198 results about "G penicillin" patented technology

The invention relates to a gene, mutant plasmid and engineering bacteria which have improved synthesis performance to penicillin G acylase and are obtained by a gene site-directed mutagenesis method, and mutant enzyme can also be obtained with improved synthesis performance to penicillin G acylase by fermenting and purifying the engineering bacteria. Two enzymes Kpn I and Pst I are firstly used for cutting pUC18 by the invention, then T4 polymerase is adopted to make the ends blunt, and pZ01 is obtained through self-linkage; the enzyme of EcoR I is used for cutting pZ01, and then connected with pEES102 that is also cut by the enzyme of EcoR I, thereby obtaining the recombinant plasmid pY020; the pY020 is adopted as a template plasmid, and TaKaRa MuTanBEST Kit is utilized for conducting the site-directed mutagenesis to B.megaterium PGA, thereby obtaining the mutant plasmid with improved synthesis performance to the penicillin G acylase. The mutant plasmid is transformed to bacillus subtilis to obtain the required engineering bacteria. The engineering bacteria are amplified and fermented, and the mutant enzyme with improved maximum conversion rate of 7-ADCA and the ratio of synthetic product / hydrolysate can be obtained after the engineering bacteria are purified.

The invention relates to a genetic-engineering antibacterial peptide, as well as a preparation method and application thereof. The preparation method comprises the following steps: according to the characteristics of the amino acid sequence of Magainin and Cecropin P and LL-37 which are antibacterial peptide, a novel antibacterial peptide gene is designed; the sequence of the gene is synthesized and transformed to be expressed in pichia to form antibacterial-peptide gene transformation pichia engineering bacteria, and to be expressed in a fermentor to achieve the effects of high-density cultivation and efficient expression. Fermentation broth is separated and purified to be an antibacterial peptide product which can be applied to feed and food additives and be used as injection. The antibacterial peptide has the advantages of good antibacterial activity to most gram-negative bacteria and gram-positive bacteria, in particular better effects of inhibiting and killing ampicillin-resistant bacteria and kanamycin-resistant bacteria.

The invention discloses a magnetic polymer microsphere for enzyme immobilization and a relative preparation method, which uses hydrophilic nanometer magnetic particles as magnetic material, uses vinyl compound as functional monomer, uses composite surface activator as disperser and uses inverse suspension polymerization technique to prepare the macromolecule polymer pear carrier with narrow grain distribution and superparamagnetic and hydrophilic expoy group. The immobilization penicillin acylase prepared by the magnetic carrier has high apparent activity as 330IU / g (humidity), and the immobilization enzyme can be recovered easily to be circulated by external magnetic field, thereby improving the utilization of immobilization. The preparation method has simple process, easy operation, low production cost and support for large-scale production.

The invention discloses a biological cell freezing antifreeze, and 100ml of the cell freezing antifreeze consists of the following raw materials: 0.025 to 0.3g of kelp polysaccharide, 2.0 to 3.5g of glucose, 0.5 to 1.5g of sodium citrate, 60 to 80ml of distilled water, 10 to 20ml of albumin, 5 to 18ml of glycerol and 0.05 to 0.15 million units of penicillin. The preparation method is that: the glucose and the sodium citrate are dissolved in the distilled water to prepare a base liquid; the albumin is added, the mixture is mixed evenly, and the penicillin is added to prepare a I liquid; the glycerol is added in the I liquid to prepare a II liquid; the kelp polysaccharide is added in the II liquid for even mixing; the pH value is adjusted to 6 to 7.5, then the filtration and sterilization are carried out, and the mixture is cooled until reaching the room temperature and then arranged in a refrigerator of 2 to 5 DEG C for standby. The cell freezing antifreeze is applicable to the cryopreservation of mammalian semen, somatic cells, oocytes, early embryos, organs and tissues, in particular to the cryopreservation of the semen of various mammals.

