79results about How to "Reduce separation and purification steps" patented technology

The invention discloses a gene for coding a glutamine dipeptide biosynthetic enzyme and application thereof. The nucleotide sequence of the gene is shown as SEQ ID NO.1. The invention further discloses an amino acid sequence coded by the gene, and provides a recombinant vector containing the gene, recombinant escherichia coli, and a method for performing biotransformation to synthesize the glutamine dipeptide by using the gene. The gene and the application of a recombinant bacterial strain of the gene have the advantages of high mol conversion rate, high reaction speed, easy separation, low cost and the like. In the synthesis method, the maximum mol conversion rate of the glutamine dipeptide can reach 83.3 percent; meanwhile, the thalli can be applied to the catalytic synthesis of the glutamine dipeptide again after the circulation recovery; in addition, the catalytic activity is stable. Therefore high market competitive power and application values are realized; a foundation is laid for the industrial production of the glutamine dipeptide.

The invention relates to a novel method for preparation and purifying an electrolyte lithium salt LiDFOB for a lithium ion battery, which belongs to the technical fields of novel energy materials and preparation. The method comprises the following specific steps of: (1) dissolving anhydrous dry NaBF4 and LiCl into an organic solvent; (2) adding a catalyst, heating, stirring and reflowing at the temperature of 30-80 DEG C, and fully reacting for 4-12 hours; (3) filtering, separating a solid from a liquid to obtain an organic solution of LiBF4, adding H2C2O4 and a catalyst in an inertial atmosphere, performing a magnetic stirring reaction at the temperature of 30-120 DEG C for 4-12 hours till no gas is generated, and ending reaction; and (4), filtering, evaporating a filtrate with a rotary evaporimeter till white solid particles are just formed, adding a non-polar solvent into the white solid particles, recrystallizing at a low temperature, filtering, and drying in vacuum at the temperature of 60 DEG C to obtain a high-purity LiDFOB product. The LiDFOB synthesizing method is simple, used raw materials are safe and nontoxic, an intermediate, i.e., LiBF4 is not required to be separated from a solvent, and the process flow is simplified. The LiDFOB prepared by using the process has superior performance, has a good application prospect in the field of power batteries, and is convenient to industrialize.

The invention relates to an extraction, separation and purification method of trehalose, in particular to a separation and purification method of trehalose by one-step. The method comprises the steps of: after extracting dry yeast by using water, adding a flocculating agent to adjust the pH value; and then filtering to obtain clear filtrate, and directly concentrating and crystallizing to obtain trehalose crystals. The method has simple separation and purification steps, convenience and feasibility, low cost and good effect, and provides conditions for the trehalose to be widely used in the fields of medicine, cosmetics, food preservation and the like.

The invention relates to novel engineering bacteria for producing L-asparaginaseII. An extracellular expression product of the engineering bacteria has L-asparaginaseIIprotein activity. The invention also discloses a construction method and fermentation culture conditions of the engineering bacteria, wherein recombinant plasmids pET-ASP invert colon bacillus BL21 to obtain the engineering bacteria; and the structure of the recombinant plasmids pET-ASP is shown in figure 1. The recombinant engineering bacteria are obtained by connecting the L-asparaginase II genes on expression plasmids pET22b to obtain the recombinant plasmids pET-ASP and then using the recombinant plasmids pET-ASP to invert the colon bacillus BL21. The L-asparaginase II can be produced massively by carrying out fermentation culture on the engineering bacteria. And the engineering bacteria has wide raw material source, low cost, and few separation and purification steps and high yield of the products in the fermentation liquor.

The invention provides an engineering bacterium for producing caffeine by fermentation. The engineering bacterium is a recombinant corynebacterium glutamicum which is established by guiding a key enzyme gene in a caffeine biosynthetic pathway into a corynebacterium glutamicum. The key enzyme gene in the caffeine biosynthetic pathway is over-expressed, so that the corynebacterium glutamicum is capable of producing the caffeine; and moreover, a large number of caffeine as products can be secreted out of cells, separation and purification steps are few, yield of the caffeine is high, and a flask fermentation result shows that the yield of the caffeine reaches 3.75 mg/L, so that the recombinant engineering bacterium has a good industrial application prospect, and lays a theoretical foundation for producing food safety-level caffeine by fermentation by using the corynebacterium glutamicum.

