104 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 belongs to the technical field of separation of products for synthesizing antibiotics by an enzymic method, and particularly relates to a method for separating and recycling an amoxicillin product synthesized by a one-step method based on catalysis of kluyveromyces citricacidophilus beta subunit G385 mutant penicillin acylase. An immobilized penicillin acylase mutant is used for catalyzing penicillin potassium to synthesize amoxicillin in one step, a set of method for separating amoxicillin and phenylacetic acid is developed for an obtained reaction mixture, and the technical scheme mainly comprises the following steps: firstly, separating the immobilized penicillin acylase mutant from a reaction solution through filtration; then, amoxicillin is separated out through crystallization; and then crystallizing, separating and recovering phenylacetic acid through toluene extraction, back extraction and the like. According to the separation method, the amoxicillin can be rapidly and efficiently separated, the yield of the amoxicillin is relatively high, and the average crystallization rate reaches 93.22%; meanwhile, the separation and recovery effects of phenylacetic acid are good; and the extraction agent toluene can be recycled.

The invention belongs to the technical field of medicines, and discloses a near-infrared cyanine probe for detecting penicillin G acylase as well as a preparation method and application of the near-infrared cyanine probe. According to the research, a near-infrared fluorescent probe Cy-NEO-PA responding to penicillin G acylase (PGA) is developed, so that the influence of environmental factors on the formation of an acinetobacter baumannii biological membrane is observed. Research results show that glucose inhibits the generation of PGA to enhance the formation of a biological membrane, while phenylacetic acid (PAA) stimulates the generation of PGA and inhibits the formation of the biological membrane. The observation results highlight the excellent capability of Cy-NEO-PA in accurately measuring PGA kinetics, and reveal the key effect of PGA in biological membrane development. In addition, the Cy-NEO-PA has good biocompatibility, strong active oxygen generation, efficient photo-thermal conversion and bacterial targeting ability, so that the Cy-NEO-PA becomes a promising drug for resisting bacterial infection and promoting wound healing through photo-thermal / photodynamic therapy.

The invention discloses a penicillin G acylase mutant with high synthetic activity and low hydrolytic activity and application of the penicillin G acylase mutant, and belongs to the field of bioengineering. The penicillin G acylase gene derived from Escherichia coli is subjected to site-specific mutagenesis, so that the amino acid sequence coded by the penicillin G acylase gene mutates from phenylalanine F to alanine A at the 24th site of an alpha chain, the 146th site of the alpha chain mutates from phenylalanine F to valine V, and the 386th site of a beta chain mutates from serine S to alanine A to obtain the mutant. According to the penicillin G acylase mutant alphaF24A / betaS386A disclosed by the invention, compared with the conversion rate of cefamandole synthesized by catalyzing the reaction of methyl mandelate and 3-(1-methyl-1H-tetrazole-5-yl) thiomethyl-7-aminocephalosporanic acid (7-TMCA) within 4 hours before mutation, the conversion rate of cefamandole synthesized by catalyzing the reaction of methyl mandelate and 3-(1-methyl-1H-tetrazole-5-yl) thiomethyl-7-aminocephalosporanic acid (7-TMCA) is improved by 13.70%, and the yield of a side reaction product mandelic acid is reduced by 82%. The penicillin G acylase mutant disclosed by the invention can be used for catalyzing methyl mandelate and 7-TMCA to synthesize cefamandole while greatly reducing the occurrence of side reactions, and can be applied to preparation of cephalosporin antibiotics.

The invention relates to the technical field of production of penicillin V potassium, and particularly discloses a preparation method of penicillin V potassium, which comprises the following steps: S1, under the catalysis of penicillin V acylase, enabling 6-APA and methyl phenoxyacetate to react, performing full-water-phase synthesis to obtain a penicillin V solution, filtering out the penicillin V acylase, adding dilute acid into filtrate, and crystallizing to obtain a penicillin V acid crystal; s2, dissolving the penicillin V acid crystal by using an organic solvent I to obtain a solution I, dissolving potassium isooctanoate by using an organic solvent II to obtain a solution II, mixing the solution I and the solution II, reacting and crystallizing to obtain a penicillin V potassium crystal, and filtering, washing and drying to obtain a penicillin V potassium finished product. According to the method, an enzyme method is used for replacing traditional fermentation and extraction methods to produce the penicillin V potassium, so that the process steps and energy consumption can be reduced, and the product quality is effectively improved.

