79 results about "G penicillin" patented technology

The invention provides an expansible ring enzyme mutant and an application of the expansible ring enzyme mutant in synthesis of G-7-ADCA. A series of mutants with penicillin G ring expansion activity are obtained through ancestor enzyme sequence reconstruction, directed evolution and the like, and the mutants can directly expand the ring of the substrate penicillin G to generate G-7-ADCA. Therefore, the expansive ring enzyme mutants have important application value in industry.

The invention relates to the technical field of bioengineering, in particular to a penicillin G acylase mutant, polynucleotide, an expression vector and application. The penicillin G acylase mutant is obtained by carrying out site-directed mutagenesis on a wild type penicillin G acylase gene of parent Escherichia coli, and carrying out site-directed mutagenesis on the wild type penicillin G acylase gene to obtain the penicillin G acylase mutant. Phenylalanine (F) at the 24th site of an alpha chain, proline (P) at the 383rd site of a beta chain, threonine (T) at the 384th site of the beta chain, glutamic acid (G) at the 385th site of the beta chain and serine (S) at the 386th site of the beta chain of the penicillin G acylase are introduced and mutated by a whole plasmid PCR (Polymerase Chain Reaction) technology to obtain mutants. When the penicillin G acylase mutant obtained by the invention is used for catalyzing methyl mandelate and 3-(1-methyl-1H-tetrazole-5-yl) thiomethyl-7-aminocephalosporanic acid to synthesize cefamandole, the synthesis activity is improved, and meanwhile, the side reaction rate is greatly reduced.

The invention discloses a broad-spectrum combined medicine for treating liver cancer, which comprises purified water, tripterygium wilfordii alkaloid, tripterygium hypoglaucum alkaloid, chamaejasmine, gelsmium elegans alkaloid, xanthoxylin and various B medicines, and further comprises folic acid, vitamins, copper sulfate, zinc sulfate, manganese sulfate, yeast and penicillin sodium. Through the multi-component, multi-target and multi-path design, the medicine disclosed by the invention shows a potential effect of inhibiting the progress of liver cancer and good safety in cell experiments, animal models and human individual case observation. Cellular immunity and humoral immunity can be enhanced, the treatment cost is lower than that of a traditional treatment scheme, and no pain is caused during treatment.

The application discloses a preparation method of near-infrared light enhanced response Cu, Fe-Lyz / AgNPs nanoscale enzyme. The method uses Cu, Fe doped carbon dots as a reducing agent to prepare silver nanoparticles, and then the silver nanoparticles are modified by lysozyme to prepare Cu, Fe-Lyz / AgNPs nanoscale enzyme with positive charges on the surface. The Cu, Fe-Lyz / AgNPs nanoscale enzyme has dual activities of peroxidase-like and glutathione oxidase-like and photo-thermal performance. Meanwhile, the near-infrared light can significantly enhance the POD-like activity of the nanoscale enzyme. The Cu, Fe-Lyz / AgNPs nanoscale enzyme has good antibacterial effect on escherichia coli and staphylococcus aureus. Meanwhile, based on the enhancement of the penicillin G potassium on the Cu, Fe-Lyz / AgNPs oxidase-like activity, the application establishes a rapid colorimetric detection method for the penicillin G potassium, and when the method is used for detecting the penicillin G potassium in blood plasma and milk powder samples, the detection limit is as low as 0.05 mu g / mL, and the method has high reliability and accuracy.

This invention discloses a penicillin V potassium capsule and its preparation method. The preparation method includes: preparing modified polylactic acid glycolic acid copolymer microspheres; mixing penicillin V potassium micropowder with the modified polylactic acid glycolic acid copolymer microspheres, adding fumed silica and anhydrous calcium hydrogen phosphate for further mixing to obtain a blend; granulating the blend using a dry granulation method; filling the obtained granules into capsules, polishing, and packaging; and modifying the polylactic acid glycolic acid copolymer with povidone and ethyl cellulose to obtain the modified polylactic acid glycolic acid copolymer microspheres. This invention designs polylactic acid glycolic acid copolymer microspheres that are double-modified with povidone and ethyl cellulose, resulting in better drug loading flowability, more stable mixing uniformity, and significantly improved flowability compared to penicillin V potassium powder. The addition of fumed silica and anhydrous calcium phosphate further enhances antistatic properties and reduces the powder return rate.