The invention relates to the fields of cytobiology and cell culture, and particularly relates to a breast cancer organoid culture kit. A method for realizing the kit comprises the following steps: (1)preparing, sub-packaging and using a breast cancer organoid culture medium A; (2) preparing, sub-packaging and using a breast cancer organoid culture medium B; and (3) preparing, sub-packaging and using a sample preserving fluid. The breast cancer organoid culture medium A is a DMEM / F12 culture medium containing 100 U / ml of penicillin, 0.1 mg / ml of streptomycin, 1% of HEPES and 1% of Gluta Max 100 X; the organoid culture medium B is a DMEM / F12 culture medium containing EGF, an FGF growth factor, an ALK inhibitor A83-01, Noggin, an HEPES buffer solution, Y27632 and one or more of a Wnt signalpath related secretory protein family R-Spondin 1-4; and the sample preserving fluid is a DMEM / F12 culture medium containing 100 U / ml of penicillin, 0.1 mg / ml of streptomycin and 1% of L-glutamine. The three components form the breast cancer organoid culture kit.

The invention discloses a process for preparing 6-aminopenicillanic acid, which comprises the following steps: (1) filtering the penicillin fermentation liquid, acidifying, extracting and concentrating with butanol, decoloring to obtain butyl extract of penicillin, (2) subjecting butyl extract of penicillin to back extraction with alkaline solvent, obtaining aqueous solution of penicillin salts, (3) degreasing the aqueous solution of penicillin salts, (4) loading the degreased aqueous solution of penicillin salts into penicillin acylated enzyme retort for enzyme conversion, charging 6-APA seeds, cultivating quartz, crystallizing and drying.

The invention provides a penicillin G acylase mutant for synthesis and an application thereof in the preparation of amoxicillin. Penicillin G acylase of Achromobacter xylosoxidans origin is mutated by computer aided design in connection with semi-rational design of site-saturation mutagenesis technique and enzyme engineering modification of orthogenesis, thus acquiring the penicillin G acylase mutant lower in hydrolytic activity, better in synthetic activity, higher in synthesis-hydrolysis ratio (S / H), higher in acid resistance and better in stability, and amoxicillin can be catalytically synthesized more effectively and quickly. Immobilized enzyme hydrolytic activity of the mutant SPGA-4 obtained is decreased by 8.7 times, synthetic activity is increased by 5.6 times, the S / H ratio is increased by 8 times, the activity remains at 79% for 60 min under the condition of pH 2.0, amoxicillin is catalytically synthesized by a solid method at 10 DEG C or 20 DEG C, substrates 6-APA and D-HPM are directly charged in a solid form without dissolving, reaction pH need not be controlled, substrate conversion rate is higher than 99%, continuous use is available in more than 300 batches, and good operational stability is given.

The invention relates to a preparation method of a frozen pig semen diluent. The frozen semen diluent comprises four diluent components: a diluent A comprises glucose, sodium citrate, sodium chloride and penicillin; a diluent B comprises lactose, glucose, penicillin, streptomycin and fresh yolk; a diluent C comprises glucose, sodium citrate, sodium chloride, penicillin, glycerol, coconut oil monoethanol amide, lauryl sodium sulfate, triethanolamine, hydrochloric acid and distilled water; a diluent D comprises anhydrous citric acid, caffeine g, trihydroxymethyl amino methane, sodium bicarbonate, ethylenediamine tetraacetic acid disodium salt, sodium pyruvate and glucose; the preparation steps are as follows: respectively dissolving raw materials in 100ml of double distilled water to prepare four diluents, regulating the pH values to be 6.5-7.5, and refrigerating in a refrigerator. The product disclosed by the invention is used for the freezing storage of the pig semen diluent.

The invention discloses anti-oxidative cattle frozen semen diluent. The anti-oxidative cattle frozen semen diluent is composed of the following raw materials by mass: 20-26 parts of trihydroxy methyl-aminomethane, 10-15 parts of trisodium citrate, 7-15 parts of fructose, 700-800 parts of distilled water, 230-260 parts of fresh yolk, 0.125-0.375 part of sodium selenite, 2-32 parts of Vitamin E, 90-95 parts of glycerol, 1-2 parts of penicillin potassium for injection (specification of 1.6 million units / g) and 1-2 parts of streptomycin sulfate for injection (specification of 1.0 million units / g). The preparation method of the anti-oxidative cattle frozen semen diluent comprises the steps of preparing a basic buffer solution, a basic diluent and a semen diluent; then transferring the prepared semen diluent to a condition of 4 DEG C, and balancing for 5-20 min to obtain the anti-oxidative cattle frozen semen diluent. Compared with a conventional anti-oxidative cattle frozen semen diluent, the anti-oxidative cattle frozen semen diluent provided by the invention increases semen motility rate by 24.56%, increases 4 h semen motility rate by 12.39%, increases semen perforatorium integrity rate by 38.72%, increases GSH-PX activity by 260%, and reduces semen aberration rate by 5.49%.

