31 results about "Amycolatopsis orientalis" patented technology

The invention provides ribosome resistance mutagenesis breeding vancomycin production bacterial strain and an application thereof, specifically the mutagenesis breeding method of mutagenesis bacterial strain of Amycolatopsis orientalis and the application thereof in preparing vancomycin, relating to breeding of microorganism. The method comprises the following steps: inoculating the Amycolatopsis orientalis to Gause-1 agar culture medium panel to be cultured; taking out spore and using sterile water to prepare spore suspension and coating the spore suspension in Gause-1 agar culture medium panel featuring mixing resistance of vancomycin and streptomycin to be cultured; randomly selecting produced single bacterial colony and streak-inoculating the single bacterial colony to the Gause-1 agar culture medium panel to be cultured, carrying out biological activity determination on produced mutagenesis bacterial strain; selecting mutagenesis bacterial strain with inhibition zone diameter thereof being 2mm larger than the inhibition zone diameter of original strain and inoculating the mutagenesis bacterial strain to liquid fermentation medium to be cultured; determining the content of vancomycin in fermentation liquor by HPLC method to verify the obtained vancomycin mutagenesis bacterial strain; subjecting the obtained vancomycin mutagenesis bacterial strain to 5 times of continuous passage stability inspection to obtain the vancomycin production bacterial strain.

The invention provides isolated nucleic acid compounds encoding the glycosyltransferase protein GtfC of Amycolatopsis orientalis. Also provided are vectors carrying the gtfC gene, transformed heterologous host cells for expressing the GtfC protein, and methods for producing glycopeptide compounds using the cloned gtfC gene.

The invention provides isolated nucleic acid compounds encoding the glycosyltransferase protein GtfD of Amycolatopsis orientalis. Also provided are vectors carrying the gtfD gene, transformed heterologous host cells for expressing the GtfD protein, and methods for producing glycopeptide compounds using the cloned gtfD gene.

The invention provides isolated nucleic acid compounds encoding the glycosyltransferase protein GtfB of Amycolatopsis orientalis. Also provided are vectors carrying the gtfB gene, transformed heterologous host cells for expressing the GtfB protein, and methods for producing glycopeptide compounds using the cloned gtfB gene.

The invention discloses a CRISPR/Cas9 editing system based method for gene knockout and gene knock-in in Amycolatopsis orientalis. The method introduces a CRISPR/Cas9 edited plasmid pLYNY04 into Amycolatopsis orientalis HCCB10007, realizes gene knockout and gene knock-in, simplifies genetic operation and improves the integration efficiency of the Amycolatopsis orientalis. The invention also discloses a CRISPR/Cas9 edited plasmid pLYNY04 containing a gtfD gene homologous arm; and eGFP is inserted into a target sequence of the Amycolatopsis orientalis to facilitate detection of gene editing results. The invention also discloses application of the CRISPR/Cas9 edited plasmid pLYNY04 containing the gtfD gene homologous arm.

Novel polyene polyketides, their pharmaceutically acceptable salts, prodrugs and derivatives have been found to have antibiotic activity. One method for obtaining the compounds is by cultivation of Amycolatopsis orientalis ATCC™ 43491 or a mutant or variant such as the strain IDAC-220604-1. Another method for obtaining the compounds is post-biosynthesis chemical modification of the compounds obtained by cultivation. Novel polynucleotide sequences and encoded proteins for the biosynthesis of the polyene polyketides are also presented.

The present invention provides a process for preparing vancomycin hydrochloride using a mutant strain of Amycolatopsis orientalis (accession No. KCCM-10836P) comprising the steps of i) mutating and isolating a strain of Amycolatopsis orientalis (accession No. KCCM-10836P) for producing vancomycin from mother strain of Amycolatopsis orientalis (accession No. ATCC-19795) using NTG(N-methyl-N′-nitro-N-nitrosoguanidine) in the selection medium; ii) seed culturing the isolated mutant strain of Amycolatopsis orientalis (accession No. KCCM-10836P); iii) cultivating and fermenting said mutant strain of Amycolatopsis orientalis in the fermentation medium consisting of 12˜18 (w/v) % of dextrin, 2.2˜3.8 (w/v) % of bean powder, 1.9˜2.9 (w/v) % of potato protein, 0.10˜0.14 (w/v) % of sodium chloride and a small amount of minerals; iv) filtering vancomycin using microfilter in the cultivation broth by removing mycelia; v) purifying obtained vancomycin using column system compacted with cation exchange resin, anion exchange resin and absorbent resin; and vi) crystallizing the purified vancomycin with hydrochloric acid.