This invention discloses an in vitro maturation culture medium for ferret oocytes and its preparation method, belonging to the field of animal embryo engineering technology. It comprises the following components: TCM199 basal culture medium, 50 mL fetal bovine serum, 25 IU follicle-stimulating hormone, 25 IU luteinizing hormone, 0.5 mg 17β-estradiol, 0.005 mg epidermal growth factor, 12.1 mg sodium pyruvate, 0.2 mM cysteine, 0.1 mM cysteine, 0.030 g penicillin, 0.025 g streptomycin sulfate, as well as an antioxidant complex, a combination of growth factors, and extracellular matrix components. This invention provides a comprehensive and suitable nutritional environment for ferret oocytes by precisely formulating a composite culture medium containing multiple key components. Specifically, the hormone combination at specific concentrations accurately simulates the in vivo physiological environment, effectively regulating the oocyte maturation process; multiple growth factors work synergistically to promote oocyte growth, development, and differentiation; and the addition of the antioxidant complex effectively eliminates free radicals generated during in vitro culture, reducing oxidative stress damage to oocytes.

This invention discloses a penicillin G acylase mutant SEQ ID NO:3, which can efficiently catalyze the reaction of substrate 6-APA with DHPGM to produce amoxicillin. The synthesis / hydrolysis ratio S / H reaches a maximum of 18.1. When the side chain to parent nucleus ratio is 1.05:1, the conversion rate of parent nucleus 6-APA reaches more than 99.0%. This mutant has high thermal stability, which helps to improve the efficiency of industrial production of amoxicillin.

The application discloses a kind of soft measurement modeling methods of fermentation process based on time sequence diagram network, belongs to the technical field of soft measurement of fermentation process.It includes the following steps: (1) data acquisition and integration: the penicillin fermentation process under different conditions is obtained, and the data is divided, collected and integrated;(2) data selection: the data is selected, the redundant useless data is removed and the causal diagram between variables is established;(3) modeling training: algorithm model is constructed, and learning training is carried out;(4) model prediction: the trained algorithm model is used for prediction, and the prediction result is given.The application proposes a kind of soft measurement modeling method of fermentation process based on time sequence diagram network, improves the prediction accuracy of key product quality of fermentation process;The method uses graph convolution network and long short-term memory, extracts data in time and space dimensions, increases the generalization of model;The method can accurately measure the key product quality of different fermentation processes.

The invention discloses a bimodal detection method of penicillin G based on Au-coated C3N4 nano-enzyme. The bimodal detection method comprises the following steps: step 1, measuring fluorescence intensity values F1 of reaction solutions containing penicillin G standard substances with different concentrations at 550nm under excitation of 450nm wavelength; 2, measuring absorbance values A1 of reaction solutions containing different concentrations of penicillin G standard substances at 450 nm; 3, constructing a linear regression equation of the fluorescence F and the absorbance A of the standard substance; the method is successfully applied to detection of penicillin G in an actual sample, and has good sensitivity and selectivity. And moreover, the bimodal detection method shows higher adaptability in practical application, and different test requirements can be met according to different instruments.

The application belongs to the technical field of biological medicine, and particularly relates to a penicillin G expandase mutant, polynucleotide, expression vector and use. The mutant is a mutant with a R27M point mutation in the amino acid sequence shown in the sequence table SEQ ID NO. 19. The mutant also contains a point mutation at one or more of the following positions: L50V, M73T, R139K, Y184H, Y217A, R249A and T292A. The mutant provided in the application has a significantly improved conversion rate of penicillin G into G-7-ADCA than the wild type, and has a good application prospect.

