in solvent-free system is disclosed. In particular, the process comprises (a) providing a fatty acid source, (b) stepwise adding a short-chain free alcohol, preferably methanol, or any other alcohol donor, to said fatty acid source in the presence of a lipase preparation and (c) allowing the reaction to proceed under suitable conditions, until said fatty acid source triglycerides are converted to fatty acid methyl esters (FAME). The lipase preparation comprises at least two lipases. The two lipases being separately or jointly immobilized on a suitable support and wherein at least one of the lipases has increased affinity for partial glycerides and at least one of the lipases is sn-1,3 positional specific.

The invention relates to carrier-free immobilized lipase and a preparation method thereof. The preparation method comprises the following steps of: 1, preparing enzyme solution; 2, precipitating lipase molecules by a buffer system with the final concentration of ammonium sulfate of 50 percent; 3, adding a cross-linking agent glutaraldehyde; 4, centrifuging to obtain water insoluble precipitate particles; and 5, freezing and drying the water insoluble precipitate particles to ensure the water insoluble precipitate particles to be dried completely so as to obtain a flavescent powder material, i.e. the carrier-free penicilllum expansum immobilized lipase (PEL-CLEA). In the method, the lipase molecules are cross-linked and aggregated by the cross-linking agent glutaraldehyde through the lipase under the action of ammonium sulfate precipitates, so that the purpose of immobilizing the lipase is fulfilled. The method has low cost and is easy to operate.

The invention relates to a preparation method of magnetic response composite lipase, and preparation of biodiesel by using the magnetic response composite lipase, and belongs to the technical field of preparation of biodiesel by using immobilized enzyme. According to the preparation method, magnetic nano Fe3O4 is adopted to modify lipase A with 1,3-position specificity, an immobilization technique is adopted to prepare the lipase A with the 1,3-position specificity and lipase B without position specificity into a composite enzyme double-enzyme system, as a method of composite lipase concerted catalysis is adopted, the substrate specificity of single lipase is overcome, the transesterification efficiency in preparing biodiesel by using an enzyme method is improved, recycling and repeated use of enzyme are achieved, and the enzyme activity loss in the reaction process is reduced.

The invention relates to a method for preparing lutein carboxylic ester through promoting reaction between a lutein compound containing hydroxyl and carboxylic acid or carboxylic acid anhydride by taking lipase as a catalyst. The method comprises the following steps: adding the lutein compound into a high-pressure reaction kettle, and meanwhile, adding short-chain carboxylates or carboxylic acid anhydride and a lipase preparation with a certain proportion; sealing the high-pressure reaction kettle, then pumping a pre-cooled subcritical fluid into the reaction kettle through a high-pressure pump, controlling the reaction temperature to range from 10 DEG C below zero to 80 DEG C and the pressure to be 2-10 Mpa, and enabling reaction for 0.5-12 h while stirring; and after finish of reaction, filtering a reactant through an enzyme filter arranged in the reaction kettle or a discharge channel to recycle an enzyme catalyst, then reducing the pressure to remove a reaction medium, washing the product with deionized water or a 5% hydrochloric acid solution, and performing drying under reduced pressure to obtain corresponding lutein carboxylic ester.

The invention discloses a method for catalyzing and synthesizing octenyl succinic anhydride modified starch ester through yeast show lipase, comprising the following steps of: dissolving grain starch in water; after imbibition, adding octenyl succinic anhydride and the yeast show lipase; stirring and reacting at 34-36 degrees centigrade for 1-1.5 h; and separating and purifying to obtain starch sodium ester sodium octenyl succinate. A method for preparing the yeast show lipase comprises the following steps of: transforming linearly treated recombinant plasmids into pichia pastoris GS115; inoculating the obtained transformant into a BMMY (buffered methanol-complex medium) culture medium; after inducing and culturing for 72-144 h, centrifugally collecting thallus; and washing, biologically imprinting, freezing and drying the thallus so as to obtain the yeast show lipase. By showing the lipase outside cells, the octenyl succinic anhydride modified starch ester is catalyzed and synthesized through the lipase preparation. By means of the method disclosed by the invention, the transformation efficiency is improved; furthermore, the reaction time is shortened; and the production cost is reduced.

The invention provides duck feed containing lipase. The duck feed comprises the following components, by weight, 40-50 parts of wheat, 12-15 parts of rice bran, 10-13 parts of maize germ dregs, 5-10 parts of corn, 5-10 parts of rice bran dregs, 4-7 parts of rapeseed dregs, 3-6 parts of cottonseed meal, 3-5 parts of pre-mixed materials, 0.01-0.04 part of liquid phytase, 0.01-0.03 part of xylanase, and 0.01-0.03 part of lipase. Preferably, the lipase comprises aspergillus niger lipase. The invention further comprises a preparation method of the lipase. The duck feed can improve the digestion rate of thick fat, can improve quality of feed, is stable and lasting in function and higher in activity, achieves the purpose of reducing cost through reducing adding amount of fat.