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.

The invention relates to the technical field of biological enzymes, in particular to recombinant penicillin G acylase as well as a preparation method and application thereof. The amino acid sequence of the recombinant penicillin G acylase comprises three mutation sites: a 171 site, a 647 site and a 266 site, the mutation amino acid of the 171 site is tyrosine, the mutation amino acid of the 647 site is arginine, and the mutation amino acid of the 266 site comprises one of glutamine, methionine and arginine. According to the recombinant penicillin G acylase, through three key mutation sites (F171Y, Y647R and S266Q / M / R), the catalytic efficiency of the recombinant penicillin G acylase and the binding affinity of the recombinant penicillin G acylase with a target substrate are remarkably improved on the basis of the multidimensional structural characteristics of collaborative optimization of enzyme-substrate interaction.

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 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.

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 present invention relates to an epithelial organoid culture medium and a culture method, which solve 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 include DMEM / F12 basal medium, penicillin-streptomycin solution, Glutamax additive, recombinant spondin, recombinant norrin, Y-27632, epidermal growth factor, N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid buffer solution, nicotinamide, A83-01 and CHIR99021. The additional components include Wnt-3A, ITS-X additive and hydrocortisone. The present invention also provides a culture method for epithelial organoids. The present invention can be used for the culture of epithelial organoids.

The present invention provides a composition with a solubilizing effect, a preparation method thereof, and an application thereof. The composition comprises a solubilizing agent, a glidant, and an inclusion compound, wherein the mass ratios of the components are as follows: the mass ratio of the solubilizing agent to the inclusion compound is 1:1-1:20, and the mass ratio of the glidant to the inclusion compound is 1:100-3:20. The composition of the present invention can effectively improve the solubility of macrolides, tetracyclines, and penicillins, and the composition of the present invention does not react with the active ingredients of the drugs, ensuring that after the drugs are mixed therewith, the active ingredients of the drugs are stable, and the mixture of the drugs and the composition does not agglomerate or discolor.

The invention belongs to the technical field of microorganisms, and particularly relates to salt-tolerant and high-temperature-resistant saccharomyces rouxii and application thereof.The method comprises the following steps that S1, screening of salt-tolerant saccharomycetes is conducted, specifically, 25 g of soy sauce mash samples fermented for 30 days, 45 days and 60 days are taken and added into 225 mL of normal saline, and oscillation elution is conducted through a centrifugal machine under the normal-temperature condition; the method comprises the following steps: firstly, performing gradient elution on an eluent, then performing gradient dilution on the eluent to 10 <-3 >, 10 <-4 > and 10 <-5 >, sucking 100 microliters of each gradient diluent, uniformly coating on a YPD agar solid culture medium containing 18 mg / 100 mL of table salt and 100 mg / mL of ampicillin sodium antibiotic, standing and culturing at 30 DEG C, performing repeated separation and purification, and finally selecting a single colony with typical saccharomycetes colony characteristics and maximum diameter; compared with the first soy sauce of the Cantonese-style brewed soy sauce prepared by a normal process, the yeast adding process has the advantages of more glossy color, richer sauce fragrance, ester fragrance, mellow fragrance, more prominent delicate flavor, more palatable salty, fresh and sweet taste, clarification and better wall hanging property.

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.

The invention relates to the technical field of production of penicillin V potassium, and particularly discloses a novel process for preparing penicillin V potassium, which comprises the following steps: a) carrying out flocculation treatment on penicillin fermentation liquor, and passing through drum filtering equipment; b) passing the drum filtrate through a roll-type ultrafiltration membrane device, and acidifying and crystallizing the roll-type ultrafiltration membrane to obtain penicillin V acid crystals I; c) recrystallizing the penicillin V acid crystal I to obtain a penicillin V acid crystal II; d) reacting and crystallizing the penicillin V acid crystal II and potassium isooctanoate to obtain penicillin V potassium; e) drying and storing; the molar yield and the quality index of the penicillin V potassium obtained by the novel process are greatly improved compared with those of the existing level.

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.