The invention belongs to the technical field of enzyme catalysis, and particularly relates to recombinant penicillin G acylase, encoding nucleotide and application of the recombinant penicillin G acylase. Compared with an amino acid sequence shown in SEQ ID NO. 1, the mutant has the advantages that the mutation mode is F24 beta R; or F146 [alpha] K amp; f24 [beta] R; or F146 [alpha] K amp; f24 [beta] R amp; f71 [beta] Y; or F146 [alpha] K amp; f24 [beta] R amp; f71 [beta] Y amp; n241 [beta] K amp; and G385 [beta] Y. According to the invention, wild type penicillin acylase is mutated to obtain penicillin acylase with coordinated enzymatic activity of hydrolysis and synthesis. The method can be used for synthesis production of beta-lactam antibiotics, especially for one-step preparation of amoxicillin by catalyzing penicillin potassium salt and the like, and separation of an intermediate 6-APA is avoided. The invention provides a key enzyme for efficient preparation of beta-lactam antibiotics, and greatly promotes the innovation of a beta-lactam antibiotic preparation technology.

The present disclosure relates to engineered penicillin G acylase (PGA) enzymes having improved properties, polynucleotides encoding such enzymes, compositions including the enzymes, and methods of using the enzymes.

This invention belongs to the field of product separation technology in enzymatic antibiotic synthesis, and particularly relates to a method for separating and recovering amoxicillin products from a one-step synthesis catalyzed by a Kluyveromycin β-subunit G385 mutant penicillin acylase. This application uses an immobilized penicillin acylase mutant to catalyze the one-step synthesis of amoxicillin from potassium penicillin. A method for separating amoxicillin and phenylacetic acid is developed for the obtained reaction mixture. The technical solution mainly includes: firstly, separating the immobilized penicillin acylase mutant from the reaction solution by filtration; then separating amoxicillin by crystallization; and finally, separating and recovering phenylacetic acid by toluene extraction, back-extraction, etc. This separation method can rapidly and efficiently separate amoxicillin with a high yield, with an average crystallization rate of 93.22%; simultaneously, the separation and recovery of phenylacetic acid is good; and the extractant toluene can be recycled.

The mixing and stirring device comprises a box body and a main stirring assembly, the top of the box body is open, a discharging pipe is fixedly arranged below the side wall of the box body, a positioning frame is fixedly arranged at the top of the box body, and the main stirring assembly is fixedly arranged on the positioning frame. The main stirring assembly comprises a stirring shaft rotationally connected with the positioning frame, a plurality of limiting frames circumferentially fixed to the side wall of the stirring shaft, a stirring frame fixedly connected with the limiting frames and a local stirring rod assembly capable of moving up and down in the limiting frames, and a plurality of stirring blades are fixedly arranged on the outer wall of the stirring frame; a plurality of communicating holes are formed in the stirring frame, firstly, drugs and reagents for producing penicillin powder injections are poured into the box body, the stirring shaft drives the main stirring rod assembly to rotate, the main stirring assembly stirs mixed materials, meanwhile, the local stirring rod assembly can stir the materials up and down, and the materials are stirred through the communicating holes. The stirring effect is further improved.

The invention discloses an induction culture medium with definite components and application of the induction culture medium in preparation of a hepatic fibrosis treatment medium, and belongs to the technical field of hepatic cell culture and artificial liver. The induction culture medium is composed of a serum-free basic culture medium, primary bile acid and secondary bile acid, the serum-free basic culture medium is composed of a William's E culture medium, penicillin, streptomycin, L-glutamine, insulin, transferrin, sodium selenite and dexamethasone, the primary bile acid is an FXR signal channel stimulant, and the secondary bile acid is a TGR signal channel stimulant. The hepatic fibrosis treatment medium prepared by utilizing the induction culture medium has a remarkable hepatic fibrosis relieving effect, and has the advantages of definite production materials, controllable process, high product stability and the like.