The invention discloses a magnetic core-shell type ionic liquid immobilized lipase preparation method. The preparation method includes the steps that firstly, a chemical coprecipitation method is adopted for preparing Fe<3>O<4> nanometer particles; then, the prepared Fe<3>O<4> nanometer particles serve as magnetic cores for synthesizing a Fe<3>O<4>@MCM-41 mesoporous composite material of a core-shell structure; a magnetic carrier is subjected to ionic liquid functionalized modification; finally, immobilization of the lipase is conducted by the adoption of a mesoporous core-shell type magnetic nanometer microsphere carrier modified by ion liquid. According to the magnetic core-shell type ionic liquid immobilized lipase preparation method, the magnetic carrier is modified through the ion liquid, the adaptability of the lipase to the carrier is improved, meanwhile the load of the lipase is increased, and the activity and stability of the lipase are improved; the prepared magnetic immobilized enzyme can be used for a grease ester-ester exchange reaction system with large viscosity, and the magnetic immobilized enzyme can be used for preparing modified grease in the edible oil processing industry.

The invention relates to a preparation method of an organic water flushed fertilizer. According to the preparation method of the water flushed fertilizer, oil residue is taken as the raw material; a lipase preparation, a neutral protease preparation and a phytase preparation are used for decomposing effective nutritional ingredients in the oil residue; protein hydrolysate, phytase hydrolysate and plant ash extract are used for preparing the water flushed fertilizer according to a ratio. The organic water flushed fertilizer is abundant in available nitrogen, available phosphorus and available potassium and capable of meeting the requirements on nutrients of different crops; after performing field experiment, a good yield-increasing effect is achieved.

The invention discloses an industrialized preparation method of a pawpaw enzyme preparation. According to the industrialized preparation method, plate filtration and spray drying are adopted to separate and prepare pawpaw lipase and papain; the preparation method is scientific; the process is convenient to control; material utilization rate is high. Compared with a pawpaw lipase preparation method wherein water soluble proteases in pawpaw milk are removed via repeat washing with buffer solution, the industrialized preparation method comprises following advantages: yield is high; enzyme activity recovery rate is high; pawpaw lipase and papain can be extracted at the same time; water and electricity consumption is reduced greatly; the industrialized preparation method is especially suitable for industrialized production. The invention also discloses a method used for preparing an enzymolysis milk flavor essence using the pawpaw lipase. According to the method, single cream, PBS buffer solution, and glycerin monostearate are mixed at a certain ratio, an obtained mixture is subjected full stirring emulsification at 60 to 80 DEG C, pawpaw lipase is added for enzymatic hydrolysis at 30 to 60 DEG C, and an obtained product is heated to 85 to 90 DEG C for 30min for enzyme deactivation so as to obtain the pawpaw lipase enzymolysis milk flavor essence. The pawpaw lipase enzymolysis milk flavor essence is unique in flavor, and can be used for preparing milk flavor products with unique flavor.

The present invention discloses a powdery lipase preparation which is a granulated material comprising a lipase derived from Rhizopus oryzae and / or a lipase derived from Rhizopus delemar and a soybean powder having a fat content of 5 mass % or more. A lipase activity is improved by using this powdery lipase preparation.

A process for preparation of optically active 4-halogeno-3-hydoxybutyronitrile and 4-halogeno-3-alkanoyloxybutyronitrile, which are simply and economically prepared by reacting a racemic 4-halogeno-3-alkanoyloxybutyronitrile with a microorganism such as a strain of the genus Pseudomonas or the genus Enterobacter, which has the activity stereoselectively hydrolyzing the ester portion, its culture broth or an enzyme(s) derived from the microorganism or an enzyme preparation such as a lipase preparation.

The invention relates to a method for preparing a special foliar fertilizer for organic agricultural crops. The method comprises the following steps: by taking wheat germ oil residue as a raw material, firstly, decomposing grease in oil residue by using a lipase preparation, and then decomposing protein by using a protease preparation, so as to produce amino acids and plant active substances; filtering, reserving liquid for later use, and decomposing an organic phosphorus compound in a solid part by using a phytase preparation, so as to generate available phosphorus; filtering and reserving liquid for later use; finally, mixing two parts of liquid, so as to prepare the foliar fertilizer with high amino acid content and abundant available phosphorus nutrition. The foliar fertilizer prepared by the method obtains a good yield-increasing effect by a field experiment.

The invention belongs to the technical field of polylactic acid production, particularly discloses chemically modified lipase for synthesizing L-lactide, and further discloses a preparation method of the lipase. According to the preparation method, epoxy isopropyl oleate is adopted for chemically modifying the lipase B derived from Candida antarctica, such that chemically modified lipase is obtained. The chemically modified lipase is high in enzyme activity, and the yield is remarkably improved when the chemically modified lipase is used for catalyzing to generate L-lactide.

