105 results about "Penicillin Acylase" patented technology

This invention discloses a method for preparing D-amino acid. The method comprises: derivatizing DL-amino acids to obtain N-phenylacetyl-DL-amino acids, performing enzyme-catalyzed asymmetric hydrolysis in aqueous solution to obtain N-phenylacetyl-D-amino acid, and performing chemical hydrolysis and crystallization to obtain D-amino acid. The enzyme used is immobilized penicillin acylase, which can be used for more than 100 times. The method has such advantages as high yield and high product optical purity, and is suitable for the majority of DL-amino acids resolution.

The invention relates to a method for comprehensively recovering effective ingredients in amoxicillin mother liquid prepared by an enzyme process. The method comprises the following steps: (1) concentrating the amoxicillin mother liquid, namely adjusting the pH value of the amoxicillin mother liquid prepared by the enzyme process to be 8.0-9.5, and performing nanofiltration and concentration to obtain concentrated mother liquid; (2) synthesizing amoxicillin under enzyme catalysis, namely adjusting the pH value of the concentrated mother liquid to be 5.8-7.0, and converting 6-APA (6-amino penicillanic acid) and D-methyl hydroxyphenyl glycinate into amoxicillin in the presence of immobilized penicillin acylase for synthesis; (3) preparing D-hydroxyphenyl glycine concentrated liquid by ultrafiltration and nanofiltration, namely separating after the enzyme catalysis reaction is ended to obtain amoxicillin crystals and secondary amoxicillin mother liquid, and performing ultrafiltration and nanofiltration on the secondary mother liquid to obtain the D-hydroxyphenyl glycine concentrated liquid; (4) crystallizing D-hydroxyphenyl glycine. According to the method, 6-APA and D-methyl hydroxyphenyl glycinate remained in the mother liquid are consumed through an indirect process of synthesizing amoxicillin under enzyme catalysis, the product quality of D-hydroxyphenyl glycine is improved, and the yield of D-hydroxyphenyl glycine is increased.

The invention discloses a method for the enzymatic synthesis of cefprozil in a recyclable aqueous two-phase system by using immobilized penicillin acylase. The method comprises the following steps: dissolving a cefprozil parent nucleus and an acylation reagent in an aqueous two-phase polymer solution, wherein the molar ratio of the parent nucleus to the acylation reagent is 1: (1-4); and adding immobilized penicillin G acylase, then adding a two-phase distribution conditioning agent, adjusting the pH value to 4.5-7, and reacting for 1 to 80 hours at the temperature of 5-30 DEG C, wherein the concentration of enzyme in a reaction system is 50-150U / ml. By adopting the method, the hydrolytic activity of the penicillin acylase can be effectively inhibited, thus the hydrolysis degrees of the acylation reagent and the cefprozil product can be reduced. Compared with a method using water as a medium, the method disclosed by the invention has the advantages that the product and the enzyme are distributed in two different water phases, a reactant is fully in contact with the enzyme, so that a synthesis / hydrolysis specific value is greatly increased, the yield of the cefprozil can be increased by 20%, and the highest yield can reach 78%; the adopted novel recyclable aqueous two-phase system can be recycled, so that the production cost is reduced.

The invention discloses a recovering method of effective components in amoxicillin enzymatic synthesis mother liquor by utilizing nanofiltration. The method comprises the following steps: (1) regulating pH of the mother liquor; (2) hydrolyzing amoxicillin in the mother liquor by utilizing penicillin acylase, thus only D-hydroxyphenylglycine and 6-amino-penicillanic acid (6-amino-penicillanic acid) exist in the other liquor; (3) preparing concentrated mother liquor by nanofiltration; and (4) separating and recovering the D-hydroxyphenylglycine and the 6-amino-penicillanic acid (6-amino-penicillanic acid) by isoelectric point crystallization. The method has the advantages that: the recycling rate of an inactive lateral chain can be improved regarding certain amoxicillin synthetase with highhydrolysis activity, so that the enzymatic synthesis of amoxicillin is more economical.

The invention discloses a making method of fixed penicillin acylated enzyme, which comprises the following steps: (1) grinding the porous resin with lateral functional group; sieving; drying; activating; blending with chitose solution; drying; obtaining the solid; (2) adding solid in the step (1) in the polar hydrophilic solvent; adding adjuvant to suck after decorating; washing; drying to obtain the porous resin carrier; (3) loading acylated penicillin on the porous resin carrier; obtaining the product.

The present invention discloses isolation of Penicillin Acylase (PA) from Achromobacter sp CCM 4824 expressed in recombinant strain E. coli BL21 CCM 7394 bearing the recombinant plasmid pKXIP1 and processing of PA into biocatalyst useful for the industrial synthesis of antibiotics. More particularly the invention discloses a synthesis of semi-synthetic [beta]-lactam antibiotics in the reaction mixture consisting of activated acyl-donor (D-p-hydroxyphenylglycine methyl ester or amide for Amoxicillin and Cefadroxil; D-phenylglycine methyl ester or amide for Ampicillin and Cephalexin) and nucleophile (6-APA or 7-ADCA) catalyzed by PA obtained from recombinant E. coli BL21 CCM 7394 as the biocatalyst.