The invention discloses preparation and an enzyme-linked immunosorbent assay method for heavy metal mercury polyclonal antibody in the technical field of heavy metal detection. The polyclonal antibody is prepared for the enzyme-linked immunosorbent assay by the following steps of: bonding the heavy metal mercury at one end of penicillin G sodium serving as a dual-functional chelating agent; coupling the other end with high molecular carrier proteins, namely bovine serum albumin and ovalbumin, to form immunogen MMPA-BSA and envelope antigen MMPA-OVA of mercury-chelating agent-protein; and finally emulsifying and immunizing the immunogen MMPA-BSA to prepare the polyclonal antibody. The prepared antibody has high specificity for the mercury ions, a cross reaction ratio with all other metals of less than 0.001 percent apart from the cross reaction ratio with cadmium of 7.9 percent and an adding recovery ratio of between 91.4 and 120 percent; and thus the antibody can be used for the assayof the heavy metal mercury in a water sample and can also be developed into rapid immunological detection technology for rapidly detecting the heavy metal mercury in other samples such as agricultural products and the like by combining certain pre-processing technology.

The invention relates to a penicillin G acylase mutant constructed by genetic engineering. In comparison with wild type penicillin G acylase from achromobacter (Achromobacter sp.CCM 4824), the synthesis performance of the penicillin G acylase mutant is improved substantially, the maximum synthetic product / hydrolysate value S / H reaches 22.3 and is 3.9 times that of wild type enzyme, and various beta-lactam antibiotics can be efficiently synthesized catalytically. When the ratio of side chains to mother nucleus is 1.05:1, the conversion rate of the mother nucleus 6-APA, 7-ADCA, 7-ACCA and 7-APRA reaches 99.0% or above, and the penicillin G acylase mutant has wide industrial application prospects.

The invention relates to a method for preparing penicillin-G-1-(S)-oxide. In the method, penicillin G is used as a raw material, and the oxidization of the penicillin G and the crystallization of penicillin-G-1-(S)-oxide which is the oxidization product of the penicillin G are performed at the same time. A penicillin-G-1-(S)-oxide crystal product is obtained by directly adding an oxidant into penicillin G organic phase solution, reacting and crystallizing, the oxidization reaction and the crystallization are performed at the same time, and the crystal product directly precipitates in organic solution. Compared with the conventional direct oxidization process, the method has the advantages that: the operation flow is simplified considerably; the operation is simple and convenient; the labor intensity is lowered; and the production period is shortened obviously. The process of extracting the penicillin G from the organic phase back to a water phase and the process of regulating pH value with diluted acid and crystallizing are saved, so the consumption of waster resources is reduced greatly, the discharge of waste acid liquor is reduced, and the problem of environmental pollution is alleviated obviously. The high-performance liquid chromatography (HPLC) content of the product reaches over 99.0 percent, and the weight yield of products with a main particle size over 30 mu m is over 90 percent.

The invention discloses benzopyrone-phenyl-oxazolidone compounds with the following structural general formulas shown in the specification. The compounds have better inhibiting and killing effects on various germs, and some of the benzopyrone-phenyl-oxazolidone compounds have higher bacteriostatic activity than positive control penicillihe G, kalamycin and ketoconazole, so that the benzopyrone-phenyl-oxazolidone compounds can be used for preparing anti-infective drugs. The invention also discloses preparation methods of the benzopyrone-phenyl-oxazolidone compounds.