The invention belongs to the field of chemical compounding, and particularly relates to a preparation method and application of immobilized aspergillus terreus lipase. The invention aims to solve the technical problems that free aspergillus terreus lipase is low in stability and cannot be recycled and the current ketoprofen vinyl ester hydrolysis method is low in productivity. In order to solve the technical problems, the preparation method for the immobilized aspergillus terreus lipase comprises the following steps of uniformly mixing aspergillus terreus lipase with alginate grafted methoxypolyethylene glycols (Alg-g-mPEG), and reacting with alpha-cyclodextrin (alpha-CD) to obtain the immobilized aspergillus terreus lipase. The invention also provides a method for catalyzing the hydrolysis of racemic ketoprofen vinyl ester by the immobilized aspergillus terreus lipase; the method has better stereoselectivity, the productivity is greatly improved, and the immobilized aspergillus terreus lipase can be recycled.

The invention discloses a lipase, which is formed by an amino acid sequence shown as the SEQ ID No.1. The invention further discloses a polynucleotide encoding the lipase, and an expression vector and a transformant containing the polynucleotide, as well as a method for preparing the lipase. The lipase according to the embodiment of the invention has activity which is significantly greater than that of the existing lipase, and the preparation method of the lipase according to the embodiment of the invention has higher yield.

The invention discloses a lipase gene, a recombinant expression vector, a recombinant expression strain, lipase and a preparation method thereof and a preparation method of biodiesel. The lipase geneTLLsyn is used for coding lipase, and a nucleotide sequence of the lipase gene TLLsyn is shown as SEQ ID NO: 1. According to the lipase gene TLLsyn provided by the invention, the original low-frequency codon is replaced with a high-frequency codon in pichia pastoris, and the expression quantity of the synthetic lipase gene TLLsyn in pichia pastoris cells is obviously improved. In addition, in thepreparation process of the lipase, the synthetic lipase gene TLLsyn is transferred into the pichia pastoris, and multi-copy construction of target genes is combined, so that the expression quantity ofthe synthetic lipase gene TLLsyn in the lipase is improved, and the enzyme activity of the lipase is improved. The preparation process is simple, the yield is high, and the production cost of the lipase is reduced.

The invention discloses a method for catalytically synthesizing L-ascorbyl oleate with yeast display lipase. The method comprises the following steps: dissolving L-ascorbic acid and oleic acid in an organic solvent; adding yeast display lipase to react at 50-60 DEG C for 4-8 hours under anaerobic conditions; and carrying out separation and purification to prepare L-ascorbyl oleate, wherein the yeast display lipase is prepared by transforming recombinant plasmid which is subjected to linearization into Pichia pastoris GS115, inoculating the obtained transformant in a BMMY culture medium, carrying out inducing culture for 72-144 hours, centrifuging to collect a thallus, and carrying out washing, bio-imprinting and freeze-drying on the thallus. By displaying the lipase outside the cell and utilizing the lipase preparation to catalytically synthesize the L-ascorbyl oleate, the invention improves the transformation efficiency, shortens the reaction time and lowers the production cost.

The present invention discloses a powdery lipase preparation in which the particles constituting the powdery lipase have 3,000 to 40,000 pores / mm2 on the surface thereof, and each pore has a diameter of 0.5 μm to 6 μm. This powdery lipase preparation was able to improve lipase activity without using a soybean powder.

The invention relates to a preparation method of biomineralized immobilized lipase and application of the biomineralized immobilized lipase in catalytic synthesis of OPO. The method comprises the following steps: adding lipase and a certain proportion of calcium chloride solution into a phosphate buffer solution, then standing a mixture at 1-8 DEG C to obtain immobilized lipase, and applying the immobilized lipase to catalytic synthesis of 1,3-dioleoyl-2-palmitoyl glyceride. The process is simple, the temperature stability of the prepared immobilized lipase is obviously improved, and when theimmobilized lipase is used for catalytic synthesis of the 1,3-dioleoyl-2-palmitoyl glyceride, the catalytic yield is high; and according to the invention, the thermal stability and catalytic activityof the lipase can be effectively improved.

Disclosed is an enzymatic process for the preparation of fatty acid alkyl esters, particularly fatty acids methyl esters (biodiesel) in a solvent-free microaqueous system, from a fatty acid source and an alcohol or alcohol donor, employing a robust lipase preparation that comprises at least two lipases separately or jointly immobilized on a suitable support, where one of the lipases has increased affinity to partial glycerides, another is sn-1,3 positional specific, and an optional third lipase has high selectivity towards sn-2 position of the glycerol backbone of the fatty acid source.