34 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.

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.

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 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 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 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 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.

This invention relates to the field of enzyme catalysis technology, specifically to a deep-sea-derived protein fragment and its application as a penicillin cyclase. The protein is (a1) K141 1913, composed of the amino acid sequence shown in Sequence 1 of the sequence listing. The penicillin cyclase of this invention catalyzes the reaction of the substrate PenG to generate G-7-ADCA. Compared with existing chemical synthesis methods, the G-7-ADCA synthesis method provided by this invention has advantages such as being green, environmentally friendly, and low-cost, laying a solid foundation for the large-scale industrial production of G-7-ADCA.

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

This invention relates to the field of immunology, and more particularly to a hybridoma cell line and its secreted monoclonal antibody against the metamizole residue marker 4-methylaminoantipyrine (MAA), and its applications. This invention uses 4-aminoantipyrine to react with methyl 4-bromomethylbenzoate, followed by alkaline hydrolysis to obtain a hapten (Formula I). ​​An immunogen is then synthesized and used to immunize mice. After cell fusion and screening, a hybridoma cell line capable of secreting a monoclonal antibody recognizing MAA is prepared, named Metamizole-6A4. This hybridoma cell line has a high efficiency in secreting monoclonal antibodies. The secreted monoclonal antibody can accurately detect MAA, IC50, and other markers. 50 The concentration is 4.06 ng / mL, with high specificity. Cross-reactivity with aminopyrine and its metabolites 4-aminoantipyrine, 4-formamidoantipyrine, 4-acetamidoantipyrine, sulfadiazine, norfloxacin, lincomycin, tylosin, penicillin G, streptomycin, and gentamicin is less than 5%. It can accurately detect MAA residues in food. Formula I.

A mutant penicillin acylase and uses thereof are provided. Compared to the amino acid sequence set forth in SEQ ID NO: 1, the mutant includes at least one mutation selected from the group consisting of: F146αK, F24βR, F71βY, N241βK, G385βY, and G385βR. By introducing mutations into the penicillin acylase derived from Kluyvera citrophila, the invention obtains a mutant enzyme exhibiting enhanced and well-coordinated hydrolytic and synthetic activities. It can be used for the synthesis of β-lactam antibiotics, particularly for the one-step preparation of amoxicillin from penicillin potassium salt, thereby avoiding the isolation of the intermediate 6-APA. The present invention provides a key enzyme for the efficient production of β-lactam antibiotics and is poised to significantly advance the innovation of their manufacturing technology.

The application discloses an ex vivo perfusion preservative solution for improving the curative effect of liver transplantation. 1000 mL of the ex vivo perfusion preservative solution comprises the following components: 20-60 mg of oxamic acid, 2 g of sodium penicillin, 15000-20000 U of heparin, 20 mg of N-acetyl cysteine, and HTK liquid is supplemented to 1000 mL. The ex vivo perfusion preservative solution for improving the curative effect of normal liver and fatty degeneration liver transplantation can effectively improve the liver injury caused by I / R. Compared with the HTK liquid used at present, the ex vivo perfusion preservative solution has better protection effect, and has a good application prospect for the preservation of donor liver in liver transplantation surgery, especially the fatty degeneration liver.

The invention discloses an in-vitro perfusion preserving fluid for improving the curative effect of liver transplantation. Every 1000 mL of the in-vitro perfusion preserving fluid comprises the following components: 20 to 60 mg of oxamic acid, 2 g of penicillin sodium, 15000 to 20000 U of heparin, 20 mg of N-acetylcysteine, and the balance of HTK liquid, wherein the total volume is 1000 mL. The in-vitro perfusion preserving fluid for improving the curative effect of normal donor liver and fatty degeneration donor liver transplantation can effectively improve donor liver injury caused by I / R. Compared with the existing HTK liquid, the HTK liquid has a better protection effect, and has a very good application prospect for preserving the donor liver in the liver transplantation operation, especially the fatty degeneration donor liver.

The invention relates to a culture medium for culturing human mycoplasma pneumoniae and a culture method, and belongs to the technical field of microorganisms, the culture medium for culturing the human mycoplasma pneumoniae comprises a mycoplasma culture medium and a growth promoting additive, the growth promoting additive comprises horse serum or fetal calf serum, cholesterol, penicillin sodium and an HEPES buffer solution, according to the culture medium for culturing the human mycoplasma pneumoniae, various components are simple and easy to obtain, the preparation method of the culture medium is greatly simplified, and the efficiency is improved; according to the culture method of the human mycoplasma pneumoniae, the culture mode is changed from a static state to a dynamic state, so that the culture efficiency of the human mycoplasma pneumoniae is improved, and meanwhile, the thallus yield of the human mycoplasma pneumoniae is also improved; the culture method for preparing the human mycoplasma pneumoniae is short in period, low in cost, high in culture viable organism density and high in yield, and is suitable for large-scale culture of the human mycoplasma pneumoniae.