The invention discloses a heterogeneous biocatalyst taking alpha-amino-acid ester hydrolase as basis, a preparation method thereof and application in synthesizing amino beta-lactams. The activity of catalyst synthetase is 2,200 U / g by dry weight, and the content of protein is 0.95 g / g by dry weight; and xanthomonas rubrilineans cell biomass is processed in an organic solvent under a pH gradient through low temperature effect to extract enzyme, and enzyme aggregates are deposited and cross-linked to obtain the hydrolase. The alpha-amino-acid ester hydrolase as a catalyst can be synthesized into amino penicillin and amino cephalosporin drugs through acylating a beta-lactam compound by D-phenylglycine methyl ester derivatives in water or in a mixed medium of water and an organic solvent.

The invention relates to the technical field of biosensors, especially relates to a fluorescent biosensor based on hybrid chain reaction amplification, and solves the problems in the prior art that the method for detecting ampicillin is low in specificity and sensitivity, and the cost is high. According to a biosensor for detecting the ampicillin based on an aptamer, a cyclic amplification effectis realized by the cooperation of Nb.BbcCI and a chain hybrid chain reaction, and Thioflavine-T and G-quadruplex are combined to generate fluorescence and a homogeneous reaction mixed solution. The preparation method comprises the following steps of preparing gold nanoparticles; modifying Walker and Track to the surfaces of the gold nanoparticles; mixing a labeled nanogold solution with the homogeneous reaction solution; performing hyperbranched hybrid chain reaction and fluorescent detection; carrying out high specificity detection on the target ampicillin by using the aptamer by utilizing the specific recognition of the aptamer; and using hyperbranched hybrid chain reaction amplification to achieve signal amplification.

A process for the use of low concentration levels of Penicillin G Procaine to eliminate or control the growth of unwanted or undesirable bacteria (contaminating bacteria) in the fermentation production of alcohol without inhibition of the growth or replication of the yeast.

The invention provides a mycoplasma anatis culture medium. The mycoplasma anatis culture medium is composed of a liquid culture medium and a solid culture medium. The liquid culture medium is prepared from deionized water, a 10% thallium acetate solution, PPLO broth, glucose, peptone, tryptone, 1% phenol red, a CMRL-1066 culture medium containing L-glutamine, inactivated horse serum, yeast leachate, penicillin, L-cysteine hydrochloride and nicotinamide adenine dinucleotide. Besides the situation that agar powder is added and phenol red is removed, other components of the solid culture medium are the same as those of the liquid culture medium. The culture medium is used for separating and purifying mycoplasma anatis, wherein a bacterial colony is separated from the solid culture medium, the separated bacterial colony is placed in the liquid culture medium to grow, then filtering, dilution and plate coating of liquid culture substances and purification of picked single colony multiplication are conducted, and the steps are repeated till purified mycoplasma anatis is obtained. The culture medium can promote growth of mycoplasma anatis, and the success rate of separation and purification of mycoplasma anatis is high.

The invention discloses a crynebacterium glutamicum engineering strain for producing 5-aminolevulinic acid and a construction method thereof. The construction method comprises the steps of 1, knocking out lactic dehydrogenase encoding gene idhA, and acetic acid generation genes pta-ackA, pqo and cat in the corynebacterium glutamicum, inserting a strong sod promoter in front of a phosphoenolpyruvate carboxylase encoding gene ppc, knocking out a phosphoenolpyruvate carboxykinase pck, thus acquiring the strain CB6; 2, knocking out a penicillin-binding protein encoding gene pbpla in the strain CB6, thus acquiring the strain CB7; transferring the plasmid pXA of a built over-expressed 5-aminolevulinic acid synthase gene into the strain CB7, thus acquiring the engineering strain L2. According to the engineering strain provided by the invention, the 5-aminolevulinic acid is produced in a medium in which 10g / L glucose is added as a carbon source; compared with reference strains, the engineering strain is respectively improved by 13.53% and above.

