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New compound and application thereof

A compound, separation and purification technology, applied in the biological field, can solve the problems of reduced vancomycin sensitivity and the threat of clinical anti-infection treatment, and achieve the effect of good antibacterial activity

Active Publication Date: 2010-12-29
SHANGHAI LAIYI BIOMEDICAL RES & DEV CENT +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, with the continuous use of vancomycin, the sensitivity of Staphylococcus aureus to vancomycin has decreased, which will pose a serious threat to clinical anti-infection therapy. Glycopeptide antibiotics are imminent. At the same time, the preparation of recombinant strains that can ferment and produce second-generation glycopeptide antibiotics that can improve the activity of drug-resistant strains has become an important research topic, and it is also of the same urgency.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 , strain preparation

[0030] The inventor carried out the heterologous replacement operation on the glycosyltransferase gene in the vancomycin-producing strain A. orientalis HCCB10007, that is, catching the glycosyltransferase gene from Amycolatopsis balhimyceticus NRRL B-24207 and replacing it by conjugative transfer The glycosyltransferase gene of vancomycin was obtained to obtain a new strain.

[0031] The bacterial strain was submitted to the General Microorganism Center (CGMCC) of the China Committee for Culture Collection of Microorganisms (CGMCC) located in Beijing on May 6, 2009, and the preservation number is CGMCC No.3053.

Embodiment 2

[0032] Example 2 , fermentation culture

[0033] Inoculate CGMCC NO.3053 into the seed medium and culture at 28°C and 220rpm for 40-48h. Then, under aseptic conditions, the cultured seed solution was transferred to a fermentation shaker flask with an inoculation amount of 8%, and cultured at 28° C. and 220 rpm for 115-120 h, and the fermentation liquid was harvested.

Embodiment 3

[0034] Example 3 , Separation and purification of fermentation products

[0035] 3.1. Fermentation broth pretreatment

[0036] The fermented liquid obtained in Example 2 was adjusted to pH 3-4 with HCl, the mycelia was removed by filtration, and the filtrate was collected.

[0037] 3.2. Separation and purification

[0038] With reference to the separation and purification method provided in the patent US5843437, the fermentation broth is separated and purified, and the specific separation and purification process is as follows:

[0039] Dynamically adsorb the filtrate obtained in step 3.1 with macroporous weak polar resin HP-20, carry out gradient elution with ethanol-water solution containing 0.01% hydrochloric acid, collect the ethanol-water solution (8:2) eluate, and then After activated carbon decolorization and concentration, carry out medium-pressure liquid chromatography, and then use 0.2% NH 4 h 2 PO 4 The methanol-water solution gradient elution of the buffer s...

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PUM

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Abstract

The invention provides a new compound and a preparation method and application thereof. The structure of the compound provided by the invention is shown as a formula (1). Four-potential hydroxyl of glycosyl on benzyl hydroxyl of peptide framework six-potential amino acid of the compound serves as an axial bond and the compound is obtained by fermentation and has the collection number of China General Microbiological Culture Collection Center (CGMCC) No.3053. The compound provided by the invention has high antibiotic activity, so that the compound plays a very important role in developing a new antibacterial medicament.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a new compound and its application. Background technique [0002] Vancomycin is fermented by Amycolatopsis orientalis isolated from Indonesian soil in 1956, and it is the drug of choice for the clinical treatment of methicillin-resistant Staphylococcus aureus (MRSA) infection . However, with the continuous use of vancomycin, the sensitivity of Staphylococcus aureus to vancomycin has decreased, which will pose a serious threat to clinical anti-infection therapy. Glycopeptide antibiotics are imminent. At the same time, the preparation of recombinant strains that can ferment and produce second-generation glycopeptide antibiotics that can improve the activity of drug-resistant strains has become an important research topic, and it is also of the same urgency. Contents of the invention [0003] The inventors of the present application obtained a new strain of A. orientalis...

Claims

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Application Information

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IPC IPC(8): C07K7/06C12P21/02C07K1/22A61K38/08A61P31/04C12N1/21C12R1/01
CPCC12R1/01A61K38/08C12P19/62C12P21/02A61P31/04C12R2001/01C12N1/205
Inventor 戈梅黄鹤魏维阮林高杨晟朱丽姜卫红陈代杰罗敏玉杨志钧夏兴李秋爽王天娇殷瑜金文翔杨天
Owner SHANGHAI LAIYI BIOMEDICAL RES & DEV CENT
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