The invention discloses a method for improving the yield of ECO-0501 produced by fermenting Amycolatopsis orientalis, which is achieved by interdicting the Amycolatopsis orientalis to synthesize vancomycin, wherein the synthesis of the interdicted vancomycin comprises interdicting or knocking out vancomycin synthetase genes or vancomycin precursor synthetase genes. The method for improving the yield of the compound ECO-0501 produced by fermenting the Amycolatopsis orientalis interdicts the synthesis of the vancomycin while improving the yield of the ECO-0501, thus the method is particularly favorable for the downstream separation steps.

The invention provides a method for producing chlorine-free vancomycin, which comprises the following steps: (i) interdicting a gene of encoding halogenase in amycolatopsis orientalis for producing vancomycin, constructing gene engineering bacteria for producing the chlorine-free vancomycin; and (ii) fermenting the gene engineering bacteria for producing the chlorine-free vancomycin so as to prepare the chlorine-free vancomycin. The invention also provides a method for constructing the gene engineering bacteria for producing the chlorine-free vancomycin and the gene engineering bacteria obtained through the method. The invention opens up a new approach for the preparation of the chlorine-free vancomycin.

The invention relates to a fermentation culture medium for producing vancomycin through fermentation of streptomyces orientalis and a material supplementing method. By the adding of composite modified starches, fibroin powder, casein and other substances into the fermentation culture medium, the culture quality and the fermentation unit of vancomycin fermentation liquor can be improved favorably, and the fermentation cost can be reduced; furthermore, the influence of the environment on original auxiliary material sources can be reduced to the maximum extent, so that sufficient supply of the original auxiliary material sources can be guaranteed, and efficient production of vancomycin is realized.

The present invention provides a deshydroxy vancomycin compound,a method of its preparation and a pharmaceutical composition comprising a pharmaceutically effective amount of the deshydroxy vancomycin and the use of the said composition in the preparation of drugs for the treatment of susceptible bacteria infections. The method includes the following steps: (1) preparing a concentrated vancomycin solution containing the deshydroxy vancomycin by fermentations of Amycolatopsis Orientalis with Deposit No. CGMCCNO.1183; (2) separating and purifing the concentrated vancomycin solution to obtain a refined filtrate of vancomycin hydrochloride containing the deshydroxy vancomycin by column chromatography; and (3) further separating and purifing the refined filtrate to obtain the deshydroxy vancomycin by chromatography. Wherein, separation and purification is processed by column chromatography in a gel chromatography column containing a salt-water mobile phase, separation and purification is processed by chromatography in a macroporous adsorption resin chromatography column containing a buffer-methanol mobile phase.

The invention relates to a seed culture medium for producing vancomycin through fermentation of streptomyces orientalis and a cultivation method. By the adding of composite modified starches, fibroin powder, casein and other substances into the seed culture medium, the seed culture quality and the fermentation unit of vancomycin can be improved favorably, and the fermentation cost can be reduced; furthermore, the influence of the environment on original auxiliary material sources can be reduced to the maximum extent, so that sufficient supply of the original auxiliary material sources can be guaranteed, and efficient production of vancomycin is realized.

The invention discloses a method for preparing A82846B by fermentation. The method includes extracting a fermentation broth obtained after fermentation and culture of amycolatopsis orientalis, and conducting alkali degradation on the obtained impurity waste liquid to obtain an alkali degrading solution; after the alkali degradation liquid is added to a fermentation medium, performing fermentationagain, culturing for 25-35h and adding calcium chloride to continue the fermentation; adding the alkali degrading solution in different concentrations of impurities and adding calcium chloride after the fermentation culture is performed for about 30h. The A82846B fermentation unit is greatly improved, and the composition is significantly improved.

Disclosed are a ketoreductase mutant which can be used for an efficient production of daunorubicin derivatives, a DNA encoding the mutant, a transformant prepared by introducing the DNA thereinto to produce a daunorubicin derivative, and a process of producing a daunorubicin derivative using the transformant. The ketoreductase mutant has an amino acid sequence in which one amino acid residue or two or more amino acid residues selected from the group consisting of amino acids located at positions corresponding to the 42nd, 149th, 153rd, 270th, and 306th amino acids in the amino acid sequence of a ketoreductase (EvaE) from a chlororemomycin-producing bacterium (Amycolatopsis orientalis) are substituted with another amino acid residues.

The invention discloses a method for preparing A82846B in a fermenting manner. The method comprises the following steps: extracting fermentation culturing solutions obtained by performing fermentationculturing on amycolatopsis orientalis; and performing alkali degradation on impurity waste liquid obtained by performing extraction to obtain alkali degradation liquid. The alkali degradation liquidis added into a fermentation culture medium, and then fermentation is carried out again; through adding the impurity alkali degradation liquid with different concentrations, the fermentation unit of A82846B is greatly improved, and components are obviously improved at the same time.