The invention discloses a method for clathrating tributyrin by an enzyme method, and belongs to the technical field of grease microencapsulation. According to the method, cyclodextrin synthesized by the enzyme method and tributyrin clathrated by cyclodextrin are combined. The method comprises the step of preparing the cyclodextrin by the enzyme method using CGT enzymes; the tributyrin is added inthe preparation process; after the reaction, Tween is added; and homogenizing and spray drying are performed. The effect of the finally obtained tributyrin powder is much better than the single cyclodextrin clathration effect. The method provided by the invention has the advantages that the process is simple; the cost is low; the operation is convenient; non-toxic harmless and pollution-free effects are achieved in the reaction process; no toxic reagent is remained; the clathration effect is obvious; and the better utilization of a nutritive additive of the tributyrin in the practical production is facilitated.

The invention discloses a method for preparing lithium carbonate by extracting lithium from lepidolite by adopting two-stage chloridizing roasting and an alkali liquor leaching method. The method disclosed by the invention comprises the following steps: (1) levigating the lepidolite and calcium chloride, adding the levigated lepidolite and calcium chloride into ammonium chloride solution, uniformly blending and pelletizing, so as to obtain green balls; (2) performing first-stage roasting on the green balls obtained in step (1) at the temperature of 150-300 DEG C and performing second-stage roasting on the green balls at the temperature of 500-800 DEG C, so as to obtain clinkers; (3) leaching the clinkers obtained in step (2) with water, and filtering to obtain leaching solution; (4) addingcarbonate solution into the leaching solution obtained in step (3), stirring and filtering to obtain mother solution and filter residues, and collecting the filter residues, so as to obtain the lithium carbonate. In the method disclosed by the invention, all the steps are cooperated, and a process of preparing the lithium carbonate by extracting the lithium from the lepidolite is environmentallyfriendly, low in energy consumption, low in cost and high in extracting efficiency. Therefore, the method disclosed by the invention has a broad industrial application prospect.

The invention discloses a Pt-Fe/C catalyst as well as a preparation method and application thereof. The catalyst is composed of an active component and a carrier, wherein the active component is platinum, iron and oxide of iron; the carrier is active carbon; Pt accounts for 0.5-3% of the total mass of the catalyst, and a Fe/Pt mass ratio is (0.5 to 4) : 1. The preparation method of the catalyst comprises the following preparation steps: preparing a platinum-iron active component impregnation solution, performing equal-volume loading, performing in-situ reduction, washing and drying. The catalyst is used for preparing chloroaniline by chloronitrobenzene hydrogenation though a no-solvent method; dechloridation can be effectively restrained in case of not adding a dehalogenation inhibitor, and the advantages of good reaction activity and high selectivity are achieved. Under the conditions of no solvent and auxiliaries, the dosage of the catalyst is 0.8% of the mass of chloronitrobenzene,the reaction temperature is 90 DEG C, hydrogenation pressure is 1.0 MPa, reaction can be completed with two hours, and dechlorination amount is less than 0.1%.

The invention relates to the technical field of biomedicines, and concretely relates to a method for separating and purifying bleomycin (BLM) derivatives 6'-dehydroxy-BLMs from a recombinant engineering bacterium Streptomyces flavovirens SB9026 (with the preservation number of CCTCC M2011292) fermentation liquid. The method comprises the steps of deposition separation pretreatment, D-113 and Diaion HP-20 resin adsorption elution, Silica Flash column isogradient elution, copper removing refining and the like. The method has the advantages of simplification and reduction of the separating and purifying steps, production cost reduction and recovery rate increase. The chemical structure of the bleomycin derivatives is represented by a general formula shown in the specification.

This invention provides one effective and rapid method to extract target molecule and target compound find three dimensional structure from Chinese medicine product, which uses target protein transistor stricture as base and uses X ray transistor means to filter the product for active component and for study from one new respective.

The invention discloses a preparation method of a 1,2,3-triazol-[1,5-a] quinoline compound. The method comprises the steps of making substituted methyl quinoline react with an iodine simple substancein advance, adding a solid potassium phosphate trihydrate after the methyl quinoline disappears, and adding p-methyl benzene sulfonyl hydrazide for continuing a reaction to obtain the 1,2,3-triazol-[1,5-a] quinoline compound. The preparation method has the advantages that the adopted raw materials are cheap and easy to obtain, the operation method is simple, the reaction condition is mild, a one-pot approach can be adopted, the steps are short, the yield is up to 83%, and a feasible method is provided for industrial preparation of the compound.