This invention belongs to the field of pharmaceutical technology, specifically relating to the application of glutamine in the preparation of drugs that inhibit gene mutations in *E. coli*. This application combines glutamine and ampicillin, and after multiple passages, it can inhibit drug-resistance gene mutations in *E. coli*, significantly reducing the mutation probability of drug-resistance genes, decreasing the fold increase in the minimum inhibitory concentration (MIC) of ampicillin after passage, and lowering the survival rate. Animal experiments have also demonstrated that passaged bacteria with added glutamine and ampicillin are more easily cleared by the body, achieving the effect of slowing down the formation of drug-resistance gene mutations in *E. coli* to ampicillin.

The invention belongs to the technical field of enzyme catalysis, and particularly relates to penicillin acylase for synthesizing beta-lactam antibiotics. Compared with an amino acid sequence shown in SEQ ID NO. 1, the mutant has the advantages that the mutation mode is F146 alphaK; or F146 [alpha] K amp; g385 [beta] Y; or F146 [alpha] K amp; g385 [beta] R is shown in the description. According to the penicillin acylase mutant disclosed by the invention, penicillin acylase from kluyvera citrophila (Kluyvera citrophila) is mutated, so that the penicillin acylase mutant which is strong in hydrolytic activity and synthetic activity and is coordinated in two enzyme activities is obtained. The method can be used for synthesis production of beta-lactam antibiotics, especially for one-step preparation of amoxicillin by catalyzing penicillin potassium salt and the like, and separation of an intermediate 6-APA is avoided. The invention provides a key enzyme for efficient preparation of beta-lactam antibiotics, and greatly promotes the innovation of a beta-lactam antibiotic preparation technology.

The present application relates to the field of cefoperazone acid, and discloses a preparation method of cefoperazone impurity A, which comprises the following steps: (1) performing acyl chloride treatment on oxypiperazine acid in the presence of an acyl chloride reagent to generate an acyl chloride compound of oxypiperazine acid; (2) performing silylation treatment on 7-aminocephalosporanic acid in the presence of a silylation reagent to generate a silylated compound of 7-aminocephalosporanic acid; (3) reacting the acyl chloride compound of oxypiperazine acid with the silylated compound of 7-aminocephalosporanic acid to generate an intermediate compound (I); (4) hydrolyzing the intermediate compound (I) under the action of penicillin acylase to generate an intermediate compound (II); and (5) performing esterification and cyclization reaction on the intermediate compound (II) in the presence of a first crystallization solvent, a first organic solvent and a cyclization reagent to obtain the cefoperazone impurity A. The method disclosed by the present application is simple in process, and can obtain the cefoperazone impurity A with high purity, which can be used as a control sample for cefoperazone detection.

The present disclosure relates to penicillin G acylases (PGAs), including engineered PGA enzymes, and their use in catalyzing chemical reactions associated with beta-lactam antibiotic biosynthesis.

The present application relates to a culture method and culture medium for reducing the number of epithelial organoids and in vivo tissue differential transcripts, which solves the technical problems of low growth rate and low formation rate of organoids in the prior art. The epithelial organoid culture medium contains basic components and additional components. The basic components contain DMEM / F12 basic medium, penicillin-streptomycin solution, Glutamax additive, recombinant vertebral protein, recombinant cephalopod protein, Y-27632, epidermal growth factor, N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid buffer solution, nicotinamide, A83-01 and CHIR99021. The additional components contain Wnt-3A, ITS-X additive and hydrocortisone. The present application simultaneously provides a culture method for epithelial organoids. The present application can be used for the culture of epithelial organoids.