The application discloses a goat sperm endogenous heat shock protein 70 expression inducer, a method for improving the expression level of goat sperm endogenous heat shock protein 70 and application thereof. The goat sperm endogenous heat shock protein 70 expression inducer comprises the following components: Tris, citric acid, glucose, glutamine, glycerol, a penicillin-streptomycin mixed solution, egg yolk liquid and ultrapure water. The application further provides a method for improving the expression level of goat sperm endogenous heat shock protein 70, which comprises the following steps: preparing a goat sperm membrane protective agent; collecting semen; diluting the semen with the goat sperm endogenous heat shock protein 70 expression inducer; and freezing the semen. The application further provides application of glutamine in improving the expression level of goat sperm endogenous heat shock protein 70. The application can improve the expression level of heat shock protein 70 in sperm in the freezing recovery process, effectively reduces the membrane damage of sperm, guarantees the sperm activity and the intact rate of biological membrane structure, and guarantees the normal physiological function of sperm.

The present application relates to the technical field of biological enzyme, in particular to a penicillin G acylase mutant, a preparation method and application thereof. The penicillin G acylase mutant is based on the penicillin G acylase, and one or two of the following mutations are carried out: the phenylalanine at the 274th site is mutated to alanine, and the tryptophan at the 404th site is mutated to glycine; the amino acid sequence of the penicillin G acylase is shown as SEQ ID NO. 1. It is verified through experiments that, compared with the wild enzyme, the penicillin G acylase mutant provided by the present application significantly improves the efficiency of catalyzing hydrolysis of antibiotics, and has a wide application prospect.

The invention discloses a preparation method of DL-penicillin amine, which comprises the following steps: reacting penicillin G potassium with hydrazine hydrate for ring opening, and then reacting with formaldehyde for cracking, racemization and hydrolysis to obtain the DL-penicillin amine; according to the synthesis scheme, the penicillin G potassium is directly used as a starting raw material, the DL-penicillin amine is synthesized by adopting a one-pot boiling scheme through four-step reaction, the raw materials are low in cost and easy to obtain, the operation is simple, and the method is suitable for industrial production.

This invention proposes a microneedle electrochemical biosensor and its fabrication method. The sensor comprises: a substrate and a glucose sensing electrode, a sodium penicillin sensing electrode, a sodium chloride sensing electrode, a reference electrode, and a counter electrode disposed on the substrate. Specifically: the glucose sensing electrode comprises a first graphite layer, a Prussian blue layer, and an enzyme immobilization layer stacked from bottom to top; the sodium penicillin sensing electrode comprises a second graphite layer and a β-lactamase immobilization layer stacked from bottom to top; and the sodium chloride sensing electrode comprises a third graphite layer, a signal conversion layer, and an ion recognition membrane layer stacked from bottom to top. This sensor selectively measures the concentrations of glucose, sodium penicillin, and sodium chloride, achieving high sensitivity and accuracy in detection.

The application provides a method for preparing penicillin V potassium by a spray drying method, and belongs to the technical field of drug synthesis. In the method, potassium hydroxide and a mixed solution of strong alkali and weak acid potassium salt are salted at a specific temperature, and then negative pressure spray drying is performed at a low temperature. The method has the advantages of simple process flow, no need to use a large amount of solvent, no solid waste and waste liquid, and environmental protection. In addition, high-purity penicillin V potassium can be obtained. The results of examples show that, according to the method, the penicillin V potassium obtained has the purity of C 16 H 18 N2O5S is not less than 87%, the maximum single impurity is not more than 0.8%, the total impurity is not more than 2.1%, and the polymer is not more than 0.1%.

The invention relates to the field of cell culture, in particular to a totipotent stem cell culture medium and a preparation method of totipotent stem cells. The totipotent stem cell culture medium comprises a basic culture medium and a supplementary culture medium, the basic culture medium comprises a Knock-out DMEM culture medium and Matrigel matrigel, and the supplementary culture medium comprises fetal calf serum, a penicillin-streptomycin mixed solution, GlutaMAX, non-essential amino acid, beta-mercaptoethanol, a leukocyte inhibiting factor and a 2 '-flucytidine tripolymer. According to the totipotent stem cell culture medium disclosed by the embodiment of the invention, only the 2 '-fluorocytidine tripolymer is used as a chemical small molecule additive, and other conventional stem cell culture mediums are matched to realize efficient preparation and long-term maintenance of the mouse totipotent stem cells, and the obtained mouse totipotent stem cells are excellent and stable in totipotent state.