The invention discloses an application of beta-carotene in preparation of a pig sperm cryoprotectant. The pig sperm cryoprotectant is composed of a cryoprotective basic solution, beta-carotene, fresh egg yolks and glycerin. The additive amount of beta-carotene is 1 mg-2 mg in 100 mL cryoprotective basic solution, and the additive amount of fresh egg yolks is 20 mL in 100 mL cryoprotective basic solution. 97 mL cryoprotective basic solution with addition of beta-carotene and fresh egg yolks is taken, 3 ml glycerin is added, and then 100 ml pig sperm cryoprotectant is prepared. 100 mL cryoprotective basic solution comprises 1.1g glucose, 1.48 g citric acid, 2.42 g Tris, 0.06 g penicillin sodium and 0.1g streptomycin sulphate, and the rest is double distilled water. In the application, beta-carotene is firstly used in pig sperm cryopreservation, after thawing, the pig sperm motility rate, the plasma membrane intact rate and the acrosome intact rate are 51%, 53% and 52%.

The present invention relates to a mutant prokaryotic penicillin G acylase derived from a wild-type penicillin G acylase characterized in that the mutant is having an amino acid substitution at one or more amino acid positions selected from the group consisting of amino acid positions A3, A77, A90, A144 A192, B24, B109, B148, B313, B460 and B488 according to the amino acid numbering of the Escherichia coli penicillin G acylase having the amino acid sequence depicted in SEQ ID No: 1.

The invention relates to a culture medium for human dental pulp stem cells and a preparation method for human dental pulp stem cells and belongs to the technical field of stem cell preparation. The culture medium for human dental pulp stem cells, provided by the invention, is based on an alpha-MEM culture medium and contains 5%-10% of human AB serum by volume percent, 100mu M ascorbic acid, 2mM L-glutamine, 100U / ml penicillin sodium and 100mg / ml streptomycin. The dental pulp stem cells prepared by the culture medium provided by the invention are high in yield, are free from human immunological rejection and can supply reliable seed cells for repairing and regeneration of clinical teeth.

The invention provides a preparation method of methoxybenzyl, which uses penicillin and saturated salt water containing hydric sulphate. The invention uses penicillin as raw material to obtain penicillin sulfoxide via oxidization, to obtain penicillin sulfoxide ester via esterification, to be reacted with benzene sulfinate, to open ring and generate nitrogen heterocyclic butanone sulfinic acid intermediate, and electrolyzes the saturated salt water containing hydric sulphate, to utilize generated chlorizating agent to chlorizate the intermediate, at last ammonolyzes and closes ring to generate GCLE. The invention has the advantages of high product yield, high purity, low cost, and high quality, which uses electrolysis close-ring to process chlorization in production to assure safe reaction.

The invention relates to a method for synthesizing cefprozil through a green enzymatic method. The method includes following steps: S1, adding parent nucleus 7-APRA or hydrochloride thereof into a buffer solution, and adding D-para hydroxybenzene glycinate derivative and / or D-para hydroxybenzene glycine amide under the condition that pH is 5-8; S2, adding cefprozil synthetase into the S1, reacting at temperature of 15-30 DEG C and pH of 6.5-7.8 for 1-3h, separating out separation liquid after reaction finishes, and immobilizing cefprozil synthetase to obtain a cefprozil crude product; S3, subjecting the crude product obtained in the S2 to acidolysis dissolved clarification, filtering and re-crystallizing to obtain cefprozil. Raw materials are cheap and easy to obtain, reaction is simple, hydrolysis activity of penicillin acylase can be effectively inhibited, and production cost is low; compared with conventional chemical synthesis methods, the method is simple and convenient to operate and low in cost, synthesis period is shortened, production efficiency is improved, total yield is high, controllability is high, and needs on industrial production are met.