The invention belongs to the technical field of enzyme catalysis, and particularly relates to a high-activity penicillin G acylase mutant, encoding nucleotide and application of the high-activity penicillin G acylase mutant. Compared with an amino acid sequence shown in SEQ ID NO. 1, the mutant has the advantages that the mutation mode is F71betaY; or F146 [alpha] K amp; f71 [beta] Y; or F146 [alpha] K amp; f71 [beta] Y amp; n241 [beta] K; or F146 [alpha] K amp; f71 [beta] Y amp; n241 [beta] K amp; and G385 [beta] Y. Through site-directed mutagenesis, the penicillin acylase mutant with strong hydrolytic activity and synthetic activity and coordinated two enzyme activities is obtained. The method can be used for synthesis production of beta-lactam antibiotics, especially for one-step preparation of amoxicillin by catalyzing penicillin potassium salt and the like, and separation of an intermediate 6-APA is avoided. The invention provides a key enzyme for efficient preparation of beta-lactam antibiotics, and greatly promotes the innovation of a beta-lactam antibiotic preparation technology.

The application discloses an expandase mutant SEQ ID NO:3 which can efficiently catalyze the expandase reaction of a substrate to generate phenylacetyl-7-amino deacetoxycephalosporanic acid in a reaction system with a penicillin G concentration of up to 10 mM, and can be used for industrial production of G-7-ADCA.

The invention belongs to the technical field of biology, and particularly relates to a bifidobacterium longum subsp. Longum FMBL B241768LS, a microbial inoculum and application of the microbial inoculum in hypoglycemic products, the bifidobacterium longum subsp. Longum FMBL B241768LS is preserved in China Center for Type Culture Collection on October 14, 2024, and the preservation number is CCTCC NO: M 20242173; the inhibitory activity of the compound on dipeptidyl peptidase IV is as high as 79.17%; the compound has a relatively good inhibition effect on the activity of alpha-glucosidase and alpha-amylase; the compound shows drug resistance to ciprofloxacin, ampicillin, penicillin G, clindamycin, kanamycin and polymyxin B; a good inhibition effect is achieved on pathogenic bacteria; the strain can be used for preparing medicines, fermented foods, health-care products and food additives for reducing blood sugar and inhibiting pathogenic bacteria, and has a wide application prospect.

The application discloses a culture medium and application thereof, and a method for in-vitro separation and culture of goat intramuscular precursor fat cells and adipogenic differentiation of the cells, and relates to the technical field of cell culture. The culture medium comprises the following components: 8-12 mu g / mL insulin, 8-12% fetal bovine serum in volume fraction, 95-105 U / mL penicillin, 95-105 mu g / mL streptomycin, and the like. On the basis of the prior art, the application mainly adds appropriate glycyrrhizin and pilocarpine in the culture medium, and finds that the two components are in cooperation, and the adipogenic differentiation ability of cells such as mesenchymal stem cells and precursor fat cells is significantly improved, and the application range is wide. On the basis, the application further provides a complete method for in-vitro separation and culture of goat intramuscular precursor fat cells and adipogenic differentiation of the cells, and provides a method and a way for a series of researches on goat primary intramuscular precursor fat cells in the future.

The invention belongs to the technical field of medicine synthesis, and particularly relates to a method for preparing oxacillin sodium through an enzyme method. The method comprises the following steps: taking 6-APA and 5-methyl-3-phenylisoxazole-4-carboxylic acid methyl ester as raw materials, and adding immobilized penicillin G acylase for reaction to obtain an oxacillin sodium solution; and filtering, dialyzing, concentrating and crystallizing the oxacillin sodium solution to obtain the oxacillin sodium. The method has the advantages of mild reaction conditions, reduction of the generation of wastes, low production cost, accordance with a green production process, and very good industrialization value; and a nanofiltration membrane dialysis process is introduced into the post-treatment of the reaction liquid, so that the product quality control is better guaranteed.