The invention discloses a preparation method of pyridodipyrimidine and pyridodipyrazole derivatives. The method comprises the following steps: carrying out a contact reaction on substituted methylquinoline derivatives, 6-amino-1,3-dimethylpyrimidine-2,4-dione or 3-methyl-1-phenyl-5-aminopyrazole and an iodine elementary substance to synthesize pyridodipyrimidine compounds, pyridodipyrazole compounds and derivatives thereof in one-pot. According to the preparation method, the adopted raw materials are cheap and easy to obtain, the preparation method is simple, steps are relatively short, the yield is up to 97%, and a feasible method is provided for industrially preparing the compounds.

The invention discloses a method for preparing an anti-human chorionic gonadotrophin (hCG) monoclonal antibody fragment. The method comprises the following steps of: putting activated immobilized papain into a 0.1mol/L acetic acid buffer solution with pH of 5.0, adding an anti-hCG monoclonal antibody, performing water bath reaction for 4 to 24 hours at the temperature of between 30 and 60 DEG C, and filtering the reaction solution, wherein filter cakes B are immobilized papain and recycled, and filtrate B is a hydrolysate containing the monoclonal antibody fragment; and separating and purifying the hydrolysate containing the monoclonal antibody fragment by adopting weak anion exchange chromatography, and thus obtaining the anti-hCG monoclonal antibody fragment. The method is easy to implement and has a few purification steps, and the obtained antibody fragment has high purity; and the antibody fragment prepared in the method can reach the purity of over 90 percent by one-step purification.

The invention provides a heat resistant beta-amylase-trehalose synthase fusion enzyme, an expression gene of the heat resistant beta-amylase-trehalose synthase fusion enzyme, an engineering bacterium secreting the fusion enzyme and application, and belongs to the technical field of genetic engineering. The heat resistant beta-amylase-trehalose synthase fusion enzyme comprises an amino acid sequence with SEQ ID No.1 and another amino acid sequence, and the similarity of the another amino acid sequence to the amino acid sequence with SEQ ID No.1 is 80%-100%. The expression gene of the heat resistant beta-amylase-trehalose synthase fusion enzyme comprises an amino acid sequence with SEQ ID No.2 and another amino acid sequence, and the similarity of the another amino acid sequence to the amino acid sequence with SEQ ID No.2 is 80%-100%. The obtained beta-amylase-trehalose synthase fusion enzyme has a heat resistant property, directly converts amylose into trehalose at the higher temperature, saves the complicated steps of separating and purifying endoenzyme, simplifies the production process of the trehalose, and reduces the production cost.

The invention discloses a preparation method of fumagillin amino alcohol shown as a formula I in the specification. The method comprises the following steps: (1) reducing amino acid shown in a formulaII to obtain a compound shown in a formula III; and (2) under the heating condition of a methanol solution, carrying out an ammonium salt synthesis reaction on fumagillin and the compound shown in the formula III to obtain fumagillin amino alcohol. The preparation method is short in reaction route and high in synthesis efficiency, the separation and purification steps are reduced, and mass production can be achieved. Meanwhile, the synthesized fumagillin amino alcohol is low in toxicity, can be dissolved and degraded in nature and is good in water solubility.

The invention discloses a method for preparing fumagillin amino alcohol shown as a formula I in the specification. The method comprises the following steps: (1) reducing amino acid shown in a formulaII to obtain a compound shown in a formula III; and (2) under the heating condition of a methanol solution, carrying out an ammonium salt synthesis reaction on fumagillin and the compound shown in theformula III to obtain fumagillin amino alcohol. The method is short in reaction route and high in synthesis efficiency, the separation and purification steps are reduced, and mass production can be achieved. Meanwhile, the synthesized fumagillin amino alcohol is low in toxicity, can be dissolved and degraded in nature and is good in water solubility.

The invention discloses a method for directly synthesizing cyclic carbonate from olefin under catalysis of metalloporphyrin, and relates to the technical field of catalytic synthesis. The technical scheme is characterized in that the method comprises the following steps: catalyzing olefin, carbon dioxide and an oxidant to carry out epoxidation carboxylation reaction by taking metalloporphyrin as acatalyst and ionic liquid as a cocatalyst, and preparing cyclic carbonate by a one-pot method. The specific reaction equation is as follows: the reaction conditions are mild, the catalyst preparationis simple and convenient, the catalyst consumption is low, and the target product cyclic carbonate is obtained with high conversion rate and high selectivity.