The invention discloses a method for performing high performance liquid chromatography with fluorescence detection on residues of ampicillin and amoxicillin in an aqueous product, which belongs to the field of food safety detection. The method comprises the following steps of: 1, preparing standard solution; 2, performing sampling, namely, uniformly dividing not less than 500 grams of mashed sample into two parts and storing the sample at -18 DEG C; 3, performing extraction, namely, adding 5 grams of sample into sodium dihydrogen phosphate, adding water for dissolution until the volume is fixed to be 500 milliliters, preparing 30 milliliters of extract, performing secondary homogenization and centrifugation, extracting supernate and adding a trichloroacetic acid into the supernate for later use; 4, performing purification, namely, rinsing a small solid-extraction column sequentially with methanol, distilled water and the trichloroacetic acid at the controlled speed of 20 drops per minute, performing complete pumping, and performing elution with 5 milliliters of ethanol; and 5, performing derivation, namely, performing blow-drying with nitrogen at 40 DEG C, adding the trichloroacetic acid and 7 percent formaldehyde solution into 0.5 milliliter of standard solution for uniform mixing, placing the mixed solution in a sealing way into a boiling water bath to perform heating for 30 minutes, eluting the solution with ecetonitrile after the solution is cooled until the volume is fixed to be 1 milliliter, and performing filtration for the liquid chromatography, wherein the limit of detection is less than 5 mu g / kg.

The present invention relates to a mutant prokaryotic penicillin G acylase derived from a wild-type penicillin G acylase characterized in that the mutant is having an amino acid substitution at one or more amino acid positions selected from the group consisting of amino acid positions A3, A77, A90, A144 A192, B24, B109, B148, B313, B460 and B488 according to the amino acid numbering of the Escherichia coli penicillin G acylase having the amino acid sequence depicted in SEQ ID No: 1.

The present invention relates to 7beta-alkylamido-3-(1-methyl-1-pyrrolidyl onium) methyl-3-cephalo olefine-4-carboxylate and its preparation process and the synthesizing process of cephalo pyroxime hydrochloride with the said intermediate. The present invention has mild reaction condition, no need of expensive reagent and benzyl penicillin acylase, low cost, high product purity, and simple and practical reaction path.

The invention discloses an isolated adipose protectant which comprises normal saline, DMEM solution, L-glutamine, penicillin and streptomycin according to the proportion of 500ml to 500ml to (146.1mg-584.4mg) to 125000U to 125000U. The invention further discloses a preparation method and application of the protectant. The protectant has effect of effectively reducing adipose tissue culture stem cell contamination rate. Growth rate of the stem cells during culture of the adipose tissue is higher than that of stem cells growing without protective liquid, and culture time of primary cells can be shortened, culture efficiency is improved, and cost is saved.

The invention provides a method for synthesizing penicillin G sulfoxide by using a continuous flow reactor, and the method comprises the following steps: (1) providing the continuous flow reactor which comprises a peroxidation reaction system and an oxidation reaction system; (2) respectively putting hydrogen peroxide and glacial acetic acid into a first containing bottle and a second containing bottle in the peroxidation reaction system, and enabling the hydrogen peroxide to be in contact with the glacial acetic acid so as to obtain peracetic acid; and (3) enabling the peracetic acid to enterthe oxidation reaction system through a pre-cooling module, and contacting the peracetic acid with a penicillin G potassium salt aqueous solution so as to obtain the penicillin G sulfoxide and a penicillin G sulfoxide aqueous solution. The method can be used for effectively synthesizing the penicillin G sulfoxide, and in the whole process of the method, reaction materials are easy to obtain, treatment after reaction is simple, industrial three wastes are easy to treat, the yield of a target product is high, and the method is very suitable for industrial production.

The invention discloses a novel method for preparing penicillin G sulfoxide, which comprises the following steps: a, selecting a 60000-120000 mu / g penicillin G fermentation solution, and dropwisely adding peroxyacetic acid to obtain a solution I; b, filtering the solution I to obtain a filtrate, and concentrating the filtrate through a nanofiltration membrane of which the molecular weight cut-off is 200-800Da, thus obtaining a concentrated solution; c, regulating the pH value of the concentrated solution with sulfuric acid, and filtering to obtain a penicillin G sulfoxide crude product; and d, dissolving the penicillin G sulfoxide crude product in a methanol water solution, recrystallizing, then cooling to 0-10 DEG C, filtering, washing the obtained crystals with methanol, swabbing off, and performing microwave drying on the obtained product to obtain the penicillin G sulfoxide. According to the invention, the method is simple, the process period is short, and extraction treatment can be performed without using a large amount of organic solvent. Thus, the method disclosed by the invention is low in cost and environment-friendly.