The invention relates to a method for efficiently removing penicillin G sodium in mushroom dregs by using a magnetic mushroom dreg biochar immobilized degrading enzyme. According to the method, the enzyme for degrading the penicillin G sodium is produced in an amplified mode through a 5L stirring type reactor, the working procedures comprise strain culture, shake flask first-stage culture, shake flask second-stage culture and 5L stirring type reactor fermentation, and the amplified enzyme activity reaches up to 1288.09 U / mL. Penicillin G sodium in actual fermentation mushroom dregs is removed in a 5L stirring type reactor by utilizing the mushroom dreg biochar immobilized degrading enzyme, and the removal rate of 0.1 g of immobilized enzyme on the penicillin G sodium in the fermentation mushroom dregs within 120 minutes reaches 98.72%. The invention also provides a method for extracting the residual penicillin G sodium in the mushroom dregs. The method specifically comprises three steps of extraction, purification and detection. According to the invention, a new low-cost choice is provided for efficiently repairing the pollution of the penicillin G sodium in the environment, and the treated mushroom dregs have important significance for realizing high-value utilization of hazardous wastes.

The application belongs to the technical field of enzyme catalysis, and particularly relates to a penicillin acylase for synthesizing beta-lactam antibiotics. Compared with the amino acid sequence shown in SEQ ID NO. 1, the mutation mode of the mutant is F146aK; or F146aK&G385bY; or F146aK&G385bR. The application obtains a penicillin acylase mutant with high hydrolysis activity and synthesis activity and coordinated two enzyme activities by mutating the penicillin acylase from Kluyvera citrophila. The mutant can be used for the synthesis and production of beta-lactam antibiotics, and particularly catalyzes the one-step preparation of amoxicillin from penicillin potassium salt, thereby avoiding the separation of the intermediate 6-APA. The application provides a key enzyme for the efficient preparation of beta-lactam antibiotics, and will greatly promote the innovation of the preparation technology of beta-lactam antibiotics.

This invention belongs to the field of biopharmaceutical technology, and particularly relates to a one-pot method for synthesizing amoxicillin. This invention provides a novel enzymatic synthesis route for amoxicillin, specifically including the following steps: (1) Pretreatment of immobilized penicillin G acylase – soaking PGA washed with deionized water in phosphate buffer; (2) dissolving potassium penicillin G and L-p-hydroxyphenylglycine methyl ester in a phosphate buffer and hexane system to obtain an initial reaction solution; (3) mixing the immobilized penicillin G acylase with the initial reaction solution and reacting to obtain an amoxicillin suspension; (4) filtering out the immobilized penicillin G acylase and washing with deionized water, adding hydrochloric acid until the amoxicillin suspension becomes clear, filtration, pH adjustment, cooling crystallization, washing and drying to obtain crude amoxicillin. The entire preparation process is simple, easy to operate, environmentally friendly, has good substrate conversion effect, and a high amoxicillin yield.

The utility model relates to the technical field of penicillin sodium production equipment, in particular to penicillin sodium production equipment for continuous fluid separation, which comprises a bottom plate, a support rod is fixedly mounted at the middle end of the top of the bottom plate, and a double-head motor is fixedly mounted at the middle end of an inner cavity of the support rod. The two output ends of the double-end motor are fixedly connected with threaded rods, the outer surfaces of the threaded rods are in threaded connection with movable sleeves, and the outer surfaces of the movable sleeves are fixedly connected with movable rings. Through the cooperation of the structures, the production equipment has the advantage that the resin in the resin barrel does not need to be replaced by frequently disassembling and assembling the heating equipment, and solves the problem that the existing traditional heating equipment fixed on the outer side of the strong acid type cation exchange resin barrel cannot be replaced; when the adsorption capacity of resin in the strong acid type cation exchange resin cylinder is reduced and the resin needs to be replaced, heating equipment on the outer side of the strong acid type cation exchange resin cylinder needs to be frequently disassembled and assembled, and the operation process is very tedious and labor-consuming.

The invention relates to an expansible ring enzyme mutant and application thereof in preparation of a cephalosporanic acid compound. Through technical means such as directed evolution, a series of enzyme mutants with a penicillin G ring expanding function are successfully screened out, and the mutants can efficiently convert penicillin G into G-7-ADCA. Therefore, the expansive ring enzyme mutants have important application value in industry.