The invention discloses an epoxy mesoporous molecular sieve for use in bio-enzyme immobilization and a preparation method thereof. In the invention, cyclo groups of the mesoporous molecular sieve which contains vinyl and has a cubic phase Ia3d structure and the mesoporous molecular sieve which has a surface vinyl function are oxidized by using metachloroperbenzoic acid to introduce epoxy functional groups with a shorter chain length onto the surface of the mesoporous molecular sieve while the influences of a surface functionization process on the aperture, the specific surface area and the pore volume of the mesoporous molecular sieve are reduced as much as possible, so the bio-enzyme can be directly immobilized on the surface of the mesoporous molecular sieve in a covalent bonding form without further activation, and the performance of the immobilized enzyme can be improved. The epoxy mesoporous molecular sieve can be used for the immobilization of water soluble bio-enzymes such as penicillin acylase, glucose isomerase, glucosidetransferase, parenzyme and diastase, in particular for the immobilization of the penicillin acylase; and the activity of the obtained immobilized enzyme is 8416U / g, and 90 percent of original activity of the immobilized enzyme is retained after 10 times of recycling.

The invention provides a chemical-enzyme method for preparing D-basic amino acid hydrochloride. The method comprises the following steps of: derivatizing DL-basic amino acid serving as a raw material to obtain DL-N-diphenyl acetyl basic amino acid by utilizing an acylating agent, hydrolyzing the DL-N-diphenyl acetyl basic amino acid by using immobilized penicillin acylase as a biocatalyst through non-stereo selectivity and enantioselectivity to obtain L-basic amino acid, phenylacetic acid and D-alpha-N-phenylacetyl basic amino acid; and performing acidolysis on the D-alpha-N-phenylacetyl basic amino acid to obtain phenylacetic acid and D-basic amino acid dihydrochloride, and performing treatment of epoxypropane to obtain the D-basic amino acid hydrochloride. The D-lysine hydrochloride and D-ornithine hydrochloride which are prepared by the method have the yield of more than 40 percent, and ee is more than 99 percent. The method has the characteristics of high yield and optical purity, simple process and the like, and is suitable for the large-scale production of the D-basic amino acid hydrochloride.

The invention discloses an aldehyde group mesoporous molecular sieve used for immobilization of a biological enzyme and a preparation method thereof. In the preparation method, a 60 Co-gamma ray pre-irradiation grafting acrolein technique or gamma-aldehyde propyl trimethoxy silane is used for performing functionalized modification on the surface of the mesoporous molecular sieve, an aldehyde propyl functional group with a shorter chain length is introduced to the surface of the mesoporous molecular sieve, the influences, caused by the surface functionalizing process, on the aperture, the specific area and the pore volume of the mesoporous molecular sieve are reduced as much as possible, and further activation is not needed to ensure that the biological enzyme is directly immobilized on the surface of the mesoporous molecular sieve in a covalent bonding mode, and the performance of the immobilized enzyme is improved. The aldehyde group mesoporous molecular sieve can be applied to the immobilization of water-soluble enzymes such as penicillin acylase, glucose isomerase, transglucosidase, trypsase, amylase and the like, and is particularly suitable for the immobilization of the penicillin acylase; and the obtained immobilized enzyme has the activity of 8895 U / g, and after 10 times of recycling, the immobilized enzyme maintains 92 percent of the initial activity.

The invention relates to a method for synthesizing amoxicillin and generating a byproduct of a sodium phenylacetate solution by using a semi-direct method and belongs to the technical field of medicine preparation. The method comprises the following steps: splitting benzylpenicillin potassium into 6-APA (Amino Penicillanic Acid) and phenylacetic acid under the action of penicillin acylase, extracting a splitting solution with dichloromethane to separate 6-APA from the phenylacetic acid, putting ammonia water into an extraction water phase, adjusting the pH value of a material liquid to be neutral, removing the dichloromethane, putting the solid 6-APA into the obtained 6-APA dissolving liquid, and synthesizing the amoxicillin together with D-methyl p-hydroxyphenylglycinate under the actionof amoxicillin synthetase. A sodium phenylacetate solution can be prepared by alkalizing the dichloromethane obtained by extracting the splitting solution, the sodium phenylacetate solution can be directly applied to fermentation production of penicillin, the step of recycling the phenylacetic acid is avoided, and the production cost is reduced.

The invention belongs to the technical field of biology, and discloses an aramagnetic epoxy group mesoporous molecular sieve for immobilizing biological enzymes, and a preparation method thereof. According to the invention, gamma glycidoxy propyl trimethoxy silane is utilized to introduce aramagnetic epoxy groups onto the surface of the mesoporous molecular sieve, and then a covalent bond is utilized to immobilize aramagnetic Fe3O4 nano particles and biological enzymes onto the outer surface and the inner surface of the mesoporous molecular sieve respectively, wherein the aramagnetic Fe3O4 nano particles are subjected to L-cysteine surface modification and have the particle sizes larger than the pore diameter of the mesoporous molecular sieve, as a result, specially-structured aramagnetic immobilized enzymes can be prepared and are separated from the liquid phase system easily under the action of an external magnetic field, and further, the property and separation efficiency of the immobilized enzymes are improved. The aramagnetic epoxy group mesoporous molecular sieve can be used for immobilization of water-soluble biological enzymes such as penicillin acylase, glucose isomerase, glucosylase, trypsin and amylase, the activity of the prepared aramagnetic immobilized penicillin acylase is 8800 U / g, and 94.5% of the initial activity is retained after ten times of cycle use.