43 results about "Amycolatopsis" patented technology

This invention provides an Amycolatopsis sp. strain (zhp06), and a method of using the whole cell preparation of the strain for vanillin production. The strain was deposited in China Center for Type Culture Collection on Jul. 26, 2011 with the number of CCTCC NO: M 2011265. Under high concentrations of ferulic acid substrate, the vanillin production by this method can reach more than 10 g/L. The molar conversion rate of ferulic acid is more than 50% and the purity of vanillin is from 80% to 95%. The advantage of this invention includes: repeated use of biocatalyst cells, mild biotransformation condition, low environmental pollution, short production cycle, high product purity and simple purification procedure. It has a great potential for industrial applications.

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.

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 discloses a method for converting ferulic acid to produce vanillin by immobilized amycolatopsis, and belongs to the technical field of biology engineering. Cellulosic materials such as sawdust, bagasse and the like are used as carriers, and an amycolatopsis cell is fixed by an adsorption way. An immobilized cell can be recycled for 10 times, and the mole conversion rate for converting ferulic acid into vanillin is kept at more than 50%. A resin is coupled and adsorbed in a conversion system. The output of vanillin reaches 19.65g/L, and the production strength of vanillin is 0.131g/L.h. The method is mild in conversion reaction condition, little in environmental pollution, short in production period, low in cost and easily separates and purifies converted products, so that the method has a good application prospect.

Lovastatin, pravastatin, simvastatin, mevastatin, atorvastatin, fluvastatin and cervastatin and derivatives and analogs thereof are known as HMG-CoA reductase inhibitors and are used as antihypercholesterolemic agents. The majority of them are produced by fermentation using microorganisms of different species identified as species belonging to Aspergillus, Monascus, Nocardia, Amycolatopsis, Mucor or Penicillium genus, some are obtained by treating the fermentation products using the methods of chemical synthesis (simvastatin) or they are the products of total chemical synthesis (fluvastatin, atorvastatin and cervastatin). The present invention relates to a crystalline form of the sodium salt of pravastatin, which is known by the chemical name 1-naphthaleneheptanoic acid, 1,2,6,7,8,8a-hexahydro-β,δ,6-trihydroxy-2-methyl-8-(2-methyl-1-oxobutoxy)-, mono sodium salt, which is useful as a pharmaceutical substance. The present invention further relates to the method for its preparation and isolation, to a pharmaceutical formulation containing the sodium salt of pravastatin in the crystalline form and a pharmaceutically acceptable carrier, and to a pharmaceutical method of treatment. The novel crystalline form of the sodium salt of pravastatin is useful in the treatment of hypercholesterolemia and hyperlipidemia.

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.

Crude solid vanillin-containing material is precipitated from a solution obtained by biotransformation, and purified by a process comprising contacting it with a purification fluid selected from (a) a liquefied gas whose pressure exceeds its critical pressure and whose temperature is below its critical temperature; (b) a supercritical fluid; (c) a gas. The fluid is preferably liquid carbon dioxide. The temperature is maintained below 25°. The product may be further purified by treatment with CO₂ in a fluid bed drier. The crude material is preferably one precipitated from a solution resulting from biotransformation of ferulic acid. A new strain of Amycolatopsis capable of generating high concentrations of vanillin with minimal odoriferous by-products (e.g. guaiacol) is also disclosed.

The invention discloses Amycolatopsis sp. YIM M13163 and a method for preparing efrotomycin pivalate by adopting the strain through fermentation. Amycolatopsis sp. YIM M13163 is preserved in the China Center for Type Culture Collection (CCTCC) on July 10, 2015, the preservation number is CCTCC M 2015445, and the preservation address is Wuhan University in Wuhan city of China. The Amycolatopsis sp. YIM M13163 provided by the invention can generate antibiotic, namely, efrotomycin pivalate through fermentation; meanwhile, the method for preparing efrotomycin pivalate by adopting the strain is simple, the yield is high, thus sufficient sample support is provided for the development of the medicinal value of efrotomycin pivalate, and therefore, the application prospects are wide.

The invention discloses a detection method for identifying amycolatopsis, and belongs to the field of molecular biology. The invention provides a primer pair capable of specifically identifying the amycolatopsis. The primer pair is designed based on a specific conserved gene AMETH_3452 in the amycolatopsis and homologous gene segments in the amycolatopsis and has strong specificity; a PCR method for rapidly detecting the amycolatopsis is established through optimization of PCR parameters, and it is repeatedly proved that the method is rapid and sensitive, a plurality of strains can be detected at the same time, the detection cost is low, consumed time is short, and the effect is good. A basis is provided for further research on environment detection and animal or clinical actinomycetes.

A novel biotechnological process for the preparation of nitriles, starting from amides, is described. Micro-organisms of the genus Amycolatopsis, Actinomadura or Rhodococcus are employed for this process.

The invention discloses a gene engineering strain of amycolatopsis, a construction method and application thereof, and belongs to the technical field of genetic engineering. According to a method for performing gene knockout in amycolatopsis producing vanillin by using a CRISPR/Cas9 editing system, a CRISPR/Cas9 editing plasmid pLYZYP01 containing a Vdh gene homologous arm is introduced into amycolatopsis CCTCCM 2011265 to achieve vdh gene knockout, and under the condition that 12 g/L of substrate ferulic acid is added into the obtained genetic engineering strain Amycolatopsis sp.delta vdh, the yield of vanillin reaches 9.19 g/L, and the molar conversion rate is 97.7, so that good industrial application value is shown.

The present invention is directed to a process for producing trans-4-hydroxy-L-proline, which is useful as a raw material for medicines or as an additive to foods. In the process, L-proline is converted into trans-4-hydroxy-L-proline in the presence of an enzyme source which is derived from a microorganism belonging to the genus Dactylosporangium, Amycolatopsis or Streptomyces and which catalyzes the hydroxylation of L-proline into trans-4-hydroxy-L-proline, a divalent iron ion and 2-ketoglutaric acid, in an aqueous medium, and the produced trans-4-hydroxy-L-proline is collected from the aqueous medium. In addition, the present invention is directed to a process for producing trans-4-hydroxy-L-proline, wherein the L-proline biosynthesis activity of the host cell of the transformant is reinforced.